Modification of lymphocyte response to phytomitogens by polycations and polyanions

Summary The stimulation effect on mitogen-induced lymphocyte response by polycationic compounds such as polylysine, DEAE dextran protamine and methylated albumin, is studied at different serum-protein concentrations and mitogen concentrations. It is suggested that the polar interaction between polycationic compounds and glycoproteins are of major importance for reactivity of lymphocytes to mitogens.     

Stimulation of peripheral guinea pig blood lymphocytes by autologous epidermal cells]

Using a strictly autologous (and not syngeneic) mixed culture system in the guinea-pig, it was shown that Mitomycin C treated ear epidermal cells (E) were capable of stimulating peripheral blood lymphocytes (L) from the same animal. The proliferative response of the lymphocytes as measured by 3H thymidine incorporation, is optimal on the 5th day of culture, and when the ratio of the concentrations of the two cell populations (L : E) is 1 : 1. The autoreactivity of the lymphocytes varies with the animal.     

Cell-mediated cytotoxicity by natural killer and killer cells, lipid peroxidation and glutathione

In the course of spontaneous cell-mediated cytotoxicity (SCMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) with human peripheral lymphocytes as effector cells, no lipid peroxidation occurred as measured by the production of ethane and thiobarbituric acid-reactive material. Furthermore, impairment of major cellular defense systems of target cells (K562 cells for SCMC, Chang liver cells for ADCC), by decreasing their glutathione content, had no effect on either lipid peroxidation or the cytotoxic response. These findings indicate that peroxidative damage is not a mechanism of NK and K cell-mediated cytotoxicity.     

Protein kinase C-dependent NF-κB activation is altered in T cells by chronic stress

Chronic stress has been associated with impaired immune function. In this work we studied the effect of chronic mild stress (CMS) exposure on the early intracellular pathways involved in T cells after stimulation with mitogen. We found that mitogen stimulation of T lymphocytes from CMS-exposed mice resulted in a reduction of the intracellular [Ca²⁺] rise, an impairment of growth-promoting protein kinase C (PKC) activation, a lower NF-κB activation and an increase in the inhibitory cAMP-protein kinase A (PKA) pathway activity with respect to those found in control lymphocytes. However, T cell activation with the direct PKC activator phorbol 12-myristate 13-acetate plus calcium ionophore led to a similar proliferative response in both CMS and control lymphocytes, indicating that signals downstream of PKC would not be affected by stress. In summary, our results show that chronic stress induced an alteration in T cell early transduction signals that result in an impairment of the proliferative response.Received 11 February 2005; received after revision 20 May 2005; accepted 6 June 2005     

Suppression of PHA-induced lymphocyte blastogenesis by concomitant presence of PPD in the culture

Summary PHA-induced lymphocyte blastogenic response was remarkably suppressed by the simultaneous presence of PPD in cultures of lymphocytes derived from individuals sensitized to PPD, but not affected by the presence of PPD when the cultures contained lymphocytes from an individual not sensitized to the protein. This double stimulation blastogenesis study with PHA and a specific antigen is feasible as a simple and rapid test to measure cell-mediated immunity to the antigen.Supported by United States grant USPHS Grant AM 27384.     

Inhibition of DNA biosynthesis in human lymphocytes by alpha 1-antitrypsin]

Incorporation of tritiated thymidine into human lymphocytes from tonsils is markedly inhibited by purified alpha1-antitrypsin-preparations. This inhibitory effect is observed in lymphocytes stimulated by a mitogen factor (phytohemagglutinin) as in non stimulated lymphocytes.     

Chronic overcrowding decreases cytoplasmic free calcium levels in T lymphocytes of aged CBA/CA mice

Intracellular calcium concentration is a sensitive marker of the homeostasis of living cells, and its increase is an essential step of T lymphocyte activation. Changes in the environment provoke an adaptive stress-response of the organism. In our present work we have investigated the effect of chronic overcrowding on resting and lectin-stimulated cytoplasmic free calcium concentration of splenic T lymphocytes from young and aged CBA/CA mice (50 animals total). The animals were kept under ‘normal’ (68 cm²/animal) or ‘overcrowded’ (22 cm²/animal) conditions for 3 months. Young animals showed no change in resting and stimulated calcium after overcrowding. T cells from aged mice, however, displayed significantly smaller levels of both resting and lectin-stimulated intracellular calcium concentration (p<0.01 each), as compared to those of the non-stressed, aged animals. This inadequate adaptation in the calcium metabolism of T lymphocytes may significantly contribute to the diminished immune response of the aged in stress.     

