Interferon gamma and interleukin 10 levels in preimplantation embryo culture media

Purpose The aim of our study is to elucidate whether human oocyteslembryos secrete IFN and/or IL-10 and whether the fertilization process depends on the balance between these cytokines.Methods A total of 142 embryo culture media from 24 patients were collected and the cytokine levels were tested with ELISA.Results IFN and IL-10 were detectable in 40.1% and 29.6% of culture media respectively. The difference of IFN and IL-10 levels in media from fertilized oocytes between day 1 and day 2 are significant (0.46 vs. 1.47 and 34.2 vs. 12.7, respectively). However there was no significant difference between the IFN levels of the media from fertilized and nonfertilized oocytes 0.46 vs. 0.85 at day 1 and 1.47 vs. 1.49 at day 2, as well as IL-10 levels 34.2 vs. 30.9 at day 1 and 12.7 vs. 9.58 at day 2 respectively.Conclusions Human preimplantation embryos secrete the cytokines IFN and IL-10. No effect of these cytokines on fertilization process could be shown.Presented at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, April 3–7, 1995, Vienna, Austria.     

Tumor-associated lymphocytes (TAL) are competent to produce higher levels of cytokines in neoplastic pleural and peritoneal effusions than those found in sera and are able to release into culture higher levels of IL-2 and IL-6 than those released by PBMC

This work was designed to study the proliferative response of tumor-associated lymphocytes (TAL) from neoplastic effusions against autologous tumor cells and the immunophenotype pattern of TAL from neoplastic effusions and that of PBMC of the same patients. We also compared the serum levels of the cytokines interleukin (IL) 1, 2 and 6, tumor necrosis factor- (TNF) and soluble IL-2 receptor (sIL-2R) with those present in neoplastic effusions of the same patients. Moreover, we examined the ability of TAL and peripheral blood mononuclear cells (PBMC) to produce and release the cytokines and sIL-2R and to express membrane CD25 following their stimulation with phytohemagglutinin (PHA) in vitro. Finally, we compared the cytokines/sIL-2R production and membrane CD25 expression by PHA-stimulated PBMC of the patients with neoplastic effusions with a series of 90 cancer patients without neoplastic effusions and 20 normal healthy subjects. Thirteen neoplastic pleural and eight peritoneal effusions were collected from 11 patients with primary lung cancer, 7 with primary epithelial ovarian cancer, 1 with breast cancer, 1 with pleural mesothelioma, and 1 with pancreatic cancer. The proliferative response of TAL from neoplastic effusions against autologous tumor cells was lower than the response to PHA, IL-2, and anti-CD3, but significant. The percentage distribution of CD3⁺ and CD8⁺ lymphocyte subpopulations was higher in peritoneal than in pleural effusions, while the CD16⁺ subset was higher in pleural than in peritoneal effusions. The percentage distribution of CD16⁺ was significantly lower in pleural effusions than in PBMC of patients with pleural effusions. The CD39 antigen was higher on TAL from peritoneal effusions than on PBMC of the same patients. The levels of IL-1 and sIL-2R in peritoneal effusions did not differ from those measured in the sera of the same patients, while the levels of IL-2, IL-6, and TNF were higher in the peritoneal effusions. The levels of IL-2, IL-6, TNF, and sIL-2R, but not IL-1, in pleural effusions were significantly higher than those found in the sera of the same patients. The amounts of IL-2 and IL-6 produced by TAL were generally higher than those released by PBMC. The secretion of cytokines IL-1, IL-2, and sIL2R by PHA-stimulated PBMC was lower, but IL-1 and IL-6 secretion was higher in cancer patients with neoplastic effusions than in either cancer patients without neoplastic effusions or normal subjects. The CD25 expression on PHA-stimulated PBMC derived from cancer patients with neoplastic effusions was in the same range as that of cancer patients without neoplastic effusions and normal subjects. These findings suggest that TAL may be able to produce cytokines and may be amenable to immune manipulation.Abbreviations FITC Fluorescein-isothiocyanate - IL Interleukin - mAb Monoclonal antibody - MHC Major histocompatibility complex - NK Natural killer - PBMC Peripheral blood mononuclear cells - PHA Phytohemagglutinin - TAL Tumor-associated lymphocytes - TIL Tumor-infiltrating lymphocytes - TNF Tumor necrosis factor- - sIL-2R Soluble interleukin-2 receptor     

