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|Title:||Monoclonal Analysis of Spontaneous and Induced Antibody Responses in Autoimmune New Zealand Mice|
|Author(s):||Ballard, Dean Williams|
|Department / Program:||Microbiology|
|Degree Granting Institution:||University of Illinois at Urbana-Champaign|
|Subject(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.
Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1985.
|Date Available in IDEALS:||2014-12-16|