Leukocyte mobilization from the guinea pig spleen by muscarinic cholinergic stimulation

Important interactions between the immune system and the nervous and endocrine systems have become increasingly accepted. The present results demonstrate that the cholinergic agonist carbacholine greatly increased the number of granulocytes and lymphocytes in the splenic venous blood, but not arterial blood, shortly after administration to guinea pigs. The effect was largely blocked by pretreatment with atropine. In contrast, animals treated with indomethacin had a decreased number of leukocytes in both splenic venous and arterial blood. A decrease in relative splenic weight due to carbacholine treatment was also blocked by atropine. However, cholinergic leukocyte mobilization, or that previously observed after adrenergic stimulation, may not be caused by capsule contraction since it is not accompanied by mobilization of erythrocytes. Furthermore, indomethacin, which potentiates the response of splenic smooth muscle to adrenergic stimuli, blocked the effect of noradrenaline (NA) on leukocyte mobilization.     

Leukocyte mobilization from the guinea pig spleen by muscarinic cholinergic stimulation

Important interactions between the immune system and the nervous and endocrine systems have become increasingly accepted. The present results demonstrate that the cholinergic agonist carbacholine greatly increased the number of granulocytes and lymphocytes in the splenic venous blood, but not arterial blood, shortly after administration to guinea pigs. The effect was largely blocked by pretreatment with atropine. In contrast, animals treated with indomethacin had a decreased number of leukocytes in both splenic venous and arterial blood. A decrease in relative splenic weight due to carbacholine treatment was also blocked by atropine. However, cholinergic leukocyte mobilization, or that previously observed after adrenergic stimulation, may not be caused by capsule contraction since it is not accompanied by mobilization of erythrocytes. Furthermore, indomethacin, which potentiates the response of splenic smooth muscle to adrenergic stimuli, blocked the effect of noradrenaline (NA) on leukocyte mobilization.     

T-Lymphocyte regulation of contact sensitivity: effect of thymectomy in adult mice]

6 weeks after adult thymectomy (ATx) in the Mouse, the contact sensitivity reaction to dinitrofluorobenzene (DNFB) is enhanced. 4 to 7 months after ATx, this reaction is deeply, but incompletely depressed, whereas the concomitant antibody response is not affected. These results suggest that both a suppressor, and an amplifier, T lymphocytes, the life span of which is different after ATx, are involved in the regulation of contact sensitivity. The effect of circulating thymic factor on this reaction suggests that this factor acts exclusively, at least in short treatments, on the suppressor function.     

Glucagon-like peptide-1(1-37) inhibits chemokine-induced migration of human CD4-positive lymphocytes

The present study examined the effect of GLP-1(1-37) on chemokine-induced CD4-positive lymphocyte migration as an early and critical step in atherogenesis. Pretreatment with GLP-1(1-37) reduced the SDF-induced migration of isolated human CD4-positive lymphocytes in a concentration-dependent manner. Similar effects were seen when RANTES was used as a chemokine. GLP-1(1-37)’s effect on CD4-positive lymphocyte migration was mediated through an early inhibition of chemokine-induced PI-3 kinase activity. Downstream, GLP-1(1-37) inhibited SDF-induced phosphorylation of MLC and cofilin and limited f-actin formation as well as ICAM3 translocation. Furthermore, exendin-4 inhibited SDF-induced migration of CD4-positive lymphocytes similarly to GLP-1(1-37), and transfection of these cells with GLP-1 receptor siRNA abolished GLP-1(1-37)’s action on chemokine-induced ICAM3 translocation, suggesting an effect mediated via the GLP-1 receptor. Thus, GLP-1(1-37) inhibits chemokine-induced CD4-positive lymphocyte migration by inhibition of the PI3-kinase pathway and via the GLP-1 receptor. This effect provides a potential novel mechanism for how GLP-1(1-37) may modulate vascular disease.     

