How I Survived Melanoma Skin Cancer

How To Prevent Skin Cancer

How To Prevent Skin Cancer

Complete Guide to Preventing Skin Cancer. We all know enough to fear the name, just as we do the words tumor and malignant. But apart from that, most of us know very little at all about cancer, especially skin cancer in itself. If I were to ask you to tell me about skin cancer right now, what would you say? Apart from the fact that its a cancer on the skin, that is.

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How I Survived Malignant Melanom

By The Time You've Finished Reading How I Survived Melanoma Skin Cancer Seven Survivors Tell Their Stories. You'll Feel Like A New Person, with A New, More Positive Outlook! You will learn: 1. How do I know if I have melanoma? What are the signs and symptoms? I wanted to know why the doctor was so concerned when she looked at that little mole on my forearm. What was it that looked so sinister? How worried should I be? Was the doctor over-reacting? 2. What tests will the doctor carry out to see if I have melanoma? Will they be able to tell me on the spot if there is a problem? Or will I have to wait for days, fretting about whats going on? 3. How curable is melanoma? If they do tell me its melanoma, what exactly does that mean? Is it a death sentence? Will they tell me You have 12 months to live. Get your life in order and prepare for the worst.? 4. What are the stages of the disease? The reading Id done said that there were different stages of melanoma. What are the symptoms of each stage? What are the survival rates of each stage? If I had a later stage melanoma, wouldnt I know about it? Wouldnt I actually feel like I was sick? 5. How quickly does the disease progress or spread? Should I have gone to the doctor sooner? Id noticed the mole changing over about 3 months. Was this delay critical? 6. How is melanoma normally treated? Would I have to go through chemotherapy and radiation treatment? If so, for how long? What are the odds of curing the disease using these treatments? How extensive is any surgery likely to be? How big will the scars be? 7. What are the common side effects of the treatments? Would I lose my hair? Would I become sterile? What else could I expect? 8. What alternative treatments are available? Id heard of people going on special macro-biotic diets. Id seen lots of herbal remedies on the internet. Which of these are proven and documented, and which ones are snake oil? Is it possible to combine alternative treatments with surgical other western treatments? How do I find a doctor that is open to using both alternative and western treatments? 9. What are the latest treatments being developed, and who is carrying out clinical trials of these new treatments?

How I Survived Malignant Melanom Summary


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Melanoma Microenvironment Analysis Through Gene Expression Profiling

One strategy that has been pursued to gain a broad array of information regarding the melanoma tumor microenvironment and potential associations with clinical outcome to immunotherapies is gene expression profiling. Marincola and colleagues were the first to investigate whether patterns of transcripts in the melanoma tumor site might correlate with clinical response in patients. Using cDNA arrays applied to retrospective fine needle aspirate samples, they identified a set of transcripts present in pretreatment lesions that appeared to be linked with a subsequent response to therapy. These included multiple immunologically relevant genes, including EBI3, TIA1, IRF2, and IFI27, suggesting features of interferon (IFN) responsiveness and cytolytic potential 7 . The expression of interferon-response genes suggests the possibility that innate immune signals mediated through host type I IFNs might contribute to generating a supportive tumor microenvironment for an adaptive immune response....

Post Transplant Squamous Cell Skin Cancer

Immune defences, resulting from heavy immunosuppression are also associated with an increased risk of malignancy. Genetic risk factors are also suspected. Accordingly, genetic polymorphisms associated with differential levels of cytokine production may have an important effect on tumorigenesis after organ transplantation. In this context, IL-10 plymorphism is of particular interest, since, in addition to the biological functions of IL-10 discussed in some detail above, ultraviolet-induced DNA damage, a risk factor for skin cancer, also increases production of IL-10.142 For the above reasons, Alamartine et al,84 in a French-based study, investigated possible associations between the IL-10 -1082, -819 and -592 SNPs and the occurrence of post-transplant skin cancers in a series of 70 renal transplant recipients who developed post-transplant squamous (SCC) or basal cell carcinoma (BCC), 70 healthy controls and 70 age, sex and immunosuppression-matched renal recipients without cancer....

Patient Measurements In Melanoma

The measurements in 10 melanoma patients were performed according to the same protocol as in the breast cancer patients. Because the tracer was injected intracutaneously, dose rates at short distances of 10 and 15 cm could be obtained in these patients. The injected dose in the melanoma patients amounted to 19 6 MBq 99mTc nanocolloid in a total volume of 0.4 mL saline. Due to more rapid transportation of the tracer, measurements were obtained within 30 min after injection. Results are displayed in Table 2.

Case 13 Sentinel Node Close to Primary Melanoma Site

A 42-year-old man was referred after excision biopsy of a pigmented lesion on his left midneck. Histological examination had revealed it to be a melanoma 1.8 mm in maximum thickness, invading to Clark level IV. At operation, the sentinel node demonstrated on the lymphoscintigram was found without difficulty, being both blue-stained and hot when examined with the gamma-ray detection probe (8978 counts per 10 seconds after removal). The site of the presumed second-tier node was explored, but only a nonblue node that was not hot with the probe was found. When the melanoma site was

SSL Induce Specific CTL in PBMC from Healthy Donors and from Melanoma Infiltrating Lymphocytes

Regarding TIL from melanoma patients, SSL and SM26-35 were equally effective at 20 mg mL TAA concentration, with induced CTL precursor frequencies (CTLp) on limiting dilution analysis (9) of about 20 106 (Fig. 6, panel A). However, at lower (physiological) antigen concentration, while SSLM26-35 peptide retained its full immunogenic capacity,

Metastatic Melanoma In Regional Lymph Nodes

Sentinel Lymph Node

Once patients develop metastatic melanoma in their regional nodes, prognostic factors based on the primary melanoma contribute very little to the prognostic model. For most solid tumors, including melanoma, the most powerful predictor of survival is the presence or absence of lymph node metastases. The presence of lymph node metastases decreases the 5-year survival of patients approximately 40 compared with those who have no evidence of nodal metastases. Much time, effort, and expense is placed on identifying prognostic factors based on the primary tumor, and not enough emphasis is given to identifying which patients really have signs of micrometastatic disease in their nodal basins. For instance, there are currently 26 prognostic factors (Table 1) for melanoma based on variables from the primary tumor. Yet, in multiple regression analysis performed on many collected populations in the literature, the lymph node status of the patient is the most powerful factor for predicting...

Assays For Occult Metastatic Melanoma Lymph Node Culture

Sent to pathology and half of the node placed into tissue culture. Several patients with histologically negative nodes had melanoma cells grow in culture. Detailed characterization of these cells with immunohistochemical staining, monoclonal antibody staining, and electron microscopy confirmed that the growing cells were indeed melanoma cells. Thirty-one percent of the histologically node-negative population were upstaged to Stage III with the cell culture technique. The clinical correlation between lymph node culture and disease-free survival was very good. Patients who were histologically node negative, but node positive by cell culture, had an increased recurrence rate compared with those patients with melanoma whose nodes were negative by both assays 8 . Widespread applicability was questioned with the lymph node culture technique because this would require hospitals to establish cell culture facilities and the results of the assay are not available for 4-6 weeks.

Lymphocyte Subpopulations in Melanoma Patients Treated with Dendritic Cell Vaccines

The main goal of cancer immunotherapy is to induce or boost tumor-specific effector cells able to eliminate or reduce tumor progression. In this study, we characterized lymphocyte phenotypes in melanoma patients receiving dendritic cell (DC)-based vaccinotherapy. We found that several biological markers served as unfavorable prognostic factors for patients' response to therapy. This included decrease of CD4+ and CD8+ lymphocyte levels, 10 and higher increase of CD16+CD3+CD8+ lymphocyte population, and increase of CD16+CD8+perforin+ T lymphocytes, especially in combination with decreased levels of CD16+CD8-perforin+ and CD8+CD16-perforin+ cells. Increase in CD8+CD16-perforin+ T lymphocytes with normal levels of CD16+CD8-perforin+ cells and the absence of CD16+CD8+perforin+ and regulatory lymphocytes were shown to be the positive prognostic markers for patients' response to DC vaccines.


Melanoma is one of the most common leptomeningeal cancers in adults however, it is infrequently observed in children since melanoma comprises less than 1 of childhood cancers.88'99 Leptomeningeal dissemination of melanoma most commonly occurs in children who have advanced systemic disease, especially underlying CNS parenchymal metastases. Children who have large congenital nevi rarely present with primary leptomeningeal melanoma.88'99 Both primary leptomeningeal melanoma and metastatic leptomeningeal melanoma are rapidly progressive entities for which there is no known effective therapy. In contrast, neurocutaneous melanosis (NCM), a rare phacomatosis characterized by the presence of large non-malignant melanocytic nevi in combination with an excess of melanotic cells in the leptomeninges may be asymptomatic for prolonged periods or have neurologic symptoms with or without associated malignant transformation. The prognosis for patients with neurologic symptoms is generally poor, even...

Skin Cancer

Skin Medical Terminology

Skin cancer is the most common type of human cancer. Its rate has been increasing in recent years, mainly because of the mutation-causing effects of the ultraviolet rays in sunlight. Malignant melanoma results from an overgrowth of melanocytes, the pigment-producing cells in the epidermis. It is the most dangerous form of skin cancer because of its tendency to metastasize. This cancer appears as a lesion that is variable in color with an irregular border (Fig. 21-5). It may spread superficially for up to 1 or 2 years before it begins to invade the deeper tissues of the skin and to metastasize through blood and lymph. The prognosis for cure is good if the lesion is recognized and removed surgically before it enters this invasive stage. Basal cell carcinoma constitutes more than 75 of all skin cancers. It usually appears as a smooth, pearly papule (Fig. 21-7). Because these cancers are easily seen and do not metastasize, the cure rate after excision is greater than 95 . FIGURE 21-5....

