Effect of “Re Du Qing” on the activation,proliferation and membrane fluidity of lymphocytes

Summary Effects of Chinese Medicinal Preparation “Re Du Qing” (RDQ) on the activation, proliferation and membrane fluidity of T lymphocytes from human peripheral blood Were studied by means of³H-TdR incorporation and DPH fluorescence polarization. The results showed that “RDQ” can:1) significantly inhibit the activation of T lymphocytes; 2) restrain the proliferation of activated T lymphoblasts in the presence of exogenous interleukin-2 (IL-2); and 3) increase the membrane fluidity of T lymphocytes and antagonize the decreased fluidity of lymphocyte membrane mediated by Con A or PHA. The functional abnormalities of T lymphocytes in some autoimmune diseases such as arthritis and the usefulness of RDQ in the treatment of these diseases were also discussed.     

Modulation of human T cell functions by surface sulphydryl groups: differential effects on IL-2 production and responsiveness.

An impermeable thiol blocker has been used to investigate the role of sulphydryl (SH) groups in the production of and responsiveness to IL-2 by normal human T lymphocytes. Surface SH blockade of mononuclear cells prior to incubation with mitogen (phytohaemagglutinin, concanavalin A, CD3 MoAb) had no effect on production of IL-2 but markedly impaired cellular responsiveness to exogenous IL-2. Studies using MoAbs indicated that this effect was accompanied by decreased expression of both the CD25 and p75 subunits of the IL-2 receptor. Blocking surface SH groups did not affect binding of IL-2 to p75 on unstimulated mononuclear cells, but inhibited binding to high-affinity receptors on a T lymphoma cell line. The data are consistent with the hypothesis that sulphydryl groups on the IL-2 receptor are required for its function and may be involved in the interaction of the CD25 and p75 subunits leading to generation of the high-affinity binding site. The surface thiol identified on the IL-2 receptor may be a candidate for oxidation on cells from patients with chronic inflammatory diseases such as rheumatoid arthritis and thus contribute to the aberrant function of T cells in these patients.     

子宫内膜异位症及卵巢癌腹腔IL-6、IL-8变化的临床意义

目的:探讨白细胞介素6(IL-6)、白细胞介素8(IL-8)与子宫内膜异位症(endomotriosisEMs)卵巢癌的发生发展关系。方法:采用双抗体夹心ABC-酶联免疫吸附实验(ELISA)对EMs患者30例、血清及腹腔液中IL-6、IL-8进行检测,并以卵巢癌患者10例、卵巢良性肿瘤患者30例、正常妇女血清30例做对照组。结果:EMs组与卵巢癌组患者血清及腹腔液中IL-6、IL-8水平明显高于对照组(P<0.01),EMs组与卵巢癌组对比无显著性差异(P>0.05)。结论:EMs患者腹腔液中IL-6、IL-8异常增高是腹腔免疫内环境失衡,与EMs的发生发展有关,在药物治疗中血清IL-6、IL-8可作为EMs疗效和预后的指标之一。     

利多卡因对N-甲基-D-天冬氨酸抑制大鼠肾上腺嗜铬细胞瘤细胞增殖的影响

目的 观察利多卡因对 N-甲基-D-天冬氨酸(NMDA)抑制大鼠肾上腺嗜铬细胞瘤细胞(PC12细胞)增殖的影响。方法 将体外培养的 PC12细胞分为6组，分别采用正常不含药液的培养基(C组)；含400μmol·L(-1)NMDA 的培养基(N组)；NMDA 分别混合10μmol·L~(-1)(L_1组)、10~2μmol·L~(-1)(L_2组)、10~3μmol·L~(-1)(L_3组)以及10~4μmol·L~(-1)(L_4组)利多卡因的培养基培养5d，应用流式细胞仪测定细胞 DNA 相对含量，解析细胞周期，计算 S 期细胞荧光强度占受测细胞总荧光强度的百分数为 S期分数(SPF)和 S 期与 G_2期细胞荧光强度之和与 M 期细胞荧光强度的比值[(S G_2)/M]。结果 与C 组比较，N、L_1组 SPF 和(S G_2)/M 均降低(P<0.05)，L_4组 SPF 降低(P<0.05)，而 L_2及 L_3组 SPF和(S G_2)/M 差异无统计学意义(P>0.05)。与 N 组比较，L_2、L_3及 L_4组 SPF 和(S G_2)/M 升高(P<0.05)，L_1组 SPF 升高(P<0.05)，而(S G_2)/M 差异无统计学意义(P>0.05)。结论 NMDA 可以通过抑制 PC12细胞 DNA 合成而影响细胞的增殖活性，利多卡因能拮抗 NMDA 对 PC12细胞增殖的抑制作用。     

人野生型parkin基因真核表达载体的构建及其在PC12细胞中的表达

目的 克隆人野生型parkin基因并构建真核表达载体pCDNA3．1—parkin，将重组质粒转染PC12细胞获得高表达人野生型parkin基因的PC12细胞克隆。方法 从胎脑组织中提取总RNA，用RT—PCR方法获得人野生型parkin基因的全长cDNA，插入pCR2．1—TA克隆载体中进行序列测定，测序正确后将其亚克隆至表达载体pCD—NA3．1，利用脂质体将重组质粒转染PC12细胞，经G418筛选获得抗性细胞克隆，采用RT—PCR和Western Blot方法鉴定人野生型parkin基因在PC12细胞中的过表达。结果 经限制性内切酶酶切图谱分析和DNA序列测定证实目的基因已插入重组质粒，RT—PCR和Western Blot证明经G418筛选得到的转基因PC12细胞克隆中存在人野生型parkin基因的表达。结论 成功构建了人野生型parkin基因的真核表达载体，获得了稳定表达人野生型parkin基因的PC12细胞克隆，为进一步研究parkin的生物学功能以及parkin在帕金森病发病机制中的作用奠定了良好的基础。     

