Soluble interleukin-2 receptor, interleukin-2 and interleukin-4 in sera and supernatants from patients with progressive systemic sclerosis.

We studied the sera of patients with progressive systemic sclerosis (PSS) for elevated levels of soluble interleukin-2 receptor (sIL-2R), interleukin-2 (IL-2) and interleukin-4 (IL-4). We also measured IL-2, IL-4 and B cell growth factor (BCGF) activity in supernatants of peripheral blood mononuclear cells from the same patients. The finding of elevated serum sIL-2R and IL-2, and the increased levels of IL-2, IL-4 and BCGF activity in culture supernatants indicates that T lymphocyte hyperactivity likely play a major role in PSS. The failure to detect under our experimental conditions a direct proliferative effect of recombinant IL-2 on enriched normal B cells might suggest that IL-4 is the cytokine mainly responsible of the BCGF activity recovered in PSS supernatants.     

Increased serum neopterin in patients with HIV-1 infection is correlated with reduced in vitro interleukin-2 production.

Recently we have observed that the CD4+ T cell response of peripheral blood mononuclear cells (PBMC) to soluble antigens is the first to be lost in the course of HIV-1 infection followed by the loss of response to HLA alloantigens. In this study we compared serum neopterin concentrations of individuals with early stages of HIV-1 infection (stages WR1 and WR2, Walter Reed staging system) with in vitro interleukin-2 (IL-2) production of PBMC in response to stimulation with soluble antigens (influenza A virus and tetanus toxoid) and alloantigens. Neopterin concentrations were significantly higher in HIV-1-seropositive individuals who showed deficient IL-2 production in response to recall antigens only or to all of the stimuli tested in vitro, compared with HIV-1-seropositive individuals who exhibited no CD4+ T cell defects. No difference in serum neopterin concentrations was observed between the group that was functionally deficient to soluble antigens only versus those who were unresponsive to both types of stimuli. It appears that the selective loss of the MHC self-restricted CD4+ T cell function is associated with an increase in serum neopterin levels. Neopterin concentrations are an estimate of the activation status of macrophages. We conclude that defective in vitro production of lymphokines by T lymphocytes is associated with activated macrophages in vivo.     

Effects of immunization with tumor cells double transfected with interleukin-2 (IL-2) and interleukin-12 (IL-12) genes on artificial metastasis of colon26 cells in BALB/c mice

In order to induce tumor specific cytotoxicity, the poorly immunogenic murine colon cancer cell colon26 was transfected with murine IL-2 cDNA and/or IL-12 cDNA and their anti-tumor effects were investigated. Double transfectants produced murine IL-2 and murine IL-12, the same as single transfectants. Intraperitoneal administration of double transfectants inhibited pulmonary metastasis of colon26 inoculated intravenously to a stronger degree than that of single transfectants. Splenocytes from mice administered double transfectants intraperitonealy showed higher cytolytic activity against colon26 than those from mice administered single transfectants, and also showed cytolytic activity against murine B16-BL6 melanoma. In the NK cell-depleted mice, double transfectants inhibited pulmonary metastasis from the control markedly, but could not do completely, the same as in the NK cell-reserved mice. The difference of the metastatic colonies between NK cell-depleted mice and the control was much greater than that between NK cell-depleted mice and NK cell-reserved mice. These results suggested that cytotoxic T lymphocytes might participate in this anti-tumor effect.     

Limiting disturbances of function by the regulatory peptide dalargin in burned rats infected withStaphylococcus aureus

A. V. Vishnevskii Institute of Surgery, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR D. S. Sarkisov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 110, No. 7, pp. 24–26, July, 1990.     

Role of interleukin-2 and interleukin-6 in the mitogen responsiveness of T cells from patients with ''common-variable'' hypogammaglobulinaemia.

We have assessed the ability of interleukin-2 (IL-2) and interleukin-6 (IL-6) to augment the proliferative response of T lymphocytes from 'common-variable' hypogammaglobulinaemia (CVH) patients and from normal controls, to the mitogens phytohaemagglutinin (PHA) and OKT3. We show that with cells from the control group and from those patients whose T cells respond to PHA within the control range, both IL-2 and IL-6 will significantly augment the response to OKT3. However, in those patients with a T cell defect in which the PHA response is below the control range, neither IL-2 nor IL-6 could restore the PHA or OKT3 response to normal. Responses to IL-2 or IL-6 alone were always in or above the control range.     

