Production of Interleukin 2 like Growth Factor from the Mitogen Activated Peripheral Blood Lymphocytes of Indian Major Carp, Labeo rohita

Optimum conditions for in vitro production of interleukin 2 （IL 2） like activity from the Peripheral blood lymphocytes （PBL） of an Indian major carp, Labeo rohita were studied. Culture supernatants were generated by culturing the PBL in RPMI-1640 media supplemented with Glutamine and 10% fetal bovine serum （FBS） and stimulating with two different mitogens： concanavalin A （Con A） and Phytohaemagglutinin （PHA） at different concentrations separately. Significantly （P 〈 0.01） higher proliferation response was obtained from the culture supematant stimulated with concanavalin A （Con A） at a concentration of 10 lag mLl. The effect of phorbol myristate acetate （PMA） was also studied by co-stimulating PBL with Con A and PHA separately and it was found to synergistically enhancing the stimulation index with Con A whereas the stimulation index remain unchanged with PHA. The Con A （10 μg mLl） stimulated PBL were also cultured at different cell density, incubation period and incubation temperature in order to optimize the in vitro L. rohita IL2 production. The IL2 like activity was studied by lymphocyte proliferation assay on 72 h Con A blasts using WST based assay technique. Significantly （P 〈 0.01） higher stimulation indices were obtained when the PBL were cultured at a cell density of 1 × 10^6 cells mL^-1 for 30-36 h at an incubation temperature of 30 ℃. The IL2 like activity was purified by DEAE-Sepharose anion exchange chromatography and recorded between 70-130 mM NaCI with peak activity at 110 Mm NaCI. The molecular weight of the factor responsible for IL2 like activity was found to be 15-17 KD.

血清IL-19、VASH-1、NLR在2型糖尿病肾病中的水平及临床意义

目的探讨血清白介素19(IL-19)、血管生成抑制蛋白1(VASH-1)、中性粒细胞与淋巴细胞比值(NLR)在2型糖尿病肾病(T2DN)中的水平及意义。方法选取2020年12月—2022年12月在邯郸市第一医院内分泌一科治疗的T2DN患者100例作为T2DN组,同时收集单纯2型糖尿病(T2DM)患者100例作为T2DM组,比较2组血清IL-19、VASH-1、NLR水平,并分析血清IL-19、VASH-1、NLR对T2DN的预测价值。结果T2DN组血清IL-19、VASH-1、NLR明显高于T2DM组(t/P=8.374/<0.001、8.277/<0.001、8.762/<0.001)。经二元Logistic回归分析,IL-19、VASH-1、NLR升高是发生T2DN的危险因素[OR(95%CI)=1.120(1.037~1.210)、1.006(1.001~1.012)、2.586(1.165~5.740),P<0.05]。受试者工作特征(ROC)曲线分析显示,IL-19、VASH-1、NLR及三者联合预测T2DN的曲线下面积(AUC)分别为0.807、0.799、0.822、0.854,约登指数分别为0.520、0.490、0.550、0.740,IL-19在最佳截断值对应的敏感度、特异度分别为0.940、0.580,VASH-1为0.660、0.830,NLR为0.920、0.630,三者联合为0.860、0.880。结论T2DN患者血清IL-19、VASH-1、NLR水平升高,且血清IL-19、VASH-1、NLR三者联合对T2DN具有较高的预测价值。

