Differentiation of B lymphocytes

vsem vsem%compsvc.FRC.MASS.EDU at MADMAX.FRC.MASS.EDU
Tue Mar 7 20:09:52 EST 1995

   Hello everyone!  I have a little immunological riddle.

   A certain Dr. in interested in using differntiation markers to study
hematopaiesis.  In particular of the development and differentiation of B
lymphocytes.  The Dr. begin to develope momoclonal Abs (mab) to
differentiation markers present on cells in the bone marrow.  So, some bone
marrow cell extracts were harvested.  The Dr. then harvested splenic B cells
from the immunized mice and set about to make hybridomas.  Many hybridomas
were abtained.
   Q:  What is the most efficient way to screen these hybridomas?  How would
you recommed that the Dr. initially screen her hybridomas for Mabs that will
recognize differentiation markers present on the cells?

   Q:  Having taken your advice on how to detect Mabs, the Dr. now has 5
cadidate mabs that reacted with rabbit bone marrow(bm) and that of these 5 the
Dr. wanted to find out which will detect only cells of the myloid lineage and
which will only detect only lymphoid lineage?  Using only immunohistochenical
techniques on rabbit tissues how would the Dr. do this?

   Q:  The Dr. found that 1 mabs recognizes cells of the myloid lineage and 2
of mabs of the lymphoid lineage.  What kind of experiment can you design to
tell the Dr. if thee 2 lymphocyte-specific Abs reconize cells of the
T-lineage, B- lineage, or both?

   Q:  What if it turns out that one of the lymphocyte-specific Abs recognizes
a cell type that was not clear;ly a differentiate T or B cell type.  The Dr.
suspects that perhaps this mabs is recognizing a differentiation markers
present on a lymphocyte progenitor cell.  What kind of experiment could you
use which will unequivocally prove this?

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