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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6318
pubmed:dateCreated
1991-5-13
pubmed:abstractText
Primary antigenic exposure results in an initial antibody response and the T cell-dependent induction of B-cell memory. Memory B-cell differentiation is characterized by somatic hypermutation in antibody variable region genes (V) and selection of B cells expressing high-affinity variants of this antigen receptor. Despite our current understanding of B-cell memory, the origin of memory B cells and the regulation of their differentiation remain elusive. This is largely due to the difficulties in observing and purifying this minor component of the immunized spleen. Further, molecular characterization of memory B cells requires hybridoma formation which restricts analyses to only those clones capable of fusion and does not allow isolation of cells in a normal physiological state. We have therefore developed a unique system which allows isolation and unambiguous enumeration of IgG1+ memory B cells, based on six-parameter flow cytometry, secretion of antibody in clonal cultures and analysis of clonally expressed V genes using the polymerase chain reaction. Here we report that single IgG1+ antigen-binding B cells from an early secondary immune response proliferate in lipopolysaccharide-driven microcultures and produce antigen-specific IgG1 antibodies. Individual B-cell clones in these cultures express somatically mutated heavy chain V genes, confirming their designation as memory B cells. Although isolated memory B cells undergo extensive proliferation in vitro, V gene sequence analysis of their individual progeny shows that further hypermutation does not occur.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0028-0836
pubmed:author
pubmed:issnType
Print
pubmed:day
11
pubmed:volume
350
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
502-5
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:2014051-Amino Acid Sequence, pubmed-meshheading:2014051-Animals, pubmed-meshheading:2014051-Antibody Formation, pubmed-meshheading:2014051-Antigens, pubmed-meshheading:2014051-B-Lymphocytes, pubmed-meshheading:2014051-Base Sequence, pubmed-meshheading:2014051-Cell Differentiation, pubmed-meshheading:2014051-Cells, Cultured, pubmed-meshheading:2014051-Flow Cytometry, pubmed-meshheading:2014051-Immunoglobulin G, pubmed-meshheading:2014051-Immunoglobulin Variable Region, pubmed-meshheading:2014051-Immunologic Memory, pubmed-meshheading:2014051-Mice, pubmed-meshheading:2014051-Mice, Inbred C57BL, pubmed-meshheading:2014051-Molecular Sequence Data, pubmed-meshheading:2014051-Mutation, pubmed-meshheading:2014051-Polymerase Chain Reaction, pubmed-meshheading:2014051-Receptors, Antigen, B-Cell, pubmed-meshheading:2014051-Spleen
pubmed:year
1991
pubmed:articleTitle
Molecular characterization of single memory B cells.
pubmed:affiliation
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't