Preparation and characterization of a monoclonal anti-T helper factor antibody

Sprague-Dawley rats were immunized by injection of antigen B-specific T helper factor THF) eluted from Sepharose-antigen D adsorbents. Rat spleen cells from animals immunized with THF were fused with a BALB/c tumour cell (P3x63-Ag8.653) to prepare monoclonal anti-THF antibodies. The hybrids produced...

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Bibliographic Details
Published in:Immunology 1984-04, Vol.51 (4), p.765-772
Main Authors: MALLEY, A, BRADLEY, L, SHIIGI, S
Format: Article
Language:English
Subjects:
Animals
Antibodies, immunoglobulins
Antibodies, Monoclonal - biosynthesis
Antibodies, Monoclonal - immunology
Biological and medical sciences
Enzyme-Linked Immunosorbent Assay
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Hybridomas - immunology
Immunoglobulin Idiotypes - immunology
Lymphocyte Activation
Male
Mice
Molecular immunology
T-Lymphocytes, Helper-Inducer - immunology
T-Lymphocytes, Regulatory - immunology
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Summary:Sprague-Dawley rats were immunized by injection of antigen B-specific T helper factor THF) eluted from Sepharose-antigen D adsorbents. Rat spleen cells from animals immunized with THF were fused with a BALB/c tumour cell (P3x63-Ag8.653) to prepare monoclonal anti-THF antibodies. The hybrids produced were screened for anti-THF antibodies by an enzyme-linked immunoassay (ELISA), and we shall describe the characteristics of one of the hybrid (hybridoma 6-2.2 anti-THF) antibodies produced. (i) The monoclonal hybrid 6-2.2 anti-THF antibody blocks water-soluble timothy extract-induced proliferation of timothy-specific T helper cells when these cells were preincubated with an excess of the anti-THF hybrid 6-2.2 antibody; (ii) incubation of timothy-specific T helper or T suppressor cells with an optimal dose of anti-THF 6-2.2 antibody induces significant levels of [3H]-thymidine incorporation in the absence of antigen; (iii) it binds specifically to the idiotypic determinant expressed on THFk, THFd, TSFk, and antigen B-specific IgE in an ELISA; and (iv) it has no effect upon spleen cells from mice primed with ovalbumin or Ascaris suum antigens. In addition, the monoclonal anti-THF 6-2.2 antibody cultured with normal spleen cells in mini-Marbrook chambers induced significant levels of antigen B-specific T suppressor cells. These studies indicate that the monoclonal anti-THF 6-2.2 antibody has anti-idiotypic antibody properties.
ISSN:0019-2805
1365-2567