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C J Burger

Publications and source records attributed to C J Burger.

At least 37 records · Page 2Linked to original sources
PubMed1992

Cytokines and suppressor macrophages cause tumor-bearing host CD8+ T cells to suppress recognition of allogeneic and syngeneic MHC class II molecules.

Quantitative and qualitative tumor-associated changes in T cell phenotype and function were identified in CD8+ T cells. Tumor growth changed splenic CD4+/CD8+ T cell ratios and induced the appearance of more cells with the CD8+ phenotype. In comparison to equal concentrations of normal host (NH) counterparts, tumor-bearing host (TBH) CD8+ T cells were highly suppressive to allorecognition and autorecognition. Suppression was not due to quantitative reductions in CD4+ T cells, although minor qualitative differences were observed. Suppression appeared to be mediated partly by prostaglandin E2 (PGE2). Interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) contributed to TBH CD8+ T cell-mediated suppression. Blocking studies using monoclonal antibodies (mAb) in conjunction with indomethacin suggested that cytokine networks involving IFN-gamma, IL-4, and PGE2 were disrupted during tumor growth and promoted TBH CD8+ T cell suppression. Alloresponses and autoresponses were significantly suppressed when TBH CD8+ T cells mediated these reactions simultaneously with TBH Ia- macrophages. Inhibition of PGE2 production was unable to reverse the additive suppression caused by these two cell types. These results collectively suggest that tumor-induced changes in CD8+ T cells lead to suppressed allo-recognition and autorecognition through both soluble mediator molecules and cellular interactions.

Animals↗
PubMed1992

Tumor growth changes the contribution of granulocyte-macrophage colony-stimulating factor during macrophage-mediated suppression of allorecognition.

Tumor-bearing host (TBH) macrophages (M phi) suppress T cell alloresponses, and this study suggests granulocyte-macrophage colony-stimulating factor (GM-CSF), a molecule associated with suppressive M phi activity during tumor growth, signals more immunosuppression. In the absence of M phi, GM-CSF increased T cell proliferation in response to alloantigen. However, TBH M phi-mediated suppression of allorecogntion was further induced by GM-CSF. Allogeneic mixed lymphocyte reaction (MLR) cultures, containing normal host (NH) M phi, were either unaffected or enhanced. Prostaglandin E2 (PGE2), a highly suppressive monokine that decreases alloreactivity, did not seem to be involved in the suppression caused by the TBH M phi/GM-CSF interaction. M phi-CSF (M-CSF) addition to cultures did not reverse the suppression caused by TBH M phi and GM-CSF, and inhibition of PGE2 synthesis did not change the response to M-CSF. TBH Ia- M phi, a suppressor population that predominates among splenic M phi during tumor growth, demonstrated significantly lower reactivity in the presence of GM-CSF. In contrast, alloresponses suppressed by NH Ia- M phi demonstrated higher reactivity in the presence of GM-CSF. The data collectively suggest that TBH M phi respond differently to GM-CSF, and that tumor-induced changes in GM-CSF responsiveness affect M phi accessory ability.

Animals↗
PubMed1992

Tumor growth changes responsiveness to and production of granulocyte-macrophage colony-stimulating factor during recognition of self MHC class II molecules.

Tumor growth decreases T-cell recognition of self major histocompatibility complex (MHC) class II molecules by inducing changes in splenic macrophage (M phi) phenotype and function. The current investigation shows tumor-induced alterations in autorecognition also are associated with changes in responsiveness to and production of granulocyte-M phi colony-stimulating factor (GM-CSF). In contrast to normal host (NH) M phi, tumor-bearing host (TBH) M phi failed to express higher MHC class II molecule density after exposure to GM-CSF. Autoreactive T cells stimulated by either NH or TBH M phi were suppressed by GM-CSF. Inhibition of prostaglandin E2 (PGE2) synthesis reversed M-CSF-induced suppression of autoreactivity to NH M phi and, to a lesser extent, to TBH M phi. When TBH autoreactive T cells were stimulated by TBH M phi, autoreactivity increased when GM-CSF was added and PGE2 synthesis was inhibited. Although GM-CSF can contribute to tumor-induced suppression, it did not affect the contribution of GM-CSF during autorecognition. Increased GM-CSF production was responsible, at least in part, for the TBH M phi-mediated suppression. Low concentrations of GM-CSF were produced endogenously by tumor isolates, and GM-CSF production was significantly increased when isolates were stimulated with lipopolysaccharide. Autoreactive T cells stimulated solely by TBH M phi produced more GM-CSF than autoreactive T cells stimulated by NH M phi. Cultures supplemented with several concentrations of NH or TBH M phi produced similar amounts of GM-CSF in a dose-dependent manner. Inhibition of PGE2 synthesis by NH and TBH M phi reduced GM-CSF production equally. Collectively, these results suggest that during tumor growth, responsiveness to and production of GM-CSF alters recognition of self MHC class II molecules.