Evidence for Locoregional Immune Dysfunction in Cancer

Nevertheless, TIL freshly isolated from certain tumors, especially melanoma and renal cell carcinoma (RCC), are responsive to immunologic intervention in vitro 24 . This suggests that even in the face of tumor-induced immune deviation, these lymphocytes can be salvaged and may be of clinical benefit to the patient, if an appropriate treatment is administered. Functional impairments have also been

Interference with the Induction of TAAspecific Responses

The key role of DC in cross-presentation of TAA to T cells is well known 14, 156 . Dysfunction of DC cross-presenting TAA to T cells could lead to an inadequate or biased antitumor immune response. DC not only process and present TAA to T cells but are important sources of IL-1, IL-12, IL-15, IL-18, IL-23 and Interferons, among other cytokines and chemokines. They are also rich in co-stimulatory molecules necessary as second signals or in growth factors for T-cell differentiation, proliferation and memory development 46, 186 . Therefore, if DC are depleted or unable to perform normally, the induction of TAA-specific immunity is likely to be impaired. Indeed, inhibition of DC in the presence of tumors has been reported 1, 65 . Defective maturation of DC in the tumor microenvironment may be mediated by tumor-derived vascular endothelial growth factor (VEGF) 44 . Others report that tumor-derived exosomes interfere ex vivo with differentiation of human DC from peripheral blood monocytes...

Facilitating the Search for the Sentinel Node

The discovery by Morton and co-workers caused others to search for more rapid and less technically demanding methods of locating sentinel lymph nodes. Alex and Krag 37 described the use of a labeled colloid in combination with a gamma-detecting probe to locate the sentinel node in patients with melanoma. This technique was much simpler than the original method described by Morton and colleagues. Uren and colleagues showed that the standard lymphoscintigra-phy technique, which had been in routine use to determine which node fields drained melanoma primary sites, could be modified to allow the exact location of the sentinel nodes to be marked on the skin. This information could then be used to aid in rapid surgical location of sentinel nodes 24,25 . Digital gamma cameras were found to be important in this process as they allowed dynamic studies to be performed, with lymph channels followed until they were seen entering the sentinel nodes. Because some tracer may pass onward quite...

Unexpected Lymphatic Drainage Pathways

Using lymphoscintigraphy in this way meant that patients with lesion sites previously thought to have unambiguous drainage were also studied, and this led to the discovery of several unusual and previously unknown pathways for lymphatic drainage of the skin 28-33 . It is now clear that when faced with an individual patient with a melanoma, there are, in fact, very few, if any, places on the skin of the human body which have completely unambiguous drainage, and in any event, this concept is now redundant because the location of the sentinel node has become the main application of lymphatic mapping of the skin. Several large surgical studies have now confirmed that sentinel node status is an accurate reflection of node field status in melanoma patients 34-36 .

Interference with Functions Survival of Effector T Lymphocytes

Apoptosis of immune cells is not limited to the tumor site. Apoptosis of circulating CD8+ T cells in subjects with cancer has been described in melanoma, breast, ovarian, and head and neck cancers 74, 139, 140, 190 . Studies involving TUNEL staining of TIL and Annexin V (ANX) binding to circulating T cells suggest that CD8+ rather than CD4+ T cells selectively undergo apoptosis at the tumor site and in the peripheral circulation of cancer patients 74, 190 . The proportion of Sera and body fluids of patients with melanoma, ovarian carcinoma or HNC contain membranous 50-100 nm microvesicles (MV) presumably originating from the tumor, which contain biologically active 42 kDa FasL and MHC class I molecules and mediate apoptosis of Fas+ lymphocytes at sites distant from the tumor 2, 12, 70, 163 . Upon isolation, these MV induce TCRZ degradation and DNA fragmentation in activated T cells reviewed in 196 . MV are prominent in tumor cell super-natants, but activated normal cells also produce...

Preoperative Lymphoscintigraphy

The techniques and results of cutaneous lymphoscintigraphy are discussed in detail in Chapter 3, but several aspects of the procedure need to be emphasized here because they are directly relevant to the planning and execution of surgery to identify and remove sentinel nodes. High-quality preoperative lymphoscintig-raphy is invaluable to ensure that sentinel lymphadenectomy is accomplished expeditiously and to minimize the amount of dissection of the relevant lymph node field (or fields) which must be performed. As well, lymphoscintigraphy provides preoperative identification of drainage pathways to unexpected node fields 3-5 . Some have suggested that preoperative lymphoscintigraphy is necessary only for melanomas in sites from which the lymphatic drainage might be ambiguous. It has now become clear, however, that there are few, if any, sites in the body from which lymphatic drainage pathways are able to be predicted with complete confidence 6,7 . Even in sites that appear to have...

Assess Rates for Diseases Known Not to Be Affected by the Exposure

For example, in a study of the effects of sunscreen use on risk of developing melanoma (an illustration from an oral presentation by Diana Petitti, Kaiser Permanente of Southern California), we would not expect sunscreen use to influence the risk of myocardial infarction, breast cancer, or motor vehicle injury. To determine whether our group of nonusers of sunscreen is a good counterfactual comparison group to the sunscreen users, reflecting the risk that the sunscreen users would have had in the absence of sunscreen use, we might find it useful to examine an array of causes of death that include some that should not differ due to a causal impact of sunscreen use. Even if our assumptions are incorrect in some of the diseases thought to be unrelated to sunscreen use, examination of the overall pattern of results across a range of presumably unrelated diseases would reveal whether a systematic tendency is present for exposed and unex-posed groups to differ. If, for example, we observed...

Lymphatic Mapping Of The Skin

Lymph Node Map

Colloidal 198Au was used by Fee et al. in 1978 when they studied 32 patients with melanoma 9 . This study confirmed that lymphoscintigraphy could accurately predict the node fields that potentially contained metastatic melanoma. Nine of their patients had nodal metastases, and in every case, lymph drainage from the primary site to this node field was demonstrated on lymphoscintigraphy. Around this time, other investigators were also using scintigraphy to map lymph drainage in patients with melanomas located on the trunk and elsewhere when drainage was considered uncertain (i.e., in Sappey's zones of uncertainty or in the head and neck 10,11 ). It was emphasized that lymphoscintigraphy could not predict whether lymph nodes contained metastases, but that it could identify which node fields were at greatest risk of harboring occult metastases 12 . For many years, cutaneous lymphoscintigraphy was thus used to identify which node fields received lymphatic drainage from the primary melanoma...

Systemic T Cell Immunity and T Cell Entry into Tumor Tissue

Despite these observations, numerous studies have demonstrated that tumor specific T cells can be isolated from the tumor tissues of various cancers. Among 123 tumor-infiltrating T cell (TIL) cultures from melanoma biopsies, 57 recognized HLA-A2 restricted epitopes of the highly immunogenic melanoma-associated antigens MART-1 and gp100, while much less cultures reacted to antigens presented in the context of other HLA-alleles 28 . T cell responses were frequently dominated by a single HLA allele and besides Melan-A, no dominant antigens could be characterized among a broad variety of TAAs that were simultaneously tested 29 . Multiple HLA-A2-restricted melanoma differentiation antigens were recognized by TILs from melanoma patients 30 . Interestingly, the frequencies of Mart 1-reactive HLA-A2-restricted CD8 TIL as detectable by HLA-tetramer analysis in approximately half of the freshly excised melanomas were between 1 50 and 1 200, which is similar to the frequencies of such cells in...

Ralph S Freedman md phd

Advances continue to be made in tumor immunology and in strategies to integrate the growing number of bioimmunotherapeutic molecules into the treatment of ovarian cancer, as well as other malignancies. Extensive studies have provided support for antigen-driven T-cell activation in vivo. The number of known tumor antigen epitopes is expanding, although advances in this area remain behind that of melanoma. Evidence suggests that the tumor environment is contributing to a state of in vivo immunosuppression however, in vitro experiments and laboratory correlative studies also show that immune suppressor activity might be reversible. These findings could lead to new approaches, such as the use of antibodies or cytokines to overcome the immunosuppressive effects, in addition to the more established surgical and chemotherapeutic debulking. Evidence for specific T-cell activation in vivo in ovarian cancer follows criteria that have been set for melanoma, and includes the development of...

Sources Of Error In Interpreting Sentinel Lymph Nodes

And more cohesive than melanoma cells and while strongly S-100 protein positive (Fig. 4b) will either express no HMB-45 or HMB-45 at a relatively weak level (Fig. 4c). Nevocyte clusters are often arranged around capsular vessels. Another pitfall is the presence of neural tissue within the lymph node (Fig. 5). If the nerve has associated Schwann cells, these may stain relatively strongly, and if the nerve is cut transversely, an appearance suggestive of a cluster of S-100-protein-positive melanoma cells may result. Errors may also be associated with HMB-45 staining. These are fewer than those encountered with S-100 protein, but some care is necessary. A minority of melanomas (10-15 ) are made up of cells that do not express HMB-45. In hya-linized and calcified connective tissue within lymph nodes, especially lymph nodes from the groin and internal iliac area, extracellular HMB-45 positivity may be seen and care is necessary to avoid overcalling this appearance.

Altered Phenotypes of HLA Class I in Various Types of Cancer

All of these phenotypes can be found in various types of tumor, regardless of the tissue origin or of the carcinogen inducing the tumor. Differences are observed in the distribution of the phenotypes and in the combination of molecular mechanisms leading to each phenotype. An example of such distribution in various types of solid tumors and melanoma cell lines (ESTDAB project, http ipd estdab ) (Pawelec and Marsh 2006) is presented in Table 1.

DNA chips can reveal DNA mutations and RNA expression

The emerging science of genomics must deal with two major quantitative realities. First, there are a large number of genes in eukaryotic genomes. Second, the pattern of gene expression in different tissues at different times is quite distinctive. For example, a skin cancer cell at its early stage may have a unique mRNA fingerprint that differs from that of both normal skin cells and the cells of a more advanced skin cancer.

Association of HLA Class I Alterations with Tumor Escape and Cancer Progression

Recently, we have observed an interesting association of HLA class I expression and metastatic progression in a melanoma patient, whose subcutaneous metastases responded differently to autologous tumor cell vaccination (Cabrera et al. 2007). We analyzed three progressing and three regressing metastatic lesions for HLA class I expression using immunohistochemistry, tissue microdissection, LOH analysis, and other molecular techniques. Interestingly, the progressors had low HLA class I expression and higher frequency of LOH in chromosomes 6 and 15. In addition, all of the progressing metastases were HLA-B negative. Expression of class I molecules was normal in the regressing metastases. LOH at chromosome 6 was detected in all six studied metastases, suggesting that this defect may also be found in the primary tumor, contributing to the mechanism of tumor escape and metastatic progression. Real-time quantitative PCR of the samples obtained from microdissected tumor showed lower mRNA...

