Abstract

This proposal is designed to study structural features of the antibody molecules that elicit auto-anti-idiotypic (AAI) immune responses in rabbits and to study the functioning of AAI T cells and T cell factors. Past work showed that unique reduction-sensitive idiotopes (RSID) regularly elicit natural AAI antibody responses in rabbits. The proposed research plan is designed to elicit immune responses in outbred rabbits, detect the appearance of immunoregulatory AAI antibodies by radioimmunoassay and to identify and purify the antibody clonotypes that elicited AAI responses by analytical and preparative isoelectric focusing. The purified RSID will be studied to determine which polypeptide chain bears the RSID. These chains will then be characterized for amino acid composition and amino acid sequences will be determined in order to locate the RSID in collaboration with Dr. Mike Kehoe at the Northeastern Ohio Universities College of Medicine. Functional studies of the interaction of idiotype and AAI will be done by studies of the spontaneous appearance and regulatory role of AAI T cells. These studies will be done using cryopreserved PBL taken during the response to determine if AAI T cells can affect immune reactivity of these cells. Isoelectric focusing technology will be used to purify circulating T cell receptors. These molecules will be purified and studied biochemically and serologically using monoclonal antibodies. Studies of internal image structures will be done by making monoclonal antibodies specific for rabbit b locus epitopes, making monoclonal anti-idiotype and characterizing the structural relatedness of the original epitope and the anti-idiotype. These projects exploit current biochemical and biophysical technology to gain insights into the mechanisms that are functioning in a normal immune response. The results will be relevant to understanding of pathogenesis of some autoimmune diseases, allergies, and transplantation rejection as will as characterizing new methodology for vaccine manipulation of the immune system.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI020590-03
Application #
3130323
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1984-09-30
Project End
1988-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Texas Health Science Center Houston
Department
Type
Schools of Medicine
DUNS #
City
Houston
State
TX
Country
United States
Zip Code
77225

  Publications

Knisley, K A; Rodkey, L S (1995) Affinity immunoblotting. Methods Mol Biol 51:165-70
Rodkey, L S (1995) Evaluation of antibody clonality. Methods Mol Biol 51:151-63
Seferian, P G; Rodkey, L S (1994) Identification of laboratory animals by using affinity immunoblotting. Lab Anim Sci 44:85-7
Seferian, P G; Rodkey, L S (1994) Antibody synthesis induced by endogenous internal images. Appl Biochem Biotechnol 47:213-26;discussion 226-7
Knisley, K A; Rodkey, L S (1992) Direct detection of carrier ampholytes in immobilized pH gradients using picric acid precipitation. Electrophoresis 13:220-4
Simeckova-Rosenberg, J; Rodkey, L S (1990) Induction and immunochemical properties of a novel anti-antibody. Mol Immunol 27:1119-26
Knisley, K A; Rodkey, L S (1990) Comparative studies of recycling isoelectric focusing and continuous flow electrophoresis: separation of proteins with minor charge differences. Electrophoresis 11:927-31
Rodkey, L S (1990) Free flow cell electrophoresis using zwitterionic buffer. Appl Theor Electrophor 1:243-7
Adler, F L; Adler, L T; Seferian, P G et al. (1989) Adoptive immunity transferred by naive donor cells immunized in vitro. Bone Marrow Transplant 4:663-8
Rodkey, L S (1988) Ionic binding properties of carrier ampholytes. J Chromatogr 437:147-59

Showing the most recent 10 out of 15 publications