Animals↗
PubMed1990

Tumor modulation of autoreactivity: decreased macrophage and autoreactive T cell interactions.

The autologous mixed lymphocyte reaction (AMLR) is an in vitro measure of autoreactivity, a key mechanism in immune homeostasis. In this system, macrophages (M phi) act as accessory cells to autoreactive L3T4+ T cells by presenting self-Ia and releasing soluble modulators. During tumor growth, changes occur in M phi and T cells. Tumor-bearing host (TBH) M phi have a reduced ability to act as accessory cells. In fact, TBH M phi suppressed autoreactivity by 60-70%. The decrease in TBH M phi or T-cell abilities was not due to differences in cell numbers or incubation time. Because tumor growth causes increased prostaglandin E2 (PGE2) production by M phi, indomethacin was used to assess the contribution of prostaglandins. Normal and TBH T-cell reactivity increased nearly 50% when stimulated by normal host M phi, while normal and TBH T-cell reactivity increased nearly 100% when stimulated by TBH M phi. Thus increased prostaglandin production is partly responsible for the increased TBH suppressor M phi activity and in the normal host, suppressor M phi may be responsible for maintaining immune regulation. To assess the direct role of prostaglandins in T-cell hyporesponsiveness, PGE2 was titrated into the cultures. PGE2 suppressed normal and TBH T-cell responsiveness in a dose-dependent manner. Normal host T cells were suppressed to a greater extent than TBH T cells by PGE2 (66% versus 42% suppression, respectively). Reduced Ia expression and active suppressor mechanisms are not the only mechanisms mediating hypoautoreactivity during tumor growth. TBH autoreactive L3T4+ T cells were less responsive to self-Ia; they were only 60-80% as reactive as their normal counterparts. To address whether the helper T (TH)-cell defect involved cytokines, T cells were treated with interleukin (IL)-1, IL-2, and IL-4. In all cases, the TBH T-cell response to the factors was decreased (only 60-75% as reactive as normal T cells). Because TBH M phi-mediated suppression can override the addition of IL-1, IL-2, and IL-4, indomethacin was also added with the exogenous interleukins. This coaddition significantly enhanced normal host autoreactivity above control levels while TBH autoreactivity (the combination of TBH T cells and TBH M phi) only returned to normal host unstimulated levels. Tumor growth modulates the immune response at least by (i) decreasing the accessory cell abilities of TBH M phi through decreased Ia expression and increased production of suppressive molecules such as prostaglandins; and (ii) decreasing the responsiveness to immune enhancing factors by TH cells.

Animals↗
PubMed1990

Two-color flow cytometric analysis of the expression of MAC and MHC class II antigens on macrophages during tumor growth.

Tumor-bearing host (TBH) macrophages (M phi) exhibit immune dysfunction that is concomitant with phenotypic changes. We examined M phi subpopulations by changes in the expression of surface antigens Mac-1, -2, -3, and Ia on normal and TBH peritoneal and splenic M phi. M phi were double-labeled and analyzed by flow cytometry to observe multiple expression of surface antigens. Tumor growth alters the multiple expression of these M phi markers. Peritoneal and splenic M phi had different Mac+ and Mac+Ia+ population percentages. In TBH, peritoneal M phi had decreased percentages of Mac-1+2+, Mac-1+3+, Mac-2+3+, and Mac+Ia+ M phi. This decrease correlated with functional changes in TBH M phi. In contrast, there was an increase in Mac-2-Ia- TBH peritoneal M phi. Previously undiscovered Mac-1+2-3- and Mac-1-2-3+ populations were found. In contrast to peritoneal M phi, there was an increase in the percentage of Mac-1+2+, Mac-1+3+, and Mac-2+3+ splenic TBH M phi but, like peritoneal M phi, there was a decrease in the percentage of Mac+Ia+ M phi. Also, TBH splenic M phi showed a smaller but more uniform antigen density than normal host splenic M phi. Tumor growth modulated phenotypic alterations in peritoneal and splenic M phi subpopulations. Combined with earlier functional studies of M phi subpopulations, these data suggested a relationship between changes in M phi phenotype and tumor-induced dysfunction of M phi-modulated immune activity.