Mitogenic action of neuraminidase

Summary A new effect of NCV on lymphocytes is demonstrated. This property is the capacity to act as a mitogen in and of itself. The possible mechanisms of this phenomenon are discussed.     

IL-18 in inflammatory and autoimmune disease

Inflammation serves as the first line of defense in response to tissue injury, guiding the immune system to ensure preservation of the host. The inflammatory response can be divided into a quick initial phase mediated mainly by innate immune cells including neutrophils and macrophages, followed by a late phase that is dominated by lymphocytes. Early in the new millennium, a key component of the inflammatory reaction was discovered with the identification of a number of cytosolic sensor proteins (Nod-like receptors) that assembled into a common structure, the ‘inflammasome’. This structure includes an enzyme, caspase-1, which upon activation cleaves pro-forms of cytokines leading to subsequent release of active IL-1 and IL-18. This review focuses on the role of IL-18 in inflammatory responses with emphasis on autoimmune diseases.     

Receptors for cytomegaloviruses on the surface of human lymphocytes]

The relationships between cytomegalovirus (CMV) and lymphocytes have already been noted because of: (a) the immunological abnormalities induced by this virus, and (b) activation of latent CMV in the course of lymphocyte reactions associated with anti-histocompatibility antigen immune response. The present work shows that the lymphocyte surface may have specific receptors for CMV. Cultured fibroblasts infected with DMV were incubated with lymphocytes isolated from the blood of human immune subjects. Rosettes defined by the adherence of three or more lymphocytes around a fibroblast were formed in infected preparation while no rosettes were seen with normal control fibroblasts. Approximately 1.2 per cent of lymphocytes were involved in the formation of these rosettes. Rosette formation is inhibited when infected fibroblasts have been incubated with anti-CMV antibodies prior to the addition of lymphocytes.     

Immunosuppressive effect of a human serum ferroprotein of hepatic origin, alpha 2 H globulin. Study on blastic transformation of normal lymphocytes in the presence of phytohemagglutinin]

The alpha2 H globulin, a glycoferroprotein, has been found in the serum of patients with malignant diseases. Its level varies according to the evolution of the disease. The activity of this protein has been studied on the normal lymphocytes cultivated in presence of phytohemagglutinin. This study has showed that 5 mug/ml of alpha2 H globulin can inhibit the blastic transformation. The inhibitory effect is proportional to the amount of alpha2 H globulin added as judged by the tritiated thymidin incorporation. The study of morphological aspects of the lymphocytes shows that alpha2 H globulin acts upon the synthesis of the cytoplasmic proteins.     

Differences in cytochalasin D-induced surface alterations between chronic lymphocytic leukaemic and normal lymphocytes

Summary Cytochalasin D (CD) causes an unusual surface alteration in normal lymphocytes consisting of the formation of focal irregular club-shaped cell processes. Lymphocytes from chronic lymphocytic leukaemic cases did not show this change on exposure to CD. There was either no surface change or, in some cases, clear double-membrane lined vesicles were formed and appeared to be discharged from the cell. This difference in response may be related to the changes in cell membranes known to occur in malignant transformation.The study was supported by the National Cancer Institute of Canada.     

Mode of action of a soluble restricted factor, a suppressor of the allogeneic proliferative response in man

Appearance of "suppressor cells" is induced by in vitro hyperimmunization of lymphocytes against allogeneic cells, incompatible for one HLA-DR antigen. These "suppressor cells under certain conditions, release in the culture medium, "suppressor factors" of the in vitro allogeneic proliferative response in Man. They are not immunoglobulins and act in a non specific way towards the stimulators. Only one of them is restricted to some individuals. This is shown when either responders or stimulators are incubated for different periods, with the "suppressor factors" prior to the primary mixed lymphocyte reaction (MLRI). The beneficial effect of transfusions on kidney graft survival, could be, in part, explained by a suppressor mechanism, analogous to the one described in vitro.