Vascular endothelial cells enhance T cell responses by markedly augmenting IL-2 concentrations

Cultured human endothelial cells (EC), dermal fibroblasts (DF), and blood monocytes were compared for their effects on IL-2 concentration measured in the medium of both unfractionated peripheral blood mononuclear cells (PBMC) and highly enriched CD4+ T cell populations maximally stimulated by the polyclonal mitogen phytohemagglutinin (PHA). EC, but not DF or blood monocytes, could markedly augment IL-2 concentrations, up to 30-fold or more. This action of EC could not be replaced by fixed EC, EC-conditioned medium, or recombinant IL-1. Antibody to CD2 but not to CD18 blocked the EC effect. The augmented IL-2 concentrations generated in the presence of EC appeared biologically active in that the addition of living EC conferred a proliferative advantage to PBMC at suboptimal PHA concentrations, an effect which could be mimicked by exogenous IL-2. We propose that EC augmentation of IL-2 synthesis may contribute to the relatively unique ability of EC to stimulate a primary allogeneic response in vitro and may function in vivo to boost T cell responses to limiting quantities of antigen.     

Lysozyme-induced inhibition of the lymphocyte response to mitogenic lectins

Both human lysozyme (HL) and hen egg white lysozyme (HEWL) inhibited the proliferative response of peripheral blood lymphocytes to T cell mitogens such as the lectins phytohemagglutinin and concanavalin A. This inhibition was observed both when HL or HEWL was added to the lymphocyte cultures in combination with phytohemagglutinin or concanavalin A and when lymphocytes were pretreated with either lysozyme and extensively washed prior to culture with mitogens. Under both conditions, the effects were strictly dose dependent; the lysozyme concentrations yielding maximal inhibitory effect were 5 micrograms/ml for HL and 1 microgram/ml for HEWL, while both lower and higher concentrations were less effective. Specific antilysozyme rabbit sera completely prevented the inhibitory effects of both HL and HEWL on the proliferative response of lymphocytes to phytohemagglutin or concanavalin A. Chitotriose (a lysozyme inhibitor) caused a strong reduction in the inhibitory effects of the two lysozymes on the lymphocyte response to either lectin. HL and HEWL also were found to markedly inhibit the polyclonal B cell proliferation and differentiation induced by pokeweed mitogen and T cells. A less marked inhibition was also obtained when T cells, but not B cells, were pretreated with HL or HEWL. Again, as in the experiments with T cell mitogens, the effects were dose dependent and 5 micrograms/ml HL and 1 microgram/ml HEWL proved to be the most effective concentrations. The possible mechanisms by which lysozyme inhibits the lymphocyte response to mitogenic lectins are considered and discussed. The enzymatic activity seemed to perform an essential function, as shown by the loss of effect when the heat- or trypsin-inactivated lysozymes were used and by the fact that only the enzymatically active compound, among certain semisynthetic derivatives of HEWL, inhibited the lymphocyte response to the mitogens. However, the cationic properties of the lysozyme molecule appeared to be essential too, since enzymes with a similar specificity of action showed effects similar to those observed with HL or HEWL only when they carried a strong positive charge. It is suggested that lysozyme, which is naturally secreted by monocytes and macrophages, might interact with lymphocyte surface receptor sites and participate in the complex mononuclear phagocyte-lymphocyte interactions and in the modulation of lymphocyte activation.     

Effect of catalase on the proliferation of human lymphocytes to phorbol myristate acetate