Animals↗
PubMed1990

Modulation of alloreactivity by Mac-1+, -2+, and -3+ macrophages from normal and tumor-bearing hosts: flow cytofluorometrically separated macrophages.

Macrophages (M phi) are multifunctional cells that regulate humoral and cellular immune responses. Our studies of tumor-induced M phi-mediated dysfunction used M phi subsets which were defined by their Mac-1, Mac-2, and Mac-3 surface markers. To measure the accessory activity of M phi for T cell alloreactivity, thioglycollate-elicited peritoneal M phi from normal and tumor-bearing hosts (TBH) were labeled with anti-Mac-1, -2, or -3 antibodies and separated by flow cytofluorometry. The separated Mac-1+, -2+, and -3+ M phi were called sorted M phi, while unseparated M phi were designated unsorted M phi. Both M phi types were added to mixed lymphocyte reaction (MLR) cultures at concentrations ranging from a low of 2% M phi to a high of 20% M phi. The low concentration of unsorted normal host M phi caused a 31% suppression of alloreactivity. Suppression reached 68% when high concentrations of unsorted normal host M phi were added to the MLR cultures. Unsorted TBH M phi reduced alloreactivity by 64% and 86% at low and high concentrations, respectively. When separated into subpopulations, normal host Mac-1+ M phi reduced alloreactivity by 48% and 81% when added at low and high concentrations, respectively. TBH Mac-1+ reduced alloreactivity by 31% and 59% at low and high concentrations, respectively. There were no differences in the suppression caused by normal or TBH Mac-2+ M phi, and by normal or TBH Mac-3+ M phi. Indomethacin treatment did not effect the suppression caused by Mac-1+ M phi, suggesting that proataglandin E2 was not involved. Indomethacin treatment did reduce suppression mediated by Mac-2+, -3+, and unsorted M phi. Mac-2+ M phi dramatically enhanced alloreactivity at low concentrations with normal host Mac-2+ M phi providing greater enhancement of alloreactivity than TBH Mac-2+ M phi. The division of M phi into subpopulations on the basis of Mac antigens suggested that Mac-1+ and -3+ M phi played a major role in immunosuppression in the normal host, while Mac-3+ M phi were more active in TBH immunosuppression. Because no one population of sorted TBH M phi was more suppressive than sorted normal host M phi, we suggest that tumor-induced immunosuppression may involve a network of suppressor M phi.

Animals↗
PubMed1990

Normal and tumor-bearing host splenic macrophage responses to lipopolysaccharide.