miR-760/CPEB2轴对急性淋巴细胞白血病细胞凋亡和增殖的影响

目的探究miR-760对急性淋巴细胞白血病(ALL)凋亡和增殖的影响和机制。方法采用实时荧光定量PCR(qRT-PCR)检测miR-760和细胞质多腺苷酸化元件结合蛋白2(CPEB2)相对表达。Ball-1细胞分为不同组:miR-NC组(转染miR-NC)、miR-760mimic组(转染miR-760 mimic)、si-NC组(转染si-NC)、si-CPEB2组(转染si-CPEB2)、oe-NC组(转染oe-NC)、oe-CPEB2组(转染oe-CPEB2)、miR-760 mimic+oe-NC组(共转染miR-760 mimic+oe-NC)和miR-760 mimic+oe-CPEB2组(共转染miR-760 mimic+oe-CPEB2)。流式细胞术检测细胞凋亡,Western blot检测Bax、c-caspase-3、t-caspase-3、Ki67、PCNA和CPEB2相对表达量。双荧光素酶报告实验验证miR-760和CPEB2靶向结合。结果与人B淋巴母细胞HMy2.CIR比较,ALL细胞系Ball-1、Reh和JurkatE6中miR-760水平表达下降,CPEB2 mRNA和蛋白水平表达上升(P<0.05)。与miR-NC组比较,miR-760 mimic组细胞凋亡率、Bax和c-caspase-3水平表达上升,而Ki67和PCNA水平表达下降(P<0.05)。生物信息学方法预测miR-760靶向调控CPEB2,双荧光素酶报告实验验证CPEB2是miR-760的靶标。与si-NC组比较,si-CPEB2组细胞凋亡率、Bax和c-caspase-3水平表达上升,而Ki67、PCNA水平表达下降(P<0.05)。上调CPEB2表达能逆转miR-760过表达对细胞凋亡率、Bax、ccaspase-3的促进和对Ki67、PCNA的抑制(P<0.05)。结论miR-760靶向调节CPEB2,诱导细胞凋亡、抑制细胞增殖,为ALL治疗提供新选择。

蜂胶补充治疗对2型糖尿病合并肺部感染患者效果、炎性因子及免疫功能的影响

目的 探究蜂胶补充治疗对2型糖尿病(T2DM)合并肺部感染的效果。方法 选择2021年3月—2022年9月收治的T2DM合并肺部感染90例,随机分为观察组、对照组各45例。对照组予以常规胰岛素、抗感染治疗,观察组在对照组基础上给予酒制蜂胶辅助治疗。比较2组临床效果及临床症状、体征改善时间及胰岛素用量,分析2组治疗前后血糖代谢及波动、血清炎性指标、免疫功能指标水平变化。结果 观察组总有效率[93.33%(42/45)]高于对照组[75.56%(34/45)](P<0.05)。观察组胰岛素用量少于对照组,血糖达标时间、体温恢复时间、肺部啰音消失时间及肺部阴影吸收时间均短于对照组(P<0.05)。治疗后,观察组空腹血糖、餐后2 h血糖、C反应蛋白、降钙素原及白细胞介素-6水平低于对照组,自然杀伤细胞、CD4+/CD8+、CD4+及CD3+高于对照组(P<0.05)。2组不良反应总发生率比较差异无统计学意义(P>0.05)。结论 蜂胶补充治疗对T2DM合并肺部感染效果较好,可快速缓解临床症状,有效调节糖代谢状态,降低炎性因子,改善机体免疫功能。

孕早期监测BAFF、AMH、IL-2预测复发性流产患者流产再发生的临床价值

目的探究孕早期监测B淋巴细胞活化因子(BAFF)、抗米勒管激素(AMH)、白介素-2(IL-2)对复发性流产患者流产再发生的临床预测价值。方法选取2019年9月至2023年6月上海健康医学院附属崇明医院收治的100例复发性流产患者作为研究对象并纳入观察组,根据患者早期妊娠情况分为非流产组(n=65)和流产组(n=35);选取同期同院100例产检健康的孕妇作为对照组。比较各组血清BAFF、AMH、IL-2水平,分析BAFF、AMH、IL-2水平预测流产再发生的临床价值。采用Logistic回归分析影响复发性流产患者流产再发生的因素。结果观察组BAFF、IL-2水平比对照组高,AMH水平比对照组低(P<0.05)。流产组BAFF、IL-2水平比非流产组高,AMH水平比非流产组低(P<0.05)。流产组孕次、流产≥3次比例高于非流产组(P<0.05)。绘制受试者工作特征(ROC)曲线发现,BAFF、AMH、IL-2水平预测患者发生再流产的曲线下面积(AUC)分别是0.809、0.821、0.803,三者联合检测的AUC为0.919,均优于各自单独检测(Z=1.920、1.974、2.298,P<0.05)。多因素Logistic回归发现,BAFF、IL-2是影响患者发生再流产的危险因素,AMH为保护因素(P<0.05)。结论复发性流产患者BAFF、IL-2水平升高,AMH水平降低,且三者联合检测有助于早期预测流产再发生,具有一定的诊断价值。

