TRACING B-CELL DEVELOPMENT IN HUMAN GERMINAL-CENTERS 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 ummutated antibodies in the dark zone of the other germinal centre may represent the initial founder cells.