Phorbol esters have been documented to stimulate the proliferation of human blood mononuclear cell cultures. In addition, these agents are also known to stimulate the production and release of reactive oxygen species by monocytes. We demonstrated previously that H2O2, one of these oxygen metabolites, impairs the proliferative capacity of human blood lymphocytes. Therefore, in these experiments, we determined whether or not the H2O2 released by monocytes after activation by PMA modifies the proliferation of lymphocytes to this agent. Human blood mononuclear cells (80% lymphocytes and 20% monocytes) were incubated with PMA, and lymphoblastic transformation (LBT) was quantitated at 3 and 5 days by pulsing the cultures with thymidine. Initial experiments established that the concentration of PMA required for optimal LBT was 50 ng/ml. We then demonstrated that this concentration of PMA also induces a burst in hexose monophosphate shunt activity and H2O2 production of mononuclear cells as indicated by the enhanced oxidation of 14C-glucose and 14C-formate, respectively. The amount of H2O2 released into the medium was substantial. Our measurements indicate that the concentration of H2O2 could reach values as high as 0.008 mM during the first 2 hr of the cultures. The addition of catalase to PMA-treated cultures in concentrations sufficient to scavenge the H2O2 released by the monocytes was associated with an enhanced thymidine uptake (mean 79%). These results indicate that the hydrogen peroxide released by the monocytes modifies the response of lymphocytes to the PMA. Paradoxically, mononuclear cell cultures depleted of monocytes also had a lower proliferation to PMA than mononuclear cell cultures. This observation indicates that monocytes also produce factors required for lymphocyte proliferation to PMA such as an interleukin. In contrast, to PMA cultures, catalase did not alter the proliferation of mononuclear cell cultures stimulated by PHA. We previously documented that PHA does not stimulate an immediate burst in the oxidative metabolism of mononuclear cultures. Therefore, the effect of catalase in these two culture systems appears to correlate with the capacity of the mitogen to stimulate the oxidative metabolism of mononuclear cells. These observations suggest that the release of reactive oxygen species by monocytes may modify the response of lymphocytes to antigens both in vitro and in vivo.     

The role of monocytes in the effects of β-endorphin and selective agonists of μ-and δ-opiate receptors on the proliferative activity of peripheral blood lymphocytes

The effects of β-endorphin under the conditions of naloxone hydrochloride blockade of opiate receptors, as well as the effects of the selective agonists of μ-and δ-receptors DAGO and DADLE and the effects of melanocyte-potentiating factor (MPF), on the in vitro proliferative response of lymphocytes were studied. The dose-effect dependence indicated stimulating effects of β-endorphin, DAGO, and DADLE on the proliferative response in the presence of phytohemagglutinin (PHA). The tetrapeptide MPF, which is the C-terminal sequence of β-endorphin, had almost no effect on the proliferative activity of lymphocytes. β-Endorphin, naloxone, and the μ-and δ-receptor selective agonists enhanced the proliferative response of lymphocytes in an unfractionated cell culture, whereas β-endorphin, naloxone, and DAGO suppressed the proliferative activity of lymphocytes in the mononuclear fraction purified of monocytes. In both cases, the naloxone blockade of opiate receptors enhanced rather than eliminated the β-endorphin effect.     

Lack of specificity in the mechanisms involved in the enhancement of the concanavalin A driven human T lymphocyte stimulation by beta-endorphin: studies on activation marker expression, cell cycle and interleukin release.

We report on the effects of a physiological concentration of Beta-Endorphin (BE) (10(-12)M) on Concanavalin A (ConA) stimulated human peripheral blood T-lymphocytes and monocytes. We evaluated the effect of timing of BE addition to the culture medium on thymidine uptake, the kinetics of expression of activation markers (CD69, CD25 and CD71) on CD4+ and CD8+ lymphocytes, and of class II MHC antigens on CD14+ cells (monocytes), the kinetics of interleukin-1 (IL-1), interleukin-2 (IL-2) and interferon gamma (IFN-gamma) release, and the cell cycle. Data show that BE is able to influence T lymphocyte only when added together with ConA at the beginning of culture, suggesting its major activity is on the early phases of the T cell response. BE did not increase the amount of class II MHC antigens on monocytes and did not preferentially stimulate CD69, CD25 and CD71 antigen expression on either CD4+ or CD8+ lymphocytes. After 24 hours, the relative proportions of CD4+ and CD8+ lymphocyte in S and G2-M phases were not affected by BE, although the opioid did augment the number of cells in the proliferative compartments of the cell cycle, S and G2-M, indicating an actual increase in the number of cells committed to proliferation. BE did not consistently influence the amount of IL-1, IL-2 and IFN-gamma found in the supernatant of ConA stimulated cultures. The mechanism of the enhancing effect on the proliferative response of normal human lymphocytes to ConA by BE, does not seem to be selective for or unique to specific lymphocyte subsets.     

