Increased interleukin-1 and modulation of interleukin-2 production by murine macrophages and lymphocytes treated with LF 1695

Previous results have demonstrated that lectin-induced T cell proliferation was potentiated or suppressed by LF 1695, a synthetic immunomodulator, depending on the dose used. Therefore the activity of this compound was investigated on murine IL-1 and IL-2 production. Adherent peritoneal cells, incubated with LF 1695, could secrete high levels of IL-1 with only a slight elevation in intracellular IL-1. This effect apparent at 5 and 10 μg/ml was linked to a transient state of activation. At low doses, LF 1695 increased IL-2 production by Con A-stimulated spleen cells. A decrease was found at higher doses only when cells were preincubated 20 h with the compound. In murine macrophages stimulated either by A 23187 or LPS PGE₂ synthesis was inhibited by LF 1695 even at low doses. However, supernatant LTB₄ level was increased in LF 1695-treated culture with a time-dependent effect. Therefore modulation of lectin-induced T cell proliferation by LF 1695 may be IL-2 production-mediated. Inhibition of the cycloxygenase and stimulation of the lipoxygenase pathway of arachidonic acid metabolism may be responsible for this pattern of activity.     

Redirection of cytokine production by lymphocytes from women with pre-term delivery by dydrogesterone

PROBLEM: To study the ability of dydrogesterone to modulate the production of pro-inflammatory and anti-inflammatory cytokines by lymphocytes from women undergoing pre-term delivery (PTD). METHOD OF STUDY: Peripheral blood mononuclear cells (PBMC) from 18 subjects undergoing PTD were stimulated with the mitogen phytohemagglutinin in the presence and absence of progesterone and dydrogesterone. The levels of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-4, and IL-10 in culture supernatants were then estimated by enzyme-linked immunoabsorbant assay. Cytokine production in the presence and absence of progesterone and dydrogesterone were compared. RESULTS: The exposure of PBMC to dydrogesterone resulted in a significant inhibition in the production of the pro-inflammatory cytokines IFN-gamma and TNF-alpha and a significant increase in the levels of the anti-inflammatory cytokine IL-4, resulting in a substantial shift in the ratio of Th1/Th2 cytokines. CONCLUSION: Dydrogesterone induces a shift in cytokine bias, by inhibiting pro-inflammatory cytokine production and increasing anti-inflammatory cytokine production.     

Effect of interleukin-2 and mitogen on in vitro immunoglobulin production by peripheral blood lymphocytes from patients with selective IgM deficiency

Immunoglobulin production by mitogen and recombinant interleukin-2-(rIL-2)-stimulated lymphocytes from IgM-deficient patients was studied. The findings were that subnormal serum IgM levels did not necessarily predict defective in vitro IgG or IgM production, lymphocytes from some IgM-deficient patients exhibited defective T cell function, and rIL-2 did not enhance defective in vitro immunoglobulin production.     

In vitro cytokine production in patients with iron deficiency anemia

The in vitro production of interleukin (IL)-1beta, IL-2, IL-6, IL-10, and tumor necrosis factor alpha (TNFalpha) by peripheral blood mononuclear cells (PBMC) from 20 patients with iron deficiency anemia (IDA) was examined before and after iron supplementation and compared to values obtained for PBMC from healthy controls. A significant decrease in IL-2 production was observed in IDA patients, whereas the secretion of the other cytokines did not differ from that of controls. Addition of iron to the culture medium did not affect the secretion of IL-2 and IL-1beta, but caused an increase in IL-6, IL-10, and TNF-alpha production. Since a deficiency in IL-2 production plays a role in the pathogenesis of certain infectious and malignant diseases, the results of the present study may explain in part the increased susceptibility to infections observed in patients with IDA.     

