Transcript expression of two Iglambda rearrangements and RAG-1/RAG-2 in a mature human B cell producing IgMlambda islet cell autoantibody

Abstract

A human B cell clone, EBV-MB91, producing IgMlambda islet cell autoantibody (ICA), obtained by Epstein-Barr virus (EBV) transformation of peripheral CD5- surface Ig+ B cells from a Type 1 diabetic child, and an EBV-MB91-derived hetrohybridoma, HY-MB91, were analyzed for rearranged Ig genes. Both EBV-MB91 and HY-MB91 contained and expressed a unique IgH chain rearrangement (unmutated VH5-51-D6-19-JH5) but contained and expressed two Iglambda chain rearrangements: (i) Vlambda1-4-Jlambda3-Clambda3, which encoded the Iglambda chains (pI, 8.0) of IgMlambda-ICA, showing few mutations but consistent with Ag-driven selection according to the multinomial probability model; and (ii) Vlambda4-1-Jlambda3-Clambda3, with more mutations but inconsistent with antigen-driven selection and involving stop codons that precluded Iglambda synthesis. HY-MB91 showed a progressive loss of IgMlambda-ICA secretion, which was coupled with transcripts of the aberrant Vlambda4-1-Jlambda3-Clambda3 predominating (1.7-fold) over those of Vlambda1-4-Jlambda3-Clambda3. EBV-MB91 also showed the loss of IgMlambda-ICA secretion, associated with cell death. RAG-1 and RAG-2 transcripts occurred in EBV-MB91 but not in HY-MB91, indicating that the former but not the latter might have been able to exhibit V(D)J recombinase activity. Data show that a mature nonmalignant human B cell clone producing IgMlambda-ICA can express RAG-1/RAG-2 transcripts. That the aberrant Vlambda4-1-Jlambda3-Clambda3 was a nonproductive rearrangement occurring at the pre-B cell stage cannot be excluded. However, the hypothetical possibility that one of the two rearrangements corresponded to a secondary rearrangement occurring in the mature B cell represented by the EBV-MB91 clone might also be considered and is discussed.