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In vitro studies on the structure and rearrangement process at the human immunoglobulin lambda locus
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In vitro studies on the structure and rearrangement process at the human immunoglobulin lambda locus
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In vitro studies on the structure and rearrangement process at the human immunoglobulin lambda locus
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Dissertation
In vitro studies on the structure and rearrangement process at the human immunoglobulin lambda locus
1995
Overview
The efficiency of the adaptive immune system is dependent on the diverse repertoire of antigen receptors expressed on the surface of mature B lymphocytes, surface immunoglobulins (sIg). For a B lymphocyte to produce a virtually unique sIg receptor, it relies on a process of site-specific recombination to create the genes coding for this receptor. Rather than being specified in the germline, these are created through the somatic assembly of individual members of various germline gene segment families (V, D and J gene segments), a process commonly referred to as V(D)J recombination. This thesis describes studies on the control and regulation of V(D)J recombination at one of the human Ig loci, the Ig$\\lambda$ light chain locus. Chapter 2 describes a clonal cell culture system derived from a human B lymphoma cell line. This system, established from a single sIg$\\sp+$ B cell, is shown to exhibit sIg variation as a result of continuous rearrangement activity at the Ig$\\lambda$ locus. A correlation between Ig$\\lambda$ gene rearrangement, upregulated RAG expression and C$\\lambda$ germline transcription is observed. In chapter 3 the contribution of somatic hypermutation to this sIg variation is investigated and found to be undetectable. Chapter 4 shows that the process of Ig$\\lambda$ gene rearrangement is quite conservative, with an absence of N nucleotide insertions as well as a complete lack of deletions in the 5$\\sp\\prime$ end of the J$\\lambda$ gene segment, restricting junctional diversity to infrequent deletions in the 3$\\sp\\prime$ end of the V$\\lambda$ gene segment. This chapter also describes an unusual sIg receptor which lacks the majority of the IgL constant region, sIg$\\Delta$CL. Chapter 4 also provides an example of a functional V$\\lambda$ gene segment which is infrequently rearranged compared to other V$\\lambda$ gene segments. It is shown that, compared to a frequently rearranged V$\\lambda$ gene segment, this V$\\lambda$ gene segment is transcribed at significantly reduced levels in the germline. This is likely due to a mutated octamer motif in the promoter of this V$\\lambda$ gene segment. In chapter 5 this V$\\lambda$ gene segment is described in some detail and shown to belong to a previously unidentified V$\\lambda$ gene family, V$\\lambda$X.