Stimulation and inhibition of human T cell subsets by wheat germ agglutinin

Wheat germ agglutinin (WGA), a tetravalent lectin, has both stimulatory and inhibitory effects on human T lymphocytes. It has been suggested that these actions are related and that WGA selectively stimulates a suppressive subset of T cells. We studied the ability of WGA to stimulate and inhibit subpopulations of human peripheral blood mononuclear cells (PBMC) known to have helper or suppressor activity. Fresh human PBMC were depleted of either T4+ or T8+ cells by using antibody-mediated complement lysis. The resultant cell populations were stimulated with WGA, and the proliferative response was assessed by [3H]thymidine incorporation, IL 2 receptor expression, the ability to elaborate IL 2 in culture supernatants, and the susceptibility to inhibition by the monoclonal antibody anti-Tac. Similar experiments with cells from a WGA-responsive continuous T cell culture were also performed. WGA inhibited phytohemagglutinin (PHA)-induced proliferation of PBMC depleted of either T4+ or T8+ cells. WGA also inhibited PBMC that had been depleted of adherent cells and Ia+ cells and then induced to proliferate with a combination of TPA and PHA. Our findings indicate that WGA induces IL 2-dependent proliferation in a small proportion of both T4+ and T8+ lymphocytes. We also provide evidence that the inhibitory activity of WGA is not mediated by a T4+, T8+, or Ia+ cell, suggesting that WGA acts directly on the proliferating cell rather than selectively stimulating a suppressive subpopulation.     

Influence of sex hormones on Coxsackie B-3 virus infection in Balb/c mice

Background and “spontaneous” proliferation are terms often used for the proliferative activity normally exhibited by peripheral blood mononuclear cells (MNC) in vitro. In this report, we show that Interleukin-2 (IL-2) added to unfractionated MNC but not to isolated T or non-T cells significantly increased their proliferative activity. The cells responding to IL-2 stimulation from MNC were OKT3 positive lymphocytes. In addition, treatment of MNC with either a monoclonal anti-HLA-DR antibody (in the absence of C′) or Cyclosporin-A strongly suppressed the “background” whereas treatment of MNC with the 3A1 monoclonal anti-human T cell antibody did not modify “spontaneous” proliferation of these cells. IL-2 could not restore or increase the proliferative activity of MNC exposed to the anti-HLA-DR antibody or Cyclosporin-A while the T cell growth factor significantly enhanced proliferation of MNC cultured in the presence of the OKT4 antibody. Taken together these results strongly suggest that IL-2 responding T cells from MNC become sensitive to IL-2 by interacting with HLA-DR antigens on B lymphocytes and/or monocytes contained in MNC (resting T cells are Dr^?). By a similar mechanism we have previously shown that T cells acquire responsiveness to IL-2 in the autologous mixed lymphocyte reaction (AMLR). Since all the cells that participate in AMLR are present in MNC, we postulate that a “mini” AMLR taking place within MNC may explain the “spontaneous” proliferation of peripheral blood mononuclear cells.     

