IL—2、IL—12、IL—10在慢性病毒性肝炎治疗中的应用概况

对于病毒感染,至今尚无特效药物。近年来,免疫学治疗愈来愈受到人们的重视,以细胞因子为基础的免疫治疗更受人们的青睐。本文就近年来研究较多的IL－2、IL－12、IL－10三种细胞因子在慢性病毒性肝炎治疗中应用的进展作一综述。     

IL—2,IL—4基因转染后肿瘤细胞表面ICAM—1分子的表达及其作用

为了进一步研究细胞因子基因转染后肿瘤细胞体内致瘤原性、免疫原性变化的分子机制，将ＩＬ－２、ＩＬ－４基因转染入Ｂ１６黑色素瘤细胞，观察了其体内接种后致瘤原性变化，通过ＦＡＣＳ法检测了其细胞表面粘附分子－１（ＩＣＡＭ－１）的表达水平，分析了其细胞表面ＩＣＡＭ－１表达水平以及它们与肿瘤细胞对ＬＡＫ、ＣＴＬ等免疫效应细胞杀伤敏感性变化的关系。结果表明，ＩＬ－２、ＩＬ－４基因转染后Ｂ１６细胞体内致瘤原性明显     

IL—2基因转染逆转绒癌耐药细胞系多药耐药性

用RT－PCR的方法克隆人的IL－2基因,将IL－2基因插入真核表达载体pcDNA3.1( )中,通过阳离子脂质体将IL－2基因转染用足叶乙甙（VP－16）诱导建立的绒癌耐药细胞系JEG－3／VP16,用G418筛选含IL－2 DNA片段的单个细胞克隆,检测转染前后细胞对5－氟尿嘧啶（5－Fu),氨甲喋呤（MTX）,VP－16,更生霉素（KSM）,紫杉醇（TAXOL）耐药指数的变化,用RT－PCR的方法测定转染前后细胞IL－2和多种耐药基因包括肺耐药蛋白（LRP）,多药耐药相关蛋白（MRP）,谷胱甘肽转移酶（GST－π）,二氢叶酸还原酶（DHFR）和多药耐药基因（MDR－1）的mRNA表达情况。结果表明 转染IL－2的细胞中用RT－PCR的方法检测到IL－2 mRNA表达,转染IL－2基因的细胞耐药指数降低,MDR－1 mRNA表达转阴,而其它耐药基因的表达无明显改变。     

IL2受体及其抗体

<正> 自从1976年Morgan等人发现TCGF(1979年改称IL2)以来,人们很快证实了它在激活和增强免疫反应过程中有着极其重要的生物学作用。它不仅诱导T杀伤细胞的产生,是T细胞克隆生长因子,而且可以激活B细胞和LGL细胞的增殖反应,并使后者转化成有效的 NK、LAK细胞,同时参与介导T和B细胞的其它生物学功能。人们对于IL2的这些生物学作用的机制产生了极大的兴趣。1979年,Bonnard等人发现激活的T细胞均能吸收IL2这一现象,从而提出了“IL2受体(IL2R)”假设。直到1981     

人重组IL—1受体拮抗剂在大肠杆菌的高效表达

利用ＰＣＲ技术从分裂素刺激的人外周血细胞ｃＤＮＡ文库扩增出人白细胞介素－１受体拮抗剂（ｈＩＬ－１ｒａ）的成熟分子编码区ｃＤＮＡ，将其克隆化插进高效表达的质粒载体ｐＫｐＬ－３α后，在大肠杆菌中高效表达出ｒｈＩＬ－１ｒａ，其Ｎ端序列与预期结果一致。经ＳＤＳ－ＰＡＧＥ分析，ｒｈＩＬ－１ｒａ的表达量占菌体蛋白的３０％以上。体外试验证明，该表达产物对单核细胞衍生的ＩＬ－１活性有明显抑制作用。     

强直性脊柱炎及类风湿关节炎患者IL—2及IL—2受体测定

应用流式细胞仪检测强直性脊柱炎(AS)及类风湿关节炎(RA)患者外周血淋巴细胞(PBL)表面IL-2受体(IL-2R)及经PHA刺激、体外培养产生的IL-2。结果表明,AS患者PBL表达IL-2R无明显增加,而RA患者则明显高于正常人及AS患者(P<0.01);AS及RA患者PBL体外培养产生IL-2明显多于正常人(P<0.01),提示IL-2异常在AS及RA患者免疫功能失调中起重要作用。     

Preactivation with IL-12, IL-15, and IL-18 Induces CD25 and a Functional High-Affinity IL-2 Receptor on Human Cytokine-Induced Memory-like Natural Killer Cells

