犬蛔虫（Toxocaracanis）对感染鼠的细胞免疫和IL－1、IL－2免疫调节作用的影响

应用犬蛔虫（Ｔ．ｃａｒｎｉｓ）的感染性虫卵口饲感染Ｃ３Ｈ／ＨｅＮ鼠，体外培养后观察脾细胞的免疫学变化。感染第１－４周的脾细胞用ＣｏｎＡ刺激，Ｔ细胞的增殖反应，ＩＬ－２的诱生均明显地受到抑制，而且感染鼠脾细胞抑制了正常鼠脾细胞对ＣｏｎＡ的反应。感染鼠的脾细胞经ＳｅｐｈａｄｅｘＧ－１０过柱后，Ｔ细胞则不显示被抑制作用。表明影响Ｔ细胞被抑制作用的是感染鼠的巨噬细胞。相反，感染第１－４周的鼠所产生的ＩｇＧ和ＩｇＭ等抗体的Ｂ细胞活性增强；用ＬＰＳ刺激巨噬细胞诱生的白介素（ＩＬ）ＩＬ－１也增多。     

细胞因子与哮喘发病机理的关系

研究分设３组，哮喘发作组、哮喘缓解组和正常对照组各２０例，外周血ＩＦＮ－ｒ测定采用微量细胞病变抑制法。ＩＬ－４分泌细胞阳性率测定采用ＡＰＡＡＰ法。Ｃｏｎ－Ａ诱导Ｔｓ细胞活性测定采用Ｓｍｉｔｈ改良法。血清ＩｇＥ测定采用ＥＬＩＳＡ法。试验结果如下：（１）哮喘发作组外周血Ｔ淋巴细胞检测ＩＬ－４分泌细胞阳性率（７．６±３．５％）明显高于哮喘缓解组（３．８±２．０％），Ｐ＜０．０１，更高于正常对照组（１．８±０．５％），Ｐ＜０．０００１。在体外用ＰＨＡ诱导检测产生ＩＰＮ－Ｖ的能力，发现哮喘发作组（７３２．…     

中华麦饭石对肝癌大鼠免疫功能的作用

给Ｗｉｓｔａｒ大鼠喂养ＤＡＢ饲料诱发肝癌，同时饮用１０％中华麦饭石浸液共２６ｗｋ，在实验第８、１８和２６ｗｋ，用脾细胞增殖法和间接免疫荧光法检测白细胞介素－２（ＩＬ－２）活性和Ｔ淋巴细胞亚群。结果显示，实验组ＩＬ－２活性和辅助Ｔ细胞抑制Ｔ细胞比值３匀显著高于对照组（Ｐ〈０．０５￣０．００１），在第１８和２６ｗｋ，实验组总Ｔ细胞和ＴＨ细胞显著高于对照组（Ｐ〈０．０５￣０．０１），Ｔｓ细胞显著低于对照     

通过维持微环境IL－2水平抑制肿瘤形成

为研究肿瘤微环境中ＩＬ－２对肿瘤生长的抑制效应，用转基因方法，将携带ｍＩＬ－２ｃＤＮＡ基因的牛乳头瘤病毒载体（ＢＣＭＧ）转染小鼠Ｂ１６瘤细胞，获稳定持久分泌ｍＩＬ－２的Ｂ１６细胞株（ｍＩＬ－２＋Ｂ１６）。动物实验结果显示：ｍＩＬ－２＋Ｂ１６细胞的致瘤性明显下降，但体外生长率与其亲代Ｂ１６细胞相比较无区别。用ｍＩＬ－２＋Ｂ１６细胞诱导的小鼠腹腔渗出细胞（ＰＥＣ）对ＹＡＣ－１靶细胞和Ｂ１６靶细胞的体外杀伤能力比对照组ＰＥＣ（Ｂ１６诱导的ＰＥＣ）高一倍以上（Ｐ＜０．０１）。表明提高微环境中的ＩＬ－２水平能提高抗肿瘤免疫细胞对肿瘤细胞的杀伤能力，从而抑制肿瘤的形成。     