术前NLR、IL-35及MTA2与前列腺癌患者术后复发及转移的相关性

目的探讨术前中性粒细胞与淋巴细胞比值(NLR)、白细胞介素-35(IL-35)及转移相关蛋白2(MTA2)对前列腺癌患者术后复发及转移的预测价值。方法选取2019年6月-2022年6月在河北北方学院附属第二医院接受手术治疗的160例老年前列腺癌患者作为研究对象,根据术后有无复发或转移分为发生组(68例)和未发生组(92例)。比较两组一般资料,采用logistic回归分析NLR、IL-35、MTA2与前列腺癌患者术后复发及转移风险的相关性,ROC曲线评估各指标对前列腺癌患者术后复发及转移风险的预测价值。结果两组TNM分级、Gleason评分,NLR、IL-35及MTA2比较,差异均有统计学意义(P<0.05)。Logistic回归分析显示,在校正多个混杂因素后,NLR、IL-35水平及MTA2评分均是前列腺癌患者术后复发及转移发生的独立危险因素(P<0.05)。ROC曲线显示,NLR、IL-35水平及MTA2评分对前列腺癌患者术后复发及转移具有一定的预测价值。结论NLR、IL-35水平及MTA2评分对前列腺癌术后发生复发及转移具有一定的预测价值,对早期前列腺癌术后复发及转移的高危患者的判断和改进患者预后具有重要的临床价值。

外周血CD4^(+)T、CD8^(+)T细胞及免疫活化分子CD38、IL-2R表达水平在Graves眼病患者中的临床意义

目的研究Graves眼病(GO)患者外周血CD4^(+)T、CD8^(+)T细胞及免疫活化分子CD38、IL-2R水平的临床意义。方法选取2022年1月至2023年1月我院收治的80例Graves病(GD)患者为GD组,65例GO患者为GO组,根据CAS评分将GO组患者分为活动期组(26例)和非活动期组(39例),另选50例体检中心健康者为对照组。比较各组血清甲状腺功能、甲状腺相关抗体及外周血各细胞因子水平,分析GO患者外周血各细胞因子水平与甲状腺功能、甲状腺相关抗体、CAS评分的相关性,并分析外周血各细胞因子对GO疾病活动性的预测价值。结果GO组FT3、FT4低于GD组,高于对照组,GD组高于对照组(P<0.05);GO组TSH高于GD组,低于对照组,GD组低于对照组(P<0.05);GO组、GD组TGAb、TPOAb、TRAb、CD4^(+)T高于对照组(P<0.05);GO组、GD组CD8^(+)T低于对照组(P<0.05);GO组CD4^(+)/CD8^(+)T、CD4^(+)CD38^(+)T、CD8^(+)CD38^(+)T、IL-2R高于GD组和对照组,GD组高于对照组(P<0.05)。GO患者活动期组CD4^(+)/CD8^(+)T、CD4^(+)CD38^(+)T、CD8^(+)CD38^(+)T、IL-2R高于非活动期组(P<0.05)。CD4^(+)/CD8^(+)T、CD8^(+)CD38^(+)T与CAS评分呈正相关(P<0.05);CD4^(+)CD38^(+)T与TRAb、CAS评分呈正相关(P<0.05);IL-2R与FT4、TRAb、TPOAb、CAS评分呈正相关(P<0.05)。CD4^(+)/CD8^(+)T、CD4^(+)CD38^(+)T、CD8^(+)CD38^(+)T、IL-2R联合检测预测GO患者疾病活动性的AUC值为0.930,高于外周血四者单独检测(AUC=0.736、0.749、0.668、0.699,P<0.05)。结论外周血CD4^(+)/CD8^(+)T、CD4^(+)CD38^(+)T、CD8^(+)CD38^(+)T、IL-2R联合预测GO活动性的临床价值更高。

