Immunochemical and molecular analysis of beta 2 ---> 6 fructosan binding monoclonal antibodies bearing the A48 regulatory idiotype.
Item
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Title
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Immunochemical and molecular analysis of beta 2 ---> 6 fructosan binding monoclonal antibodies bearing the A48 regulatory idiotype.
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Identifier
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AAI8821066
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identifier
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8821066
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Creator
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Barak, Zahava Tova.
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Contributor
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Adviser: Constantin A. Bona
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Date
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1988
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Language
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English
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Publisher
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City University of New York.
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Subject
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Engineering, Biomedical
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Abstract
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The overall goal of this study was to investigate the immunochemical and molecular properties of monoclonal antibodies expressing A48 regulatory idiotype. The experimental strategy devised was to activate A48-UPC10 regulatory idiotype bearing clones subsequent to administration of syngeneic antiidiotypic reagents and then immortalize those clones as hybridomas. Hybridomas secreting antibodies were selected for the expression of A48-idiotype by sensitive radioimmunoassay.;Regulatory idiotypes are autoimmunogenic, shared by antibodies exhibiting different antigen specificities, and recognized by regulatory T cells which could contribute to their dominant expression.;In order to fully appreciate the implications of immmunoregulation through regulatory idiotypes, I performed a complete immunochemical and molecular characterization of antibody molecules bearing the A48-UPC10 regulatory idiotype.;The V{dollar}\sb{lcub}\rm H{rcub}{dollar} region expressed by A48, UPC10 and the hybridomas in this work derives from the V{dollar}\sb{lcub}\rm H{rcub}{dollar}441-1 germline gene which belongs to the small X-24 V{dollar}\sb{lcub}\rm H{rcub}{dollar} gene family. Northern-blot analysis of mRNA extracted from the hybridomas revealed that in addition there is a preferential usage of the V{dollar}\sb{lcub}\rm L{rcub}{dollar} gene belonging to the V{dollar}\sb{lcub}\rm K{rcub}{dollar}10 family located on chromosome 16 in the mouse, like the V{dollar}\sb{lcub}\rm L{rcub}{dollar} used by UPC10 and MOPC-173 A-48RI bearing myelomas.;Surprisingly, the antigen binding specificities manifested by those hybridomas are different from that of A48 and UPC10, which exhibit {dollar}\beta\sb{lcub}2\to6{rcub}{dollar} fructosan binding activity. They included specificities to {dollar}\beta\sb{lcub}2\to6{rcub}{dollar} fructosan, {dollar}\beta\sb{lcub}2\to6{rcub}{dollar} and {dollar}\beta\sb{lcub}2\to1{rcub}{dollar} fructosan, branch point determinant in bacterial levan, and non-fructosan determinants. These data which show heterogeneous antigen binding specificities and yet preferential pairing of heavy and light chains, suggest differences in the sequence of genes derived from the same germline gene.;The sequencing data presents a random usage of J{dollar}\sb{lcub}\rm H{rcub}{dollar} and J{dollar}\sb{lcub}\rm L{rcub}{dollar} segments, and the variety of D segments used by the heavy-chain genes, suggest no possible contribution of J{dollar}\sb{lcub}\rm H{rcub}{dollar}, J{dollar}\sb{lcub}\rm L{rcub}{dollar} or D segments to the regulatory idiotype expression or antigen binding specificity. Differences are found in the sequences of the hypervariable regions of the V{dollar}\sb{lcub}\rm H{rcub}{dollar} and the V{dollar}\sb{lcub}\rm L{rcub}{dollar} genes, but nevertheless they still derive from a unique V{dollar}\sb{lcub}\rm L{rcub}{dollar} germline gene, the V{dollar}\sb{lcub}\rm K{rcub}{dollar}10 and a unique V{dollar}\sb{lcub}\rm H{rcub}{dollar} germline gene belonging to the X-24 germline gene family.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
