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      Tracing B cell development in human germinal centres by molecular analysis of single cells picked from histological sections.

      The EMBO Journal
      Adult, Amino Acid Sequence, B-Lymphocytes, cytology, Base Sequence, Cell Differentiation, Child, DNA Primers, chemistry, Gene Rearrangement, B-Lymphocyte, Genes, Immunoglobulin, Humans, Immunoglobulin Heavy Chains, genetics, Immunoglobulin Variable Region, Lymph Nodes, Micromanipulation, Molecular Sequence Data, Mutation, Sequence Alignment, Sequence Homology, Nucleic Acid, Spleen

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          Abstract

          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.

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