Assays For Occult Metastases Rtpcr

A study was initiated to develop a highly sensitive method to detect micrometas-tases by examining lymph nodes for the presence of tyrosinase messenger RNA (mRNA) 9 . The assay is based on the biosynthetic pathway of melanin. It is known that tyrosine is converted to melanin in the melanocyte or melanoma cell. The key is that the first two steps of the synthesis are catalyzed by the enzyme tyrosinase. Tyrosinase is a mono-oxygenase that catalyzes the conversion of tyro-sine to 3,4-dihydroxyphenylalanine (DOPA) and of DOPA to dopaquinone. Tyro-sinase is one of the most specific markers of melanocytic differentiation. All cells of the body will have the gene for tyrosinase, but only cells that are actively producing pigment, such as melanoma cells or melanocytes, will express the mRNA for the tyrosinase gene. If this gene product is found in the lymph node preparation, in the peripheral blood, or in the bone marrow, then that finding is good evidence that metastatic melanoma cells are...

Rtpcr Assay In Peripheral Blood

The development of new molecular biology techniques has provided a means by which molecular markers present at low copy numbers can be detected with a sensitivity higher than that of immunohistochemistry. In 1991, Smith et al. proposed for the first time that melanoma cells could be detected in the peripheral blood using RT-PCR to target any message from the tyrosinase gene 10 . Several other groups have pursued this lead, but the results have been extremely variable. The most striking discrepancies occur in patients with Stage IV melanoma, in which the incidence rates at finding RT-PCR-positive circulating cells varies from 0 to 100 13,14 . In a series from Germany 13 , 100 of the patients with Stage IV melanoma had RT-PCR evidence of circulating melanoma cells, whereas a report from Kunter showed that only 28 of the patients with Stage IV melanoma were marker positive 14 . In addition, in the latter series, a positive peripheral blood RT-PCR assay was associated with progression of...

Aire and Thymic Selection

Such spatially secluded antigens as cancer testis (CT) antigens, of which more than 40 have now been identified, are encoded by genes normally expressed in the human germ line but are also expressed in melanoma and carcinomas of the bladder, lung, and liver. These immunogenic proteins are being vigorously pursued as targets for therapeutic cancer vaccines (Van Der Bruggen, Zhang, Chaux, Stroobant, Panichelli, Schultz, Chapiro, Van Den Eynde, Brasseur, and Boon 2002 Simpson, Caballero, Jungbluth, Chen, and Old 2005). It is very important that the array of promiscuously expressed self-antigens in mTECs includes well-known targets for cancer immunotherapy, such as a-fetoprotein, tyrosinase, and gp100. Therefore, intrathymic selection makes the immune system tolerant to tumor-associated antigens. Gene expression in normal tissues may result in tolerance of high-avidity CTL, leaving behind low-avidity CTL that cannot provide effective immunity against tumors expressing the relevant target...

Insights into Mechanisms of Immune Resistance in the Tumor Microenvironment through Molecular Profiling

Abstract Many patients with melanoma show spontaneous T cell responses against tumor antigens, and induction or amplification of these T cells responses can frequently be achieved through vaccination or adoptive T cell transfer. However, tumor responses as measured by tumor shrinkage remain infrequent. These observations have argued for analysis of the tumor microenvironment in metastatic melanoma for potential mechanisms of resistance to immune effector function at the level of the target tumor site. This review discusses two categories of regulation at the level of the tumor microenvironment, chemokine-mediated migration of effector T cells and active immune suppression, that have been identified through gene expression profiling of human specimens. Melanoma cell-intrinsic apoptosis also is discussed. The identification of these mechanisms points toward new strategies of intervention to consider for improving the clinical efficacy of T cell-based immunotherapy for cancer, and also...

Chargebased Targeting Of Liposomes

Cationic liposomes loaded with drugs have been investigated for therapeutic efficacy in preclinical models. In a model of amelanotic hamster melanoma grown in a dorsal skinfold window chamber, cationic liposome-encapsulated paclitaxel effectuated strong inhibition of tumor growth (66). At a dose of 5mg kg body weight of paclitaxel in cationic liposomes, tumor volume was approximately 6-fold, 6-fold, and 10-fold lower as compared to Taxol, empty cationic liposomes, or buffer-treated animals, respectively, and significantly delayed local lymph node metastasis. The observation that control treatments (paclitaxel and empty cationic liposomes) provide a modest therapeutic effect on their own may suggest that the antitumor effects for paclitaxel-loaded cationic liposomes represent a merely additive effect. Nevertheless, it may also be the result of angiogenic EC delivery of pacli-taxel by cationic liposomes. The focus of a subsequent study was the mechanism of action of paclitaxel-loaded...

Early CLS Formation Requires Activation of Apoptotic Pathways

To determine the molecular mechanisms underlying the phenomenon of VM in the tumor, we prepared six cell lines from the surgical species of patients with disseminated melanoma (Mikhailova et al. 2005). Melanoma status was confirmed by im-munocytochemistry for the expression of four melanoma-associated markers MelanA, S100, tyrosinase, and high-molecular weight antigen (HMW). Furthermore, using the short-term Matrigel assay, we showed that five cell lines with high invasive potential were engaged in CLS formation. In the presence of growth factors, melanoma cells were induced to align and form CLS in cultures. A gradual decrease in cell number during the CLS formation could readily be observed in in vitro cultures. Cell counts revealed that only 65-70 of cells were viable 24 h later, suggesting that apoptosis might be an essential part of the VM phenomenon. Both morphological data and the PI staining suggested that apoptosis may be responsible for the decrease in cell numbers (Figure...

CLS Formation Requires Activation of Apoptotic Pathways Mediated by Mitochondrial Cytochrome c Release and Caspase3

There are two major mechanisms mediating apoptosis intrinsic and extrinsic. Cell surface Fas-receptor signaling upon FasL binding triggers caspase-8 activity and caspase cascade (Schulze-Osthoff et al. 1998). Involvement of this pathway in CLS formation by melanoma cells was ruled out in our studies, since antagonistic anti-Fas antibody did not block the in vitro tube-like organization process and CLS formation (Figure 3A). Fas and FasL expression on melanoma cells was also analyzed by flow cytometry and neither Fas nor FasL was seen during CLS formation (data not shown). These findings suggest that extrinsic apoptotic pathways did not participate in CLS formation. Figure 3. Pathways of capillary-like structure (CLS) formation. (A) Fas-FasL croslinkage does not modify CLS formation. (A1) Cells treated with anti-Fas blocking antibody (A2) Control, non-treated cells. (B) Flow cytometry analysis of melanoma cells stained with PI. (B1) Control cells were detached from plastic 10 h after...

IL10 Gene Polymorphisms and Cancer

In the following consideration of the studies summarised in Table 2, cutaneous malignant melanoma (CMM), prostate (PC) and breast cancer (BC) are considered first, since angiogen-esis is crucial for the development of these tumors89 and indeed the extent of angiogenesis correlates with the probability of metastasis and or prognosis in these malignancies.90-96

Regulation of Migration into Tumor Metastases

A striking difference between clustered melanoma metastases was the presence or absence of a lymphocyte gene expression signature. It is critical to perform confirmatory assays to determine whether this difference is truly reflected at the level of T cell involvement. In this case, differential presence of CD8+ T cells was confirmed by immunohistochemistry. In some instances, T cells were extremely abundant and were uniformly distributed throughout the tumor mass. This result suggests that some tumors have the capability to recruit T cells and others do not. One might imagine that even if a patient developed circulating tumor antigen-specific T cells as measured in the blood, if homing into tumor sites did not occur then tumor regression would be unlikely to follow. Another implication of this result is, assuming a subset of these T cells is antigen specific as we and others have observed in similar cases previously 9-11 , then it seems likely that the tumor microenvironment has...

Intraoperative Lymphatic Mapping Using Vital Blue

In 1994, Giuliano and co-workers published their initial work of intraoperative lymphatic mapping with a vital blue dye 26 . A 1 isosulfan blue dye solution (Lymphazurin, Hirsh Industries, Inc., Richmond, VA, U.S.A.) was used to identify the sentinel node in this study of 172 patients (174 lymph node basins). The sentinel node was identified in 66 of patients overall, but the identification rate improved as the investigator became familiar with the nuances of the technique. The 59 rate of sentinel node detection in the first 87 cases increased to 72 in the next 87 cases and the detection rate reached 78 by the last 50 cases in this series. The technique was modeled after the melanoma model, a cutaneous tumor, but had to be adapted to breast cancer, which is a parenchymal tumor. This led to significant changes in the technique. In this study, there were five false-negative cases reported in which the pathology of the sentinel node did not accurately predict the status of the axilla....

When the Wall Comes Tumbling Down

The fact that this form of pneumonia appeared in a cluster of five men who had no apparent reason to be immunosuppressed was so striking that the physicians immediately reported it to the Centers for Disease Control (CDC) in Atlanta. Barely one month later it was reported that 26 male homosexual patients seen during a short time period in another state had P. carinii pneumonia and or Kaposi's sarcoma, a rare form of skin cancer. The UCLA cluster, upon examination, also turned out to be gay. In a matter of months, new cases belonging to this syndrome numbered in the hundreds, and then the thousands, and it had acquired a name acquired immune deficiency syndrome (AIDS). Thus began a journey into the unknown, a journey whose end is still beyond our vision.

Resistance at the Level of the Tumor Cells Themselves

Melanoma in general is quite resistant to killing by cytotoxic chemotherapy and conventional doses of radiation. These clinical observations suggest that mechanisms of resistance to cell death are likely highly active in this tumor type. Several observations in our gene expression profiling experiment support this contention. Highly expressed in many melanoma tumors was the gene encoding anti-apoptotic protein survivin, which has been shown to be correlated with poor clinical outcome in other series 46 . Many melanoma tumors were found to overexpress the Notch transcriptional target Heyl, arguing that Notch signaling is constitutive in those tumors. Notch signaling has been shown to support survival of melanoma tumor cells in vitro, with inhibition of Notch processing leading to melanoma cell apoptosis 47 . An additional factor found to be overexpressed was the serine protease inhibitor SerpinA3. Molecules in the Serpin family have been shown to mediate resistance of target cells to...