IL-2基因修饰成纤维细胞对肝癌荷瘤小鼠的治疗实验

目的研究双顺反子逆转录病毒载体介导ＩＬ－２基因转导成纤维细胞对小鼠肝癌的治疗作用。方法利用双顺反子逆转录病毒载体ｐＧＣＥＮ／ＩＬ－２将ＩＬ－２基因转导小鼠成纤维细胞ＮＩＨ３Ｔ３，然后将分泌ＩＬ－２的成纤维细胞与６０Ｇｙγ射线照射的肝癌Ｈ２２细胞皮下植入三天前５×１０５或２．２×１０５肝癌Ｈ２２细胞皮下注射的ＢＡＬＢ／ｃ鼠。结果分泌ＩＬ－２的成纤维细胞治疗组能明显增加荷瘤小鼠的生存率（Ｐ＜０．０２５），抑制肿瘤生长（Ｐ＜０．０５），在肿瘤植入第７０天无肿瘤形成的小鼠再次注射１×１０６Ｈ２２细胞仍然不发生肿瘤。５１Ｃｒ释放试验表明来自照射Ｈ２２与分泌ＩＬ－２成纤维细胞混合液注射的小鼠脾细胞与５１Ｃｒ标记的Ｈ２２细胞共培养后，５１Ｃｒ特异释放明显高于对照组（Ｐ＜０．０５）。结论实验结果表明ＩＬ－２基因修饰的成纤维细胞能诱导针对肝癌的特异免疫反应。     

通过维持微环境IL－2水平抑制肿瘤形成

为研究肿瘤微环境中ＩＬ－２对肿瘤生长的抑制效应，用转基因方法，将携带ｍＩＬ－２ｃＤＮＡ基因的牛乳头瘤病毒载体（ＢＣＭＧ）转染小鼠Ｂ１６瘤细胞，获稳定持久分泌ｍＩＬ－２的Ｂ１６细胞株（ｍＩＬ－２＋Ｂ１６）。动物实验结果显示：ｍＩＬ－２＋Ｂ１６细胞的致瘤性明显下降，但体外生长率与其亲代Ｂ１６细胞相比较无区别。用ｍＩＬ－２＋Ｂ１６细胞诱导的小鼠腹腔渗出细胞（ＰＥＣ）对ＹＡＣ－１靶细胞和Ｂ１６靶细胞的体外杀伤能力比对照组ＰＥＣ（Ｂ１６诱导的ＰＥＣ）高一倍以上（Ｐ＜０．０１）。表明提高微环境中的ＩＬ－２水平能提高抗肿瘤免疫细胞对肿瘤细胞的杀伤能力，从而抑制肿瘤的形成。     

环核苷酸及［Ca~（2＋）］i介导β－内啡肽刺激大鼠巨噬细胞IL－1的分泌

本实验初步观察β－ＥＰ刺激ＩＬ－１分泌的第二信使，培养的大鼠腹腔巨噬细胞，分别加入β－ＥＰ（１）组、β－ＥＰ＋ＮＬＸ（２）组、β－Ｅｐ＋ｆｏｒｓｋｏｌｉｎ（３）组、β－ＥＰ＋亚甲兰（１）组、β－ＥＰ＋尼凡地平（５）组、β－ＥＰ＋ｆｏｒｓｋｏｌｉｎ＋亚甲兰＋尼凡地平（６）组、主理盐水（７）组。２４ｈ后收集巨噬细胞及上清液检测有关指标，发现：第１组细胞内ｃＡＭＰ显著降低，ｃＧＭＰ、［Ｃａ￣（２＋）］ｉ明显升高，上清液呈强烈的ＩＬ－１活性；第２、６、７组细胞内ｃＡＭＰ、ｃＧＭＰ、［Ｃａ￣（２＋）］无明显变化，上清液无ＩＬ－１活性；第３、４、５组上清液们有较强的ＩＬ－１活性，而细胞内ｃＡＭＰ、ｃＧＭＰ及［Ｃａ￣（２＋）］ｉ呈不同变化，提示：ｃＡＭＰ，ｃＧＭＰ及［Ｃａ￣（２＋）］ｉ可能是β－ＥＰ刺激巨噬细胞分泌ＩＬ－１的第二信使。     

糖皮质激素对大鼠脾淋巴细胞表达IL－2受体的调节

采用荧光标记的抗ＣＤ２５单抗，观察了地塞米松对大鼠脾淋巴细胞表达低亲和力ＩＬ－２受体（ＣＤ２５抗原）的影响。结果显示，在ＣｏｎＡ激活的淋巴细胞培养体系中加入地塞米松后，低亲和力ＩＬ－２受体的表达明显受到抑制；表达ＣＤ２５抗原的阳性细胞率亦明显减少。采用￣（１２５）Ｉ－ｒＩＬ－２结合分析实验，进一步研究了地塞米松对高亲和力ＩＬ－２受体的影响。结果证实，１０ｎｍｏｌ／Ｌ地塞米松使高亲和力ＩＬ－２受体的数量由１０．０±０．７３降至６．５１±１．８２ｆｍｏｌ／１０￣７细胞（ｎ＝４），但亲和力无显著变化。糖皮质激素的上述调节作用可能是其抑制免疫功能的重要分子机制之一。     

