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Lymphotoxin and interferon production by rosette-separated human peripheral blood leukocytes
346
WORKSHOP
ON
LYMPHOCYTE
MEDIATORS
nonspecific factor or, alternatively, multiple nonspecific factors which need to li~ld a1~1)r~~l~riately matching B-cells to be functional. “Allogcnric Factor Like” Activity Produced by T-Cells Reactive to I-loptclt-~foriili~,d SyrlH. WAGNER, A. STARZINSKY-POWITZ, K. PFIZENMAIER, J. BIER, grncic MHC-Stwwctwes. AND M. R~LLINGHOFF, Institut for Medical Microbiology, D-6500 Ma&/W. Germany,
Hochhaus am Augustuplatz. Recently, we established a system which allows the sensitization of T-cells against TNPor FITC (fluorescin-isothio-cyanat)-conjugated syngeneic stimulator cells in viva. When such ifi &o-sensitized T-cells are transferred into b vitro culture conditions, they proliferate and differentiate into hapten-specific cytotoxic T-lymphocytes (CTL). The supernatant of such cultures, in turn, allows an effective response of B-cells (anti-@-treated splenic cells of nu-nu mice) toward the thymus-dependent antigen SRBC. Thus, T-cells reactive in vivo to hapten-modified syngeneic major histocompatibility complex (MHC) structures release a factor, iti vitro, in the presence of which the requirement for T-cells in the in vitro response to SRBC can be bypassed. Whether or not this finding bears any relation to the generation of autoantibodies during a state of drug allergy remains to be established. Lylnphotoxin kocytes.
and Interferon Production by Rosette-Sebarated Humalt Perijhcral Blood LeuG. R. KLIMPEL, J. H. DEAN, K. D. DAY, P. B. CHEN, AND D. 0. LUCAS, De-
partment of Microbiology, Arizona Medical Center, Tucson, Arizona; Immunology, Litton Bionetics, Inc., Kensington, Maryland; and NIDR, tutes of Health, Bethesda, Maryland.
Department of National Insti-
Human peripheral blood leukocytes and isolated T-cell-enriched and B-cell-enriched populations were studied for their ability to produce lymphotoxin (LT) and interferon (IF) and for their proliferative response following stimulation with phytohemagglutinin (PHA) and staphylococcal phage lysate (SPL). Sheep erythrocyte-rosette-forming cells (E-RFC) were separated from nonrosetting cells by sedimentation on Ficoll-hypaque gradients. PHA induced LT production and cell proliferation primarily in E-RFC-enriched populations. In contrast, SPL induced LT production in the E-RFC-depleted population. PHA stimulated little cell proliferation in the E-RFC-depleted population, but induced high levels of IF in both populations. Further, the two populations showed different dose responses to PHA for the production of IF. SPL stimulated cell proliferation and induced IF production in both populations. Therefore, the potentials to produce LT and IF appear to be inherent in both cell populations, with their in vitro expression being dependent upon the nature of the stimulus. Studies on. T-Lymphocyte Immune Interferon. LOIS B. EPSTEIN, CHRISTINE SHAUL YATZIV, DELLA GOLDBLATT, AND MIRIAM D. SEELIG, University
R. NEUMANN, of California,
San Francisco, California. Previous studies have shown that the production of the antiviral protein, interferon (IF), by human T-lymphocytes in response to mitogenic or specific bacterial or viral antigenic stimulation in vitro can be used as a measure of the effector competence of the T-cells. The type of IF produced as a mediator of cellular immunity in these situations is called Type II or immune IF to distinguish it from Type I or classical viral induced IF. Our recent studies have shown the following. (1) Adherent cells other than macrophages, i.e., mesothelial cells, can augment the production of IF by T-lymphocytes in vitro. (2) The nature of the supporting adherent cell in the in vitro system can influence the physical characteristics of the IF produced (i.e., stability to low pH and heat). (3) Lysosomal enzyme activity of the supporting adherent cells is not correlated with their ability to augment T-lymphocyte IF production. (4) A micro-macrophage-lymphocyte culture system can now be used for serial studies of T-lymphocyte IF production. (5) The inhibition of actinomycin-resistant viral RNA synthesis can be used as the basis for a new microassay for IF, but, in contrast to conventional virus plaque reduction assays, it is less sensitive to immune than to classical IF.
