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Recall that B-cells from neonatal spleen and adult marrow are susceptible to tolerance induction via deletion or editing strategies we discussed in class. However, if only early B-cell differentiation stages are susceptible to tolerance induction, consider this potential problem: Memory responses and long-lived plasma cells contain somatically mutated antibodies, that were selected for higher affinity. These mutations occur during primary responses, yielding altered CDR's. Since the mutations occur in functionally "mature" B-cells, how is any autoreactivity generated by these mutations eliminated from the memory B-cell pool?
Outline an experiment to establish whether mature B-cells that are the immediate progenitors of memory cells transit a "second window" of tolerance susceptibility while undergoing somatic mutation during initial antigen challenge. Be sure to indicate:
• how you would identify/isolate arising memory clones or their progenitors;
• how you would assess tolerance susceptibility and/or induction
• what your control and comparison groups would be.• Describe the result you would expect it there is such a second window of tolerance susceptibility.
What effect do you think EBV would have on B cell function. What phenotypes would you expect in the F2, and in what actual numbers (not ratios) will you expect to find out them.
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If our goal was to measure the Absorbance of DCPIP (blue substance), why did we set the spectrophotometer to measure Absorbance at ~620nm, the orange part of the visible spectrum?
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Enzymes greatly decrease the activation energy. Describe two of the ways they do this and be specific in terms of chemical mechanisms.
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