The aim of the proposed studies is to analyze, at the molecular level, the fine specificity of immune response gene function in mice. Specifically, we will attempt to map the hypervariable regions on the T cell receptor Alpha and Beta genes that determine specificity for antigen and MHC restriction, respectively. Furthermore, we will design a model system to test whether an antigen-MHC molecule interaction takes place during antigen presentation. A more precise understanding of the immune response phenomenon is essential in order to gain direct insight in possible mechanisms of immune disorders causing human diseases, such as some types of cancer and AIDS (acquired immune deficiency syndrome). The proposed studies are based on our detailed analysis of the genetic restriction of T cell recognition of heterologous species of insulin at the cellular level. We have already established libraries of T cell hybridomas with precisely defined antigen epitope specificity and Ia epitope restriction. These T cell hybridomas will be used to clone the cDNAs coding for the T cell receptor Alpha and Beta chain and to study the function of specifically mutagenized Ia molecules. We will prepare cDNA libraries from T cell hybridomas with slightly different Ia epitope and/or antigen epitope recognition. We then will sequence the variable region of the Alpha and Beta gene coding for the T cell receptor in the various T cell lines. With gene transfection experiments we will test the requirement for a functional receptor molecule. The possibility exists that MHC restriction and antigen recognition can be scrambled by combining an Alpha chain of a specific T cell clone with a Beta chain from a T cell clone with diffeent antigen and/or Ia epitope recognition. Finally, we will analyze the mechanism of antigen presentation in conjunction with an Ia molecule; namely we will test Ia molecules that have been altered with site specific mutagenesis at a potential Ia-insulin interaction site. Ideally, only insulin presentation should be affected by such an alteration, while recognition by allo- or autoreactive clones should be left intact.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI023031-01
Application #
3134867
Study Section
Immunobiology Study Section (IMB)
Project Start
1985-12-01
Project End
1990-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Tufts University
Department
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02111
Munoz, E; Zubiaga, A M; Huber, B T (1992) Interleukin-1 induces c-fos and c-jun gene expression in T helper type II cells through different signal transmission pathways. Eur J Immunol 22:2101-6
Munoz, E; Zubiaga, A; Huang, C et al. (1992) Interleukin-1 induces protein tyrosine phosphorylation in T cells. Eur J Immunol 22:1391-6
Molina, I J; Huber, B T (1991) Regulation of macrophage activation markers by IL-4 and IFN-gamma is subpopulation-specific. Cell Immunol 134:241-8
Munoz, E; Zubiaga, A M; Huber, B T (1991) Tyrosine protein phosphorylation is required for protein kinase C-mediated proliferation in T cells. FEBS Lett 279:319-22
Zubiaga, A M; Munoz, E; Huber, B T (1991) Superinduction of IL-2 gene transcription in the presence of cycloheximide. J Immunol 146:3857-63
Flentke, G R; Munoz, E; Huber, B T et al. (1991) Inhibition of dipeptidyl aminopeptidase IV (DP-IV) by Xaa-boroPro dipeptides and use of these inhibitors to examine the role of DP-IV in T-cell function. Proc Natl Acad Sci U S A 88:1556-9
Munoz, E; Zubiaga, A M; Sims, J E et al. (1991) IL-1 signal transduction pathways. I. Two functional IL-1 receptors are expressed in T cells. J Immunol 146:136-43
Zubiaga, A M; Munoz, E; Huber, B T (1991) Production of IL-1 alpha by activated Th type 2 cells. Its role as an autocrine growth factor. J Immunol 146:3849-56
Merrow, M W; Huber, B T (1991) rIL-4 differentially regulates rIL-2-induced murine NK and LAK killing in CD8+ and CD8- precursor cell subsets. Int Immunol 3:551-61
Munoz, E; Beutner, U; Zubiaga, A et al. (1990) IL-1 activates two separate signal transduction pathways in T helper type II cells. J Immunol 144:964-9

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