Immune Mediated Tumor Destruction In vivo

Since activated CD8+ T cells possess direct cytotoxic capacities and are responsible for most immune-mediated tumor regression, it has been assumed that their cyotoxic potential is a vital component for the promotion of tumor regression in vivo. However, the recent availability of a variety of gene-targeted or mutant mice has allowed for this hypothesis to be tested in well-defined tumor models. Surprisingly, in a variety of mouse tumor models including the B16 melanoma, MCA-101 sarcoma (Winter et al. 1999), MCA-205 sarcoma (Peng et al. 2000), and the mouse renal cancer Renca (Seki et al. 2002), it has been demonstrated that tumor rejection by the transfer of activated T cells could occur in the total absence of perforin. This despite the fact that the perforin-dependent granule-mediated lytic pathway is usually the predominant cytotoxic pathway employed by activated CD8+ T cells in vitro. This suggests that granule-mediated cytotoxic effects may not be as crucial for tumor rejection...

Discussion Of Phantom And Patient Measurements

The dose rates as measured in melanoma patients were not significantly different from those in breast cancer patients (Student's -test, p > 0.5). The higher dose It can be assumed that, in clinical practice, the surgeon's hands will be at an average distance of 10 cm from the tumor. The dose rate to the surgeon's hands, with a breast tumor at a depth of 2 cm, will not exceed 11.7 j,Sv h 10 MBq. In the case of a melanoma, this will be 9.8 iSv h 10 MBq.

Exposure Of The Surgeon

The total radiation dose received by a surgeon working in a community hospital as a result of sentinel node biopsies in melanoma probably will be minor. In the first place, the incidence of melanoma is far less compared with breast cancer. Second, the time required for excision of the injection site in melanoma is less than for excision of a breast tumor. Moreover, it remains to be established which melanoma patients benefit from sentinel lymphadenectomy 20 .

Lymphatic Mapping In Other Neoplasms

With the increased experience with sentinel node technology in melanoma and breast cancer, investigators have begun to apply this technique to the staging of other solid neoplasms. Bilchik et al. reported their experience with the universal application of lymphatic mapping at the John Wayne Cancer Institute 26 . The investigators evaluated their experience with lymphatic mapping in Merkel cell tumors, squamous cell carcinoma of the head and neck, thyroid cancer, gastrointestinal malignancies (including small bowel, pancreas, and colon primaries), and vulvar neoplasms. They found the technique to be feasible for solid tumors other than breast and melanoma. Lymphatic mapping may ultimately replace conventional dissection with more accurate staging. Other investigators have confirmed the feasibility of the technique in colon and oral squamous cell carcinoma 27,28 . However, there have been no larger prospective studies in malignancies other than melanoma and breast cancer. At this point,...

Use of Liposomes for Localizing the Sentinel Lymph Node

In the last decade, cancer surgeons have become very interested in methods to definitively localize the sentinel lymph node. The sentinel lymph node is the first lymph node that receives lymphatic drainage from the site of a primary tumor. The sentinel node is much more likely to contain metastatic tumor cells than other lymph nodes in the same region. It is believed that the initial draining lymph node (sentinel node) of a tumor may reflect the status of the tumor's spread to the remaining lymphatic bed. Localization of the sentinel lymph node and its close histological assessment following its removal from the body was initially developed as a prognostic indicator in patients with malignant melanoma (51). If no cancer cells are found in the sentinel node on pathologic examination, the prognosis for the patient is greatly improved. After many detailed studies validating the effectiveness of this approach for patient prognosis and as a method to guide future therapy of melanoma...

Incidence and Distribution of Cancers

The incidence of specific cancers varies significantly across the world, possibly due to different environmental factors. For example, the incidence of melanoma is 150-fold greater in Australia than in Japan, prostate cancer occurs 70 times more frequently in the U.S. than in China, and liver cancer is 50 times more prevalent in China than in Canada. At the same time, the incidences of certain malignancies such as leukemia, ovarian cancer, and breast cancer are similar in most countries. Even within these general differences, the use of cancer maps has, in certain instances, disclosed some dramatic variations that are likely to be related to local socioeconomic or environmental factors.

The Immune System and Cancer

Our thinking about cancer has changed remarkably in recent years. Bone cancer still looks different from brain cancer skin cancer is still treated differently than lung cancer. But the current focus is on understanding what causes cancer in the first place, and here the emphasis is on what cancers have in common, rather than on what makes them different. Cancer can be contributed to by external agents radiation, chemicals, and some viruses or by mistakes made within a cell when it reproduces its DNA. And indeed, any cell or tissue in the body can give rise to a tumor. But ultimately every cancer is a disorder of DNA. All cancer cells share one single, common feature they have their lost ability to regulate DNA synthesis and cell division. And the regulatory elements governing these processes lie in the DNA itself in our genes.

Case 1 Normal Drainage Pattern

Nodular melanoma with ulceration (Clark level IV, Breslow thickness 5.0 mm). Wide local excision and sentinel node biopsy were planned. Hematoxylin and eosin (H& E) staining and immunohistochemistry (S-100, HMB-45) revealed no melanoma deposits in the two sentinel nodes. This patient demonstrates a typical drainage pattern. A melanoma on the leg usually drains to two sentinel nodes caudally from Poupart's ligament, whereas a melanoma on the arm drains more often to a single sentinel node in the axilla. Drainage on the trunk and in the head-and-neck area is more difficult to predict. 2. Nieweg OE, Jansen L, Kroon BBR. Technique for lymphatic mapping and sentinel node biopsy for melanoma. Eur J Surg Oncol 1998 24 520-524. A 56-year-old man presented with a melanoma in the right flank. A diagnostic excision with a narrow margin was performed. Histological examination showed a superficial spreading melanoma (Breslow thickness 1.7 mm, Clark level III).

Targeting Of Antisense Oligonucleotides

Oligonucleotide-liposomes can be targeted to specific cells via covalently coupled antibodies to cell surface markers. Neutral lipid-coated cationic liposomes encapsulating antisense oligonucleotides against c-myb and targeted to disialoganglioside, GD2, were much more effective in inhibiting the growth of neuroblastoma cells compared to nontargeted liposomes and free oligonucleotides (119). These liposomes also had significant antitumor effects in vivo, but part of this effect could be attributed to the immunostimulatory effect of CpG sequences in the oligonucleotide (120). GD2-targeted coated cationic liposomes containing anti-c-myc oligonucleotides inhibited melanoma cell proliferation, the development of microscopic metastases, and tumor growth (121). Although antibody coupling to PEG-DSPE has the advantage that the targeting ligand protrudes out from the liposome surface, the inhibitory effect of this polymeric lipid on transfection (124,125) should be taken into consideration....

Pathology and FollowUp

All three lymph nodes were free of disease by H& E and immunohistochemistry staining (HMB-45, S-100). Six months later, the patient came to the clinic because he felt a lump in the neck. A 1-cm rubbery node was palpated in the right supraclavicular fossa. Fine-needle aspiration cytology revealed melanoma cells. A radical right neck dissection was performed after a search for other sites of disease (including computed tomography scans of chest, neck, and brain liver ultrasound bone scintigraphy blood chemistry) revealed no other tumor locations. Two supraclavicular nodes contained melanoma deposits the other 28 nodes in the specimen were free of disease. A distant skin metastasis was discovered 9 months later. The patient died from brain metastases 1 year after the neck dissection.

Tumor Cell Evasion Of Killer Cells

Tumor cells may evade killer cell destruction by the production of FasL that induces apoptosis in adversarial TILs, CTLs, and NK cells expressing the Fas marker. Constitutive expression of FasL (CD95L) has been described in NK lymphoma, large granular lymphocytic leukemia, melanoma, hepatocellular he-

Immunosuppressive Agents

Purine antimetabolites such as azathioprine are associated with an increased risk for lymphomas and non-melanoma skin carcinomas in renal transplant recipients. The possibility of a similar increased incidence of malignancy in rheumatic disease patients treated with azathioprine remains to be firmly established. An increased incidence of non-Hodgkin's lymphoma and cervical carcinoma in SLE patients on azathioprine has been suggested. There is no apparent relationship between the amount and duration of cytotoxic therapy and the development of malignancy. 3. Cyclosporine, although not mutagenic, is associated with lymphoproliferative disorders in up to 3 of renal transplantation patients treated with concomitant azathioprine and prednisone. Whether patients with rheumatic diseases who are taking cyclosporine, with or without methotrexate, have an increased risk for lymphoma and brain or skin cancers remains to be determined. In one review of more than 1,000 RA patients treated with...

Functional CTL Capture Membrane Fragments from Tumors

In a recent study we postulated that activated human T cells will generate an IS-like interaction with tumor cells similarly to that described with dendritic cells 61 and that as a consequence membrane fragments will be acquired by T cells. Using fluorochrome-tagged melanoma cells or peptide-loaded T2 cells, we showed that antigen-specific CD8+ T cells incorporate membrane fragments from tumor cells and fuse them with their outer cytoplasmic membrane (Fig. 8.1). T cells that captured labeled membranes were detectable by flow cytometry and sorted by FACS Fig. 8.2 Membrane capturing CD8+ T cells show increased cytotoxicity against peptide pulsed targets and autologous melanoma cells. Cytotoxic activity of membrane capturing (cap+) or non-capturing (cap-) T cells against peptide-pulsed targets and melanoma cell lines. (A) gp100154-162-reactive A2-restricted CD8+ T cells were co-incubated with Dil-labeled 624mel melanoma cells (A2+ gp100+) and sorted by FACS into cap+ and cap- populations...

Overcoming Tolerance Leads to Tumor Immunity

Initially, it was demonstrated by many groups that conferring costimulatory ligands to T cells makes tumors more immunogenic (Townsend and Allison 1993 Zheng et al. 2006). Subsequently, it was shown that blocking negative signals favors more efficient T cell activation and can result in a potent antitumor response. Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) binds to B7.1 2 on APC with higher affinity than CD28 (Linsley et al. 1994) and delivers an inhibitory signal to T cells that serves to block IL-2 expression and cell cycle progression (Krummel and Allison 1996). Transient in vivo blockade of CTLA-4 at the time of tumor vaccination can enhance vaccine potency and antitumor immunity against melanoma and prostate cancer (van Elsas et al. 1999 Hurwitz et al. 2000). We and others previously reported that treatment of a mouse model of prostate cancer (TRansgenic Adenocarcinoma of the Mouse Prostate TRAMP model) with a GM-CSF-expressing cell-based...