羧甲基茯苓多糖对HPBL分泌IL—2,TNF,IL—6,IFN—γ的调节作用

用ＣＭＰ培养外周血淋巴细胞（ＨＰＢＬ）２４、３６、４８、７２ｈ采样检测的ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ效价分别可达１３．６±４．３，４１．９±２．０，１８３７．４±４６４．３，１０３７．９±２１１．０Ｕ／ｍｌ，分别比无ＣＭＰ的细胞培养对照组的效价高０．８，７．４，０．５，１０．９倍（Ｐ＜０．０１），说明ＣＭＰ具有ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ的诱生剂功能。由ＣＭＰ预处理ＨＰＢＬ后经ＰＨＡ和／或ＣｏｎＡ促诱生组的ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ效价分别比无ＣＭＰ的ＰＨＡ和／或ＣｏｎＡ刺激的相应常规诱生组高１．２～２．８，０．５～１．１、０．５～０．８、０．４～０．６倍（Ｐ＜０．０１），尤以ＣＭＰ＋ＰＨＡ＋ＣｏｎＡ促诱生细胞因子效果最佳（Ｐ＜０．０１），说明ＣＭＰ又具有ＩＬ－２、ＴＮＦ、ＩＬ－６、ＩＦＮ－γ促诱生效应。     

IL－2和sIL－2R在哮喘发病中的意义探讨

为了探讨ＩＬ－２和ｓＩＬ－２Ｒ在哮喘发病中的意义，对３６例哮喘患者外周血单个核细胞（ＰＢＭＣ）诱生ＩＬ－２水平和血浆ｓＩＬ－２Ｒ水平进行了检测，同时以支气管炎患者与正常人作对照。结果表明，ＰＢＭｃ诱生的ＩＬ－２活性哮喘组高于正常对照组（ｐ＜０．０５）；血浆ｓＩＬ－２Ｒ水平哮喘组高于支气管炎和正常组（ｐ＜０．０１），后两者差异无显著性。以上结果表明，哮喘发病中存在着Ｔ细胞的活化，ＩＬ－２／ＩＬ－２Ｒ在哮喘发病中起作用。     

The extended IL-10 superfamily: IL-10, IL-19, IL-20, IL-22, IL-24, IL-26, IL-28, and IL-29

Cytokines are involved in virtually every aspect of immunity and inflammation. A cascade of responses evolves after cytokine activation, although optimal function might ultimately involve several complementary cytokines. Understanding the function of individual cytokines is complicated because their role can vary depending on the cellular source, target, and phase of the immune response. In fact, numerous cytokines have both proinflammatory and anti-inflammatory potential, with the contrasting outcome observed being determined by the immune cells present and their state of responsiveness to the cytokine. These issues make the study of cytokine biology daunting, particularly so for IL-10 and IL-10-related genes. The IL-10 superfamily is highly pleiotropic. These genes are linked together through genetic similarity and intron-exon gene structure. Significant commonality exists not only through shared receptors but also through conserved signaling cascades. However, its members mediate diverse activities, including immune suppression, enhanced antibacterial and antiviral immunity, antitumor activity, and promotion of self-tolerance in autoimmune diseases.     

IL-10 subfamily members: IL-19, IL-20, IL-22, IL-24 and IL-26

It has been reported that the CD4+ T cell is a very important source of interleukin 10 (IL-10), while CD8+ cells produce low amounts. IL-10 exerts several immune stimulating, as well as inhibitory effects. There are at least five novel human IL-10 family-related molecules: IL-19, IL-20, IL-22, IL-24, and IL-26. Activated T cells produce IL-19, IL-22 and IL-26, while IL-24 is produced by activated monocytes and T-cells. IL-20 induces cheratin proliferation and Stat-3 signal transduction pathway, while IL-22 induces acute-phase production by hepatocytes and neonatal lethality with skin abnormalities reminiscent of psoriasic lesions in humans. In addition, IL-22 mediates inflammation and binds class II cytokine receptor heterodimers IL-22 RA1/CRF2-4. This cytokine is also involved in immuno-regulatory responses. IL-26 (AK155) is a novel cytokine generated by memory cells and is involved in the transformed phenotype of human T cells after infection by herpes virus. All these new IL-10 subfamily member cytokines are strongly involved in immune regulation and inflammatory responses.     

