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PMID: 9153293 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Analysis of the human VH gene repertoire. Differential effects of selection and somatic hypermutation on human peripheral CD5(+)/IgM+ and CD5(-)/IgM+ B cells.

The Journal of clinical investigation ·Vol. 99 ·No. 10 ·1997-05-15 ·Pages 2488-501

Brezinschek HP, Foster SJ, Brezinschek RI, Dörner T, Domiati-Saad R, Lipsky PE

Abstract

To analyze the immunoglobulin repertoire of human IgM+ B cells and the CD5(+) and CD5(-) subsets, individual CD19(+)/ IgM+/CD5(+) or CD5(-) B cells were sorted and non-productive as well as productive VH gene rearrangements were amplified from genomic DNA and sequenced. In both subsets, the VH3 family was overrepresented largely as a result of preferential usage of a small number of specific individual family members. In the CD5(+) B cell subset, all other VH families were found at a frequency expected from random usage, whereas in the CD5(-) population, VH4 appeared to be overrepresented in the nonproductive repertoire, and also negatively selected since it was found significantly less often in the productive compared to the nonproductive repertoire; the VH1 family was significantly diminished in the productive rearrangements of CD5(-) B cells. 3-23/DP-47 was the most frequently used VH gene segment and was found significantly more often than expected from random usage in productive rearrangements of both CD5(+) and CD5(-) B cells. Evidence for selection based on the D segment and the JH gene usage was noted in CD5(+) B cells. No differences were found between the B cell subsets in CDR3 length, the number of N-nucleotides or evidence of exonuclease activity. Somatically hypermutated VHDJH rearrangements were significantly more frequent and extensive in CD5(-) compared to CD5(+) IgM+ B cells, indicating that IgM+ memory B cells were more frequent in the CD5(-) B cell population. Of note, the frequency of specific VH genes in the mutated population differed from that in the nonmutated population, suggesting that antigen stimulation imposed additional biases on the repertoire of IgM+ B cells. These results indicate that the expressed repertoire of IgM+ B cell subsets is shaped by recombinational bias, as well as selection before and after antigen exposure. Moreover, the influences on the repertoires of CD5(+) and CD5(-) B cells are significantly different, suggesting that human peripheral blood CD5(+) and CD5(-) B cells represent different B cell lineages, with similarities to murine B-1a and B-2 subsets, respectively.

MeSH Terms
Adult Amino Acid Sequence Animals Antigens, CD/blood Antigens, CD19/blood B-Lymphocyte Subsets/immunology B-Lymphocytes/immunology CD5 Antigens/blood Gene Rearrangement Genes, Immunoglobulin HLA-D Antigens/genetics Humans Immunoglobulin Heavy Chains/genetics Immunoglobulin M/blood Immunoglobulin Variable Region/genetics Male Mice Middle Aged Molecular Sequence Data Polymerase Chain Reaction
Chemicals
Antigens, CD Antigens, CD19 CD5 Antigens HLA-D Antigens Immunoglobulin Heavy Chains Immunoglobulin M Immunoglobulin Variable Region
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Brezinschek H P
Department of Internal Medicine and Harold C. Simmons Arthritis Research Center, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75235, USA.
Foster S J
Brezinschek R I
Dörner T
Domiati-Saad R
Lipsky P E
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Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI119433
Published
1997-05-15
Pages
2488-501
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC508090
Subset
IM
Grants
PHS HHS · A131229 · United States
Databases
GENBANK
AF022810, Z80363, Z80364, Z80365, Z80366, Z80367, Z80368, Z80369, Z80370, Z80371, Z80372, Z80373, Z80374, Z80375, Z80376, Z80377, Z80378, Z80379, Z80380, Z80381, Z80382, Z80383, Z80384, Z80385, Z80386, Z80387, Z80388, Z80389, Z80390, Z80391
External Links
PubMed source DOI: 10.1172/JCI119433 PMC Full Text View journal details
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