History of Photodynamic Therapy

The treatment of superficial nonmelanoma skin cancers and actinic keratoses (AKs) with lasers and light sources has recently entered a new era in dermatology with the advent of 20 5-aminolevulinic acid (ALA), a potent photosen-sitizer. This photosensitizer has demonstrated an effective ability to interact with lasers and light sources of appropriate wavelengths to selectively destroy the lesions in question. The PDT is a treatment modality which involves the use of a photosensitizer, a light source which fits the absorption spectrum of the pho-tosensitizer, and molecular oxygen, which when stimulated will destroy a specific target tissue. To be effective in the process, the photosensi-tizer must be able to preferentially penetrate more into the targeted tissue than the surrounding skin. ALA has been shown to be absorbed very well by actinically damaged skin, skin cancer cells, and by the pilosebaceous glands of the skin. The photosensitizer may be given exogenously or formed...

Liposomes in Cancer Immunotherapy

We constructed SSL containing immunodominant HLA-A2.1 restricted CTL epitopes derived from Mart-1 melanoma TAA (32,33) and we showed that antigenic peptides encapsulated in SSL display a significantly higher capacity to resist plasma enzymatic hydrolysis, to stimulate T-cell proliferation, and to induce specific CTL responses than their soluble (S) counterparts in vitro.

Cancer testes antigens CTAs

These are antigens found on a range of epithelial tumours, on placental tissues and in the testes. Although found on melanomas, these antigens are distinct from MMDAs in that the genes coding for these antigens, althought silent, are found in a wide variety of tissues. MAGE, BAGE and GAGE MAGE-1 (melanoma antigen) was the first human tumour antigen characterised. The gene is one of a family of at least 12 found on the long arm of chromosome 12. Although present, the genes are silent in a wide range of tissues. The exception is in the testes and placenta. In the testes the antigen is found on spermatocytes but since these cells do not express MHC Class I, MAGE-1 cannot be recognised by Tcyt cells. As well as being expressed on melanomas, antigens coded for by the MAGE family of genes are found on a proportion of breast and ovarian tumours, bladder, head and neck, gastric and oesophageal carcinomas, and sarcomas. BAGE and GAGE were identified using the same melanoma cell line as that...

Immunorecognition of Mart1 Epitopes by Specific CTL

The capacity of SSL-based antigenic preparations to sensitize target cells to the killing by CTL recognizing both M26-35 and M27-35, was tested (Fig. 1). NA-8 melanoma cells were preincubated in the presence of TAA or control peptides in soluble form or included in SSL. Maximal killing by specific CTL was detected upon incubation with 0.4 mg mL of soluble M27-35, but significantly lower (0.016-0.001 mg mL) concentrations of M26-35. Encapsulation into SSL did not modulate the targeting capacity of M26-35. In contrast, inclusion of M27-35 in SSL resulted in a fivefold improvement of their sensitizing capacity, with maximal killing observed at 0.08 mg mL TAA concentration.

MHC ClassI Independent Inhibition of NK Cells

Characterization of NK cells from the lymph-nodes of melanoma patients revealed enhanced expression of the CD66a (CEACAM1) inhibitory receptor, and this inhibitory receptor reduced NK cytotoxicity 82 . A 10-year follow-up study established that CEACAM1 expression correlated with worse survival of melanoma patients 83 . We have also demonstrated that the tumor microenvironment is involved in this inhibition of NK cytotoxicity. NK cells isolated from lymph nods infiltrated with CEACAM1-postive tumors express CEACAM1, whereas NK cells isolated from lymph nods of CEACAM1-negative tumors did not express CEACAM1. The homophilic interaction between CEACAM1 of NK cells and tumor cells lead to inhibition NK cytotoxicity 82 . We also investigated the molecular interactions of CEACAM1 with other CEACAM family members, and revealed that in addition to its homophilic engagement, CEACAM1 also recognize the CEA protein, which is found on many tumors 84 . Such interactions between CEA and CEACAM1...

Cell Technologies in Immunotherapy of Cancer

Tumor growth is accompanied by active immune reactions even on the early stages. Vaccine therapy implies the use of single antigen or combination of antigens, either with or without adjuvants, for the modulation of immune response. N.N. Petrov Institute of Oncology joined the field of antitumor vaccine therapy and related cellular technologies in 1998. The following activities are held (1) Optimization of the preparation of autologous and allogeneic antitumor vaccines and development of tumor cell culture bank for the experiments on allogeneic vaccination. (2) Clinical evaluation of autologous vaccine therapy by (a) bone marrow precursors of dendritic cells (DCs), which are loaded with tumor lysates (b) genetically modified tumor cells (c) intact tumor cells used in combination with various adjuvants (BCG, IL-1P, and IL-1P combined with low doses of cyclophosphamide) in patients with disseminated melanoma, metastatic kidney cancer, and colorectal cancer. Total 117 patients...

Results and Discussion

Due to genetic instability, tumor mass becomes highly heterogeneous during neoplastic progression. This heterogeneity is also applicable to the expression of specific antigens. Natural tumor cell selection leads to the changes in antigenic portrait and appears to be the main mechanism of evasion from the immune system surveillance. Tumor evolution is a serious obstacle for the efficacy of cancer vaccines (Moingeon 2001). If the treatment duration is long, or if the allogeneic vaccination is foreseen, long-living cultures are employed. In the latter case, cells cultivated ex vivo may significantly change the pattern of antigen expression. The divergence of antigenic properties of ex vivo cultivated cells and patients' metastases may seriously compromise the clinical efficiency of tumor vaccination. Our experiments have shown that melanoma cells may lose significant amount of antigens when cultivated in vitro for a long time. Indeed, while antigenic portrait was well preserved during...

Suppression of NK by Immuneregulatory Cells

Regulatory T cells (Treg) inhibition of NK functions was demonstrated in vitro and in vivo using anti-CD25 mAb against Tregs. In this model, CD25-depletion was also demonstrated to enhance NK activity in vitro 144, 145 , suggesting that Tregs potentially inhibit NK cells. In mice with FoxP3 defect NK proliferation is enhanced 146 , whereas adoptive Tregs transfer inhibit NK functions 147 . Similarly, human CD4+ CD25+ T cells were shown to impair NK cytotoxicity in vitro 148, 149 . Moreover, investigation of human gastrointestinal stromal tumor patients whom did not respond well to Gleevec demonstrated a higher Tregs numbers that were correlated with lower NK activity 150 . In agreement with these results, melanoma patients demonstrated increased NK functions if Tregs levels were low 151 . Tregs inhibition of NK activity was observed in a lung carcinoma model, where CD25-depletion enhanced the NK1.1+ cytotoxicity and resistance to tumor metastasis 152 . The rejection of allogeneic...

Most Tumor Antigens Are Not Unique to Tumor Cells

Several growth-factor receptors are expressed at significantly increased levels on tumor cells and can serve as tumor-associated antigens. For instance, a variety of tumor cells express the epidermal growth factor (EGF) receptor at levels 100 times greater than that in normal cells. An example of an over-expressed growth factor serving as a tumor-associated antigen is a transferrin growth factor, designated p97, which aids in the transport of iron into cells. Whereas normal cells express less than 8,000 molecules of p97 per cell, melanoma cells express 50,000-500,000 molecules of p97 per cell. The gene that encodes p97 has been cloned, and a recombinant vaccinia virus vaccine has been prepared that carries the cloned gene. When this vaccine was injected into mice, it induced both humoral and cell-mediated immune responses, which protected the mice against live melanoma cells ex

Manipulation of CoStimulatory Signals Can Enhance Immunity

Several research groups have demonstrated that tumor immunity can be enhanced by providing the co-stimulatory signal necessary for activation of CTL precursors (CTL-Ps). When mouse CTL-Ps are incubated with melanoma cells in vitro, antigen recognition occurs, but in the absence of a co-stimulatory signal, the CTL-Ps do not proliferate and differentiate into effector CTLs. However, when the melanoma cells are transfected with the gene that encodes the B7 ligand, then the CTL-Ps differentiate into effector CTLs. These findings offer the possibility that B7-transfected tumor cells might be used to induce a CTL response in vivo. For instance, when P. Linsley, L. Chen, and their colleagues injected melanoma-bearing mice with B7+ melanoma cells, the melanomas completely regressed in more than 40 of the mice. S. Townsend and J. Allison used a similar approach to vaccinate mice against malignant melanoma. Normal mice were first immunized with irradiated, B7-transfected melanoma cells and then...

Is There an Indication for Sentinel Lymphadenectomy

For more than a century, the value of elective lymph node dissection in patients with melanoma has been debated 3-6 . Now that the method of lymphatic mapping and sentinel lymphadenectomy offers the opportunity to reliably identify regional nodes with microscopic involvement 1 , this debate has shifted to the issue of ''selective'' lymph node dissection (i.e., completion removal of regional lymph node basin only if there is microscopic involvement of the sentinel node). Several retrospective studies have provided evidence suggesting that this ap proach has a beneficial effect. Patients who underwent node dissection at the stage of microscopic involvement had a survival advantage of approximately 20 , compared to patients who underwent dissection of clinically detected involved nodes 7 . In addition, a prospective randomized trial of the WHO Melanoma Program showed that patients with truncal melanoma, whose microscopically involved nodes were removed electively, fared significantly...

Interleukin10 in Cancer Models

B1 6 melanoma B1 6 and spontaneous melanoma lung metastases A375 human melanoma subcutaneous and pulmonary metastases IL-10 on adaptive immune effector cells remains to be convincingly demonstrated. It has been suggested that IL-10 may enhance susceptibility of target cells to NK cell lysis by reducing the surface expression of major histocompatibility antigens.32,33 IL-10 also increases the recruitment of macrophages and neutrophils34 and inhibits cancer growth by hampering tumor an-giogenesis and invasiveness through induction of metalloproteinase inhibitors.23,24,34,35 In a colon carcinoma mouse model, the transfection of tumor cells with IL-10 reduces the malignant potential in the context of a predominant Th2-type immune response.29 Transfection of mouse mammary adenocarcinoma34,36 ovarian carcinoma37 and melanoma38 cell lines with IL-10 elicits loss of tumorigenicity and increases immunogenicity accompanied by the induction of a strong lymphocyte and antibody-dependent immune...

