Abstract

Hybridoma technology will be employed to study the fine specificities and idiotypic relatedness of human lupus autoantibodies. We will prepare hybridomas by fusion of mutagenized human myeloma cells with B lymphocytes from the blood of patients with systemic lupus erythematosus. Hybridoma products will be screened for their ability to bind DNA, histones, and cardiolipin. We will characterize the fine specificities of the hybridoma autoantibodies by means of competitive immunoassays. We will also test the ability of monoclonal lupus autoantibodies to bind to neuronal cells and to lymphocytes. Anti-idiotypic antibodies will be prepared by immunization of rabbits with the hybridoma autoantibodies. We will then analyze idiotypic relatedness among a panel of hybridoma autoantibodies and measure, by immunoassay, serum levels of the idiotypes in lupus patients and their immediate relatives. Idiotypes on lymphocyte surfaces will be sought with fluorescein-tagged anti-idiotypic sera. Finally, fine specificities and idiotypic relatedness of autoantibodies from patients with drug-induced lupus will be studied and compared with those from patients with spontaneous lupus.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI026450-02
Application #
3140196
Study Section
Experimental Immunology Study Section (EI)
Project Start
1987-09-30
Project End
1992-08-31
Budget Start
1988-09-01
Budget End
1989-08-31
Support Year
2
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Tufts University
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02111

  Publications

Rubinstein, D B; Schwartz, R S; Guillaume, T et al. (1994) Germline complexity, restriction fragment length polymorphism, and coding region sequences of the human VH7 gene family identified with family-specific FR3 segment oligonucleotides. Mol Immunol 31:713-21
Stewart, A K; Huang, C; Stollar, B D et al. (1993) High-frequency representation of a single VH gene in the expressed human B cell repertoire. J Exp Med 177:409-18
Rubinstein, D B; Symann, M; Guillaume, T (1993) Oligonucleotide probes to the 5' end of the framework 3 (FR3) gene segment detect polymorphisms of VH gene sequences encoding biologically important amino acid residues. Scand J Immunol 37:33-8
Rubinstein, D B; Symann, M; Stewart, A K et al. (1993) Restriction fragment length polymorphisms and single germline coding region sequence in VH18/2, a duplicated gene encoding autoantibody. Mol Immunol 30:403-12
Sanford, D G; Stollar, B D (1992) Assay of anti-DNA antibodies. Methods Enzymol 212:355-71
Huang, C; Stewart, A K; Schwartz, R S et al. (1992) Immunoglobulin heavy chain gene expression in peripheral blood B lymphocytes. J Clin Invest 89:1331-43
Stewart, A K; Huang, C; Long, A A et al. (1992) VH-gene representation in autoantibodies reflects the normal human B-cell repertoire. Immunol Rev 128:101-22
Stollar, B D (1991) Autoantibodies and autoantigens: a conserved system that may shape a primary immunoglobulin gene pool. Mol Immunol 28:1399-412
Stollar, B D (1990) The biochemistry and genetics of DNA and anti-DNA antibodies. Clin Rheumatol 9:30-8
Sabbaga, J; Pankewycz, O G; Lufft, V et al. (1990) Cross-reactivity distinguishes serum and nephritogenic anti-DNA antibodies in human lupus from their natural counterparts in normal serum. J Autoimmun 3:215-35

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