WORKSHOP
ON
LYMPHOCYTE
MEDIATORS
nonspecific factor or, alternatively, multiple nonspecific factors which need to li~ld a1~1)r~~l~riately matching B-cells to be functional. “Allogcnric Factor Like” Activity Produced by T-Cells Reactive to I-loptclt-~foriili~,d SyrlH. WAGNER, A. STARZINSKY-POWITZ, K. PFIZENMAIER, J. BIER, grncic MHC-Stwwctwes. AND M. R~LLINGHOFF, Institut for Medical Microbiology, D-6500 Ma&/W. Germany,
Hochhaus am Augustuplatz. Recently, we established a system which allows the sensitization of T-cells against TNPor FITC (fluorescin-isothio-cyanat)-conjugated syngeneic stimulator cells in viva. When such ifi &o-sensitized T-cells are transferred into b vitro culture conditions, they proliferate and differentiate into hapten-specific cytotoxic T-lymphocytes (CTL). The supernatant of such cultures, in turn, allows an effective response of B-cells (anti-@-treated splenic cells of nu-nu mice) toward the thymus-dependent antigen SRBC. Thus, T-cells reactive in vivo to hapten-modified syngeneic major histocompatibility complex (MHC) structures release a factor, iti vitro, in the presence of which the requirement for T-cells in the in vitro response to SRBC can be bypassed. Whether or not this finding bears any relation to the generation of autoantibodies during a state of drug allergy remains to be established. Lylnphotoxin kocytes.
and Interferon Production by Rosette-Sebarated Humalt Perijhcral Blood LeuG. R. KLIMPEL, J. H. DEAN, K. D. DAY, P. B. CHEN, AND D. 0. LUCAS, De-
partment of Microbiology, Arizona Medical Center, Tucson, Arizona; Immunology, Litton Bionetics, Inc., Kensington, Maryland; and NIDR, tutes of Health, Bethesda, Maryland.
Department of National Insti-
Human peripheral blood leukocytes and isolated T-cell-enriched and B-cell-enriched populations were studied for their ability to produce lymphotoxin (LT) and interferon (IF) and for their proliferative response following stimulation with phytohemagglutinin (PHA) and staphylococcal phage lysate (SPL). Sheep erythrocyte-rosette-forming cells (E-RFC) were separated from nonrosetting cells by sedimentation on Ficoll-hypaque gradients. PHA induced LT production and cell proliferation primarily in E-RFC-enriched populations. In contrast, SPL induced LT production in the E-RFC-depleted population. PHA stimulated little cell proliferation in the E-RFC-depleted population, but induced high levels of IF in both populations. Further, the two populations showed different dose responses to PHA for the production of IF. SPL stimulated cell proliferation and induced IF production in both populations. Therefore, the potentials to produce LT and IF appear to be inherent in both cell populations, with their in vitro expression being dependent upon the nature of the stimulus. Studies on. T-Lymphocyte Immune Interferon. LOIS B. EPSTEIN, CHRISTINE SHAUL YATZIV, DELLA GOLDBLATT, AND MIRIAM D. SEELIG, University
R. NEUMANN, of California,
San Francisco, California. Previous studies have shown that the production of the antiviral protein, interferon (IF), by human T-lymphocytes in response to mitogenic or specific bacterial or viral antigenic stimulation in vitro can be used as a measure of the effector competence of the T-cells. The type of IF produced as a mediator of cellular immunity in these situations is called Type II or immune IF to distinguish it from Type I or classical viral induced IF. Our recent studies have shown the following. (1) Adherent cells other than macrophages, i.e., mesothelial cells, can augment the production of IF by T-lymphocytes in vitro. (2) The nature of the supporting adherent cell in the in vitro system can influence the physical characteristics of the IF produced (i.e., stability to low pH and heat). (3) Lysosomal enzyme activity of the supporting adherent cells is not correlated with their ability to augment T-lymphocyte IF production. (4) A micro-macrophage-lymphocyte culture system can now be used for serial studies of T-lymphocyte IF production. (5) The inhibition of actinomycin-resistant viral RNA synthesis can be used as the basis for a new microassay for IF, but, in contrast to conventional virus plaque reduction assays, it is less sensitive to immune than to classical IF.