In vitro interleukin-1 (IL-1) production in thymic hyperplasia and thymoma from patients with myasthenia gravis

Using a biological test, we measuredin vitro thymic epithelial cell IL-1 activity of 14 thymomas and 8 hyperplasia from myasthenic patients. Our results demonstrate that thymic epithelial cells from hyperplastic thymuses spontaneously produce high amounts of IL-1 compared to controls, while cells from thymomas produce very low amounts of IL-1. After LPS stimulation the difference between patients and controls is no longer significant. Immunohistochemistry studies demonstrate a limited number of IL-1-positive cells on sections from normal thymuses and a variable number of IL-1-positive cells on diseased thymuses, not clearly correlated to thein vitro production. In hyperplastic thymuses an association is found between the level of hyperplasia andin vitro IL-1 production, suggesting a role for IL-1 in the expansion of thymic lymphoid follicles. In thymoma, the low IL-1 production and the loss of corticomedullary organisation raise the possibility that some autoreactive clones could emerge from the lymphocyte pool, present in the abnormal tumoral microenvironment.     

Is schizophrenia caused by excessive production of interleukin-2 and interleukin-2 receptors by gastrointestinal lymphocytes?

Excessive production of interleukin-2 (IL-2) and IL-2 receptors (IL-2R) by gastrointestinal (GI) T-lymphocytes is hypothesized as the cause of schizophrenia. It is based on: 1) IL-2 given to human volunteers can cause all the symptoms of schizophrenia; 2) GI lymphocytes in nonhuman primates produce much more IL-2 and IL-2R when stimulated than peripheral blood lymphocytes; 3) the GI tract is the largest lymphoid 'organ' in the body. The hypothesis appears to: 1) explain the protective effect of rheumatoid arthritis on schizophrenia; 2) make mechanistically plausible the findings on wheat and schizophrenia; 3) be consistent with and explain many of the known immunological abnormalities in schizophrenia.     

Prostaglandin E2 depresses natural cytotoxicity by inhibiting interleukin-1 production by large granular lymphocytes.

Enriched large granular lymphocytes treated with varying concentrations of prostaglandin E2 (PGE2) for varying time periods showed considerably reduced natural cytotoxicity against K-562 target cells. The same cells when activated by lipopolysaccharide, produced substantially less interleukin-1 (IL-1) as compared to cells not treated with PGE2. It is concluded that the inhibition of natural killer (NK) cell activity produced by PGE2 is due to inhibition of IL-1 production by these cells.     

Short-term oral administration of ginseng extract induces type-1 cytokine production

Ginseng radix (Panax ginseng C.A. Meyer) is a popular herbal medicine used as a major ingredient in tonic recipes in eastern Asian countries. In our study, male BALB/c mice were treated orally with various doses of ginseng root extract for 5 consecutive days. The extract reduced the serum level of IgG but elevated the level of IgA. Under in vitro condition, the lipopolysaccharide-stimulated spleen cells from the ginseng-treated mice also showed a significant decrease in IgG production but an increase in IgA production. The serum level and production of IgM was unaffected. The interleukin-2, interferon-γ (Th1-type cytokines), and interleukin-10 (Tr1-type cytokine) production by Con A-stimulated spleen cells from the ginseng-treated mice showed an upregulation relative to the control group. However, the production of interleukin-4 (Th2-type cytokine) showed no significant change. The activity of natural killer cells was increased in the ginseng group, but the percentages of T-lymphocytes (CD3⁺) and CD4⁺8^-, CD4^-8⁺ subset were reduced. Thus, short-term oral administration of ginseng extract appears to enhance Th1-type cytokine production.     