沉默MAL2对乳腺癌细胞增殖和药物敏感性的影响

目的探讨沉默髓鞘和淋巴细胞蛋白2(MAL2)基因对乳腺癌MDA-MB-231和MCF-7细胞增殖和药物敏感性的影响,同时研究乳腺癌细胞对阿霉素(DOX)和顺铂(DDP)的耐药机制。方法利用数据库分析MAL2基因在正常组织、癌旁组织和乳腺癌组织中的表达及与预后的相关性;采用蛋白印迹(Western blot)法检测MAL2在人正常乳腺上皮细胞MCF-10A和乳腺癌MDA-MB-231、MCF-7及MDA-MB-468细胞中的蛋白表达;利用转染干扰序列沉默乳腺癌MDA-MB-231和MCF-7细胞中MAL2的表达,根据序列不同分为对照组(siRNA-NC组)和实验组(siRNA-MAL2-1组、siRNA-MAL2-2组及siRNA-MAL2-3组);选取沉默效果最好的实验组(siRNA-MAL2-3组)序列进行慢病毒构建及实验,沉默乳腺癌MDA-MB-231和MCF-7细胞中MAL2的表达,分为sh-NC组(序列同siRNA-NC组)和sh-MAL2组(序列同siRNA-MAL2-3组),采用Western blot法检测MAL2的蛋白表达,CCK8法和平板克隆形成实验检测细胞的增殖能力;CCK8法、流式细胞术及Western blot检测沉默MAL2基因的表达后乳腺癌细胞对DOX和DDP的敏感性;Western blot检测p-AKT/m-TOR通路相关蛋白[磷酸化蛋白激酶B(p-AKT)、雷帕霉素靶蛋白(m-TOR)、磷酸化雷帕霉素靶蛋白(p-mTOR)]的表达情况。结果生物信息学分析显示MAL2在乳腺癌组织中高表达、且其高表达和患者预后不良相关(P<0.05),MAL2在乳腺癌细胞系中高表达(P<0.01);与siRNA-NC组相比,siRNA-MAL2-3组的MAL2蛋白沉默效果较好(P<0.01);感染慢病毒实验结果表明,与sh-NC组比较,sh-MAL2组MAL2表达被抑制(P<0.05);与sh-NC组相比,sh-MAL2组细胞增殖能力无变化(P>0.05)、对DOX和DDP的敏感性增强(P<0.05)、p-AKT和p-mTOR蛋白表达下调(P<0.05)。结论沉默MAL2对乳腺癌细胞增殖能力没有影响,但可促进乳腺癌细胞对DOX和DDP的敏感性,其机制可能与AKT/m-TOR通路相关蛋白被抑制有关。

Interleukin-2 expression and glioma cell proliferation following Vaccinia vector gene transfection in vivo

BACKGROUND： The effectiveness of gene therapy is closely related to the efficiency of vector transfection and expression. OBJECTIVE： This study was designed to transfect a human brain glioma cell line with recombinant Vaccinia virus expressing the interleukin-2 （rVV-IL-2） gene, and to observe IL-2 expression and glioma cell proliferation potential after transfection. DESIGN： Experimental observation. SETTING： Department of Neurosurgery, Shenyang Military Area Command of Chinese PLA. MATERIALS： The rVV-IL-2 vectors were obtained through homologous recombination and screening in the Second Military Medical University of Chinese PLA. The human brain glioma cell line and IL-2-dependent cells were produced by the Second Military Medical University of Chinese PLA. Human IL-2 was produced by Genzyme Corporation. METHODS： At passage day l, Veto cells were amplified l ; 1 for virus and cells. A human brain glioma cell line was transfected using amplified Vaccinia viral vectors at varying multiplicities of infection （MOI）. At 2, 4, 6, 8, 12, and 24 hours post-transfection, superuatant was collected to determine by MTT assay IL-2 expression levels in IL-2 dependent cells. The transfected and non-transfected cells were divided into 4 groups, namely MOI1 ： 1, MOI 5 ： 1, MOI 10 ： 1, and control groups. MAIN OUTCOME MEASURES： IL-2 expression at different time points after transfection of human brain glioma cells with varying MOI of Vaccinia viral vectors; in vitro proliferation capacity of human brain glioma cells among the 4 groups. RESULTS： IL-2 expression was detectable 4 hours after Vaccinia viral vector transfection and reached 300 kU/L by 8 hours. There was no significant difference in the proliferating rate of human brain glioma cells among the 4 groups （P 〉 0.05）. CONCLUSION： Vaccinia viral vectors can transfect human brain glioma cells in vitro and express high levels of IL-2. Vaccinia virus and high IL-2 expression do not influence the proliferation rate of human brain glioma cells in vitro.

