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. 2006 Sep 5;52(3):227–233. doi: 10.1007/s10616-006-9003-x

In vitro immunization can elicit the expansion of diverse repertoire of B cells from peripheral blood mononuclear cells

Shin-ei Matsumoto 1, Makiko Yamashita 1, Yoshinori Katakura 1,2,✉, Eri Noguchi 2, Yoshihiro Aiba 1, Akira Ichikawa 3, Kiichiro Teruya 1,2, Sanetaka Shirahata 1,2
PMCID: PMC3449408  PMID: 19002881

Abstract

We previously developed an in vitro immunization (IVI) protocol of human peripheral blood mononuclear cells (PBMC) for generating antigen-specific human antibodies. In order to clarify whether IVI protocolinduces antigen-specific B cell responses in PBMC, we analyzed family gene usage and sequence of the variable region gene of immunoglobulin heavy chain (VH gene) of the antibody produced from the in vitro immunized PBMC. Sequence homology analyses of VH gene demonstrated that a larger repertoire of B cells can be sensitized with mite-extract than with cholera toxin B subunit and rice allergen. Further, antigen-specific B cells were efficiently expanded by using CpG oligodeoxynucleotide as adjuvant. These results suggest that appropriate combination of sensitizing antigen and adjuvant is primarily important for expansion of antigen-specific B cells in IVI protocol.

Keywords: CpG oligonucleotide, Human monoclonal antibody, In vitro immunization, Peripheral blood mononuclear cells, Variable region gene

Glossary

CTB

cholera toxin B subunit

FBS

fetal bovine serum

IL

interleukin

IVI

in vitro immunization

LLME

l-leucyl-l-leucine methyl ester

mAb

monoclonal antibodies

MDP

muramyl dipeptide

ME

mite extract

PBMC

peripheral mononuclear cells

RA

rice allergen

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