Extent of Completion Lymph Node Dissection

A question that remains is Is it always necessary to clear the entire lymphatic basin after the removal of an involved sentinel node In the series reported so far, an additional 10-20 of involved nodes were found in the definitive regional node dissection specimen 1,20-22 . An interesting observation was made by the group from Moffitt Cancer Center in Tampa 23 . In a series of 88 patients, they found that all those with involved nodes beyond the positive sentinel node(s) had a primary melanoma thickness of greater than 1.5 mm in thickness. These results suggest that the disease in sentinel-node-positive patients with a primary tumor of less than 1.5 mm in thickness may be confined to the sentinel node and, therefore, that they may not necessarily require further lymph node dissection.

Sentinel Lymphadenectomy as a Selection Criterion for Adjuvant Treatment Strategies

In this respect, the relevance of the RT-PCR technique for the detection of tyrosinase or other specific markers for the presence of melanoma cells in the sentinel lymph node has to be elucidated 9,27 . In one report, RT-PCR positivity in histologically negative sentinel nodes was associated with a recurrence rate of 10.6 , compared to a rate of 2.3 in RT-PCR-negative patients 28 . This suggests that RT-PCR is of practical value in melanoma staging and therefore could be a selection criterion for entering patients in adjuvant treatment schedules.

Breast Cancer Technical Aspects

The procedure of lymphatic mapping and sentinel lymphadenectomy is more difficult to perform in patients with breast cancer than in patients with melanoma. The injection site, for instance, of the tracers cannot always be determined as precisely as it can for melanoma, due to deeper tumor localization. This may result in reduced sensitivity and selectivity of the method. Injections into 33 and around 34-36 the breast tumor have been applied some authors have even advocated injections into the subcutaneous tissue 37 or into the skin overlying the tumor 38 . The theoretical rationale for these latter two sites is based on the fact that the embryologic origin of the mammary gland is an appendix of the skin. An interesting observation in this regard is that a radiopharmaceutical injected around the tumor and blue dye injected intradermally both drained to the same axillary nodes 38 . The absence of internal mammary drainage after intra-cutaneous injection, however, casts doubt on the...

How do we give special respect for human embryos

Rather than making absolute judgments, we are comfortable with shades of gray. For example, it is against the law to exceed the speed limit, but we typically have more tolerance for breaking this law than for armed robbery. In research with human subjects, an important principle is beneficence (National Commission for the Protection of Human Subjects of Biomedical and Behavioral Research, 1979), to do no harm but for testing new chemotherapeutic agents for melanoma, we may ask research subjects to accept the risk of harm in exchange for the possibility of finding a new and more effective treatment for an aggressive cancer. In animal research, our society has accepted the principle that research can be conducted on animal subjects, but this research does not occur without restriction. By Federal regulations, all animal research is subject to review by an Institutional Animal Care and Use Committee (Office of Laboratory Animal Welfare, 2002). A central principle...

Inappropriate Expression of Class Ii Mhc Molecules Can Sensitize Autoreactive T Cells

Other evidence suggests that certain agents can induce some cells that should not express class II MHC molecules to express them. For example, the T-cell mitogen phytohemag-glutinin (PHA) has been shown to induce thyroid cells to express class II molecules. In vitro studies reveal that IFN-7 also induces increases in class II MHC molecules on a wide variety of cells, including pancreatic beta cells, intestinal epithelial cells, melanoma cells, and thyroid acinar cells. It was hypothesized that trauma or viral infection in an organ

Consent For Ultraviolet Light Type Bphototherapy

Uvb Narrowband Burned

The side effects to ultra-phototherapy B are, during treatment the psoriasis can sometimes get temporarily worse before getting better. The skin may itch and get red due to overexposure (sunburn). The long-term risk in developing skin cancer(s) from long-term exposure to UVB is unknown. Also, long-term exposure can cause freckling and loss of skin elasticity. During the course of therapy, your skin will be evaluated. The major light sources being used for the treatment of psoriasis are the BClear (Lumenis) and the Xtrac (PhotoMedix). Clinical trials with the XTrac, a 308-nm excimer laser, have shown significant clearing of psoriatic plaques. Feld-man et al. (Feldman et al. 2002) reported on a multicenter analysis with 124 patients at five centers. Seventy-two percent of patients demonstrated 75 or greater clearance in 6.2 treatments or less. Eighty-four percent achieved improvement of 75 or better after ten treatments. Other investigators also showed significant clearance using the...

Selection of Controls from the Study Base

Obtaining perfectly coherent case and control groups from the same study base guarantees that there will be no additional selection bias introduced in the case-control sampling beyond whatever selection bias may be inherent in the underlying cohort. The failure to do so, however, does not automatically produce selection bias it just introduces the possibility. In a cohort study, the ultimate purpose of the unexposed group is to estimate the disease risk of the exposed group absent exposure. In a case-control study, the purpose of the controls is to generate an accurate estimate of the exposure prevalence in the study base that gave rise to the cases. Given this goal, by good fortune or careful planning, a control group that is not coherent with the cases may nevertheless generate a valid estimate of exposure prevalence in the study base that gave rise to the cases. If, for example, the exposure of interest in a case-control study of melanoma among women were natural hair color...

Medical Terminology Case Studies - Skin


K.B., a 32-year-old fitness instructor, had noticed a tiny hard lump at the base of her left nostril while cleansing her face. The lesion had been present for about 2 months when she consulted a dermatologist. She had recently moved north from Florida, where she had worked as a lifeguard. She thought the lump might have been triggered by the regular tanning salon sessions she had used to retain her tan because it did not resemble the acne pustules, blackheads, or resulting scars of her adolescent years. Although dermabrasion had removed the obvious acne scars and left several areas of dense skin, this lump was brown-pigmented and different. K.B. was afraid it might be a malignant melanoma. On examination, the dermatologist noted a small pearly-white nodule at the lower portion of the left ala (outer flared portion of the nostril). There were no other lesions on her face or neck. _ 3. Which skin cancer is an overgrowth of pigment-producing epidermal cells d. melanoma

MDA5 Recognizes Poly IC

MDA-5 is structurally related to RIG-I, as it also contains two CARD domains and a helicase domain. MDA-5 was originally identified as a type I IFN-induc-ible molecule mediating cell cycle arrest and apoptosis in melanoma cells (hence the name melanoma differentiation antigen 5) (Kang et al. 2002, 2004 Kovacsovics et al. 2002). A first indication of a role for MDA-5 in virus recognition came from the observation that a paramyxoviral protein that mediated immune evasion bound to MDA-5 (Andrejeva et al. 2004). In overexpression experiments, MDA-5 was shown to bind poly I C, and enhanced the interferon response to poly I C as well as several viruses. Conversely, siRNA mediated knock-down blocked type I IFN induction in response to these stimuli (Yoneyama et al. 2005). MDA-5 was then shown to play an essential role in the detection of Picornaviruses such as encephalomyocarditis virus (EMCV) or Theiler's virus (Gitlin et al. 2006 Kato et al. 2006). In addition, mice deficient in MDA-5 were...

Purposes Of Lymphoscintigraphy Preparations And Injection Technique

The purpose of lymphoscintigraphy for lymphatic mapping is to demonstrate the lymphatic drainage pathway of the neoplasm to be more precise to indicate the drainage basin, to determine the number of lymph nodes that are on a direct drainage pathway, to differentiate these first-tier nodes from subsequent nodes, and to locate sentinel nodes outside the usual nodal basins. The request form for lymphoscintigraphy should describe the disease, its location, and prior management. Relevant parts of the medical history of the patient should be mentioned. For instance, prior inguinal hernia surgery may prevent drainage of a lower abdominal wall melanoma to that groin. The purpose of the study is stated on the form. The surgeon should inform the patient of the reason for the scintigra-phy and outline how it is done so that the patient knows what to expect.

Expanding DC In Vivo With Flt3L Followed by Administration of a Tumor Associated Antigen and TLR Agonist

Maintenance of the B16 melanoma B16 cells are maintained in RPMI 1640 supplemented with 10 FBS, 5 mM glutamine, penicillin, and streptomycin. B16 cells grow in an adherent fashion in tissue culture flasks and need to be passaged at a 1 10 ratio approximately twice per week (see Notes 4 and 5). As B16 is 3. Injection of B16 melanoma cells 3.2. Vaccination of B16 Melanoma-Bearing Mice With a Mixture of TRP2 Peptide and CpG Following DC Expansion With Flt3-L

Intramedullary Neoplasms

Dumbbell Lesions Neurofibromatosis

Noma account for the majority of the cord lesions. These are more common in the thoracic spinal cord and are atypical in the lumbar area. They may expand the cord, show contrast enhancement, and not uncommonly show areas of central necrosis. High T2 signal is common, and increased T1 signal may be seen with melanoma due to hemorrhage. Spinal leptomeningeal metastases or drop metastases are neoplastic deposits in the subarachnoid space more commonly seen in the lumbar spine where they are frequently visualized in the dependent portion of the thecal sac, hence the term drop metastases. Leptomeningeal metastatic disease accounts for the majority of malignant extramedullary, intradural masses. These include CNS and non-CNS etiologies. Primary CNS neoplasms include medulloblastomas, ependymomas, glioblastomas, ana-plastic astrocytomas, choroid plexus tumors, and pineal gland tumors such as germinomas, pineocytomas, and pineoblastomas. Systemic malignancies such as lung, breast, and...

Tie2 Expressing Monocytes Mediated Immunity Escape

Wolfel, T., Klehmann, E., Muller, C., Schutt, K. H., Meyer zum Buschenfelde, K. H., and Knuth, A. Lysis of human melanoma cells by autologous cytolytic T cell clones. Identification of human histocompatibility leukocyte antigen A2 as a restriction element for three different antigens. J Exp Med, 170 797-810, 1989. 4. Marincola, F. M., Rivoltini, L., Salgaller, M. L., Player, M., and Rosenberg, S. A. Differential anti-MART-1 MelanA CTL activity in peripheral blood of HLA-A2 melanoma patients in comparison to healthy donors evidence for in vivo priming by tumor cells. J Immunother, 19 266-277, 1996. 26. Minev, B. R. Melanoma vaccines. Semin Oncol, 29 479-493, 2002. 28. Parmiani, G., Castelli, C., Rivoltini, L., Casati, C., Tully, G. A., Novellino, L., Patuzzo, A., Tosi, D., Anichini, A., and Santinami, M. Immunotherapy of melanoma. Sem Cancer Biol, 13 391-400, 2003. 59. Panelli, M. C., Wang, E., Phan, G., Puhlman, M., Miller, L., Ohnmacht, G. A., Klein, H., and Marincola, F. M. Genetic...