STUDY OF T－LYMPHOCYTE SUBSETS AND INTERLEUKIN－2 AND INTERLEUKIN－2 RECEPTOR OF RESPIRATORY SYNCYTIAL VIRUS PNEUMONIA

T-Lymphocyte subsets and humoral immune and the activity of IL-2 and IL-2R in respiratory syncytial virus (RSV) pneumonia in 26 cases were tested. The result showed in the patients with RSV pneumonia the averages of T3 and T4 were 37.56±1.46% and 27. 15±8. 02% respectively,They were significantly lower than 53.4 ±9.2% and 35.5±7.7% of averages of T3 and T4 in healthy controlled group (both. P< 0. 001 ), the average of T3 was 22. 73±7.06%, it was lower an 26. 7±6. 3 % of T8 in controlled group (P<0. 02 );the ratio of T4/T8 was 1. 245±0. 399 ,there was no significant difference from 1. 35 ±0. 17 of the ratio in controlled group (P > 0. 1). The mean value of IgG was 1. 177± 0. 3685g/L, it was significantly lower than 1. 427± 0. 498g/L of that in controlled group(P < 0. 005). The mean values of IgA and IgM were 0. 1136±0.0393g/L and 0. 768±0. 353g/L respectively, they were significantly lower than 0. 2706 ±0. 876g/L and 0. 122±0. 061g/L of IgA and IgM in controlled group. The activity of IL-2 and IL-2R were 17. 46 ±5. 79%, and 28. 32 ±5. 924% respectively, they were significantly lower than 30. 22 ±14. 55% and 39. 53±8. 61 % of those in healthy group (both P < 0. 001). The severe the pneuumonia, the greater the lowering of IL-2 and IL-2R. These about results suggested that RSV could greatly suppress the immune function of the patients, inducing secondary immunodeficiency, leading to repeated breather and asthma.

帕米膦酸二钠联合复方苦参注射液对非小细胞肺癌患者外周血CEA、CA125、IL-2、IL-4和T淋巴细胞亚群的影响

目的探讨帕米膦酸二钠单药及联合复方苦参注射液治疗非小细胞肺癌,对其外周血癌胚抗原(CEA)、糖类抗原125(CA125)、血清细胞因子白介素-2(IL-2)、白介素-4(IL-4)及T淋巴细胞亚群的影响。方法选取2018年1月至2022年3月句容市人民医院收治的非小细胞肺癌患者50例。采取分别抽样法分组,即单药帕米膦酸二钠干预的单药组(n=25)和帕米膦酸二钠联合复方苦参注射液干预的联合治疗组(n=25)。观察两组临床指标。结果联合治疗组的客观缓解率、临床获益率及生活质量均高于单药组,差异有统计学意义(P<0.05);联合治疗组的CEA、CA125水平低于单药组,差异有统计学意义(P<0.05);联合治疗组IL-2水平高于单药组、IL-4水平低于单药组,差异有统计学意义(P<0.05);联合治疗组CD4^(+)、CD4^(+)/CD8^(+)水平高于单药组,差异有统计学意义(P<0.05);联合治疗组的不良反应总发生率低于单药组,差异有统计学意义(P<0.05)。结论帕米膦酸二钠联合复方苦参注射液治疗非小细胞肺癌,可调节炎性因子水平,增强抗肿瘤活性及免疫功能,减轻炎性反应,不良反应发生率低。