抗CD3单抗增强激活骨髓抗白血病效应的研究

目的：为了研究增强ｒＩＬ－ ２ 激活的骨髓细胞（激活骨髓ＡＢＭ）的抗白血病效应。方法：应用抗ＣＤ３ 单抗与ｒＩＬ－ ２ 联合作用体外激活骨髓细胞,采用ＭＴＴ比色分析法测定不同条件下ＡＢＭ杀伤肿瘤细胞活性。结果：ｒＩＬ－ ２、抗ＣＤ３ 单抗＋ ｒＩＬ－ ２ 体外诱导３ ｄ 的ＡＢＭ 杀伤活性分别为５６．４％ ±９．０％ ,６５．８％ ±９．２％ ,两组相比差异显著（Ｐ＜ ０．０１）;体外诱导７ ｄ 细胞增殖倍数分别为３．７,６．０ 倍。结论：在合理的诱导条件下,能够增强ＡＢＭ 的杀伤活性,扩大ＡＢＭ 的增殖效应,从而使有限的骨髓细胞在短期内达到有效的治疗剂量     

恶性肿瘤放疗前后血清可溶性白细胞介素2受体含量变化的临床意义

目的　探讨血清可溶性白细胞介素２ 受体（ｓ Ｉ Ｌ２ Ｒ）水平与恶性肿瘤患者的病期及疗效的关系。方法　采用双抗体夹心 Ｅ Ｌ Ｉ Ｓ Ａ 法检测１５９ 例恶性肿瘤患者放疗前后血清ｓ Ｉ Ｌ２ Ｒ 水平。结果　恶性肿瘤患者放疗前血清ｓ Ｉ Ｌ２ Ｒ 水平明显高于正常对照组（ Ｐ＜ ００５）;放疗后血清ｓ Ｉ Ｌ２ Ｒ 水平明显低于放疗前（ Ｐ ＜ ０００１）;晚期患者（Ⅲ＋ Ⅳ期）不论是放疗前或放疗后 ｓ Ｉ Ｌ２ Ｒ 水平均明显高于早期患者（Ⅰ＋ Ⅱ期）（ Ｐ ＜ ００５）;各类恶性肿瘤之间血清ｓ Ｉ Ｌ２ Ｒ 水平无显著性差异（ Ｐ ＞００５）。结论　ｓ Ｉ Ｌ２ Ｒ水平在各种恶性肿瘤中的表达无特异性;检测恶性肿瘤患者ｓ Ｉ Ｌ２ Ｒ 放疗前后水平,是对病情估计和治疗疗效评价的一项参考指标。     

冠心病病人淋巴细胞CR1表达及血清sIL—2R水平的变化

目的探讨冠心病病人外周血淋巴细胞补体Ⅰ型受体（ＣＲ１）表达及血清可溶性白细胞介素－２受体（ｓＩＬ－２Ｒ）浓度的变化。②方法采用淋巴细胞花环试验和酶联免疫吸附法（ＥＬＩＳＡ）,检测了７２例冠心病病人和６３例健康人淋巴细胞ＣＲ１花环率（Ｌ－ＣＲ１Ｒ）及血清ｓＩＬ－２Ｒ浓度变化。③结果冠心病病人Ｌ－ＣＲ１Ｒ明显降低,ｓＩＬ－２Ｒ浓度明显增高,与对照组比较差异有极显著性（ｔ＝６．４１４,７．８０６,Ｐ＜０．００１）。冠心病病人Ｌ－ＣＲ１Ｒ与ｓＩＬ－２Ｒ呈负相关（ｒ＝－０．８１５,Ｐ＜０．００１）。不稳定心绞痛（ＵＡ）、急性心肌梗死（ＡＭＩ）和陈旧性心肌梗死（ＯＭＩ）病人Ｌ－ＣＲ１Ｒ和ｓＩＬ－２Ｒ比较,差异亦具有极显著性（Ｆ＝７．８６０,１１．５７９,ｑ＝６．６２７～１０．５５０,Ｐ＜０．００１）,且以ＡＭＩ病人的变化最明显。④结论外周血淋巴细胞ＣＲ１表达和血清ｓＩＬ－２Ｒ浓度异常与冠心病病情变化有密切关系。     