Lipopolysaccharide (LPS) was used to assess the responses of normal and tumor-bearing host (TBH) macrophages (M phi) to activation signals. M phi were incubated with LPS for either 3 hr or 24 hr and then assayed for phenotypic, functional, or cell-cycle changes. A 3-hr LPS treatment had no significant effect on M phi phenotype. In contrast, the 24-hr LPS treatment caused a significant decrease in the percentage of normal host Mac+ and Ia+ M phi. In the TBH, a 24-hr LPS treatment caused an increase in the percentage of Mac-1+ and -2+ M phi and a decrease in the percentage of Mac-3+ and Ia+ M phi. When normal host M phi were plated for 24 hr (control), there was an increase in Mac-1+ and -2+ M phi and a decrease in Ia+ M phi. The 24-hr TBH control showed a decrease in Mac-1+ and Ia+ M phi. To assess functional changes, LPS-treated normal and TBH M phi were added to the allogeneic mixed lymphocyte reaction (MLR) or the autologous MLR (AMLR). There were no significant differences in M phi accessory activity after a 3-hr LPS treatment. A 24-hr LPS treatment of normal host M phi also had no effect. A 24-hr LPS treatment of TBH M phi led to a significant decrease in allogeneic T-cell reactivity, and even the 24-hr TBH control showed significant suppression of T-cell responsiveness. In the AMLR, a measure of autoreactive T-cell responsiveness, a 3-hr LPS treatment had no affect on normal host M phi but led to increased accessory ability in TBH M phi. TBH M phi, however, were still less than 50% as responsive as normal host M phi even after LPS treatment. The 24-hr LPS treatment caused a significant decrease in normal or TBH M phi accessory activity. A 24-hr plating of normal host M phi decreased their accessory ability. In addition to the phenotypic and functional changes after LPS treatment, M phi cell-cycle kinetics were also investigated. The percentages of normal host M phi in G0/G1 were not changed significantly after a 3-hr LPS treatment. In contrast, M phi plated for 24 hr with or without LPS had a decreased percentage of cells in G0/G1. Normal host M phi showed little change in total RNA levels after a 3-hr treatment but had increased RNA levels after a 24-hr treatment.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Tumor-induced variations in a high molecular weight inhibitory monokine.

A soluble inhibitor of T cell proliferation was demonstrated in splenic and peritoneal macrophage (M phi) culture supernatants and was variably glycosylated during tumor development. This factor(s) inhibited mixed lymphocyte reaction- (MLR) and concanavalin A-induced T cell proliferation and was not prostaglandin E2 (PGE2), as demonstrated by size (nondialyzable, found in a 30 kilodalton [kd] ultrafiltration retentate and in fractions corresponding to greater than 67 kd on S-200 Sephacryl chromatography) and time course of activity in the MLR. Concentrated supernatants were PGE2-free and yet inhibited proliferation in the MLR. Isoelectric focusing (IEF) revealed normal and tumor-bearing host (TBH) concentrated M phi supernatants' major peaks of inhibitory activity differed in charge, with a pI of 6.5-7.6 for normal hosts and 4.0-6.0 for TBH. Activity in TBH M phi supernatants was found primarily in fractions eluting from hydroxylapatite at 0.3 M sodium phosphate buffer, pH 7.3, was resistant to proteolytic enzymes, but was sensitive to neuraminidase. In contrast, inhibitory activity in normal host M phi supernatants eluted from hydroxylapatite at 0.6 M sodium phosphate buffer and was not sensitive to neuraminidase. Thus, variable glycosylation (presence and absence of sialic residues) could account for the charge difference seen in IEF and may have a bearing on tumor-induced hyporesponsiveness. Kinetic addition of supernatants to the MLR revealed PGE2 may be required for inhibitory activity to be manifested early (0 and 24 h) but not if the high molecular weight (mw) inhibitor was added late (48 and 72 h post initiation). Both normal and TBH M phi supernatants suppressed the generation of interleukin 2 (IL 2) with a dose- and time-dependent difference. Cell-cycle analysis of mitogen-stimulated cells treated with normal and TBH M phi supernatants revealed that TBH M phi supernatants enhanced cell-cycle progression when measured early, but that both normal and TBH M phi supernatants suppressed the number of cells in S phase when measured late in the assay. The production of a high mw inhibitor by both normal and TBH M phi could suggest a homeostatic mechanism, which was upset by increased PGE2 production and production of an inhibitor containing sialic acid, tipping the balance in favor of immunosuppression.

Animals↗
PubMed1987

Reduced immune responsiveness and lymphoid depletion in mice infected with Ehrlichia risticii.

