Skip to main content
Immunology logoLink to Immunology
. 1984 Apr;51(4):765–772.

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

A Malley, L Bradley, S Shiigi
PMCID: PMC1454565  PMID: 6231242

Abstract

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.

Full text

PDF
765

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brandt C. J., Deppe L. B., Malley A. Antigen B-specific helper factor from timothy grass pollen: isolation and partial characterization. Immunology. 1981 Oct;44(2):373–379. [PMC free article] [PubMed] [Google Scholar]
  2. Fairchild S. S., Malley A. Mouse B and T lymphocyte responses to purified timothy pollen antigens in vitro. J Immunol. 1976 Dec;117(6):2137–2142. [PubMed] [Google Scholar]
  3. Fairchild S. S., Malley A. Response of mouse splenic lymphocytes to timothy pollen antigens in a microculture system. J Immunol. 1975 Dec;115(6):1533–1537. [PubMed] [Google Scholar]
  4. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  5. Mackler B., Malley A., Amkraut A. A. Antigen mediated transformation of rhesus lymphocytes in immediate and delayed hypersensitivity. Int Arch Allergy Appl Immunol. 1971;41(5):765–777. doi: 10.1159/000230567. [DOI] [PubMed] [Google Scholar]
  6. Malley A., Begley D. E., Forsham A. T-cell-priming characteristics of modified timothy grass pollen antigen B1. Int Arch Allergy Appl Immunol. 1980;62(3):276–284. doi: 10.1159/000232523. [DOI] [PubMed] [Google Scholar]
  7. Malley A., Begley D., Forsham A. Chemical modification of timothy grass pollen antigen B--I. Loss of antibody binding properties. Mol Immunol. 1979 Nov;16(11):929–934. doi: 10.1016/0161-5890(79)90093-2. [DOI] [PubMed] [Google Scholar]
  8. Malley A., Brandt C. J., Deppe L. B. Preparation and characterization of the anti-idiotypic properties of rabbit anti-timothy antigen B helper factor and anti-mouse timothy IgE antisera. Immunology. 1982 Feb;45(2):217–225. [PMC free article] [PubMed] [Google Scholar]
  9. Malley A., Deppe L. B., Brandt C. J. Induction and characterization of suppressor T cells and soluble factors with modified timothy grass pollen AgB. Int Arch Allergy Appl Immunol. 1981;65(2):129–137. doi: 10.1159/000232748. [DOI] [PubMed] [Google Scholar]
  10. Malley A., Dresser D. W. Anti-idiotype regulation of timothy grass pollen IgE antibody formation. I. In vivo induction of suppressor T cells. Immunology. 1982 Jul;46(3):653–659. [PMC free article] [PubMed] [Google Scholar]
  11. Malley A., Harris R. L., Jr Biologic properties of a non-precipitating antigen from timothy pollen extracts. J Immunol. 1967 Oct;99(4):825–830. [PubMed] [Google Scholar]
  12. Pease L., Claflin J. L. Clonal regulation in the response to phosphorylcholine. II. Heterogeneity among T15 idiotype-positive antibodies in inbred and wild mice. Eur J Immunol. 1981 Aug;11(8):662–667. doi: 10.1002/eji.1830110813. [DOI] [PubMed] [Google Scholar]
  13. Petit C., Gilbert M., Sommé G., Leclercq L., Mazié J. C., Dorf M. E., Thèze J. Analysis of a major rat idiotype associated with Anti-GAT antibodies. Mol Immunol. 1982 Sep;19(9):1139–1147. doi: 10.1016/0161-5890(82)90324-8. [DOI] [PubMed] [Google Scholar]
  14. Wysocki L. J., Sato V. L. "Panning" for lymphocytes: a method for cell selection. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2844–2848. doi: 10.1073/pnas.75.6.2844. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES