We intend to investigate the basic mechanisms of cellular and humoral regulation of mouse immune interferon (IFN-gamma) production. The helper and suppressor lymphocytes that regulate IFN-gamma production will be characterized along with the IFN-gamma-producing lymphocytes. Primary T-lymphocyte lines will also be studied as possible producers and regulators of IFN-gamma production. The growth factors interleukins-1 (IL-I) and 2 (IL-2) will be investigated as possible mediators of cellular regulation of IFN-gamma production. Phorbol esters, IL-2, and polypeptide hormones such as vasopressin are cell activators and may help in IFN-gamma production through a common second messenger (cyclic GMP?). Further improvement of IFN-gamma purification and characterization will be done by lectin affinity chromatography, antibody affinity chromatography, gel electrophoresis, and N-terminal amino acid analysis. Monoclonal antibodies will be produced to IFN-gamma to improve purification and for further characterization of IFN-gamma in terms of biological activity and relationship to other lymphokines. Purified IFN-gamma should be ideal for an objective assessment of its possible range of biological activities, among which are: (1) immunoregulation; (2) antitumor and anticellular activity; (3) regulation of differentiation; and (4) antiviral properties. The mechanism of action of IFN-gamma at the molecular level will be studied (if time permits) through assessment of its membrane effects, intermediary metabolism effects, and cellular protein kinase activities. The long-term goal is to understand natural regulatory functions of IFN-gamma and to be able to manipulate IFN-gamms activity in order to obtain desirable immunological, antiviral, and antitumor activities of this lymphokine. Considerable progress has been made with regards to regulation of production of IFN-gamma. (IL-2) can replace helper cell requirement for IFN-gamma production. Dibutyryl cyclic GMP, arachidonic acid and its lipooxygenase products (such as leukotrienes), the neuropeptides: arginine, vasopressin, and oxytocin, and the tumor promoter phorbol myristic acetate can replace IL-2 requirement. IL-2 help in IFN-gamma production may involve release of second messenger signals arachidonic acid and diacyglycerol from membrane phospholipids, activation of guanylate cyclase, and activation of protein kinases such ss protein kinase C. The produced IFN-gamma was shown to cause B cell maturation. Antibodies of predetermined specificity were produced to IFN-gsmme and should help in purification and characterizstion. (HF)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA039048-02
Application #
3177742
Study Section
(SSS)
Project Start
1984-04-01
Project End
1986-11-30
Budget Start
1985-04-01
Budget End
1986-11-30
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Florida
Department
Type
Schools of Medicine
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Abdullah, N A; Torres, B A; Basu, M et al. (1989) Differential effects of epidermal growth factor, transforming growth factor-alpha, and vaccinia virus growth factor in the positive regulation of IFN-gamma production. J Immunol 143:113-7
Torres, B A; Johnson, H M (1988) Arginine vasopressin (AVP) replacement of helper cell requirement in IFN-gamma production. Evidence for a novel AVP receptor on mouse lymphocytes. J Immunol 140:2179-83
Johnson, H M; Torres, B A (1988) A novel arginine vasopressin-binding peptide that blocks arginine vasopressin modulation of immune function. J Immunol 141:2420-3
Pontzer, C H; Torres, B A; Vallet, J L et al. (1988) Antiviral activity of the pregnancy recognition hormone ovine trophoblast protein-1. Biochem Biophys Res Commun 152:801-7
Johnson, H M; Russell, J K; Torres, B A (1988) Structural basis for arachidonic acid second messenger signal in gamma-interferon induction. Ann N Y Acad Sci 524:208-17
Russell, J K; Torres, B A; Johnson, H M (1987) Phospholipase A2 treatment of lymphocytes provides helper signal for interferon-gamma induction. Evidence for second messenger role of endogenous arachidonic acid. J Immunol 139:3442-6
Johnson, H M; Russell, J K; Torres, B A (1986) Second messenger role of arachidonic acid and its metabolites in interferon-gamma production. J Immunol 137:3053-6
Johnson, H M; Vassallo, T; Torres, B A (1985) Interleukin 2-mediated events in gamma-interferon production are calcium dependent at more than one site. J Immunol 134:967-70
Johnson, H M; Torres, B A (1985) Peptide growth factors PDGF, EGF, and FGF regulate interferon-gamma production. J Immunol 134:2824-6
Johnson, H M; Torres, B A (1985) Mechanism of calcium ionophore A23187-induced priming of bone marrow-derived macrophages for tumor cell killing: relationship to priming by interferon. Proc Natl Acad Sci U S A 82:5959-62

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