Tracing B cell development in human germinal centres by molecular analysis of single cells picked from histological sections.
Germinal centres are areas of intense B lymphocyte proliferation inside primary B cell follicles in spleen and lymph nodes. Rearranged V genes from single human B cells, isolated from histological sections of two such structures by micromanipulation, were amplified and sequenced. Cells from the follicular mantle were clonally diverse and largely expressed germline V genes. Germinal centres were dominated by a few large B cell clones dispersed throughout these structures and exhibiting intraclonal diversity by ongoing somatic hypermutation. Pronounced counterselection of replacement mutations seen in one of the germinal centres may indicate a late phase of the germinal centre reaction. A polyclonal population of activated B cells expressing unmutated antibodies in the dark zone of the other germinal centre may represent the initial founder cells.
Küppers
R
R
Institute for Genetics, University of Cologne, Germany.
Zhao
M
M
Hansmann
M L
ML
Rajewsky
K
K
D000328:Adult; D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000166:cytology*; D001483:Base Sequence; D002454:Cell Differentiation; D002648:Child; D017931:DNA Primers / Q000737:chemistry; D015322:Gene Rearrangement, B-Lymphocyte; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008198:Lymph Nodes / Q000166:cytology*; D008846:Micromanipulation; D008969:Molecular Sequence Data; D009154:Mutation; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013154:Spleen / Q000166:cytology*
D017931:DNA Primers; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region