LncRNA DLEU2对宫颈癌细胞生物学行为、miR-21和Wnt/β-catenin信号通路的影响

目的 探讨长链非编码RNA(lncRNA)淋巴细胞白血病缺失基因2(DLEU2)对宫颈癌细胞生物学行为的影响,并基于miR-21和Wnt/β-连环蛋白(β-catenin)信号通路探讨其作用机制。方法 将宫颈癌HeLa细胞分为对照组、DLEU2干扰组、DLEU2干扰空载组、DLEU2过表达组、DLEU2过表达空载组。除对照组外,给予其他组细胞转染相应的质粒。转染24 h后,检测各组的细胞增殖活性、细胞凋亡情况及凋亡相关蛋白表达水平、细胞周期及细胞周期相关因子的mRNA表达水平、细胞侵袭数量及上皮细胞-间充质转化(EMT)相关蛋白表达水平、miR-21表达水平、Wnt/β-catenin的蛋白及mRNA表达水平。结果 (1)与对照组相比,DLEU2过表达组的细胞增殖活性降低、细胞凋亡率增加、B细胞淋巴瘤2(Bcl-2)蛋白表达水平下调、Bcl-2相关性蛋白X和Caspase-3蛋白表达水平上调(P<0.05)。(2)与对照组相比,DLEU2过表达组的G_(1)期和S期细胞比例降低、G_(2)期细胞比例升高、细胞周期相关因子的mRNA表达水平下调(P<0.05)。(3)与对照组相比,DLEU2过表达组的细胞侵袭数减少,神经型钙黏蛋白表达水平下调,上皮型钙黏蛋白表达水平上调(P<0.05)。(4)与对照组相比,DLEU2过表达组的miR-21表达水平及Wnt3a、β-catenin的mRNA和蛋白表达水平均下调。(5)DLEU2干扰组的上述指标变化均与DLEU2过表达组相反(P<0.05),而两个空载组与对照组相比差异无统计学意义(P>0.05)。结论 DLEU2可能通过调控miR-21及抑制Wnt/β-catenin信号通路活性,从而影响宫颈癌细胞的存活、死亡、EMT和侵袭等生物学行为。

弥漫大B细胞淋巴瘤患者血浆IFN-γ、IL-2及外周血T淋巴细胞亚群、NK细胞与其临床分期的关系

目的探讨弥漫大B细胞淋巴瘤(DLBCL)患者血浆干扰素γ(IFN-γ)、白细胞介素-2(IL-2)水平及外周血T淋巴细胞亚群、自然杀伤(NK)细胞与临床分期的关系。方法纳入河南中医药大学第三附属医院2020年12月至2022年12月收治的DLBCL患者198例,根据临床分期分成Ⅰ期组(31例)、Ⅱ期组(56例)、Ⅲ期组(59例)、Ⅳ期组(52例)。比较4组血浆IFN-γ、IL-2及外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平,分析各指标与DLBCL分期的相关性。结果Ⅲ期组、Ⅳ期组的血浆IFN-γ、IL-2水平低于Ⅰ期组、Ⅱ期组(P<0.05)。Ⅲ期组、Ⅳ期组外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平低于Ⅰ期组、Ⅱ期组(P<0.05)。DLBCL患者IFN-γ、IL-2与CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平呈正相关(P<0.05)。DLBCL患者的血浆IFN-γ、IL-2及外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平与临床分期呈负相关(P<0.05)。IFN-γ、IL-2、CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平增高是控制临床分期增高的保护因素(P<0.05)。结论DLBCL患者的临床分期与血浆IFN-γ、IL-2及外周血T淋巴细胞亚群、NK细胞水平相关,且血浆IFN-γ、IL-2水平与T淋巴细胞亚群、NK细胞存在相关性,上述指标有望对DLBCL进展风险进行预测。

Role of the IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to Aspergillus fumigatus infection

AIM: To investigate the expression of interleukin(IL)-33 in the cornea and human corneal epithelial cells(HCECs) exposed to Aspergillus fumigatus(A. fumigatus), and to determine the function of IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to A. fumigatus infection.METHODS: The mRNA and protein expression of IL-33 in HCECs and mice corneas were examined by quantitative real-time reverse transcription polymerase chain reaction(q RT-PCR) and Western blot analysis, respectively. IL-33 expression was also detected in cornea samples from healthy donors and patients with fungal keratitis with immunohistochemistry. The cultured HCECs were treated with inactive A. fumigatus hyphae at various concentrations with or without recombinant human IL-33 protein, soluble recombinant ST2 protein, specific ST2 neutralizing antibody, or the mitogen-activated protein kinase(MAPK) p38 inhibitor SB203580 for evaluation of the expression and activation of IL-33/ST2/p38 signaling in the regulation of proinflammatory cytokines. The production levels of IL-6 and IL-1β were determined by qR T-PCR and enzymelinked immunosorbent assay(ELISA). The proliferation of HCECs was determined by a Cell Counting Kit-8(CCK8) assay and cell count.RESULTS: IL-33 expression levels increased in the corneal tissues of patients with fungal keratitis and in mice corneas of experimental A. fumigatus infection,as well as in HCECs with infection of A. fumigatus. A. fumigatus strongly stimulated HCECs-generated proinflammatory cytokine(IL-6 and IL-1β) production at both the mRNA and protein levels. This production of proinflammatory mediators stimulated by A. fumigatus was further stimulated by IL-33 and was prevented by soluble ST2 protein or ST2 neutralizing antibody. Moreover, IL-33 naturally promoted the p38 phosphorylation induced by A. fumigatus, which was suppressed by soluble ST2 protein. The MAPK p38 inhibitor SB203580 also inhibited the A. fumigatus-induced proinflammatory cytokine production. IL-33 administration for 48 h and 72 h promoted the proliferation of HCECs, which was attenuated by treatment with soluble recombinant human ST2 protein.CONCLUSION: A. fumigatus elevates IL-33 expression in human and mice corneas and HCECs. Thus, IL-33/ST2/p38 signaling may play an important role in amplifying the immune response of corneal epithelial cells to A. fumigatus infection. Besides, IL-33 promotes the cell proliferation of HCECs via its receptor ST2. These findings suggest a novel autocrine mechanism of amplification of the fungalinduced inflammatory response in the corneal epithelium, highlighting a potential therapeutic target for fungal keratitis.