The histopathology of the thymus and spleen and the response of spleen cells to mitogenic stimuli were evaluated in Sprague-Dawley CF-1 mice infected with Ehrlichia risticii. Intraperitoneal injection of 10(4) or 10(6) E. risticii-infected U-937 cells into mice resulted in 100% morbidity and partial mortality. Thymic atrophy became significant between 1 and 2 weeks postinfection and remained for the duration of the study. The atrophy appeared associated with antecedent destruction and rarefaction of lymphocytes, resulting in the loss of corticomedullary demarcation. Splenomegaly was prominent; significantly increased weights were detected 7 days postinfection. Histopathologic examination revealed rarefaction of lymphocytes around central arteries, the presence of necrotic debris in histiocytes, and replacement of erythropoiesis by granulopoiesis in the red pulp. Marked and acute reduction of in vitro proliferative responses of spleen cells to concanavalin A (ConA) and phytohemagglutinin were observed in mice infected with 10(4) or 10(6) E. risticii-infected U-937 cells. Interleukin-2 activity in the supernatant of ConA-stimulated spleen cells was also severely reduced. Both changes were time- and dose-dependent and were not associated with decreased spleen cell viability. Neither morbidity nor mortality occurred in mice infected with 10(2) E. risticii-infected U-937 cells. Although there was temporal reduction in phytohemagglutinin-driven lymphocyte proliferation, reduction in neither ConA-driven lymphocyte proliferation nor interleukin-2 activity was observed with this dosage. All E. risticii-inoculated mice seroconverted between days 18 and 25, as detected by the indirect fluorescent-antibody procedure. The findings indicate for the first time the hypoimmune responsiveness and histopathologic changes in lymphoid organs associated with E. risticii infection.

Animals↗
PubMed1986

An improved data analysis method for interleukin 2 microassay.

Development of the interleukin 2(IL 2) microassay, coupled with the use of highly purified or recombinant factors has allowed a detailed examination of the mechanism of action of this important biological response modifier. However, probit analysis of the microassay data does not allow inherent error of the system to be approximated nor can units of activity be assessed for significance. A computer program was developed to analyze the validity of each regression line and to generate 95% confidence intervals around each line. This program employs analysis of variance, linear regression analysis and the parallel line assay to fix confidence intervals for each IL 2 unit value. The use of recombinant IL 2 as an immunomodulator in clinical settings warrants a more precise statistical method to evaluate normal fluctuations of this factor than currently in use. The development of such a method is presented here.

Biological Assay↗
PubMed1986

Taenia taeniaeformis: inhibition of mitogen induced proliferation and interleukin-2 production in rat splenocytes by larval in vitro product.

Splenocytes from rats infected with Taenia taeniaeformis showed an early decreased proliferative response to the mitogen concanavalin A with larval growth. When larvae culture supernatant (in vitro product) was added to normal rat splenocytes, there was a decrease in the proliferative response to concanavalin A and phytohemagglutinin. The ability of infected rat splenocytes to produce interleukin-2 was decreased with larval growth. Addition of in vitro product to culture medium significantly depressed interleukin-2 production by normal as well as infected rat spleen cells. Culturing normal rat splenocytes with in vitro product for 3 days induced a suppressor cell population. When these cultured cells were admixed with fresh normal rat splenocytes plus concanavalin A, the proliferative response of the fresh cells was significantly reduced. The present results suggest that in vitro product secreted by larvae in hepatic cysts causes subversion of the cellular immune response in the host. Induction of a suppressor cell population could suppress interleukin-2 production which may lead to the inhibition of differentiation and proliferation of specific cytotoxic T lymphocytes.

Animals↗
PubMed1984

Interleukin 2 (IL-2) activity during tumor growth: IL-2 production kinetics, absorption of and responses to exogenous IL-2.

A temporal study assessed the relationship between fibrosarcoma growth and immunologic encumbrance due to the inability of BALB/c mouse splenocytes to elaborate the lymphokine Interleukin 2 (IL-2). Nylon-wool fractionation and antiserum treatments suggested the existence of a mildly nylon-wool-adherent, anti-Lyt 2-sensitive tumor-induced suppressor T (Ts) cell which significantly decreased IL-2 activity. Absorption investigations indicated that ligand-activated tumor-bearing host (TBH) spleen cells were less receptive to IL-2 than their normal counterparts. When splenocytes were antiserum treated before absorption, removal of Lyt 2+ (suppressor T) cells resulted in greater IL-2 absorption by the remaining cells. Purified IL-2 only partially restored suppressed TBH spleen cell mitogen- or alloantigen-induced blastogenesis; whereas, normal host reactivity was significantly augmented. The collective data suggest that TBH spleen cells were capable of producing IL-2 and of responding to the IL-2 amplification signal when tumor-induced Ts cells were depleted.

