Development of human-human hybridoma from anti-Her-2 peptide-producing B cells in immunized NOG mouse.

Full Abstract

OBJECTIVE:
Numerous monoclonal antibodies have been developed for the purpose of medical treatments, including cancer treatment. For clinical application, the most useful are human-derived antibodies. In this study, we tried to prepare designed antigen-specific antibodies of completely human origin using immunodeficient mouse.

METHODS:
Nonobese diabetic/severe combined immunodeficient/IL-2 receptor gamma null mouse (NOG) mouse was used to reconstitute the human immune system with umbilical cord blood hematopoietic stem cells (CB-NOG mouse) and to prepare human-derived Her-2-epitope-specific antibodies. Hybridoma lines were prepared by fusing the human myeloma cell line Karpas707H.

RESULTS:
Serum of immunized NOG mouse contained human-derived immunoglobulin M (IgM) antibodies specific for a short peptide sequence of 20 amino acids, including the epitope peptide of apoptotic Her-2 antibody CH401. Hybridoma lines were successfully prepared with spleen B cells obtained from the immunized CB-NOG mouse. One of these cell lines produced human IgM against the epitope peptide that can recognize surface Her-2 molecule.

CONCLUSION:
We could produce human-derived IgM antibody against Her-2 epitope peptide in CB-NOG mouse, succeeding in generation of human hybridoma-secreting IgM against a given peptide.

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Author information

Author/s: Kametani, Yoshie (Y); Shiina, Masashi (M); Katano, Ikumi (I); Ito, Ryoji (R); Ando, Kiyoshi (K); Toyama, Kanae (K); Tsukamoto, Hideo (H); Matsumura, Takuya (T); Saito, Yuki (Y); Ishikawa, Dai (D); Taki, Takao (T); Ito, Mamoru (M); Imai, Kohzoh (K); Tokuda, Yutaka (Y); Kato, Shunichi (S); Tamaoki, Norikazu (N); Habu, Sonoko (S);

Affiliation: Department of Immunology, Tokai University School of Medicine, Kanagawa, Japan.

Journal and publication information

Publication Type: Journal Article

Journal: Experimental hematology (Exp Hematol), published in Netherlands. (Language: eng)

Reference: 2006-Sep; vol 34 (issue 9) : pp 1240-8

Dates: Created 2006/08/30; Completed 2006/10/31;

PMID: 16939817, status: MEDLINE (last retrieval date: 12/26/2008)

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Animals
Antibodies, Monoclonal - immunology; therapeutic use
B-Lymphocytes - cytology; immunology
Cord Blood Stem Cell Transplantation - methods
Epitopes, B-Lymphocyte - immunology; pharmacology
Humans
Hybridomas - cytology; immunology
Immunization

Immunization
Immunoglobulin M - immunology; therapeutic use
Mice
Mice, Mutant Strains
Neoplasms - drug therapy; genetics; immunology
Peptides - immunology; pharmacology
Receptor, erbB-2 - immunology
Transplantation Chimera - genetics; immunology

Associated Chemicals: Antibodies, Monoclonal (0) ; Epitopes, B-Lymphocyte (0) ; Immunoglobulin M (0) ; Peptides (0) ; Receptor, erbB-2 (EC 2.7.1.112)

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