组胺对T细胞IL-2产生及增殖活性影响的实验研究

目的:了解组胺对 CD4 和 CD8 T细胞IL-2产生和细胞增殖活性的影响。方法:密度梯度离心及吸附法分离PBMC和PBLC,采用抗CD4 和CD8 抗体分别制备CD8 和CD4 T细胞进行培养,然后采用ELISA法和MTT比色法测上清液IL-2含量及增殖活性。结果:①组胺 CD4 （CD8 ）培养上清液中IL-2水平及MTT增殖指数与T细胞自然培养孔比较明显降低（P<0.05）。②组胺 CD4 （CD8 ） 西咪替丁培养孔上清液中IL-2水平及 MTT增殖指数明显高于未加西咪替丁孔（P<0.05）。③CD4 T细胞自然培养孔上清液中IL-2水平显著高于CD8 T细胞自然培养孔。结论:组胺可抑制T细胞IL-2产生及增殖。西咪替丁可阻断组胺对T细胞的抑制作用。CD8 T细胞也可产生IL-2,但其功能较 CD4 T细胞为低。     

Glutathione increases interleukin-2 production in human lymphocytes

It is known that glutathione (GSH) has an immunological effect on several features of the immune system. The present study investigated the effects of GSH on interleukin-2 (IL-2) production from normal human peripheral blood lymphocytes (PBL). The results showed that both exogenous GSH and 2-mercaptoethanol (2-ME) significantly increased intracellular GSH levels after PBL were incubated with both agents. IL-2 production from PBL was markedly increased at the presence of exogenous GSH (0.5–8 mmol/1) or 2-ME (12.5–50 μmol/1) which corresponded to 1.57–2.82 nmol/10⁶ cells and 1.41–1.80 nmol/10⁶ cells of intracellular concentrations of GSH, respectively. However, IL-2 production seemed to reach a steady level when exogenous GSH concentrations in cell culture were between 2 and 8 mmol/1. The findings also showed that there was a positive correlation between the IL-2 concentrations and intracellular GSH levels. This study indicated that both exogenous GSH and 2-ME were able to elevate intracellular GSH levels and the increased intracellular GSH could increase IL-2 production in vitro. It is suggested that GSH may exert its effects on the immune system via the regulation of IL-2 synthesis.     

In vitro cytokine production by mononuclear cells exposed to bone cement extracts

The authors evaluated the ability of bone cement to modify the profile of pro-inflammatory cytokines secreted by the immune cells. Peripheral blood mononuclear cells (PBMC) collected from healthy individuals were cultured with cement extracts and tested to assess the release of IL-1beta, TNFalpha, GM-CSF and IL-6 in both unstimulated and PHA-stimulated PBMC. The cytokine release of unstimulated PBMC was very poor, and in particular the IL-1beta was undetectable: the addition of cement extract increased both TNFalpha and GM-CSF release and decreased IL-6, sometimes significantly. The most recurrent observation in PHA-stimulated PBMCs exposed to bone cement extract was the increase in both IL-1beta and IL-6 release, while both the mean concentration and the index of release of TNFalpha and GM-CSF were changeable. In conclusion our results showed that leachable components of some bone cements can induce in vitro the release of pro-inflammatory cytokines which are known to be involved in the bone resorption associated with aseptic loosening of hip prostheses. These findings allowed us to identify materials endowed with the highest inflammatory power.     

Immunoglobulin and cytokine production by neonatal lymphocytes.

Growth and differentiation of cord blood B cells were studied using T cell-depleted populations. In the absence of in vitro activation, cord blood B cells proliferated in response to cytokines including interleukin-2 (IL-2) and interleukin-4 (IL-4); anti-mu-stimulated cord B cells had a lesser response to IL-2 than adult cells. IgM synthesis by cord blood B cells was enhanced by interleukin-6 (IL-6) and decreased by IL-2. In cultures activated by Staphylococcus aureus Cowan I (SAC), cord blood B cells produced lesser increases in IgM than adult B cells regardless of the cytokine added. Cord blood B cells produced no IgG or IgA with any cytokine preparation with or without SAC activation. Supernatants of cord blood T cells pulse-stimulated with phytohaemagglutinin and phorbol myristate acetate contained less IL-2 and IL-6 and had less growth and differentiation activity than adult T cell supernatants. The results confirm a limited cord blood B cell response and also suggest a limitation in production of B cell stimulatory lymphokines by cord blood T cells.     

