title: In vitro immunization can elicit the expansion of diverse repertoire of B cells from peripheral blood mononuclear cells.
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.
authors.affiliation: Graduate School of Systems Life Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan.
authors.forename: Shin-Ei
authors.initials: SE
authors.lastname: Matsumoto
authors.forename: Makiko
authors.initials: M
authors.lastname: Yamashita
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Katakura
authors.forename: Eri
authors.initials: E
authors.lastname: Noguchi
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Aiba
authors.forename: Akira
authors.initials: A
authors.lastname: Ichikawa
authors.forename: Kiichiro
authors.initials: K
authors.lastname: Teruya
authors.forename: Sanetaka
authors.initials: S
authors.lastname: Shirahata