Elevation of plasma thioredoxin levels in HIV-infected individuals

Thioredoxin (Trx), a ubiquitous protein intimately involvedin redox and protein disulfide reductions, has been shown tobe released from cells and to have cytokine-like activities.In addition, Trx has been implicated in the redox regulationof immunological responses and shown to be deficient in tissuesfrom AIDS patients. In studies presented here, plasma Trx levelswere measured by ELISA in plasma samples from HIV-infected individuals(n = 136) and HIV-negative controls (n = 47). To account forthe release of Trx into plasma due to hemolysis, the Trx measurementswere corrected according to the level of hemoglobin in the plasmasample. Data presented show that, in contrast to tissue Trxlevels, corrected plasma Trx levels are significantly higherin HIV-infected individuals than in controls (P < 0.0001).Furthermore, {small tilde}25% of the HIV-infected individualsstudied have plasma Trx levels greater than the highest levelfound in controls (37 ng/ml). Detailed multiparameter FACS analysisof peripheral blood mononuclear cells (PBMC) from the infectedindividuals demonstrates that those with higher plasma Trx levels(37 ng/ml or greater) tend to have lower overall CD4 counts.In addition, increases in plasma Trx levels correlate with decreasesin monochlorobimane staining (indicative of lower intracellularglutathione levels in PBMC) and with changes in surface antigenexpression (CD62L, CD38 and CD20) that occur in the later stagesof HIV infection. These correlations suggest that elevationof plasma Trx levels may be an important component of advancedHIV disease, perhaps related to the oxidative stress that oftenoccurs at this stage.     

肺结核病人红细胞免疫功能及T淋巴细胞亚群的变化

①目的探讨红细胞免疫功能及Ｔ淋巴细胞亚群变化与肺结核的关系。②方法对４４例肺结核病人和４４例健康人的红细胞免疫功能、血清循环免疫复合物、Ｔ淋巴细胞亚群、ＮＫ细胞水平的变化进行了检测，并与对照组比较。③结果肺结核病人红细胞Ｃ３ｂ受体花环率（Ｅ－Ｃ３ｂＲＲ），ＣＤ４，ＣＤ４／ＣＤ８，ＮＫ细胞水平明显降低，ＣＤ８水平明显升高，与对照组相比，差异均有极显著意义（ｔ＝９．７６～２２．２４，Ｐ均＜０．０１）；复治组较初治组变化更明显（ｔ＝４．２７～１５．３２，Ｐ均＜０．０１）。Ｅ－Ｃ３ｂＲＲ与ＣＤ４，ＣＤ４／ＣＤ８，ＮＫ细胞水平呈正相关（ｒ＝０．２６９２～０．５４７９，Ｐ＜０．０５，０．０１），与ＣＤ８呈显著负相关（ｒ＝－０．３７４，Ｐ＜０．０１）。④结论红细胞免疫功能、Ｔ淋巴细胞亚群及ＮＫ细胞的异常与肺结核的发病及病情变化有一定关系。     

Development of intestinal epithelial cell lines using a transgenic mouse

The epithelial cells of the colonic mucosa of the animal have proved impossible to culture using standard tissue culture techniques. Immortalization of adult colonic epithelial cells has been unsuccessful due to the lack of DNA synthesis in these cells once they are isolated from the tissue. Recently an unique transgenic mouse bearing a temperature sensitive mutant of the known immortalizing gene, SV40 large T has become available. The advantage of this mouse is that the SV40 large T gene is expressed in every cell. Active immortalizing protein is produced in each cell at the permissive temperature. We have used colonic mucosa from these mice to initiate cultures of epithelial cells from the colon of adult mice. The cells grow readily at the permissive temperature but die within 7 days at the non-permissive temperature. The methods used to develop these cultures are described.     

脐血CD34~+细胞体外短期培养扩增研究

为寻找更有效的体外扩增脐血CD34 + 细胞的造血细胞因子组合 ,采集健康产妇脐带血 ,用免疫磁珠法分选CD34 + 细胞。采用SCF、FLT3 L、TPO和IL 34种具有早期作用的细胞因子的不同组合进行脐血CD34 + 细胞短期无血清液体培养 ,观察培养前后有核细胞、CD34 + 细胞、CD34 + /CD38- 细胞、CFU GEMM、CFU GM和BFU E数量的变化。结果在 3种不同的细胞因子组合中 ,同时应用SCF、FLT3 L、TPO和IL 34种细胞因子培养 7d的扩增效果最好。突出的发现是在这种条件下CD34 + /CD38- 细胞亚群达到平均 1 97.9倍的扩增效果。提示 :SCF、FLT3 L、TPO和IL 34种细胞因子是脐血CD34 + 细胞体外扩增理想的细胞因子组合     