Enhancement of in vitro lipopolysaccharide-stimulated interleukin-1 production by levamisole

The production of interleukin-1 (IL-1) by the P388D1 mouse macrophage cell line and by adherent peritoneal exudate cells (PMs) was examined. In vitro IL-1 production by P388D1 cells treated with lipopolysaccharide (LPS) was enhanced by coculture with levamisole (0.1 to 10 microM). Oral administration of levamisole (3 mg/kg) to mice also resulted in potentiation of in vitro IL-1 production by thioglycollate-elicited peritoneal macrophages in response to in vitro LPS stimulation. Potentiation was approximately twofold. IL-1 production in the absence of LPS by either the P388D1 cells or the PMs was nil, and levamisole did not directly stimulate IL-1 production in these cases. IL-1 activity in the culture supernatants was measured by thymocyte comitogenic assays. The immunochemical identify of the thymocyte comitogenic activity as IL-1 alpha was confirmed by neutralization with a specific goat anti-mouse IL-1 alpha antiserum. These results suggest that one mechanism by which levamisole acts to normalize and restore immune responses may be enhancing the signals which enable activated macrophages to secrete IL-1.     

In vitro production of IgE by lymphocytes from a patient with hyperimmunoglobulinaemia E, eosinophilia and increased lymphocytes carrying surface IgE.

The peripheral blood lymphocytes of a patient with massive hyperimmunoglobulinaemia E were used for in vitro studies. The serum IgE ranged from 140,000-210,000 u/ml. Peripheral blood lymphocytes had approximately 7% of cells staining for surface IgE. When these cells were cultured in vitro, IgE was produced as measured by the double antibody radioimmunoassay technique. The total IgE produced ranged from 140 to 484 units per 24 hr in different cultures. IgE production was greatest in the first 24 hr of culture and declined progressively thereafter. Some cultures still had measurable IgE at 48 hr. If the lymphocytes staining for surface IgE were the cells producing the IgE, it was estimated that between 1-7 and 2-8 molecules per cell per second were produced. No definite effect of concanavalin A, pokeweed mitogen or phytohaemagglutinin on in vitro IgE production could be demonstrated under the conditions of these experiments.     

Perimenstrual alterations in type-1/type-2 cytokine balance of normal women.

BACKGROUND: Perturbations of the type-1/type-2 cytokine balance play a role in the pathogenesis of many diseases. Several immune-based diseases, such as asthma, have significant clinical exacerbations during specific intervals of the menstrual cycle and are associated with oral contraceptive pills (OCRs). The mechanism for these changes is not known, but may involve alterations in the type-1/type-2 cytokine balance. OBJECTIVE: To determine if the type-1/type-2 cytokine balance in healthy women changes during a regular menstrual cycle. METHODS: Peripheral blood mononuclear cells from 14 healthy women (seven taking monophasic OCPs) obtained during the perimenstrual interval (3 days prior to 4 days after the onset of menses) and the mid-cycle interval (days 13 to 16) were stimulated with PHA. Supernatants were analyzed for type-1 (IFN-gamma) and type-2 (IL-10) cytokines. RESULTS: During the perimenstrual interval PBMC produced less IFN-gamma and more IL-10, resulting in a decreased IFN-gamma: IL-10 ratio compared with the mid-cycle interval. The perimenstrual decrease in the IFN-gamma: IL-10 ratio was observed in women not taking OCP, but not in women taking OCP. Furthermore, the OCP group had a lower mid-cycle IFN-gamma: IL-10 ratio compared with the control group. Finally, subjects reported increased levels of distress during the perimenstrual interval compared with the mid-cycle interval. CONCLUSIONS: These data suggest that healthy women have a perimenstrual shift in the type-1/type-2 cytokine balance toward a type-2 response that is blunted in women taking OCP.     