Absorption↗
PubMed1984

Interleukin 3 activity in tumor-bearing hosts: decreased splenocyte production of and responsiveness to IL 3.

A kinetic study assessing the relationship between tumor growth and the ability of BALB/c mouse splenocytes to produce Interleukin 3 (IL 3) indicated a concomitant decrease in IL 3 activity with tumor growth. Tumor-bearing host (TBH) splenocytes produced 600 pmoles/hr/10(8) cells of IL 3 activity at Day 0 but only 62 pmoles/hr/10(8) cells by Day 28 post tumor cell inoculation. Nylon wool fractionation (to remove adherent suppressor cells) did not restore IL 3 activity. Addition of purified IL 3 to mitogen proliferation assays showed that IL 3 alone was mitogenic for normal host but not TBH splenocytes. In concert with concanavalin A (Con A) and phytohemagglutinin, IL 3 augmented in vitro normal host splenocyte responsiveness but significantly further suppressed it in the TBH. An absorption assay indicated that fresh cells had acceptors to remove IL 3 from supernatants. Con A-induced normal or TBH blast cells lost their ability to absorb IL 3. Intravenous inoculation of purified IL 3 into normal and TBH resulted in further suppression of TBH splenocyte mitogen-induced blastogenesis. The exacerbation of TBH spleen cell reactivity by IL 3 may be due to a tumor-induced feedback inhibition mechanism further suppressing cellular differentiation critical to cytotoxic T lymphocyte maturation.

Animals↗
PubMed1983

Level of macrophage induction during tumor growth: primed or activated?

The activation level of thioglycolate (TG)-elicited peritoneal macrophages (Mphi) from normal and fibrosarcoma-bearing BALB/c mice was investigated. Lipopolysaccharide was used to distinguish in vitro levels of Mphi activation. The results of cytotoxicity assays, which measured nonspecific killing of P815 tumor cells by activated Mphi, indicated that tumor-bearing host Mphi were primed in situ while normal host Mphi remained in a resting state. This level of Mphi induction may contribute to the resulting tumor-bearing host T cell immune hyporesponsiveness.

Animals↗
PubMed1983

Conversion of normal host splenocytes to suppressor cells by tumor-induced suppressor T-cell-derived factor(s): cyclophosphamide treatment reverses inhibitory activity.

Suppressor T (Ts) cells (or their factors) that arise in BALB/c mice as a consequence of fibrosarcoma cell growth can be adoptively transferred and can recruit new regulatory cells in vitro. In vivo temporal studies indicated that Ts cells significantly inhibited blastogenesis in normal host splenocytes as early as 4 h after adoptive transfer and lasted as long as 5 days. Suppressor T-cell-derived factor(s) did not suppress until 24 h after in vivo administration, and effects were protracted beyond time periods observed with Ts cells. Supernatants containing suppressor factor(s) not only significantly inhibited normal host spleen cell proliferation but also induced and/or recruited cells to become suppressive. Cyclophosphamide treatment of tumor-bearing hosts restored their in vitro spleen cell blastogenic ability and abolished in vitro suppression by Ts cells or their factors. This was confirmed in vivo by passive transfer experiments. Macrophage removal seemed to augment cyclophosphamide's ability to eliminate suppression.

Animals↗
PubMedDate not supplied

Interferon-gamma reduces tumor-induced Ia- macrophage-mediated suppression: role of prostaglandin E2, Ia, and tumor necrosis factor-alpha.