酸性成纤维细胞生长因子的分离纯化及活性鉴定

目的：从牛脑中分离纯化酸性成纤维细胞生长因子（aFGF)并鉴定生物活性，方法：根据DenisGOSPODAROW-ICZ的方法，新鲜牛脑匀浆，三步硫酸铵盐析。最后沉淀物溶解透析后予离子交换层析及琼脂糖胶亲和层析，促3T3细胞DNA合成率鉴定生物活性。结果：用1.0mol/LNaCl的缓冲液洗脱得到分子量为13600的多肽，得率为610μg/kg牛脑，氨基酸组成分析与已知的aFGF相同，促3T3细胞DNA合成的最低浓度为0.5ng/ml，最大效应浓度50ng/ml，ED50值为8ng/ml。结论：aFGF具有强烈的促细胞增殖作用。     

逆转录病毒载体介导的外源性主要组织相容性抗原复合体基因转移及其在T细胞的表达

目的 建立一个在胸腺内表达外源性主要组织相容性抗原复合体(MHC)抗原的实验模型,为下一步研究作准备。方法 应用逆转录病毒载体介导的基因转移技术,首次将外源性MHC基因转移到T淋巴表达并应用聚合酶链反应(PCR)及反转录聚合酶链反应(RT-PCR)检测转染T细胞DNA及mRNA。结果 外源性MHC基因已整合到靶细胞染色体DNA并有效地转录;单克隆抗体免疫荧光染色流式细胞仪检测显示在转染T细胞膜有外源性MHC分子表达;转染效率为46.2％。结论 外源性MHC基因可以转移到T细胞稳定表达,为今后的研究提供了经验。     

Antisperm antibodies and lymphocyte subsets in semen—not a simple relationship

The significance of white blood cells in the ejaculate remains a matter of controversy. Several authors have suggested that such cells are important in the modulation of an antisperm antibody response, i.e. a predominance of suppressor/cytotoxic to helper/inducer T cells may prevent the development of antisperm antibodies. In order to examine this relationship further we have documented the white blood cell types, with emphasis on the T-lymphocyte populations, in the ejaculates of men from infertile couples with and without antisperm antibodies; the latter group was divided further into two groups--vasovasostomized men and idiopathic men. All seven of the men without antisperm antibodies had a predominance of suppressor/cytotoxic T cells to helper/inducer T cells in the ejaculate. However, only in some of the men with antibodies was there a predominance of T-helper/inducer cells. It is clear that the relationship between antisperm antibodies and seminal leucocytes is therefore not as straightforward as has been proposed.     

Interactions between peripheral blood CD8 T lymphocytes and intestinal epithelial cells (iEC)

Intestinal intraepithelial lymphocytes (iIEL) are primarily CD8 cells and most of them have a CD28^? phenotype, the phenotype of effector cytotoxic T cells. We asked whether the predominance of CD8⁺ CD28^? T cells in the gut may result from peripheral blood T cells preferentially migrating to the iIEL compartment and adhering to iEC. Compared with CD4 cells, adhesion of resting CD8⁺ T cells to iEC cell lines was significantly higher. Adhesion could be blocked with a MoAb to gp180, a molecule expressed on iEC which is known to interact with CD8/lck. No significant difference in the level of adhesion was observed between CD8⁺ CD28⁺ and CD8⁺ CD28^? T cells. Thus CD8 cells may preferentially migrate to the iIEL compartment, but loss of CD28 expression could occur in situ after migration. Consistent with this hypothesis, the CD8⁺ CD28^? cells became enriched after co-culturing T cells with iEC cell lines and primary iEC. Induction of the CD8⁺ CD28^? phenotype in cord blood and adult T cells was observed in co-cultures with iEC and also with mitogens and superantigens. In the latter case, CD28 down-modulation was seen specifically in the Vβ subset targeted by the superantigen, indicating that loss of CD28 expression is a direct result of T cell receptor (TCR)-mediated stimulation. The combined results suggest that CD8⁺ CD28^? T cells are antigen experienced T cells, and that they may have a survival advantage in the presence of gut epithelial cells in vitro. This may contribute to the predominance of CD8⁺ CD28^? T cells in the iIEL compartment.     

小鼠胸腺和脾脏细胞的体外转化培养

建立了小鼠脾脏、胸腺细胞体外转化微量培养系统,确定了最佳实验条件.点蜡法充CO_2,简便易行,效果好.RPMI-1640培养基内补充胎牛血清,其浓度确定为在脾细胞培养为10%,在胸腺细胞培养为20%.脾细胞培养最适细胞密度为2～5×10~6/ml,Con A最适量为2～4μg/培养;胸腺细胞培养细胞最适密度为1×10~7/ml,Con A最适量为2～4μg/培养.两种已知免疫活性的药物在脾细胞培养体系有明确肯定的反应.     

高原地区小儿病毒性心肌炎T细胞亚群检测及临床意义

本文对２８例小儿病毒性心肌炎患者进行了Ｔ细胞检测，发现患儿外周血Ｔ细胞亚群有明显改变。提示：小儿病毒性心肌炎患者均有不同程度的一过性细胞免疫功能低下。而体液免疫功能正常，我们在临床给与一般治疗的同时，佐以胸腺因子治疗，明显改善症状，取得良好疗效。