Proliferation induced by Plasmodium falciparum antigen and interleukin-2 production by lymphocytes isolated from malaria-immune individuals.

Affinity-purified Plasmodium falciparum soluble antigens (SPAg) isolated from in vitro cultures of the parasite were shown to be relatively free of nonspecific polyclonal activators. To determine the presence of lymphocytes with specificity against SPAg in the peripheral blood of malaria-immune individuals, the proliferative response and the interleukin-2 (IL-2) production of SPAg-activated mononuclear cells (MNCs) from individuals unexposed, sensitized, and immune to malaria were measured. It was found that MNC isolated from malaria-immune individuals proliferated in response to SPAg and that this activation resulted in measurable IL-2 production in 5 of 10 MNC cultures. MNC isolates from most unexposed individuals did not respond to SPAg. To establish which cells responded to SPAg, different subpopulations of MNCs were tested. Only T helper cells were found to respond, and they responded only when cocultured with monocytes. The finding of parasite-specific T helper cells in the blood of malaria-immune individuals and the fact that some of these cells were able to produce IL-2 in vitro support the hypothesis that in malaria the cellular part of the protective immune response is initiated by immune T cells. These cells may activate nonspecific effector cells (i.e., macrophages) that eliminate the parasite.     

Enhancement of interleukin-1 and interleukin-2 production by soluble glucan

Soluble glucan, a beta-1,3-linked polyglucose, is a biologic response modifier effective in the therapy of experimental neoplasia, infectious diseases and immunosuppression. Interleukin-1 (IL-1) and interleukin-2 (IL-2) are endogenous immunomodulators which are essential for effective immune responsiveness. In view of its broad spectrum of immunobiological activity, the ability of glucan to enhance the production of IL-1 and IL-2 was evaluated. Splenic IL-1 and IL-2 secretion as well as plasma IL-1 and IL-2 levels were determined in Sprague-Dawley rats receiving glucan (100 mg/kg, i.p.) at intervals ranging from 12 days to 1 h prior to collection of splenocytes and plasma. Glucan (100 mg/kg) was also injected either s.c., i.p. or i.v. on days -4, -3 and -2 prior to harvesting splenocytes on day 0. Splenic macrophage IL-1 production was initially elevated 12 h following glucan injection and was maintained for a 5 day period. IL-2 secretion by splenic lymphocytes was enhanced 6 h post-glucan and remained elevated for an additional 9 days. Plasma IL-1 activity was elevated 12 h post-injection, while IL-2 activity in plasma was enhanced at 1 h post-glucan. Peak IL-1 and IL-2 activity in plasma occurred 9 and 12 days, respectively, following glucan administration. With regard to route of administration, IV glucan was most effective in inducing lymphokine production. This study demonstrates that: (1) glucan will enhance IL-1 and IL-2 production and (2) elevations in lymphokine production can be maintained up to 12 days post-glucan.     

Increased interleukin-2 production by human peripheral blood lymphocytes treated with low doses of dexamethasone

Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 116, N ^o 7, pp. 68–70, July, 1993.     

Contribution of interleukin-3 to antigen-induced Th2 cytokine production

Short-term stimulation of mouse spleen cells in vitro with interleukin (IL)-3 induces the secretion of the Th2 cytokines IL-4 and IL-6. Non-B/non-T cells were the target of this IL-3 effect. However, during long-term antigen-dependent culture, T cells are the major source of IL-4 and IL-6. The addition of IL-3 to such cultures also led to a significant increase in IL-4 and IL-6 production. This Th2 cytokine secretion was amplified by the addition of irradiated non-B/non-T cells at the initiation of culture, and was inhibited by anti-IL-4 antibodies. These findings suggest that IL-3 induces the rapid release of IL-4 and IL-6 by non-B/non-T cells, thereby creating an immune milieu conducive to the development of antigen-specific IL-4 and IL-6-secreting Th2 cells.