Anti-DNA antibodies are diagnostic and prognostic markers for SLE, although mechanisms of their production are poorly understood. Attempts to stimulate anti-DNA by direct DNA immunization in normal animals have until recently been only minimally successful. Recent evidence from this laboratory indicates, however, that bacterial DNAs are more immunogenic than mammalian DNAs and are able to stimulate an effective anti-DNA response in normal mice that resembles the anti-DNA reactivity of lupus. I propose, therefore, to explore this new model of SLE anti-DNA production by utilizing hybridoma, T cell cloning, and molecular biologic techniques to elucidate further the immune response to nucleic acid antigens and the role of DNA in the induction of the anti-DNA of lupus. The following aims are proposed: 1) to compare immunochemical properties of monoclonal anti-DNA antibodies derived from immunized normal mice and autoimmune MRL-lpr/lpr mice and thereby assess similarities between induced and spontaneous anti- DNA antibodies; 2) to study the response of T cells to DNA immunization by proliferation assays and the ability of these T cells to provide help to anti-DNA producing B cells. DNA specific T cell clones will also be prepared as aberrant T cell helper function has been implicated in the pathogenesis of SLE; 3) to evaluate the renal disease induced by DNA immunization including the study of bound antibodies eluted from affected kidneys given the pathogenicity of anti-DNA antibodies in lupus nephritis. Dr. Gilkeson has worked in this line of research for two years developing the methods necessary to complete these studies. Funding of this grant will provide the time and salary support to continue an already productive line of research and to allow for the development of independence in the study of autoimmunity. Current expertise in this laboratory and Duke University Medical Center provides a supportive atmosphere for learning and growth as a clinical investigator. The Division of Rheumatology, with support provided by a program project grant for arthritis research, is actively involved in the study of autoimmune diseases. There is a commitment in the Division to develop young investigators, providing them with the support necessary to pursue a career in academic medicine. In this environment, studying the immunopathogenesis of induced anti-DNA responses will be a fruitful learning experience lending new insight into anti-DNA, the hallmark of lupus autoantibody production.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08AR001847-03
Application #
3079305
Study Section
Arthritis and Musculoskeletal and Skin Diseases Special Grants Review Committee (AMS)
Project Start
1990-07-01
Project End
1995-06-30
Budget Start
1992-07-01
Budget End
1993-06-30
Support Year
3
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Duke University
Department
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705
Wloch, M K; Clarke, S H; Gilkeson, G S (1997) Influence of VH CDR3 arginine and light chain pairing on DNA reactivity of a bacterial DNA-induced anti-DNA antibody from a BALB/c mouse. J Immunol 159:6083-90
Wloch, M K; Alexander, A L; Pippen, A M et al. (1997) Molecular properties of anti-DNA induced in preautoimmune NZB/W mice by immunization with bacterial DNA. J Immunol 158:4500-6
Lefkowith, J B; Di Valerio, R; Norris, J et al. (1996) Murine glomerulotropic monoclonal antibodies are highly oligoclonal and exhibit distinctive molecular features. J Immunol 157:1297-305
Gilkeson, G S; Ruiz, P; Pippen, A M et al. (1996) Modulation of renal disease in autoimmune NZB/NZW mice by immunization with bacterial DNA. J Exp Med 183:1389-97
Wloch, M K; Alexander, A L; Pippen, A M et al. (1996) Differences in V kappa gene utilization and VH CDR3 sequence among anti-DNA from C3H-lpr mice and lupus mice with nephritis. Eur J Immunol 26:2225-33
Gilkeson, G S; Pippen, A M; Pisetsky, D S (1995) Induction of cross-reactive anti-dsDNA antibodies in preautoimmune NZB/NZW mice by immunization with bacterial DNA. J Clin Invest 95:1398-402
Gilkeson, G S; Bernstein, K; Pippen, A M et al. (1995) The influence of variable-region somatic mutations on the specificity and pathogenicity of murine monoclonal anti-DNA antibodies. Clin Immunol Immunopathol 76:59-67
Pyun, E H; Pisetsky, D S; Gilkeson, G S (1993) The fine specificity of monoclonal anti-DNA antibodies induced in normal mice by immunization with bacterial DNA. J Autoimmun 6:11-26
Gilkeson, G S; Bloom, D D; Pisetsky, D S et al. (1993) Molecular characterization of anti-DNA antibodies induced in normal mice by immunization with bacterial DNA. Differences from spontaneous anti-DNA in the content and location of VH CDR3 arginines. J Immunol 151:1353-64
Robertson, C R; Gilkeson, G S; Ward, M M et al. (1992) Patterns of heavy and light chain utilization in the antibody response to single-stranded bacterial DNA in normal human subjects and patients with systemic lupus erythematosus. Clin Immunol Immunopathol 62:25-32

Showing the most recent 10 out of 11 publications