Leptomeningeal Spread Sclc

The incidence of LM from melanoma is about 23 .4 The exact incidence in genitourinary (GU) cancers (renal, bladder and prostate) is not reported but 6 of all LM patients have GU malignancies, and the incidence rate may be increasing.12'32'74'115'134 Gastrointestinal (GI) cancers were once believed to be a major cause of LM but are now rarely encountered.38 LM from gynecological cancers (ovarian, cervical, and fallopian tube adenocarcinoma) and head and neck cancers are also seen.1'7'95'128 Rare solid tumors that metastasize to the leptomeninges include thyroid cancer,10 retinoblastoma,55 neuroblastoma,65 neuroendocrine tumors,36 carcinoid,79 sarcoma,62'134 and squamous cell of the skin or larynx.11,120'140

The Immunological Synapse

Lipid Raft Detection Snom

Fig. 7.1 Measurement of MHC I homoasso-ciation on OCM-3 uveal melanoma cells. The solid line denotes the distribution of FRET efficiency values between MHC I molecules measured by flow cytometry on a cell-by-cell basis. MHC I was targeted by Cy3- and Cy5-conjugated L368 mAbs specific for p2-micro-globulin (p2m, light chain of MHC I). The Fig. 7.1 Measurement of MHC I homoasso-ciation on OCM-3 uveal melanoma cells. The solid line denotes the distribution of FRET efficiency values between MHC I molecules measured by flow cytometry on a cell-by-cell basis. MHC I was targeted by Cy3- and Cy5-conjugated L368 mAbs specific for p2-micro-globulin (p2m, light chain of MHC I). The Clustered cell-surface distribution and anomalous diffusion of MHC II glyco-proteins as well as their heteroassociation with MHC I were also reported in numerous cell types, including APCs 77,87-89 . Atomic force and electron microscopic data showed that MHC II molecules form homoclusters not only on the nanometer...

Classical and Non Classical HLA Class I Antigens and NKCell Activating Ligands in Malignant Lesions

More recently, the non-classical HLA class I antigens, HLA-G, have been unexpectedly reported to be expressed both on cell lines and in surgically removed malignant lesions 48 . Although results in the literature conflict, there is a general agreement that malignant transformation of cells may be associated with the appearance of HLA-G. The frequency of this phenomenon markedly varies among different tumor types, as it ranges from a maximum of about 40 in cutaneous lymphoma, clear cell renal carcinoma and ovarian carcinoma to a lack of detection in uveal melanoma and laryngeal carcinoma. It is also noteworthy that the frequency of HLA-G expression is lower in cell lines than in surgically removed malignant lesions. Although one cannot exclude that this difference is due to technical reasons, such as differences in the sensitivity of the assays used or mis-scoring of HLA-G-bearing macrophages as tumor cells, we favour the possibility that this difference reflects biological...

Technique Of Intraoperative Gammaray Detection Probe Localization

Rheumatoid Lymph Nodes

The development of the hand-held, high-resolution gamma-ray detection probe, which provides real-time, rapid counting in the surgery room, has allowed the development of a much simpler technique for localizing bone lesions for biopsy. The gamma probe has been quite successfully used intraoperatively for ra-dioguided identification of sentinel lymph nodes in melanoma and breast cancer 13,14 . The gamma probe has been subsequently adapted for use at the H. Lee Moffitt Cancer Center for intraoperative localization of areas of increased uptake of a 99mTc-labeled tracer in ribs and the sternum to guide the surgeon in the open biopsy of suspicious bones 3 .

Clinical Aspects Immunity

AIDS is acquired by infection with HIV, which attacks certain T cells. These cells have a specific surface attachment site, the CD4 receptor, for the virus. HIV is spread by sexual contact, use of contaminated needles, blood transfusions, and passage from an infected mother to a fetus. It leaves the host susceptible to opportunistic infections such as pneumonia caused by the protozoon Pneumocystis carinii thrush, a fungal infection of the mouth caused by Candida albicans and infection with Cryptosporidium, a protozoon that causes cramps and diarrhea. It also predisposes to Kaposi sarcoma, a once-rare form of skin cancer. It may also induce autoimmunity or attack the nervous system.

Animal Model Approach to Testing

Host defense to melanoma is mediated by NK cells, macrophages, and some T cells (Parhar and Lala, 1987). Using an artificial tumor metastasis model in syngeneic C57BL 6 mice, mice are challenged with different number oftumor cells (Fidler etal., 1978). Because cells are injected into the tail vein, the cells lodge in the lung, which is the first capillary bed encountered. Two endpoints are usually measured. The proliferation rate of cells in the lung can be measured by injecting fluorodeoxyuridine, which blocks de novo nucleotide synthesis, followed by 125I iododeoxyuridine. The latter nucleotide is incorporated into the DNA of newly divided cells (White, 1992). As a second endpoint, the number of tumor nodules per lung can be determined. Tumor cells form characteristic black nodules on a background of white or yellow lung tissue. Up to 200-250 nodules can be enumerated in each lung section. Immune responses to the melanoma tumor are time dependent....

Use Of Immunohistochemistry

Nerves Groin Area

It is absolutely essential that all lymph nodes be examined by immunohistology using antibodies to S-100 protein and HMB-45, unless the node contains overt tumor on gross inspection or review of H& E-stained slides. The antibody MART-1 (melanoma-associated antigen recognized by T lymphocytes) (Melan-A) can be substituted for HMB-45. Immunohistology will always increase the frequency of sentinel lymph nodes found to contain tumor. The proportion of sentinel nodes that require immunohistology to identify occult tumor decreases as pathologists gain experience in evaluating sentinel nodes. This is the pathologist's equivalent of the surgeon's learning curve 24 . S-100 protein is a highly robust marker for melanoma cells, staining virtually 100 of melanomas 14,16 . We look for epithelioid, oval, or spindle-shaped cells (usually located in the subcapsular sinus) that show S-100 protein positivity in both the cytoplasm and the nucleus (Figs. 2a-2c). There are other cells within the lymph...

New Approaches for Monitoring CTL Activity in Clinical Trials

Release Assay

We have developed a modification of the ELISPOT assay that measures Gran-zyme B (GrB) release from cytotoxic T lymphocytes (CTLs). The GrB ELISPOT assay is a superior alternative to the 51Cr-release assay since it is significantly more sensitive and provides an estimation of cytotoxic effector cell frequency. Additionally, unlike the IFN-y ELISPOT assay, the GrB ELISPOT directly measures the release of a cytolytic protein. We report that the GrB ELISPOT can be utilized to measure ex vivo antigen-specific cytotoxic-ity of peripheral blood mononuclear cells (PBMCs) from cancer patients vaccinated with a peptide-based cancer vaccine. We compare the reactivity of patients' PBMCs in the GrB ELISPOT, with reactivity in the tetramer, IFN-y ELISPOT and chromium (51Cr)-release assays. Differences in immune response over all assays tested were found between patients, and four response patterns were observed. Reactivity in the GrB ELISPOT was more closely associated with cytotoxicity...

Intraoperative Use Of A Gamma Probe

Intraoperative Gamma Probe

Although the estimated position of each sentinel node will ideally have been marked on the overlying skin at the time of preoperative lymphoscintigraphy, it is useful to confirm its location with a gamma probe before making a skin incision. This is particularly valuable when the position of the patient on the operating table differs from the position they were in when the lymphoscintigram was obtained. The surgeon may occasionally decide, for example, to perform an axillary sentinel lymphadenectomy with the patient in a lateral position so that the subsequent wide excision of a primary melanoma site on the back can be performed without having to reposition and redrape the patient, whereas the preoperative lymphoscintigram was performed in routine fashion with the patient in the supine position, with the arm extended at a 90 angle to the body. Checking the position of the sentinel node with a gamma probe prior to skin incision can also be very useful in situations where anatomical...

Physiological functions

Yoo et al. (1997) studied the effect of bLF and bLFcin on inhibition of metastasis in murine tumor cells, B16-BL6 melanoma and L5178Y-ML25 lymphoma cells, using experimental and spontaneous metastasis models in syngeneic mice. Subcutaneous (s.c.) administration of apo-bLF (1 mg mouse) and bLFcin (0.5 mg mouse) 1 day after tumor inoculation significantly inhibited liver and lung metastasis of L5178Y-ML25 cells. However, apo-hLF and holo-bLF at the dose of 1 mg mouse failed to inhibit tumor metastasis of L5178Y-ML25 cells. Similarly, the s.c. administration of apo-bLF as well as bLFcin, but not apo-hLF and holo-bLF, 1 day after tumor inoculation resulted in significant inhibition of lung metastasis of B16-BL6 cells in an experimental metastasis model. Furthermore, in vivo analysis for tumor-induced angiogenesis, both apo-bLF and bLFcin inhibited the number of tumor-induced blood vessels and suppressed tumor growth on day 8 after tumor inoculation. However, in a...

Choroid plexus tumors

Leptomeningeal metastases may also occur in a variety of childhood solid tumors including retinoblastoma,82 neuroblastoma,8386 rhabdomyosarcoma,87 melanoma,88'89 and Ewing's sarcoma.90 91 These metastases occur most commonly when advanced systemic disease is present, either at the time of initial diagnosis or when the primary tumor has recurred.

In Vitroactivated Lak And Til Cells

Melanocytes Photomicrograph

Photomicrographs of cultured normal melanocytes (top) and cultured cancerous melanoma cells (bottom) in the presence (left) and absence (right) of tumor necrosis factor (TNF-a). Note Photomicrographs of cultured normal melanocytes (top) and cultured cancerous melanoma cells (bottom) in the presence (left) and absence (right) of tumor necrosis factor (TNF-a). Note Tumors contain lymphocytes that have infiltrated the tumor and presumably are taking part in an antitumor response. By taking small biopsy samples of tumors, one can obtain a population of these lymphocytes and expand it in vitro with IL-2. These activated tumor-infiltrating lymphocytes are called TILs. Many TILs have a wide range of antitumor activity and appear to be indistinguishable from LAK cells. However, some TILs cells have specific cytolytic activity against their autologous tumor. These tumor-specific TILs are of interest because they have increased antitumor activity and require 100-fold lower levels of IL-2 for...