中药肾衰宁对IL—1刺激后体外培养的人肾小球系膜细胞增殖的影响

目的：观察了具有降逆泄浊、益气活血作用的中药复方制剂肾衰宁灌肠液（以下简称肾衰宁,SSN）,对IL—1刺激后体外培养的人肾小球系膜细胞增殖的影响,冀以从细胞生物学水平探讨肾衰宁防治慢性肾功能衰竭（CRF）的作用机制。方法：按经典方法培养、鉴定人肾小球系膜细胞,分五组,分别加入不同刺激物。按MTT比色法,用酶标仪测定吸光度,记录OD值并计算抑制率。结果：IL—1可明显刺激人肾小球系膜细胞增殖,SSN大鼠血清对IL—1刺激后人肾小球系膜细胞增殖有明显的抑制作用,其抑制程度与药物浓度有一定的量效关系、即随药物浓度的增加对系膜细胞增殖所产生的抑制作用也相应增加。结论：含肾衰宁的大鼠血清对IL—1刺激后肾小球系膜细胞的增殖有明显的抑制作用。提示肾小球系膜细胞是肾衰宁发挥治疗作用的重要靶细胞。     

Tumour necrosis factor alpha and interleukin 2 in normal and infected human seminal fluid

The presence of cytokines such as the tumour necrosis factoralpha (TNF) and interleukin 2 (IL2) in human spermatozoa isstill to be defined. The aim of this study was to measure theconcentration of both soluble factors in seminal fluid. Datafrom normal semen samples (n = 24) confirmed the presence ofIL2 (258 ± 84 fmol/ml corresponding to 953 ± 369fmol/total volume of ejaculate) and TNFa (62.2 ± 16.4fmol/ml corresponding to 231.3 ± 86 fmol/total volumeof ejaculate). A significant positive correlation (r = 0.59;P < 0.01) was observed between the TNFa and the IL2 concentrations.The concentrations of these cytokines were not related to spermparameters. In contrast, IL2 concentrations (196.9 ±60.4 fmol/ml; 686.2 ± 236.7 fmol/total volume of ejaculate)evaluated in 16 seminal fluids with identified bacterial agentswere lower than in the control group, whereas TNF concentrations(68.6 ± 12.3 fmol/ml; 241.3 ± 78.9 fmol/totalvolume of ejaculate) were not significantly different from thecontrols. Further studies are needed to determine the potentialrole of these cytokines in the physiology of semen and theirusefulness as indicators of reproductive pathology.     

Stability of Interleukin 1β (IL-1β) in Aqueous Solution: Analytical Methods,Kinetics, Products,and Solution Formulation Implications

The thermal stability of IL-1 in aqueous solution as a function of temperature (5–60°C), pH (2–9), buffer (acetate, citrate, tris, and phosphate), and cyroprotectants (sugars, HSA) was investigated in this study. The analytical methodologies included RP-HPLC, SEC, ELISA, IEF-PAGE, SDS-PAGE, and bioassay. The degradation and inactivation of IL-1 at or above 39°C were attributed to autoxidation of the two cysteine residues in the denatured protein, followed by hydrophobic/covalent aggregation and precipitation. At or below 30°C, IEF- and SDS-PAGE results suggest a possible deamidation reaction. The difference in mechanism of degradation precludes the prediction of formulation shelf life from accelerated temperature data. Nonetheless, the good stability observed at 5°C suggests that a solution formulation may be feasible for IL-1.     

Immunocytochemical analysis of interferon-1β production by human monocytes

All-Union Research Institute of Highly Pure Biological Preparations, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR V. D. Belyakov). Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 9, pp. 278–280, September, 1991.