Monoclonal Analysis of Spontaneous and Induced Antibody Responses in Autoimmune New Zealand Mice
Ballard, Dean Williams
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https://hdl.handle.net/2142/71169
Description
Title
Monoclonal Analysis of Spontaneous and Induced Antibody Responses in Autoimmune New Zealand Mice
Author(s)
Ballard, Dean Williams
Issue Date
1985
Department of Study
Microbiology
Discipline
Microbiology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Health Sciences, Immunology
Abstract
New Zealand BW mice spontaneously develop lymphocytic disorders and express autoantibodies in association with an autoimmune disease similar to human SLE. A comparative monoclonal analysis of spontaneous and experimentally-induced antibody responses in autoimmune BW mice was facilitated by hybridoma technology. Various immunochemical and physicochemical parameters of generated monoclonal antibodies were examined to provide a qualitative functional and structural description of such etiologically distinct antibody repertoires in the context of inherent cellular and humoral abnormalities characteristic of this strain.
Ten monoclonal anti-DNA autoantibodies, derived from nonmanipulated BW mice and specific for various forms of DNA, were found to contain either IgG2a or IgG2b heavy chains related by a cross-reactive allotypic determinant. Binding studies, which employed defined DNA restriction fragments and synthetic nucleic acids, indicated that: (1) both helical conformation and nucleotide composition were involved in antibody recognition of duplex and single-stranded DNA, and (2) anti-DNA antibodies formed stable complexes with short synthetic oligonucleotides (<15 bases). Serological analyses of the expression of distinct idiotypic determinants possessed by purine and pyrimidine specific antibodies indicated the potential for an extensive clonal repertoire.
Thirty-eight monoclonal anti-fluorescein antibodies were derived from immunized BW mice to evaluate various aspects of induced antibody expression, including isotype and affinity distribution. Monoclonal IgM antibody 18-2-3, produced from an initial BW cell fusion, exhibited low-temperature insolubility and an unusually high hapten-binding affinity (K(,A)) of 2.9 x 10('10) M('-1). Insolubility at low temperature was reversible in the presence of fluorescyl ligand, indicative of active site involvement in the mechanism of 18-2-3 cryoprecipitation. All other IgM and IgG proteins were restricted in hapten-binding affinities (K(,A) < 7 x 10('7) M('-1)) and possessed normal solubility properties, suggesting that 18-2-3 was derived from a relatively rare B cell progenitor. Relative subclass frequencies for 30 monoclonal IgG proteins were consistent with those reported for polyclonal thymic-dependent responses in normal strains, thus providing no evidence that T cells involved in regulation of IgG subclass expression are compromised in autoimmune BW mice.
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