Ovarian follicular concentration of IL-12, IL-15, IL-18 and p40 subunit of IL-12 and IL-23

BACKGROUND: The aim of the study was to determine the presence of interleukin (IL)-12, IL-15, IL-18 and p40 subunit of IL-12/IL-23 in follicular fluid from spontaneous cycles and the relation between the concentration of selected cytokines and IVF-embryo transfer outcome. METHODS: IVF-embryo transfer and enzyme immunoassay (EIA) (R&D Systems, Minneapolis, MN, USA and MBL, Nagoya, Japan) were used. RESULTS: Follicular fluid of women included in the IVF-embryo transfer procedure contained common p40 subunit of IL-12/IL-23 (median 70.1 pg/ml), IL-15 (median 1.3 pg/ml) and IL-18 (median 38.2 pg/ml). There was a significant negative correlation between follicular fluid concentrations of IL-15 and IL-18 (R=-0.392, P=0.003). Significantly higher concentrations of common p40 subunit of IL-12/IL-23 (median 79.8 pg/ml) were found in the follicular fluid taken from follicles containing oocytes, when compared with those without an oocyte (median 44.5 pg/ml, P=0.006). Patients who achieved clinical pregnancy had significantly decreased concentration of IL-15 (median 0.8 pg/ml) compared with patients without successful IVF-embryo transfer outcome (median 1.4 pg/ml, P=0.047). CONCLUSION: Follicular fluid collected from spontaneous cycles contains detectable levels of p40 subunit of IL-12/IL-23, IL-15 and IL-18. Increased concentrations of p40 subunit of IL-12/IL-23 in follicles containing oocytes suggest an important role of this cytokine in reproduction. Possible negative value of IL-15 as a predictor of IVF-embryo transfer success remains to be determined.     

IL-1, IL-18, and IL-33 families of cytokines

Summary: The interleukin-1 (IL-1), IL-18, and IL-33 families of cytokines are related by mechanism of origin, receptor structure, and signal transduction pathways utilized. All three cytokines are synthesized as precursor molecules and cleaved by the enzyme caspase-1 before or during release from the cell. The NALP-3 inflammasome is of crucial importance in generating active caspase-1. The IL-1 family contains two agonists, IL-1α and IL-1β, a specific inhibitor, IL-1 receptor antagonist (IL-1Ra), and two receptors, the biologically active type IL-1R and inactive type II IL-1R. Both IL-1RI and IL-33R utilize the same interacting accessory protein (IL-1RAcP). The balance between IL-1 and IL-1Ra is important in preventing disease in various organs, and excess production of IL-1 has been implicated in many human diseases. The IL-18 family also contains a specific inhibitor, the IL-18-binding protein (IL-18BP), which binds IL-18 in the fluid phase. The IL-18 receptor is similar to the IL-1 receptor complex, including a single ligand-binding chain and a different interacting accessory protein. IL-18 provides an important link between the innate and adaptive immune responses. Newly described IL-33 binds to the orphan IL-1 family receptor T1/ST2 and stimulates T-helper 2 responses as well as mast cells.     

IL-4、IL-13蛋白表达及抗IL-4、IL-13人源单链抗体的筛选

目的:表达IL-4和IL-13蛋白,从人源单链抗体文库中分别筛选抗IL-4和抗IL-13单链抗体.方法:采用RT-PCR从健康志愿者外周血单核细胞(PBMC) mRNA中扩增IL-4和IL-13 cDNA;构建硫氧还蛋白融合表达载体,转化大肠杆菌BL21,IPTG诱导表达并对表达产物进行纯化鉴定.以生物素化的IL-4和IL-13为抗原从前期构建的人源抗体文库中采用噬菌体展示技术分别筛选抗IL-4和抗IL-13人源单链抗体(scFv).结果:扩增的IL-4 cDNA大小为280 bp,表达的融合蛋白大小为27 kD左右.扩增的IL-13 cDNA大小为252 bp,表达的融合蛋白大小为25 kD左右.分别以生物素化的IL-4和IL-13蛋白为抗原,采用噬菌体展示技术对人源抗体文库进行3轮富集后,分别有大约37％的scFvs与IL-4有结合特性,有约27％的scFvs与IL-13有结合特性.筛选了4株分别与IL-4和IL-13结合能力强的单链抗体进行了Westem blot鉴定和测序.结论:成功筛选到抗IL-4和抗IL-13人源性单链抗体.     