Influence of recombinant interleukin-2 on the course and pathomorphology of intraperitoneal staphylococcal infection in mice]

Resistance to the lethal doses of the infectious agent (3 DL100) develops in mice 6 days after the intraperitoneal non-lethal dose of staphylococcus in combination with recombinant interleukin-2 (RIL-2). This effect may be due to the activation of T- and B-dependent zones of the regional lymph nodes and spleen, activation and proliferation of the liver stellate reticulo-endotheliocytes, enhancement of the phagocytic activity of the circulating neutrophil granulocytes. These structural and functional mechanisms may be due to both direct RIL-2 effect in combination with an antigenic stimulation and indirect effect through other interleukins produced by lymphocytes activated by RIL-2.     

Effect of interleukin-2 on peripheral blood mononuclear cell cytokine production and the hepatic acute phase protein response

This study investigated the ability of recombinant interleukin-2 (IL-2) to modulate the ability of peripheral blood mononuclear cells (PBMCs) to stimulate an acute phase protein response in isolated human hepatocytes. The effect of IL-2 on the production of tumour necrosis factor-alpha (TNF) and interleukin-6 (IL-6) by PBMCs isolated from patients with gastrointestinal cancer, multiple organ failure, and healthy controls was also studied. The ability of supernatants from IL-2-treated PBMCs to elicit an acute phase response in hepatocytes was then investigated. IL-2 had no effect on IL-6 or TNF production by PBMCs isolated from any group in the presence or absence of bacterial lipopolysaccharide (LPS). Despite this, preincubation of PBMCs with IL-2 significantly reduced the potential of LPS-stimulated PBMC supernatants to stimulate production of alpha1 antichymotrypsin, alpha1-acid glycoprotein, and C-reactive protein by hepatocytes. These observations were not due to a direct effect of IL-2 on hepatocyte acute phase protein production. These findings suggest that in this model IL-2 may modulate PBMC-induced acute phase protein production through an IL-6 and TNF-independent pathway.     

Effects of interleukin-2 and interleukin-2-activated cells on in vitro myelopoiesis.

Lymphokine-activated killer (LAK) cells from human peripheral blood mononuclear cells cultured with recombinant interleukin-2 (IL-2) have been used clinically in adoptive immunotherapy for cancer patients. To study the influence of LAK cells and IL-2 on haematopoiesis, an in vitro assay system for colony formation of granulocyte-macrophage progenitor cells (GM-CFC) was used. LAK cells from cultures of either human peripheral blood (PB) or human bone marrow (BM) mononuclear cells were both inhibitory to allogeneic BM-derived GM-CFC. Inhibitory activity could be transferred with supernatants from co-cultures of LAK cells and BM targets, but also from the IL-2 activated PB- or BM-derived cells alone. The inhibitory activity from the initially non-cytotoxic/non-inhibitory BM population was rapidly induced by IL-2 activation, and preceded the generation of cytotoxic LAK cells in the culture. These experiments show that inhibition of haematopoietic progenitor cells by IL-2 is not dependent on generation of cytotoxic LAK cells, but rather the result of IL-2-induced cytokine production. We conclude that the synergistic action of interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) may contribute to inhibition, but that also other cytokines are responsible for the observed inhibition of BM-derived GM-CFC.     

Exogenous interleukin-16 inhibits antigen-induced airway hyper-reactivity, eosinophilia and Th2-type cytokine production in mice

BACKGROUND: IL-16 has been described as a natural soluble CD4-ligand with immunosuppressive effects in vitro. However, little is known about the effect of IL-16 on immune responses in vivo. OBJECTIVE: In the present study, we examined the effect of IL-16 administration in a murine model of allergic asthma. Next, we determined whether these effects were mediated by modulation of CD4+ T lymphocytes. METHODS AND RESULTS: Intraperitoneal administration of IL-16 completely inhibits antigen-induced airway hyper-responsiveness and largely decreases the number of eosinophils in bronchoalveolar lavage fluid (> 90%) and airway tissue of ovalbumin-sensitized and challenged mice. Firstly, it appears that thoracic lymph node cells isolated from in vivo IL-16-treated ovalbumin-challenged animals produce less IL-4 (77%) and IL-5 (85%) upon antigenic re-stimulation, when compared to vehicle-treated mice. Secondly, pre-incubation of lymphocytes with IL-16 in vitro reduces antigen-induced proliferation (55%) and Th2-type cytokine production (IL-4; 56%, IL-5; 77%). Thirdly, the presence of IL-16 during priming cultures of TCR transgenic T cells (DO11.10), reduces IL-4 (33%) and IL-5 (35%), but not IL-10 and IFNgamma levels upon re-stimulation. CONCLUSION: It can be concluded that IL-16 has potent immunosuppressive effects on a Th2dominated allergic airway response.     

