The overall objective is to determine the extent to which intratumoral macrophages in regressing and progressing tumors are activated for tumor cell killing. Monoclonal antibodies will be produced as a means of attaining this objective. The rationale behind the research is that activators cause the expression of new components within and on the surface of macrophages. Antigenic determinants on some of these components should be sufficiently specific to constitute markers for the individual stages that lead to the fully activated, cytolytic state. Our preliminary results, as well as those from other laboratories, support the validity of this hypothesis. We will accomplish our objective by attaining the following specific aims: (i) Identify rat monoclonal antibodies that will detect marker phenotypes that are specific for individual stages of mouse macraphage activation. Immunization with isolated proteins that have been associated with activation stages by 2-dimensional electrophoretic analysis will be the primary approach. For selected purposes immunization with intact macrophages that have been treated to be in a given stage of activation may also be needed. (ii) Use the stage phenotypes to monitor the development and loss of activation stages in vitro (macrophages cultured from bone marrow) and in vivo, in regressing and progressing Moloney sarcomas. (iii) Determine the effects of an immunomodulator on the state of macrophage activation in progressively growing tumors. Various agents, such as recombinant gamma interferon, will be injected either intratumorally or systemically until an efficacious immunomodulator is identified, i.e., one that has a distinct anti-tumor effect. If needed to produce an anti-tumor effect, the soluble immunomodulator will be incorporated in liposomes before injecting it. The proposed research is important because it will allow intralesional analysis of the state of macrophage activation for the first time. It is expected that the approach will be equally applicable to the analysis of macrophages in granulomas and other inflammatory responses against infectious agents, a fact that increases the importance that the technology will potentially have.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA031199-07
Application #
3169500
Study Section
Experimental Immunology Study Section (EI)
Project Start
1981-03-01
Project End
1987-11-30
Budget Start
1987-04-01
Budget End
1987-11-30
Support Year
7
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of Florida
Department
Type
Schools of Veterinary Medicine
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Gao, J J; Filla, M B; Lorsbach, R B et al. (2000) Prolonged exposure of mouse macrophages to IFN-beta suppresses transcription of the inducible nitric oxide synthase gene: altered availability of transcription factor Stat1alpha. Eur J Immunol 30:1551-61
Huang, J; Roby, K F; Pace, J L et al. (1995) Cellular localization and hormonal regulation of inducible nitric oxide synthase in cycling mouse uterus. J Leukoc Biol 57:27-35
Phillips, T A; Kujubu, D A; MacKay, R J et al. (1993) The mouse macrophage activation-associated marker protein, p71/73, is an inducible prostaglandin endoperoxide synthase (cyclooxygenase). J Leukoc Biol 53:411-9
Lowenstein, C J; Alley, E W; Raval, P et al. (1993) Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon gamma and lipopolysaccharide. Proc Natl Acad Sci U S A 90:9730-4
Lorsbach, R B; Murphy, W J; Lowenstein, C J et al. (1993) Expression of the nitric oxide synthase gene in mouse macrophages activated for tumor cell killing. Molecular basis for the synergy between interferon-gamma and lipopolysaccharide. J Biol Chem 268:1908-13
Pinson, D M; Phillips, T A; Pace, J L et al. (1991) Activation-associated marker proteins: peptide mapping and their expression in macrophage cell lines. Biochem Biophys Res Commun 176:882-6
Chen, T Y; Bright, S W; Pace, J L et al. (1990) Induction of macrophage-mediated tumor cytotoxicity by a hamster monoclonal antibody with specificity for lipopolysaccharide receptor. J Immunol 145:8-12
MacKay, R J; Pace, J L; Jarpe, M A et al. (1989) Macrophage activation-associated proteins. Characterization of stimuli and conditions needed for expression of proteins 47b, 71/73, and 120. J Immunol 142:1639-45
Jarpe, M A; Hayes, M P; Russell, J K et al. (1989) Causal association of interferon-gamma with tumor regression. J Interferon Res 9:239-44
Pace, J L (1988) Synergistic interactions between IFN-gamma and IFN-beta in priming murine macrophages for tumor cell killing. J Leukoc Biol 44:514-20

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