病毒白细胞介素-10转基因在造血干细胞中的表达

目的　观察逆转录病毒转病毒白细胞介素 10 (vIL 10 )基因在体内的表达。方法　用MSCVneo vIL 10重组体在体外转导CBA (H 2 K)小鼠的造血干细胞 (HSCs) ,给经致死照射(90 0rads)的 2 0只同基因CBA(H 2 K)小鼠注入经MSCVneo vIL 10转染的HSCs ,2× 10 6HSCs/只。酶联免疫吸附测定 (ELISA)、逆转录 聚合酶链反应 (RT PCR )、Westernblot分析vIL 10的表达。结果　移植MSCVneo vIL 10转染HSCs的 2 0只小鼠 ,移植后 8周用ELISA检测 ,其中 15只小鼠血清的vIL 10浓度为 :2 70～ 13 40ng/L ,5只小鼠血清的vIL 10为阴性。 12周后有 2只小鼠vIL 10测不出 ,13只小鼠长时间表达vIL 10达 6个月。对照组小鼠血清vIL 10均为阴性。RT PCR和Westernblot证实小鼠的器官均有vIL 10的mRNA和蛋白的表达。结论　逆转录病毒能有效地将vIL 10基因导入造血干细胞并在体内长时间表达。     

IL-2在移植免疫中的研究进展

随着器官移植手术技术的逐渐成熟,移植后免疫排斥反应已成为制约器官移植的瓶颈。IL-2在免疫系统中起着关键作用,而且与免疫排斥和免疫耐受关系密切,因此,更好的理解 IL-2在免疫和自身耐受中的生理作用,对临床治疗中控制IL-2信号,获得更好治疗效果有指导意义。     

IL-6对人精子顶体反应影响机制的初步探讨

目的:探讨IL-6对人精子顶体反应(AR)的影响机制。方法:采用BAEE/ADH法测定精子顶体酶的活性,以及通过FITC-PSA法检测精子顶体反应。结果:IL-6可诱导精子顶体酶及超氧化物歧化酶(SOD)的活性,促进精子顶体反应;胞外Ca2+单独不能诱导精子顶体反应,且没有胞外Ca2+的参与,IL-6也不能诱导精子顶体反应;蛋白激酶C(PKC)抑制剂calphC能逆转IL-6诱导的精子顶体反应。结论:IL-6对精子顶体反应有一定的促进作用,可能通过诱导精子的顶体酶和SOD活性等途径来实现,在此作用中,也涉及了PKC的激活,且还需要外源性Ca2+的参与。     

A hole in the T cell repertoire specific for a pigeon cytochrome c related peptide associated with amino acid substitutions on I-Ab molecules.

When C57BL/10(B10) mice were immunized with a pigeon cytochrome c related peptide, 50V (AEGFSYTVANKNKGIT), two helper T cell populations with different specificity were activated. A major T cell population reacted with a 50V analog, 50V54A (AEGFSYTVANKAKGIT), more potently than with the immunogen, 50V, in a heteroclitic fashion, whereas the other minor T cell population responded only to 50V. By contrast, when bm12 mice were immunized with 50V, the minor T cell population responding only to 50V could hardly be demonstrated. The apparent deletion of the minor T cell population in bm12 mice seems to be attributable to negative selection under the influence of I-Abm12 molecules, since the minor T cell population was undetectable in both I-Ab and I-Abm12 restricted T cells from (B10 x bm12)F1 mice. Thus, three mutant points on the I-A molecule in bm12 mice appear to be involved in the seemingly negative selection of the certain T cell repertoire. The present finding demonstrates that a T cell repertoire generated under the influence of a MHC product (Ab) on one parental strain is eliminated by a different MHC product (Abm12) on the other parental strain of F1 cross. The mechanism underlying the apparent negative selection is discussed.     

Effect of gamma-interferon on IL-1 alpha, beta and receptor antagonist production by normal human keratinocytes

Abstract In inflammatory dermatoses. activated T cells produce inter-feron-gamma (IFN-γ), which interacts with keratinocytes and contributes to the direct activation of these cells by inducing, among other factors, the expression of HLA-DR antigens and intercellular adhesion molecule-1. However, the action of IFN-γ on epidermal cell cytokine production is not known. Our aim was to assess the effect of IFN-γ on the production of IL-1 by normal human keratinocytes cultured in low calcium medium (MCDB153). In comparison with controls, the addition of nontoxic IFN-yγ concentrations (50-500 U/ml) to cell cultures induced a significant increase of IL-1α and IL-1β production predominantly after 100 U/ml treatment in the cell extracts as well as in the supernatants at 24h and 48h. The production of the antagonist. IL-1RA, was also enhanced and the effect of the critical concentration (100 U/ml) was more evident. However, the absence of a characteristic dose response could not be explained by an antiproliferative effect of high IFN-γ concentrations (250 and 500 U/ml) on cultured keratinocytes or by the induction of the nuclear stress protein, Hsp72. two phenomena known to down-regulate IL-1 biosynthesis. In conclusion, the modifications in keratinocyte IL-1 production under IFN-γ stimulation can contribute to activate the epidermal cells and thus involve them in the local immune response.