Interaction between IL10 and Antigen Presenting Cells

In several tumor models, IL-10 expressed within the tumors could drive rejection and elimination of the tumor, whereas the EBV expressed homologue, vIL-10 promoted tumor growth.58,116 In some models, IL-10 can promote the generation of nominally tolerogenic regulatory T cells.117 In this setting, tumor-infiltrating DCs (TIDCs) are largely immature, and are able to present tumor antigen, but they are refractory to stimulation with a combination of LPS, IFN-y, and anti-CD40 antibody. DC paralysis in this setting can be reversed by CpG plus anti-IL-10 R treatment. This combination, but not CpG alone, had a potent therapeutic antitumor effect and induce immune memory.118 In most tumor settings, IL-10 production has interestingly been associated with improved outcome in murine models and in predicting melanoma responsiveness to IL-10 therapy.119

The dosage is 100 mg twice daily Mycophenolate Mofetil Cell Cept

Gastrointestinal effects, including nausea, diarrhea, mucosal hemorrhage, and ulceration, can occur. Leukopenia and infections, especially by opportunistic organisms, are also potential problems. In the renal transplant population, there may be an increased risk for lymphoproliferative and non-melanoma skin carcinomas.

Immunotherapy for Cancer

The production of human interferons by genetically engineered bacteria has made large amounts of these substances available for the experimental treatment of cancer. Thus far, interferons have proven to be a useful addition to the treatment of particular forms of cancer, including some types of lymphomas, renal carcinoma, melanoma, Kaposi's sarcoma, and breast cancer. They have not, however, proved to be the magic bullet against cancer (a term coined by Paul Ehrlich) as had previously been hoped. The research team next identified a subpopulation of lymphocytes that had invaded solid tumors in mice. These tumor-infiltrating lymphocyte (TIL) cells were allowed to replicate in tissue culture, whereupon they were reintroduced into the mice with excellent results. Recently, the same techniques were used to treat an experimental group of people with metastatic melanoma, a cancer that claims the lives of 6,000 Americans annually. The patients were first given conventional chemotherapy and...

Sustainedrelease and infusional intrathecal chemotherapy

As mentioned previously, most available intrathecal agents have short pharmacologic half-lives (T Vi), which limit their efficacy. Prolonged drug exposure has been forwarded as a method to increase the cytotoxicity of such agents in CSF. It has been shown that some cell lines may be resistant in short exposure conditions, but demonstrate sensitivity with prolonged exposure. For example, in the human SO-M19 melanoma line, there was no inhibition of tritiated thymidine incorporation with short (2 hour) exposures despite concentrations of up to 4 ug ml ara-C, whereas low concentrations (1 ug ml) over a longer duration of 48 hours produced 90 inhibition of thymidine incorporation.7

The Tumor Microenvironment and the Induction and Function of Tumor Specific T Cells

Tumor Microenvironment Inflammatory

The tumor microenvironment regulates antitumor immunity not only at the level of induction of systemic T cell responses but also regulates the infiltration and function in situ of tumor specific effector T cells. Numerous studies have demonstrated that tumor-infiltrating T cells exert reduced functional activity after their re-isolation from tumor tissue. They fail to be activated by TCR+ anti-CD28 stimulation under conditions that fully activate peripheral blood T cells 46 . TIL from colorectal carcinoma and from melanoma are anergic, express low TCR, perforin and Fas-L 47 and are deficient in perforin-mediated cytolytic activity due to defective microtubule organizing center mobilization and lytic granule exocytosis 48 . This is due to a combination of regulatory mechanisms in the tumor microenvironment and the lack of essential proinflammatory cytokines. T cell anergy due to insufficient B7 costim-ulation, extrinsic suppression by regulatory cell populations, inhibition by ligands...

Tumor Microenvironment under the Magnifying Glass A New Challenge to Cancer Immunologists

The theory of immunological surveillance against cancer has been introduced half a century ago by McFarlane Burnet and Lewis Thomas. Despite extensive research to validate and generalize this concept, approaches to intervene in cancer by immunological means has led to more disappointments than successes, more doubts than convictions. Is this concept still valid If the question addresses the possibility that cancer cells induce protective immunity sufficient to enact tumor rejection, the answer is no. But what about manipulating innate and adaptive, cellular and soluble immune components to fight tumor progression and metastasis A pessimist's answer would allude to the ample evidence demonstrating that most malignant cells are non-immunogenic and do not become non-self' targets. The optimist will highlight cases of melanoma and non-Hodgkin's lymphoma that can be managed by active and passive immunotherapy. Realistically, however, one has to admit that approaches to induce specific...

Cytokines and chemokines chemotaxis

Isolation Monocytes Protocol

Chemotaxis is defined as the directional locomotion of cells sensing a gradient of the stimulus. Chemotaxis has been extensively studied with leukocytes that are 'professional migrants', but a variety of cell types including fibroblasts, melanoma cells, keratinocytes, and vascular endothelial cells exhibit directional locomotion in vitro. Two main techniques have been used to measure migration in vitro migration under agarose and chemotaxis across porous membranes. While the former approach may more closely resemble the in vivo conditions, the latter is easier to quantitate and allows analysis of directional versus random locomotion. We will therefore focus on the description of migration through a porous membrane. Both a classic modified Boyden chamber assay (7) and a micromethod (8, 9) will be described. Protocol 1 describes the use of a micro-method for assessing leukocyte chemotaxis. A schematic representation of the micro chemotactic chamber is shown in Figure 1.

Modulation of Tumor Recognition by NK Cells

NKG2D is involved in the prevention of tumor spread and tumor formation in mice 43 . Its human ligands, ULBPs, MICA and MICB proteins are expressed by several tumors. Numerous studies reported on the frequent expression of MICA and MICB, both on primary lesions and on metastases of melanomas, leukemic cells, gliomas and various carcinomas 61 . The ULBPs were also expressed by the same tumors, albeit at a low frequencies 61 . NKG2D-ligands are specifically induced by genotoxic stress, which contributes significantly also for tumor formation 62 . Recently, the BCR ABL human oncogene was shown to induce MICA expression in leukemia cells 63 . With this regard it is important to mention that chronic contact with NKG2D-ligand expressing target cells might be unsafe to the NK cells as it can reduce the expression of NKG2D and other receptors and consequently reduce cytotoxicity 64 . Other proteins which can impair NK activity are also secreted in the tumor microenvironment. For example,...

Pollution Effects of Chemicals

The effect of the punctuated ozone layer on animals is yet to be fully understood. It is known that the high energy level of UV radiation can damage biological molecules, including the genetic material deoxyribonucleic acid (DNA), causing mutation. In small quantities, UV light helps the skin of humans and many animals produce vitamin D, and causes tanning. However, in large doses, UV light causes sunburn and premature aging of skin, skin cancer, and cataracts, a condition in which the lens of the eye becomes cloudy. Due to UV radiation's ability to penetrate, even animals covered

The Presence of NK Cells Inside Tumors

One of the major difficulties in the detection of NK cells in tumors is the lack of a single NK specific marker. The classical NK marker in human (CD56) is expressed also on NKT cells, and is absent in mice. To identify NK cells in the mouse the DX5 or NK1.1 mAb are used and these two receptors are also not specific to NK cells only, or are expressed only in certain mouse strains 4 . The only NK specific marker known to date is the NKp46 in human, or NCR1 in mouse. Indeed, NK cells were recently identified in Renal-Cell-Carcinoma patients by using antibodies against the NKp46 receptor 14 . Developing additional antibodies to the human NKp46 and the mice NCR1 will facilitate easier detection of NK cells in the future. Several studies reported on the presence and functions of NK in tumors 15 . For example, intratumor NK cells were found in squamous cell lung cancer, gastric and colorectal carcinomas 16-18 and these studies suggested that the presence of NK cells inside the tumor is an...

Miscellaneous Tumors of the Sellar Region

An additional spectrum of rare and peculiar tumors of the parasellar region are the subjects of isolated case reports. These include fibrosarcoma (99), gloman-gioma (100), cavernous hemangioma (93,101-103), primary lymphoma (104), primary melanoma (105,106), myxoma (107), osteogenic sarcoma (108), and pseudotumor (109).

Blue Dye Mapping The Concept

Sentinel Lymph Node Mapping

Radiological lymphography (see Fig. 4 in Chapter 2). In the mid-1980s, Morton and his colleagues at the John Wayne Cancer Institute began studies using blue dye which were destined to provide new insights into the clinical relevance of lymphatic drainage pathways for malignancies such as melanoma and breast cancer. First in animal experiments and then in man, it was shown that blue dye traveled rapidly from any given injection site in the skin to a ''sentinel'' lymph node 13,14 . It was postulated that tumor cells would travel along the same lymphatic channels and lodge first in a sentinel node. If a sentinel node was found to be free of micrometastatic disease, it could therefore be assumed that the entire node field was likely to be disease-free and that full regional lymph node dissection was unnecessary. The accuracy of sentinel node status as an indicator of regional lymph node status has since been confirmed by several other histological studies involving sentinel...

NK Cells and Macrophages Are Important in Tumor Recognition

Excise Tumor Dendritic Cells

One of the best-studied tumor-immunity models is melanoma. Melanoma has evolved as a model system for several reasons. First of all and paradoxically, most human cancers are difficult to establish in tissue culture, making it difficult to develop in vitro systems for experimental manipulation. Melanoma is relatively easy to adapt to tissue culture, which has led to the identification of several tumor-associated antigens, some of which are unique to melanoma (see Table 22-5). These observations are enhanced by the ability to create cDNA libraries (see Chapter 23) from tumor cells. The cDNAs can be transfected into target cells expressing the appropriate MHC molecules and then used as targets for CTL-mediated killing. Once CTL reactivity is recognized, the transfected cDNA can be isolated and identified as a potential tumor antigen. The ability to isolate genes encoding tumor-associated antigens provides us with the opportunity to use these proteins as immunogens for the induction of...