Restricted VH gene usage in lamina propria B cells producing anticolon antibody from patients with ulcerative colitis

Nagamu Inoue, Mamoru Watanabe, Toshiro Sato, Akira Okazawa, Motomi Yamazaki, Takanori Kanai, Haruhiko Ogata, Yasushi Iwao, Hiromasa Ishii, Toshifumi Hibi

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Abstract

Background & Aims: Autoimmune responses against colonic epithelium may play a role in the development of colonic inflammation associated with ulcerative colitis (UC). In this study, we established and characterized B-cell lines and clones that produced anticolon antibody from inflamed colonic mucosa of UC subjects. Methods: B-cell lines were generated through Epstein-Barr virus transformation of lamina propria lymphocytes (LPLs) from colonic mucosa and peripheral blood lymphocytes, and these lines were screened for the production of anticolon antibodies. B-cell lines were then cloned by limiting dilution culture, and messenger RNA expression of immunoglobulin heavy-chain variable region (VH) was assessed. Results: VH gene families used in B-cell lines established from LPLs of normal controls were diverse, and B-cell lines from UC LPLs expressed a restricted VH3 family usage. All 15 clones from UC used a restricted VH3 gene family, whereas diverse VH gene families were used by 24 clones from normal controls. The analysis of nucleotide sequences indicated that these clones were derived from various germline gene segments. Conclusions: The restricted VH gene usage in anticolon autoantibodies producing B-cell clones suggests that a particular antigenic stimulus contributes to the pathogenesis of UC.

Original languageEnglish
Pages (from-to)15-23
Number of pages9
JournalGastroenterology
Volume121
Issue number1
Publication statusPublished - 2001

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Ulcerative Colitis
Mucous Membrane
B-Lymphocytes
Clone Cells
Antibodies
Cell Line
Lymphocytes
Genes
Immunoglobulin Heavy Chains
Autoimmunity
Human Herpesvirus 4
Autoantibodies
Antibody Formation
Epithelium
Inflammation
Messenger RNA

ASJC Scopus subject areas

  • Gastroenterology

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Restricted VH gene usage in lamina propria B cells producing anticolon antibody from patients with ulcerative colitis. / Inoue, Nagamu; Watanabe, Mamoru; Sato, Toshiro; Okazawa, Akira; Yamazaki, Motomi; Kanai, Takanori; Ogata, Haruhiko; Iwao, Yasushi; Ishii, Hiromasa; Hibi, Toshifumi.

In: Gastroenterology, Vol. 121, No. 1, 2001, p. 15-23.

Research output: Contribution to journalArticle

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AU - Inoue, Nagamu

AU - Watanabe, Mamoru

AU - Sato, Toshiro

AU - Okazawa, Akira

AU - Yamazaki, Motomi

AU - Kanai, Takanori

AU - Ogata, Haruhiko

AU - Iwao, Yasushi

AU - Ishii, Hiromasa

AU - Hibi, Toshifumi

PY - 2001

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N2 - Background & Aims: Autoimmune responses against colonic epithelium may play a role in the development of colonic inflammation associated with ulcerative colitis (UC). In this study, we established and characterized B-cell lines and clones that produced anticolon antibody from inflamed colonic mucosa of UC subjects. Methods: B-cell lines were generated through Epstein-Barr virus transformation of lamina propria lymphocytes (LPLs) from colonic mucosa and peripheral blood lymphocytes, and these lines were screened for the production of anticolon antibodies. B-cell lines were then cloned by limiting dilution culture, and messenger RNA expression of immunoglobulin heavy-chain variable region (VH) was assessed. Results: VH gene families used in B-cell lines established from LPLs of normal controls were diverse, and B-cell lines from UC LPLs expressed a restricted VH3 family usage. All 15 clones from UC used a restricted VH3 gene family, whereas diverse VH gene families were used by 24 clones from normal controls. The analysis of nucleotide sequences indicated that these clones were derived from various germline gene segments. Conclusions: The restricted VH gene usage in anticolon autoantibodies producing B-cell clones suggests that a particular antigenic stimulus contributes to the pathogenesis of UC.

AB - Background & Aims: Autoimmune responses against colonic epithelium may play a role in the development of colonic inflammation associated with ulcerative colitis (UC). In this study, we established and characterized B-cell lines and clones that produced anticolon antibody from inflamed colonic mucosa of UC subjects. Methods: B-cell lines were generated through Epstein-Barr virus transformation of lamina propria lymphocytes (LPLs) from colonic mucosa and peripheral blood lymphocytes, and these lines were screened for the production of anticolon antibodies. B-cell lines were then cloned by limiting dilution culture, and messenger RNA expression of immunoglobulin heavy-chain variable region (VH) was assessed. Results: VH gene families used in B-cell lines established from LPLs of normal controls were diverse, and B-cell lines from UC LPLs expressed a restricted VH3 family usage. All 15 clones from UC used a restricted VH3 gene family, whereas diverse VH gene families were used by 24 clones from normal controls. The analysis of nucleotide sequences indicated that these clones were derived from various germline gene segments. Conclusions: The restricted VH gene usage in anticolon autoantibodies producing B-cell clones suggests that a particular antigenic stimulus contributes to the pathogenesis of UC.

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