Natural killer (NK) cells are effector lymphocytes that are under clinical investigation for the adoptive immunotherapy of hematologic malignancies, especially acute myeloid leukemia. Recent work in mice has identified innate memory-like properties of NK cells. Human NK cells also exhibit memory-like properties, and cytokine-induced memory-like (CIML) NK cells are generated via brief preactivation with IL-12, IL-15, and IL-18, which later exhibit enhanced functionality upon restimulation. However, the optimal cytokine receptors and signals for maintenance of enhanced function and homeostasis after preactivation remain unclear. Here, we show that IL-12, IL-15, and IL-18 preactivation induces a rapid and prolonged expression of CD25, resulting in a functional high-affinity IL-2 receptor (IL-2Rαβγ) that confers responsiveness to picomolar concentrations of IL-2. The expression of CD25 correlated with STAT5 phosphorylation in response to picomolar concentrations of IL-2, indicating the presence of a signal-competent IL-2Rαβγ. Furthermore, picomolar concentrations of IL-2 acted synergistically with IL-12 to costimulate IFN-γ production by preactivated NK cells, an effect that was CD25 dependent. Picomolar concentrations of IL-2 also enhanced NK cell proliferation and cytotoxicity via the IL-2Rαβγ. Further, after adoptive transfer into immunodeficient NOD-SCID-γ_c^−/− mice, human cytokine–preactivated NK cells expand preferentially in response to exogenous IL-2. Collectively, these data demonstrate that human CIML NK cells respond to IL-2 via IL-2Rαβγ with enhanced survival and functionality, and they provide additional rationale for immunotherapeutic strategies that include brief cytokine preactivation before adoptive NK cell transfer, followed by low-dose IL-2 therapy.     

人红细胞对LAK细胞DNA合成和IL-2受体表达影响的研究

本文用~3H-TdR掺入法、APAAP免疫染色法和MTT法研究人红细胞(RBC)对LAK细胞DNA合成、IL-2受体(IL-2R)表达及杀伤活性的影响。人RBC对LAK细胞DNA合成有抑制作用,对LAK细胞IL-2R表达和杀伤活性有促进作用。随RBC浓度增加,抑制作用和促进作用均明显加强。     

IL-2 regulation of soluble IL-2 receptor levels following thermal injury.

In the immunosuppressed burn patient serum levels of both IL-2 and a soluble form of IL-2 receptor alpha (sIL-2R alpha) are significantly elevated. Strikingly, the production of these markers by the in vitro activated patients' cells is decreased. This study examines the role of IL-2 in the decreased production of the sIL-2R alpha in vitro in patients with major burns (n = 18, 30 to greater than 70% total body surface area). Peripheral blood mononuclear cell (PBMC) cultures from patients with highly elevated serum sIL-2R alpha, and from healthy controls (n = 12) were activated with concanavalin A (Con A) at initiation. In patients' cultures mitogen-induced increments of sIL-2R alpha levels were significantly lower. There was a significant negative correlation (r = 0.64, P less than 0.001) between a high serum sIL-2R alpha level and a decreased lectin-induced sIL-2R alpha release in vitro. Low levels of sIL-2R alpha in patients' samples were not normalized by increasing the number of T lymphocytes. Also exogenous rIL-1 was without effect, whereas rIL-3 increased sIL-2R alpha release in some cultures. However, sIL-2R alpha levels were significantly increased in patients' cultures by (i) addition of exogenous IL-2; (ii) removal of adherent cells; (iii) addition of cyclooxygenase inhibitor, indomethacin; (iv) bypassing cell surface activation by the combination of the calcium ionophore A23187 and the phorbol ester 12-o-tetradecanoyl acetate. The cyclic AMP-elevating drug, forskolin, abrogated the ability of exogenous IL-2 to increase sIL-2R alpha production. Thus, in the burn patient, the reduced in vitro sIL-2R alpha release appears to relate to abnormalities in IL-2 production and action mediated through its functional surface receptor. Elevated levels of sIL-2R alpha in vivo may, therefore, reflect systemic activation of T lymphocytes in response to biologically active IL-2.     