Single nucleotide polymorphism of interleukin-1β associated with Helicobacter pylori infection in immune thrombocytopenic purpura

To examine the role of genetic factors in development of immune thrombocytopenic purpura (ITP) in association with Helicobacter pylori infection, gene polymorphisms within the loci for human leukocyte antigen class II, interleukin (IL)-1β (−511), tumor necrosis factor-β (+252), immunoglobulin (Ig)G1 heavy chain (+643), and Igκ light chain (+573) were determined in 164 adults with ITP and 75 healthy controls. Of these gene polymorphisms, the IL-1β (−511) T allele was less frequently detected in H.   pylori -infected than in H.   pylori -uninfected (58% vs 81%, P  = 0.01, odds ratio = 0.31) ITP patients diagnosed before age 50. These findings suggest that a single nucleotide polymorphism within the IL-1β (−511) may affect susceptibility to early-onset ITP associated with H.   pylori infection.     

Changes in the interleukin-1 and interleukin-2 generation in duodenal ulcer patients during cimetidine treatment

The effect of cimetidine treatment on the generation of interleukin-1 (IL-1) and interleukin-2 (IL-2) was studied in 11 duodenal ulcer patients. The results obtained were compared with those for untreated healthy subjects. The drug was administered intravenously in a dose of 200 mg four times a day for 8 days. The investigations were performed before, during and 1 wk after cimetidine therapy. IL-1 generation was determined by the ability of supernatants from 2-day cultured adherent cells stimulated by lipopolysaccharide to enhance proliferation of PHA-stimulated mice thymocytes. IL-2 generation was determined by the ability of supernatants from 2-day cultured, PHA-stimulated mononuclear cells to proliferate autologous 17-day cultured T cells. In all ulcer patients IL-1 generation diminished during cimetidine treatment (P less than 0.005). It continued to decrease in 4 subjects and increased in the other 7 ones following drug withdrawal. All the values were higher than those in healthy controls. IL-2 activity in ulcer patients was similar to that in healthy subjects and it increased significantly in all ulcer patients following the onset of the treatment (P less than 0.005) and decreased nearly to the initial values 1 wk after termination of the treatment (P less than 0.005). The present studies indicate that cimetidine, a selective histamine H2-receptor antagonist, deeply changes mechanisms of immunoregulation in patients with duodenal peptic ulcer.     

Chromatofocusing as a tool for the characterization and partial purification of human interleukin-2

Human interleukin-2 (IL2) has been characterized and partially purified with a sequence of chromatofocusing, gel filtration and SDS-PAGE analysis. IL2 when tested in a [³H]thymidine incorporation assay by human IL2-dependent T cells, appeared to have a MW of 25,000 as determined by Ultrogel ACA 54 gel filtration. Chromatofocusing of an 80% ammonium sulfate precipitate from crude conditioned medium yielded 4 peaks of activity corresponding to fractions of pH 7.65, 7.28, 6.72 and 6.58. Neuraminidase treatment of IL2 prior to chromatofocusing reduced its charge heterogeneity to a single peak of activity at pH 7.63. IL2 which had been treated with neuraminidase, purified by chromatofocusing, radioiodinated and further separated by gel filtration was subjected to SDS gel electrophoresis. We observed a band, migrating in the 15,000 region which only occurred in the active fractions and which we tentatively identified as IL2. These findings indicate that the purification procedure described is appropriate to the characterization and preparation of quantities of human IL2.     