IL-21 and IL-21 receptor

Interleukin (IL)-21 is a new member of the type I cytokine superfamily. Although it is most homologous to IL-15, it has a unique receptor chain, IL-21R, that pairs with the γ-common cytokine receptor chain. The first experiments examining the biology of the IL-21 pathway reveal that it is a cytokine with effects on natural killer (NK) cells, T cells, and B cells. Mice deficient in the IL-21 R have also been made, and are being examined for the effects of the IL-21/IL-21R pathway in vivo. Here we summarize our current knowledge of this new cytokine pathway, and its role in innate and adaptive immunity.     

The biological paths of IL-1 family members IL-18 and IL-33

Cytokines are key mediators of the immune system, and few have been more thoroughly studied than those of the IL-1 family. IL-1α and IL-1β are the founding members and now celebrate 25 years since their cloning. In that time, IL-1-directed research has illuminated many aspects of cytokine biology and innate immunity. The family is now recognized to include 11 total members, including IL-18 and IL-33, which are the topic of this review. These two inflammatory cytokines are expressed broadly, and their actions influence a variety of physiologic responses involved in inflammation and immunity. The purpose of this article is not to provide an exhaustive review of IL-18 and IL-33 but rather, to summarize what is known about their key functions and to provide perspective on their similarities and differences.     

Polymorphisms of IL-1B, IL-1RN, IL-2, IL-4, IL-6, IL-10, and IFN-gamma genes in the Korean population

Cytokines play a crucial role in regulating the immune and inflammatory responses. The collective influence of several cytokines can regulate immune responses as complex as those underlying allograft rejections or autoimmune diseases. Polymorphisms in the regulatory regions of the cytokine genes may influence their expression. Therefore, the polymorphisms of cytokine genes are potentially important as genetic predictors of the disease susceptibility or clinical outcome. In 311 unrelated healthy Korean individuals, we investigated the polymorphisms of cytokine genes (interleukin-1 [IL-1], IL-2, IL-4, IL-6, IL-10, and interferon-gamma [IFN-gamma]), which had been previously reported to be associated with a number of immune diseases, transplant complications, and direct or indirect influences on the level of expression and production. And we also compared the results to those published for other populations. The genotype distributions were consistent with the assumption of the Hardy-Weinberg equilibrium, with the exceptions of IL-1B +3954 and IL-6-174 polymorphisms. The polymorphisms examined in this study were almost similar to that observed in Asian populations. There were significant differences of the polymorphisms, except for IL-4 receptor alpha +1902, between Korean and other populations. Comparing the alleles associated with higher level of expression and production, IL-1B +3954*T, IL-2-330*G, and IL-4-590*T alleles were significantly higher, and IL-1RN*A2, IL-10-1082*G, and IFN-gamma*2 alleles were lower in Koreans than other populations. Especially in IL-6 promoter -174 polymorphism, we found only the G allele associated with higher plasma IL-6 levels. In haplotype analysis of IL-10 promoter polymorphisms, the GCC haplotype, associated with higher expression of IL-10, was significantly lower in Koreans. These results may be helpful for understanding transplant-related complications, immune or autoimmune diseases, and malignant diseases in the Korean population.     

Regulation of T cells and cytokines by the interleukin-10 (IL-10)-family cytokines IL-19, IL-20, IL-22, IL-24 and IL-26

The family of IL-10-related cytokines includes several human members, IL-19, IL-20, IL-22, IL-24 and IL-26, and a series of herpesviral and poxviral paralogs. Some of these cytokines share common receptor subunits. In this study, we investigated the effects of these cytokines on naive T cell differentiation, antigen-specific T cell suppression, survival ad expression of surface markers in comparison to IL-10 and cytomegalovirus (CMV)-IL-10. Human CD45RA(+) T cells were stimulated in the presence of IL-10-family cytokines in sequential 12-day cycles. After three to four cycles of stimulation, IL-10 and CMV-IL-10 led to increased IFN-gamma and IL-10 but decreased IL-4 and IL-13. Interestingly, long-term exposure of T cells to IL-19, IL-20 and IL-22 down-regulated IFN-gamma but up-regulated IL-4 and IL-13 in T cells and supported the polarization of naive T cells to Th2-like cells. In contrast, neutralization of endogenous IL-22 activity by IL-22-binding protein decreased IL-4, IL-13 and IFN-gamma synthesis. The antigen-specific suppressor activity of IL-10 and CMV-IL-10 was not observed for any of the other IL-10-family cytokines. These data demonstrate that IL-19, IL-20 and IL-22 may participate in T cell-mediated diseases by distinct regulation of T cell cytokine profiles.