Role of alpha chain-IL-2 complex in the formation of the ternary complex of IL-2 and high-affinity IL-2 receptor

Using anti-Tac (anti-alpha chain) and 2R-B (anti-beta chain) antibodies, we studied the roles of IL-2 receptor subunits (alpha and beta chains) in the formation of IL-2 and high-affinity IL-2 receptor complex, which is the initial event of IL-2 induced T cell growth. High-affinity IL-2 binding which was undetectable in the presence of 2R-B antibody at 4 degrees C became fully detectable when examined at 37 degrees C, which explained the lack of inhibition by 2R-B antibody of IL-2-induced proliferation of the cells expressing high-affinity IL-2 receptor. We further studied the mechanism of the 'reappearance' of high-affinity IL-2 binding in the presence of 2R-B antibody. The addition of IL-2 to the cells preincubated with radiolabeled or fluorescence-labeled 2R-B antibody resulted in a marked decrease in the antibody bound to the cells expressing high-affinity IL-2 receptor at 37 degrees C. This decrease was blocked by the presence of anti-Tac antibody, which inhibited IL-2 binding to alpha chain, but not by 7G7/B6 antibody, which recognized a non-IL-2 binding site of its chain. Furthermore, the decrease in cell-bound 2R-B antibody was not due to the internalization of beta chain-2R-B antibody complex, because the amount of cell-bound Mik-beta3 antibody recognizing a non-IL-2 binding epitope of beta chain remained unchanged, nor to the inhibition by simple competitive binding of IL-2 molecules to beta chain as judged from comparative studies of competitive binding inhibition. Taking these data together, the reappearance of high-affinity IL-2 binding was considered to be caused by the replacement of 2R-B antibody at the IL-2 binding site of beta chain by alpha chain-mediated IL-2, and it was strongly suggested that alpha chain-IL-2 complex has a key role in the formation of the ternary complex of IL-2 and high-affinity IL-2 receptor. alpha chain may function as a dimension converter of IL-2 to effectively deliver IL-2 molecules to a relatively small number of beta chains in the dynamics of the formation of high-affinity IL-2 binding in T cells.     

IL-12 as Well as IL-2 Upregulates CCR5 Expression on T Cell Receptor-Triggered Human CD4+ and CD8+ T Cells

The expression of chemokine receptors on leukocytes is related to their activation state. However, the exact mechanism underlying the induction of each chemokine receptor is poorly understood. Here, we investigated how CCR5, a chemokine receptor implicated in T cell trafficking and HIV infection, is induced in human T cells. CCR5 was marginally detected on a freshly prepared human peripheral blood mononuclear cell (PBMC) population. Long-term (8-day) stimulation of PBMC with IL-2 resulted in high levels of CCR5 expression on T cells. IL-12 failed to induce CCR5 on T cells in such a directly stimulated PBMC population. Stimulation of PBMC T cells with anti-CD3 plus anti-CD28 induced detectable albeit very low levels of CCR5 along with the induction of IL-12 receptor. However, these TCR-triggered T cells expressed much higher levels of CCR5 when stimulated with IL-12. Although IL-2 also induced CCR5 expression, CCR5 expression was more potent in IL-12 than IL-2 stimulation. These results indicate that, in addition to IL-2, IL-12 plays an important role in the induction of CCR5 expression on T cells, particularly TCR-triggered T cells.     

病毒性心肌炎患者血清sIL-2R、IL-6和IL-8水平变化及临床意义

作者观察了50份柯萨基B组病毒（Cox B）性心肌炎患者血清IL-6、 IL-8和sIL-2R的水平变化,及与病毒血症的关系。发现病毒性心肌炎患者血清的IL-8和IL-6水平均显著高于正常对照组,其中血清Cox B抗原和特异性IgM抗体均阳性组（20例）的IL-8和IL-6含量均明显高于仅检出特异性IgM抗体组（30例）,且IL-8含量升高的程度与检测抗原的阳性强度是显著正相关。然而,两组间及与正常对照组间sIL-2R的含量均无明显差异。认为血清IL-8水平升高是病毒性心肌炎急性期的一个重要指标,并间接反映机体处于病毒血症或病毒抗原血症。     

IL-2基因修饰的肿瘤细胞对小鼠体内巨噬细胞数量和功能的影响

目的　探讨IL - 2基因修饰的肿瘤细胞对小鼠体内巨噬细胞 (M)数量和功能的影响 .方法　应用腺病毒载体介导的小鼠IL - 2基因 (Ad -mIL - 2 )修饰CT2 6小鼠结肠腺癌细胞 (CT2 6 -mIL - 2 )后皮下接种小鼠 ,计数小鼠腹腔M的数量 ,观察其吞噬功能 ,混合淋巴细胞反应法 (MLR)测定其抗原提呈能力 ,MTT法检测其杀伤活性 .结果　mIL - 2基因修饰的CT2 6细胞 (CT2 6 -mIL - 2细胞 )皮下接种后 ,小鼠腹腔M数量显著增加 ,吞噬能力明显增强 ,抗原提呈能力提高 ,并具有较强的杀伤活性 .结论　CT2 6 -mIL - 2细胞分泌的IL - 2能有效地激活M ,这可能是CT2 6 -mIL - 2细胞体内致瘤性下降的原因之一 .