Interleukin-1 production by mononuclear cells from rheumatoid synovial effusions

The polypeptide interleukin-1 (IL-1) is a cytokine that may mediate inflammation and connective tissue damage in rheumatoid arthritis (RA). We examined cytokine production by normal blood and by rheumatoid synovial mononuclear cells with sensitive (picomolar) assays. The assays were immunolabeling and immunoblotting with rabbit anti-IL-1 beta sera, and proliferation of the murine D10 cell line to IL-1. Little or no cytokine was detected in rheumatoid joint fluid or in exudate mononuclear cells from patients with acute rheumatoid flares. The mononuclear cells could be induced to make IL-1 upon stimulation with lipopolysaccharide (LPS). The responsive cells were monocytes, since all could be double-labeled with anti-IL-1 and the monocyte-specific CD14 antibody. More than 80% of the synovial fluid monocytes made IL-1 beta after 24 hr in 2 ng/ml LPS. Other agents failed to induce IL-1 from enriched populations of monocytes including interferon gamma (IFN-gamma), poly (I/C), phorbol myristate acetate (PMA), concanavalin A (Con A), phytohemagglutinin (PHA), and anti-CD3 antibodies. Relatively high levels of dendritic cells (DC) were present in RA effusions, but these did not produce IL-1 in response to any of the above stimuli. Blood dendritic cells also did not make IL-1, whereas blood monocytes responded comparably to synovial exudate cells. The data indicate that rheumatoid exudate monocytes make very little IL-1 during acute flares of arthritis and that this cytokine is primarily a macrophage rather than a dendritic cell product.     

Immunoregulatory functions of paf-acether. I. Effect of paf-acether on CD4+ cell proliferation

Paf-acether or platelet-activating factor (1-0-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is a phospholipid mediator of inflammation initially described as a potent platelet-aggregating compound. It is newly formed by a variety of cells including monocytes and is now recognized as a major mediator of cell-cell interactions. The present studies were undertaken to determine whether paf-acether could modulate T cell function. We found that addition of paf-acether to CD4+ cells cultured with phytohemagglutinin markedly inhibited the proliferative response in a dose-dependent manner. Maximal inhibition occurred when paf-acether was present during the first 24 hr of cell culture and the presence of paf-acether did not alter the kinetics of CD4+ cell proliferation. Importantly, the mechanism by which paf-acether inhibited the proliferative response was not related to inhibition of interleukin 2 (IL-2) secretion since the amount of IL-2 in cultures was not altered and addition of exogenous IL-2 failed to restore the CD4+ cell proliferative response. Further, as judged by indirect immunofluorescence, paf-acether did not inhibit IL-2 receptor expression. Taken together, these data indicate that paf-acether interferes with some processes leading to CD4+ cell proliferation. This new role for the chemically defined monokine paf-acether emphasizes the potential role of inflammatory lipid mediators in the regulation of T cell response.     

Expression of interleukin 2 and the interleukin 2 receptor in aging rats

Lymphocytes of aged animals exhibit a marked decrease in proliferative capacity in response to mitogen stimulation when compared to those of younger animals. In humans and mice the decreased proliferation is due at least in part (i) to the inability of lymphocytes to synthesize sufficient interleukin 2 (IL-2) and (ii) to decreased expression of IL-2 receptors (IL-2R) on the surface of aged lymphocytes. We compared proliferative abilities, IL-2 production, and IL-2R expression in splenocyte cultures of 4- to 5- and 22- to 24-month-old Fischer 344 rats stimulated with either concanavalin A (Con A) or A23187 and phorbol myristate acetate (PMA). Proliferation was significantly decreased in aged lymphocytes (30-50%) with both treatment protocols. However, unlike mice and humans we observed no difference in IL-2 activity, IL-2 mRNA levels, or IL-2R cell surface expression of lymphocytes from young and aged rats stimulated with either Con A or A23187 and PMA. These results indicate that factors other than decreased expression of IL-2 and IL-2R are responsible for the diminished proliferative capacity of aged rat lymphocytes following mitogen stimulation.     

Suppression of human mononuclear cell response by Helicobacter pylori: Effects on isolated monocytes and lymphocytes