Lamprey buccal gland secretory protein-2 (BGSP-2) inhibits human T lymphocyte proliferation

Lamprey is a representative of the agnathans, the most ancient class of vertebrates. Parasitic lampreys secrete anticoagulant from their buccal glands and prevent blood coagulation of host fishes. We identified a buccal gland secretory protein-2 （BGSP-2） from a buccal gland cDNA library of Larnpetrajaponica. The full-length BGSP-2 gene was cloned and the recombinant BGSP-2 protein was generated. The role of BGSP-2 on lymphocyte proliferation was studied by examining its effects on human T lymphocytes. We found that lamprey BGSP-2 was able to effectively block the proliferation of T cells in vitro by inducing G1/S cell cycle arrest. Furthermore, it inhibited the proliferation of hmnan T lymphocytes stimulated by phytohemagglutinin （PHA） at a minimum concentration of 0.1μg/ml. Our data suggest that lamprey BGSP-2 is able to block the mitosis of human T lymphocytes at the G1/S point, and has the potential of anti-proliferative effect on PHA-activated T lymphocytes.

Ribavirin contributes to eradicate hepatitis C virus through polarization of T helper 1/2 cell balance into T helper 1 dominance

The mechanism of action of ribavirin(RBV) as an immunomodulatory and antiviral agent and its clinical significance in the future treatment of patients with hepatitis C virus(HCV) infection are reviewed.RBV up-regulates type 1 and/or 2 cytokines to modulate the T helper(Th) 1/2 cell balance to Th1 dominance.Examination of co-stimulatory signaling indicated that RBV down-modulates inducible co-stimulator on Th cells,which contributes to differentiating na?ve Th cells into Th2 cells while reducing their interleukin-10 production.The effects on T-regulatory(Treg) cells were also investigated,and RBV inhibited the differentiation of na?ve Th cells into adaptive Treg cells by downmodulating forkhead box-P3.These findings indicate that RBV mainly down-regulates the activity of Th2 cells,resulting in the maintenance of Th1 activity that contributes to abrogating HCV-infected hepatocytes.Although an interferon-free treatment regimen exhibits almost the same efficacy without serious complications,regimens with RBV will be still be used because of their ability to facilitate the cellular immune response,which may contribute to reducing the development of hepatocellular carcinogenesis in patients infected with HCV.

Effect of IL-10 gene transmission on the PTg-stimulated splenocytes proliferation and Th1 cytokines production from experimental autoimminue thyroiditis rats

To investigate the effects of gene therapy with IL-10 on PTg-induced proliferation of splenocytes and Thl cytokine production from PTg-stimulated splenocytes. Methods： EAT rats were divided into four groups ：group A （PBS＋PLL） , group B （pORF＋PLL）, group C （pORFmIL10＋PLL）, and group D （pORFmIL10＋ MEM）. The substances mixed with lipofectamine were injected into the thyroid tissues of rats on the 18th dday after immunization. The rats were sacrificed at the 8th week. In vitro proliferative responses to ConA and different concentration of PTg were measured by culturing 4 ×105 splenocytes pulsed with 18.5KBq of [^3H] thymidine for the final 12h and then harvested for liquid scintillation counting. In vitro splenocytes were cultured with PTg （25 mg/L）. Thl cytokine IFN-γ,TNF-α and IL-2 were detected by ELISA. Results： The proliferative response to PTg was suppressed in group C, compared with that of group A and B （P 〈 0.05）. The levels of IFN-γ,TNF-αand IL-2 in the supernatant of PTg-stimulated splenocytes were 3548.25±779.47 pg/ml, 27.66±10.50 pg/ml and 3617.73±609.15 pg/ml, respectively, which were much lower in group C than those in group A and B（P 〈 0.01, P 〈 0.05, P 〈 0.001, respectively）. Conclusion： IL- 10 gene transmission in thyroid tissues could inhibit PTg specific proliferation of splenocytes from EAT rats and the secretion 6f Th1 cytokines from PTg-stimulated splenocytes.

