Abstract
The Ab response of a healthy adult to the first dose of a Haemophilus influenzae type b capsular polysaccharide (HibCP) conjugate vaccine was studied at the level of Ig gene usage by circulating Ab-secreting cells. Forty-one IgA and 17 IgG mRNA sequences were obtained. The major part of the response was confined to IgA Ab-secreting cells, and 72% of the IgA sequences were derived from the progeny of a single rearranged B cell. These sequences could be arranged in a genealogical tree showing multiple somatic mutations and at least two intraclonal isotype switches to IgA2. Fourteen somatic mutations were shared by this clonal progeny, indicating that extreme clonal selection had occurred early in the clonal development. Taking into account the frequency of somatic mutations and the clone size, it was evident that the responding cell population must have originated from a mutated, highly selected, and expanded population of cells existing before vaccination, i.e., memory B cells. The dominating heavy and light chains of the response were combined in a Fab that bound HibCP. It was shown that the shared heavy and light chain mutations increased the affinity for HibCP considerably, indicating that the clonal selection had been driven by affinity. Pre-existing memory cells in unvaccinated adults may explain several features of Ab responses to polysaccharide vaccines and may play a role in acquiring the ability to respond to pure polysaccharides during infancy.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adult
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Amino Acid Sequence
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Antibody Affinity* / genetics
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B-Lymphocyte Subsets / immunology*
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B-Lymphocyte Subsets / metabolism
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Bacterial Capsules
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Base Sequence
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Cell Differentiation / genetics
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Cell Differentiation / immunology
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Clone Cells / immunology
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Clone Cells / metabolism
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Gene Rearrangement, B-Lymphocyte, Heavy Chain
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Gene Rearrangement, B-Lymphocyte, Light Chain
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Haemophilus Vaccines / administration & dosage
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Haemophilus Vaccines / immunology*
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Humans
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Immunization Schedule
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Immunoglobulin A / biosynthesis*
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Immunoglobulin A / genetics
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Immunoglobulin A / metabolism
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Immunoglobulin Fab Fragments / genetics
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Immunoglobulin Fab Fragments / metabolism
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Immunoglobulin Isotypes / biosynthesis*
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Immunoglobulin Isotypes / genetics
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Immunoglobulin Isotypes / metabolism
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Immunoglobulin Variable Region / genetics
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Immunoglobulin Variable Region / metabolism
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Immunologic Memory / genetics
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Immunologic Memory / immunology*
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Lymphocyte Activation / genetics
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Lymphocyte Activation / immunology*
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Male
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Molecular Sequence Data
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Mutation / immunology
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Polysaccharides, Bacterial / administration & dosage
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Polysaccharides, Bacterial / immunology*
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Vaccines, Conjugate / administration & dosage
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Vaccines, Conjugate / immunology
Substances
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Haemophilus Vaccines
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Haemophilus influenzae type b polysaccharide vaccine
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Immunoglobulin A
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Immunoglobulin Fab Fragments
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Immunoglobulin Isotypes
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Immunoglobulin Variable Region
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Polysaccharides, Bacterial
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Vaccines, Conjugate
Associated data
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GENBANK/Z98683
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GENBANK/Z98684
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GENBANK/Z98685
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GENBANK/Z98686
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GENBANK/Z98687
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GENBANK/Z98688
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GENBANK/Z98689
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GENBANK/Z98690
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GENBANK/Z98691
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GENBANK/Z98692
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GENBANK/Z98693
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GENBANK/Z98694
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GENBANK/Z98695
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GENBANK/Z98696
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GENBANK/Z98697
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GENBANK/Z98698
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GENBANK/Z98699
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GENBANK/Z98700
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GENBANK/Z98701
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GENBANK/Z98702
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GENBANK/Z98703
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GENBANK/Z98704
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GENBANK/Z98705
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GENBANK/Z98706
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GENBANK/Z98707
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GENBANK/Z98708
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GENBANK/Z98709
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GENBANK/Z98710
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GENBANK/Z98711
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GENBANK/Z98712