Abstract Helicobacter pylori colonization of the human gastric mucosa causes a long-term, not self-limiting inflammation, suggesting that the microbe has properties to protect itself against the host immune defence system. Recently we were able to demonstrate that H. pylori suppresses the in vitro proliferative response of human peripheral blood mononuclear cells to antigens as well as to mitogens without affecting cell viability. The purpose of this study was to clarify which cell subsets of mononuclear cells are influenced by H. pylori . The use of monocytes which had been pretreated with a soluble cytoplasmic fraction of H. pylori (30 μg ml⁻¹) led to a suppressed proliferation of T cells after PHA-activation. Activation of isolated T cells with PHA and PMA revealed that the proliferative response of lymphocytes could also be inhibited independently of monocytes. The anti-proliferative effect was associated with a reduction of IL-2 receptor (CD25) expression as well as an inhibition of blastogenesis. Furthermore, the spontaneous proliferation of EBV-transformed B cell lines was suppressed in a dose-dependent manner. FACS-analysis of HLA-DR, ICAM-1 and CD14 expression on the surface of monocytes revealed an influence of H. pylori on CD14 expression at a concentration of 30 μg ml⁻¹, while the expression of HLA-DR and ICAM-1 was not affected at this concentration.     

Interleukin 2 induces proliferation of normal "resting" human T cells in the absence of other known external stimulation

Interleukin 2(IL-2) is known to stimulate the progression of activated T cells from G1 through the rest of the cell cycle. We have demonstrated that addition of purified recombinant human IL-2 (rIL-2) to fresh normal human peripheral blood mononuclear cells (PBM), which were IL-2 receptor (Tac) negative by FACS analysis, stimulated marked proliferation of the PBM. IL-2-induced proliferation was also observed with umbilical cord blood mononuclear cells. Monocyte depletion of PBM resulted in a marked reduction of rIL-2-induced proliferative response which could be restored by adding back autologous irradiated monocytes but not by interleukin 1. The T cells preincubated with rIL-2 showed a five to six times enhanced autologous mixed-lymphocyte reaction (AMLR) compared to controls. The rIL-2-induced proliferative response of PBM was inhibited in a concentration-dependent fashion by preincubation of PBM with an anti-HLA-DR framework monoclonal antibody. The proliferating cells were shown by two-color flow cytometric analysis to be primarily Leu-1+ and Leu-4+ T cells (both leu-3+ and Leu-2+ subsets); however, 6 to 19% of responding cells had surface markers for B cells or NK cells. The data demonstrate that rIL-2 can induce proliferation of "resting" human T cells. The phenomenon may be related to a monocyte-dependent AMLR which induces IL-2 receptors and IL-2 responsiveness in a subset of T cells.     

Human T cell responses to IL-1 and IL-6 are dependent on signals mediated through CD2.

We investigated the involvement of IL-1 and IL-6 in activation of resting human T lymphocytes via the Ti-Ag receptor/CD3-dependent and the CD2-dependent pathways, respectively. When lymphocytes were triggered through CD3-Ti, neither IL-1 nor IL-6 nor the combination of both cytokines was capable of inducing a proliferative response, whereas addition of monocytes or IL-2 to such a system mediated DNA synthesis and cellular mitosis. In contrast, in the presence of submitogenic concentrations of mAb directed at CD2, IL-1 and/or IL-6 produced marked comitogenic dose-dependent effects. Moreover, although the action of IL-1 was clearly dependent on expression of the IL-2/IL-2R system, proliferation to CD2 antibody plus IL-6 could not be blocked by mAb directed at the IL-2R and/or IL-4. T cell responsiveness to both IL-1 and IL-6 was facilitated in the presence of CD58-like signals as delivered by human rCD58, SRBC or a mAb (anti-T111A), which binds to an interaction site for CD58 on the human CD2 molecule. These findings indicate that CD2 and its ligand CD58 play an important role in T cell/monocyte interactions during primary immune responses by means of upregulating T cell susceptibility to monocyte-derived cytokines.     

