Generation of human anti-MUC3 IgG antibodies after in vitro immunization of naive peripheral blood B-lymphocytes |
| |
Authors: | Stavroula Baritaki Alexandros Zafiropoulos Eustathia Georgopoulou Sotirios Souris Elias Krambovitis |
| |
Institution: | (1) Department of Applied Biochemistry and Immunology, IMBB, FORTH, P.O. Box 1527, Heraklion 711 10, Crete, Greece e-mail: krambo@imbb.forth.gr Tel.: +30-81-391020; Fax: +30-81-391101, GR;(2) Department of Haematology and Histology, D.A.U. Penlelis Hospital, Athens, Greece, GR;(3) Department of Pathology, Apollonion General Hospital, Heraklion, Greece, GR |
| |
Abstract: | It has been demonstrated that IgG antibodies can be generated to self-antigen peptides as well as against viral antigens
by an antigen-specific in vitro immunization system of resting human peripheral B-lymphocytes. Using a synthetic peptide from
the consensus variable tandem-repeat region of the MUC3 mucin (TSSITTTGTTSHSTPSP) as the B cell epitope, we immunized blood
donor B-lymphocytes in vitro and tested for MUC3-specific antibodies by ELISA. After the primary activation step all antibodies
were IgM. At the end of the secondary immunization step we obtained 1.8% (21/1138) of the cultures with IgG-switched antibodies.
In a competitive inhibition ELISA using the MUC1, MUC2, MUC3, MUC4 and PIP2 peptides, only one culture (F8.1) gave satisfactory
specific inhibition. Using this antibody in fluorometric studies, it stained cells from two colon carcinoma cell lines predominantly
in the cytoplasm, whereas those from a breast cancer cell line stained predominantly the cell surface. In a preliminary immunohistological
evaluation with formalin-fixed sections, the antibody appeared to moderately stain colon sections, but not breast sections
or lymph node. This method of in vitro immunization may be a useful tool in generating IgG antibodies specific to self-antigens
and could find applications in tumour targeting and immunotherapy.
Received: 12 October 2000 / Accepted: 11 January 2001 |
| |
Keywords: | MUC3 Human antibody In vitro immunization Fluorometry Immunohistology |
本文献已被 PubMed SpringerLink 等数据库收录! |
|