Tumor growth enhances macrophage (M phi) suppressor activity by causing M phi to increase synthesis of inhibitory molecules such as prostaglandin E2 (PGE2) or decreasing their expression of up-regulatory molecules such as the class II MHC protein Ia. Although these tumor-induced changes are correlated, it is unknown whether tumor-bearing host (TBH) Ia- M phi become more suppressive by increasing their PGE2 synthesis. To assess the role of PGE2 in tumor-induced Ia- M phi-mediated suppression of CD4+ T-cell alloreactivity, unseparated (Ia(+)-enriched) or Ia(+)-depleted (Ia-) populations of murine normal host (NH) or TBH splenic M phi were added to mixed lymphocyte reaction (MLR) cultures. NH or TBH Ia- M phi were significantly more suppressive than their respective unseparated populations, and TBH Ia- M phi were more suppressive than their NH counterparts. When PGE2 production was blocked with indomethacin, TBH Ia- M phi-mediated suppression was reduced more than suppression mediated by all other M phi populations. A PGE2-specific ELISA showed more PGE2 in Ia- M phi-containing cultures than in those with whole M phi and more in cultures containing TBH Ia- M phi than in their NH counterparts. Because interferon-gamma (IFN-gamma) is a potent M phi activation molecule that regulates both Ia expression and PGE2 production, the effects of IFN-gamma on tumor-induced Ia- M phi-mediated suppression were investigated. Exogenous IFN-gamma reduced suppression mediated by all M phi populations except NH unseparated M phi. IFN-gamma suppressed alloreactivity without M phi or with NH unseparated M phi. Suppression mediated by NH or TBH Ia-, and TBH unseparated M phi was also reduced when M phi were pre-incubated with IFN-gamma before their addition to MLR cultures. IFN-gamma addition did not block Ia- M phi-mediated suppression by decreasing M phi PGE2 production. In fact, IFN-gamma addition increased PGE2 production two-fold in MLR cultures. However, IFN-gamma partly reduced suppression mediated by exogenous PGE2 added to M phi-depleted cultures. Cytofluorometric analysis showed that IFN-gamma increased the percentage of Ia+ M phi in NH and TBH Ia- M phi populations. Blocking TNF-alpha activity with anti-TNF-alpha antibodies caused IFN-gamma to suppress alloreactivity in all M phi-added cultures. Collectively, these data show that tumor-induced suppression mediated by Ia- M phi is caused by increased PGE2 synthesis.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMedDate not supplied

Normal and tumor-bearing host macrophage responses: variability in accessory function, surface markers, and cell-cycle kinetics.

Normal and tumor-bearing host (TBH) peritoneal macrophage (M phi) responses to in vitro lipopolysaccharide (LPS) treatment were measured by assessing functional and phenotypic changes. Both normal and TBH untreated M phi suppressed mixed lymphocyte reaction (MLR) reactivity at all concentrations. Normal host M phi treated with LPS for 3 h were suppressive at all concentrations. TBH M phi treated with LPS for 3 h were not suppressive in the MLR until more than 5% were added. Surprisingly, 24 h treatment of normal and TBH M phi with LPS induced cells that significantly enhanced MLR reactivity when added at 2% or 5%. These cells were not suppressive until a 20% M phi concentration was reached. LPS treatment of normal and TBH M phi changed the percentage of cells expressing the surface markers Mac-1, -2, -3, and Ia as determined by flow cytometry. Normal host peritoneal M phi treated with LPS for 3 h had decreased Mac-1 and -3 expression, but there was no change in Mac-2 or Ia. Plating for 24 h did not change the percentage of M phi expressing Mac-1, -3, or Ia but did cause an increase in Mac-2+ M phi. Treatment of normal host M phi with LPS for 24 h led to a decrease in Mac-1+ and Ia+ M phi, no change in Mac-3+ M phi, but an increase in Mac-2+ M phi. LPS treatment of TBH M phi for 3 h decreased the number of Mac-1+ M phi, but Mac-2+, -3+, or Ia+ M phi numbers did not change. Plating TBH M phi for 24 h caused a decrease in the number of Mac-1+ M phi, no change in Mac-3+ or Ia+ M phi, but an increase in Mac-2+ M phi. Treatment with LPS for 24 h led to no change in the number of Mac-1+, -3+, or Ia+ TBH M phi, but Mac-2+ M phi increased. The phenotypic and functional changes after LPS treatment led us to ask if these changes were detectable at the level of DNA and RNA. Flow cytometric analysis of acridine orange-stained M phi was used to measure DNA and RNA levels. This analysis determines M phi cell-cycle kinetics and estimates their RNA synthesis. In normal host M phi, a 3-h LPS treatment caused a decrease of cells in G0/G1 but an insignificant change in RNA levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Acridine Orange↗