Human B cell differentiation by Fc fragment. III. Effect of IL-1 and IL-2 on differentiation of human B lymphocytes induced by Fc fragments of human IgG

The human Fc fragment of IgG, when added to blood mononuclear cells in vitro, induces B cell differentiation after 6 days of culture. This activity requires the presence of T cells and monocytes. This work explores the roles of interleukin 1 (IL-1) and interleukin 2 (IL-2) in B cell differentiation induced by Fc fragments. Peripheral blood mononuclear cells (PBMC) from normal donors were examined for plasma cell differentiation following stimulation with Fc fragment (15 and 30 micrograms/ml) with or without IL-1 (6 U/ml) or IL-2 (2 U/ml). Results indicate that both IL-1 and IL-2 accelerated B cell differentiation by the Fc fragment to 3 days of culture, compared to 6 days required with the Fc fragment alone. The time required for differentiation was not further shortened when both IL-1 and IL-2 were present in culture; both IL-1 and IL-2 were able to partially induce B differentiation alone at 6 days of culture. The importance of IL-2 in B cell differentiation was further supported by the finding that antibodies specific for the IL-2 receptor blocked B cell differentiation induced by Fc fragments, with or without additional IL-1 or IL-2. The depletion of monocytes also blocked B cell differentiation and the requirement for monocytes could not be replaced by exogenous IL-1; however, Fc fragments were shown to induce monocytes to secrete IL-1 beta after 24 hr in culture. These results suggest that accelerated differentiation of B cells into plasma cells requires a double signal provided by Fc fragments and IL-1 or IL-2. Monocytes are necessary for Fc fragment-induced differentiation and cannot be replaced by either IL-1 or IL-2.     

Human colostral cells. II. Response to mitogens

The ability of colostral lymphocytes to respond to pokeweed mitogen, phytohemagglutinin, or Epstein-Barr virus was examined. None of these mitogens induced colostral cells to differentiate into immunoglobulin-containing cells, either in the absence or in the presence of mitomycin C-treated mononuclear cells or T-cell-enriched populations from peripheral blood. Cocultivation of mononuclear cells from the peripheral blood of normal adults with mitomycin C-treated colostral cells resulted in a marked suppression of the generation of immunoglobulin-containing cells in response to pokeweed mitogen. The inhibitory effect was seen in peripheral blood mononuclear cell:colostral cell ratios of 1:1, 5:1, and 10:1. However, colostral cells had little effect on the ability of peripheral blood lymphocytes to proliferate in response to phytohemagglutinin or to allogeneic stimulation.     

Phytohemagglutin-stimulated human T cell: prothymosin alpha as an accessory signal

Thymic peptide factors are known to modulate proliferation of normal human lymphocytes. In this work, we studied the effect of Prothymosin alpha (Pro alpha) on PHA-stimulated PBMC and PBLC. The observed effects of Pro alpha and thymosin alpha 1 (alpha 1) on PBMC were found to depend on the degree of cell stimulation, dose, and preincubation-time. Thymosin beta 4 (beta 4) had no effect on either cell type, regardless of the degree of stimulation, which shows that beta 4 may be used as a control peptide to work in this area. Induction of lymphoproliferation also depended on the presence of macrophages. Addition of monocytes or a cell-free monocyte culture supernatant (not containing IL-2) to the PHA-stimulated PBLC cultures resulted in T cell proliferation. Although IL-1 could not restore the PHA-induced proliferative response of isolated T cells by itself, it would enhance the helper effect of Pro alpha. Moreover, a polyclonal goat anti-human IL-2R (Tac Ag) did block the proliferative response induced by combined rIL-1 and Pro alpha, suggesting that an IL-2-dependent pathway of T cell proliferation was involved.     

5-Azacytidine induced inhibition of mitogenic response by agents that activate or augment activation of human peripheral blood T lymphocytes

Previous work from our laboratory demonstrated the mitogenic response of human peripheral blood T lymphocytes by phytohemagglutinin (PHA), phorbol myristate acetate (PMA) and ionomycin, or interleukin 2 (IL-2). Increasing levels of incorporated 5-azacytosine inhibited the action of the methyltransferase suggesting that incorporation of 5-azacytosine into DNA could be responsible for the inhibiting effect of 5-azacytidine (5-aza-CR) on DNA methylation. In this study, we first demonstrated the inhibition of mitogenic response by agents, such as PHA, PMA and ionomycin, or IL-2, that activate or augment activation of human peripheral blood T cells by treatment of the analog 5-azacytidine. Over 1 microM of 5-azacytidine, we detected significant inhibition of proliferative response and over 5 microM of 5-azacytidine toxic effect of cell viability. We found no significant change of T cell subsets after treatment of 5-azacytidine.     

