The role of the pre-B-cell receptor in regulating immunoglobulin gene rearrangement.
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The production of membrane-bound immunoglobulin (Ig) during the B-cell maturation results from Ig gene rearrangement, which is highly regulated. The heavy chain (H) rearrangement happens first and stops once a heavy chain protein is made. Light chain (L) gene rearrangement then begins.The signals that mediate the down-regulation of H chain rearrangement and the up-regulation of L chain rearrangement are mediated by the pre-B-cell receptor.Our hypothesis is that the signals to down-regulate the H chain rearrangement and the signals to up-regulate the L chain rearrangement are distinct.We are working with transgenic mice cells that have a mouse/human chimeric gene; a mouse variable region (VDJ) attached to the human g1 constant region (HUG1).The transgene HUG heavy chain (as part of pre-B-cell receptor) sends out signals to stop H chain rearrangement but does not start L chain rearrangement.Studying these signals is the main objective of this research project. Hybridomas cell lines, HYDD 1 through 16, made from 35 day old HUG transgenic mice, were thawed and screened for secreted antibodies using Enzyme-Linked Immunosorbent Assay (ELISA).The results of the ELISA assay show some changes in Ig expression from the assay that was done in the lab in 1996.The cells were also screened for intracellular antibody proteins using the Western Blotting.Future experiments include: Western Blotting to look for Ig protein chains, and crosslinking of HUG and mouse m, followed by Western Blotting to look for increase in phosphorylated tyrosine residues.