Induction of Immune Tolerance in Neonatal Mice

As previously mentioned fetal and newborn mammals have immature immune systems that do not yet have the capacity to differentiate self and nonself. This lack of maturity can be harnessed to our advantage when working with antigens, which are naturally found along with closely related (cross-reacting) substances. This technique does not guarantee success but can swing the odds in favor of the researcher producing hybridomas with the desired specificity. Situations where this methodology is used include work on viruses, bacteria, and fungi, where there may be many shared epitopes between the organism of interest and closely related species. The cross-reacting antigen used for the technique is a whole preparation of the closely related species. This technique causes the suppression of an immune response to shared epitopes, which may swamp the immune response and favors specific immunity to epitopes found only on the species of interest.

1. Obtain a "time mated" female Balb/c mouse 10 d into the pregnancy and maintain in standard cage used for mouse breeding.

2. Observe until litter are born.

3. Inject the neonates daily on d 1-5 after birth with 0.05 mL 0.5 mg/ mL of crossreacting antigen into the neck scruff. (The neonatal immune systems are maturing at this time and because the cross-reacting antigen is present they will adopt these proteins as "self" and lose the ability to mount an immune response to them.)

4. Immunize three of the animals with specific antigen of interest when they are 6 wk old using the standard monoclonal immunization protocol provided previously.

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