Proliferative response of normal and activated human lymphocytes to tetradecanoyl phorbol acetate

The mitogenicity of 12-O-tetradecanoyl phorbol-13-acetate (TPA) for normal human peripheral blood mononuclear cells was investigated. TPA was a weak mitogen giving simulation indices in the range 2.5 to 10.5 at the optimum concentration (10 ng/ml) compared with 39 to 95 for phytohemagglutinin (PHA) at its optimum concentration (1 μg/ml). No absolute requirement for a comitogen could be demonstrated, however TPA and PHA were synergistic in their action at low concentrations, and additive at optimum concentrations. Cell fractionation by rosetting with sheep erythrocytes showed that most of the proliferative response to TPA occurred in the T-cell fraction, however some proliferation of non-T cells was also observed. Surface marker studies showed that this could not have been due to residual T cells in the non-T fraction. A small number of monocytes was required for optimal proliferation of T cells in response to TPA. After a 3-day incubation with mitogen, the responding cell populations were tested for binding of a range of antibodies specific for T-cell (OKT3, OKT4, OKT8, and OKT11), “natural killer” (NK) cell (anti-Leu-7), monocyte (FMC17), and B-cell (anti-human immunoglobulin) surface markers. These experiments indicated that the responding cell types were T cells and B cells, but not NK cells or monocytes. Marked modulation of the antigen detected by OKT4, and to a lesser extent that detected by OKT3, in the presence of TPA precluded determination of which subpopulations of T cells proliferated in response to TPA. TPA was also tested for its ability to “maintain” activated T-cell blasts in a standard assay for interleukin 2 (IL-2). Mitogen-activated T cells were strongly responsive to TPA in this assay, but progressively lost responsiveness when maintained in crude IL-2 for about 2 weeks. Thus TPA does not have “maintenance” (i.e., IL-2-like) activity. However, small amounts of TPA acted synergistically with PHA in maintaining blast populations which were not responsive to TPA alone. This illustrates the importance of using long term IL-2-dependent cell lines for quantitation of IL-2 in supernatants prepared by stimulating T cells with these agents.     

Target cells for interferon production in human leukocytes stimulated by sendai virus.

Whole leukocytes, mononuclear cells, polymorphonuclear cells (PMN), MONOCYTES, PURIFIED LYMPHOCYTES, AND T (rosette-forming cells, RFC) and non-T (nonrosette-forming cells, nonRFC) lymphocytes isolated from the human peripheral blood were stimulated by Sendai virus, respectively, and examined for interferon production in their culture fluids. High levels of interferon were produced by mononuclear cells, but not by PMN. Removal of monocytes from the mononuclear cell population did not affect at all the levels of interferon produced, although it strongly suppressed interferon induction by polyinosinic-polycytidylic acid (poly IC) and mitogenic response to phytohemagglutinin (PHA) of the lymphocytes. Purified monocytes and T lymphocytes were unresponsive to the virus. In contrast, a population of purified non-T lymphocytes produced high levels of interferon. Addition of monocytes to the interferon-producing non-T lymphocytes did not affect the levels of interferon produced. No detectable levels of interferon were produced in the mixture of T lymphocytes and monocytes. It is concluded that non-T lymphocytes may be a major target for interferon induction of human leukocytes by Sendai virus.     

Cell-specific differences in the regulation of IL-6 expression by PMA.

We have studied the regulation of IL-6 expression in human blood monocytes and lymphocytes. LPS and IFN-gamma induced IL-6 gene expression with a similar qualitative profile in both cell types. Treatment of monocytes and lymphocytes with PMA resulted, instead, in different effects: monocytes accumulated IL-6 and its message, while lymphocytes were inhibited either in the absence or the presence of LPS and IFN-gamma. These results suggest that the signal transduction pathways triggered by LPS and IFN-gamma are similar in both cell types, while PMA may activate a tissue-specific pathway which leads to opposite responses.