Double potentialities of tumoricide and hematopoiesis of human bone marrow cells activated by IL-2 and IL-3

The biomodulative and hematopoietic potentialities of IL-2 and IL-3 activatedbone marrow(ABM)cells from patients with lung adenocarcinoma were studied in vitro.Human bone marrow(BM)cells could be activated by IL-2 in culture for 7d.TheseIL-2 ABM cells had higher cytolytic activities against cells of H 7402 cell line and freshautologous adenocarcinoma cells and maintained the cytotoxicities longer than IL-2 acti-vated peripheral blood lymphocytes(APBLs),a point of possible importance in adoptiveimmunotherapy for cancer patients.The IL-2 ABM cells also had similar number ofBFU-E and CFU-GM to that had fresh BM cells if 1L-3 was added 48h alter IL-2 inculture.The IL-2 and IL-3 ABM cells might be used to eliminate tumor cells and tosupply reconstitutive elements of BM for autologous bone marrow transplantation.

益气养阴活血汤辅助治疗对肾病综合征患儿TIMP-2、T淋巴细胞亚群及IL-8影响

目的分析益气养阴活血汤辅助治疗对肾病综合征患儿金属蛋白酶组织抑制因子-2(TIMP-2)、T淋巴细胞亚群及白细胞介素-8(IL-8)水平影响。方法选取2019年1月至2021年1月河南中医药大学第一附属医院收治的102例肾病综合征患儿临床资料,根据治疗方式不同分为对照组(西医常规)47例与观察组(西医常规联合益气养阴活血汤)55例。均治疗4个疗程。对比临床疗效、中医症状积分、TIMP-2、T淋巴细胞亚群、IL-8水平、肾功能[血清肌酐(SCr)、血尿素氮(BUN)、24小时尿蛋白定量(24hUTP)]、不良反应。结果观察组治疗后总有效率占比为96.36%,明显高于对照组的82.67%,差异有统计学意义(P<0.05)。观察组治疗后中医症状积分各项目评分低于对照组,差异有统计学意义(P<0.05)。治疗后观察组CD8^(+)、TIMP-2、IL-8水平均低于对照组,CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平则高于对照组,差异有统计学意义(P<0.05)。观察组SCr、BUN、24hUTP值均低于对照组,差异有统计学意义(P<0.05);两组不良反应比较差异无统计学意义(P>0.05)。结论益气养阴活血汤辅助治疗肾病综合征临床疗效显著,可有效改善患者TIMP-2、T淋巴细胞亚群及IL-8水平,缓解患者临床症状,安全性尚可。

T SUBSETS AND ANTITUMOR ACTIVITY OF LYMPHOCYTES INFILTRATING HUMAN PRIMARY BRAIN GLIOMAS

Glioma-infiltrating lymphocytes (GIL) were isolated from 9 surgical biopsy specimens of primary brain gliomas using mechanical and enzymatic digestion and discontinuous density gradent centrifugation. During cultured in the presence of interleukin-2 (IL-2) for a period of four weeks, GIL were expanded 48. 4-fold on the average, even up to 118-fold. GIL activated by IL- 2 had specific cytolytic activity against autologous glioma cells. Analysis of T subsets of GIL freshly isolated showed that CD3+ cells were 71.0±11.9%, CD4+ cells 34.2±6.1% and CD3+cells 37.0±7.6%. Ability of activated GIL to produce γ-Interferon (γ-IFN) was significantly higher than that of freshly isolated GIL and autologous peripheral blood lymphocytes (PBL). The results suggest that GIL have many advantages for an adoptive immunotherapy of patients with brain gllomas and be a new type of antitumor immune effector.