Effects of administration of recombinant human interleukin-2 in dogs

The clinical and immunohaeamatological effects of recombinant humen interleukin-2 (rhIL-2) administration were evaluated in normal dogs. Three groups of three dogs per group were administered rhIL-2 subcutaneously at a dose of 6 × 10⁴ IU, 6 × 10⁵ IU, or 6 × 10⁶ IU/kg once daily for five consecutive days. Toxic clinical signs were limited primarily to diarrhoea, the severity of which, was dose dependent, with resolution within 7 days of the last rhIL-2 injection. Marked circulating eosinophilia occurred in dogs of the two highest dose groups and transient rise in blood lymphocyte numbers occurred in dogs given the highest dose of rhIL-2. The most significant immunological effects were elevated in vitro conA and pokeweed mitogen-stimulated lymphocyte blastogenic responsiveness in the highest dose group and dose-dependent elevation of antigen-specific antibody (IgG and IgM) production. Peak relative antibody production was markedly elevated, as compared to controls, in dogs administered 6 × 10⁵ IU, 6 × 10⁶ IU rhIL-2/kg.     

Prophylactic administration of interleukin-2 protects mice from lethal challenge with gram-negative bacteria.

Prophylactic administration of recombinant human interleukin-2 (IL-2) in mice enhanced survival and produced complete recovery from an otherwise lethal acute bacterial infection. IL-2 was administered as a single intraperitoneal or intravenous bolus dose to CDI mice 18 h before challenge with a lethal dose of a clinical isolate of Escherichia coli type O2 (minimal 100% lethal dose, 6 X 10(7) CFU per mouse). At IL-2 dosages of 7 X 10(6) U/kg, 90% of treated CDI mice survived as compared to 0% for the excipient buffer control animals (P less than 0.001). This protective effect was also demonstrable in immune-deficient beige mice. The IL-2 effect was dose dependent; protection was consistently observed in mice pretreated with IL-2 at doses ranging from 1.8 X 10(6) to 7 X 10(6) U/kg. However, at 3.5 X 10(5) U/kg the protective effect was more variable. The route of administration of IL-2 was shown to play an important role; when IL-2 and challenge bacteria were given by the same route (either intravenously or intraperitoneally), protection was readily observable, but when IL-2 and challenge bacteria were given by different routes, little or no protective effect was observed. The protective effect was fully inducible as early as 1 h after IL-2 administration and was effective against various strains of gram-negative bacteria, indicating that the probable mode of action represents control of the establishment of infection by increased activity of the nonspecific host defense mechanisms. The IL-2 effect was abrogated by the administration of carrageenan, suggesting a possible role of macrophages. These data demonstrate that IL-2 may be a potentially useful adjunct for the prophylaxis of bacterial infections in both clinical and veterinary medicine.     

妊娠17～37周胎儿可溶性白介素2受体和NK细胞含量的变化

目的：探讨妊娠17～37周正常和宫内感染时胎儿外周血可溶性白介素2受体（Soluble interleuldn-2 receptor，sIL-2R）和自然杀伤细胞（Natural killer，NK）含量的变化。方法：超声引导下行脐带穿刺术，收集129例胎儿外周血，包括97例正常对照组胎儿血，32例宫内感染组（单纯疱疹病毒感染、弓形虫感染、风疹病毒感染）胎儿血，采用双色免疫荧光标记流式细胞仪技术测定胎JLPF周血NK细胞百分率，双抗体夹心酶联免疫吸附试验测定胎JLPF周血中sIL-2R的含量，分析生理状态下胎儿外周血NK细胞、sIL-2R的状况和宫内感染时NK细胞和sIL-2R含量的变化。结果：妊娠17—37周胎儿外周血NK细胞、sIL-2R含量不随孕周改变，r（NK）=-0．03，P〉0．05；r（sIL-2R）=0．167，P〉0．05，宫内感染时NK细胞含量减少，sIL-2R含量增多，与正常对照组相比差异有显著意义；t（NK）=4．29，P〈0．01；t（sIL-2R）=-5．833，P〈0．01。结论：妊娠17—37周胎儿外周血有一定量的NK细胞和sIL-2R存在，但机体免疫功能仍不完善，宫内感染时机体容易出现免疫抑制状态。     

单克隆抗体ELISA双抗体夹心法测定人白细胞介素2

本文叙述了测定人白细胞介素2(IL-2)含量的ELISA双抗体夹心法。用辣根过氧化物酶标记IL-2单克隆抗体。聚苯乙烯反应板预先用0.1％戊二醛处理,然后以单克隆抗体包被。本法批内变异系数CV=3.99％,批间变异系数CV=5.96％,标准曲线的测定范围为3.1～100u/ml,检测下限为1.55u/ml。应用本法已测定了2S例正常人和11例恶性肿瘤患者的IL-2含量。