Th1 cells inducing IFNγ response improves immunotherapy efficacy in gastric cancer

Objective: Cancer immunotherapy has made remarkable advances in recent years, but its effectiveness in treating gastric cancer is often limited by the complexity of the tumor microenvironment and the lack of effective biomarkers. This study aimed to identify effective biomarkers for immunotherapy treatment by characterizing the tumor microenvironment.Methods: We retrieved the RNA-seq data from gastric cancer patients treated with the programmed death 1(PD-1) blockade pembrolizumab. Differentially expressed genes associated with clinical outcomes were identified and further analyzed using gene ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis. Gene signature scores were calculated by single sample Gene Set Enrichment Analysis(ssGSEA). The infiltration levels of immune cells were quantified using the xCell website. Cell type enrichment analysis was performed to compare treatment response and non-response groups, and regression analysis was used to investigate the relationship between interferon gamma(IFNγ) immune response and immune cell infiltration. Biomarkers were identified using least absolute shrinkage and selection operator(LASSO) analysis.Results: Compared to normal tissues, cytokine activity and interleukin-6 production were highly activated in gastric tumors. Responders to pembrolizumab showed significantly up-regulated expression of IFNγ responserelated genes. Cell type enrichment analysis revealed that Th1 cells were significantly enriched in the tumor microenvironment of responders. Regression analysis indicated that Th1 cells induced IFNγ response more efficiently than other cell types. Using signatures of Th1 cells, stromal cells and IFNγ response, a set of eight genes were identified that effectively predicted the efficacy of immunotherapy treatment and patient prognosis.Conclusions: Th1 cells promote therapeutic efficacy of PD-1 blockade by promoting IFNγ immune response in gastric cancer. The identified biomarkers have the potential to improve the effectiveness of immunotherapy treatment for gastric cancer patients.

痛泻安肠方对肠易激综合征小鼠短链脂肪酸代谢及Th1/Th2细胞免疫平衡的影响

目的观察痛泻安肠方对肝郁脾虚证腹泻型肠易激综合征(IBS-D)小鼠模型短链脂肪酸代谢及Th1/Th2细胞免疫平衡的影响。方法30只18~22 g的雄性C57BL/6小鼠随机分为空白组、模型组、痛泻安肠方低剂量组、痛泻安肠方中剂量组、痛泻安肠方高剂量组,每组6只。采用束缚应激联合番泻叶灌胃复制IBS-D小鼠模型,造模成功后分别予相应浓度中药灌胃14 d,每日1次,通过靶向代谢组学检测小鼠粪便中短链脂肪酸含量的变化,并通过流式细胞术检测肠系膜淋巴结中CD3+CD4+INF-γ+T细胞(Th1细胞)、CD3+CD4+IL-4+T细胞(Th2细胞)的占比情况。结果与空白组相比,模型组小鼠粪便短链脂肪酸(SCFAs)相对含量显著升高(P<0.01),尤其是乙酸、丙酸及丁酸的含量均显著升高(P<0.01或P<0.05);不同剂量痛泻安肠方治疗后,粪便总SCFAs相对含量显著降低(P<0.01),乙酸、丙酸及丁酸均有不同程度降低,其中痛泻安肠方各剂量组均可显著降低乙酸和丁酸的含量,差异具有统计学意义(P<0.01),低剂量组及中剂量组可显著降低丙酸的含量,差异具有统计学意义(P<0.01)。造模后,肠易激综合征小鼠肠系膜淋巴结的Th2细胞占比明显降低,而Th1细胞占比及Th1/Th2细胞比率明显升高,差异具有统计学意义(P<0.05);治疗后,痛泻安肠方各剂量组Th2细胞占比均有升高,Th1细胞占比及Th1/Th2细胞比率降低。结论痛泻安肠方可以降低IBS-D小鼠粪便SCFAs的含量,调节小鼠Th1/Th2细胞免疫紊乱。

过敏原对过敏性鼻炎患者外周血CD4^(+)Th1 IL-18、IL-18BPa和IL-18Rα表达的影响

目的探讨过敏原对健康人(healthy control,HC)和过敏性鼻炎(allergic rhinitis,AR)患者外周血CD4^(+)Th1细胞表达IL-18、IL-18BPa和IL-18Rα蛋白水平及mRNA水平的影响。方法收集皮肤点刺试验阴性的鼻炎(AR^(-))、皮肤点刺试验阳性的鼻炎(AR^(+))患者和HC的外周血标本。流式细胞术检测过敏原对CD4^(+)Th1细胞表达IL-18、IL-18BPa和IL-18Rα蛋白水平的影响;qPCR检测过敏原对CD4^(+)T细胞表达IL-18、IL-18BPa和IL-18RαmRNA水平的影响。结果与HC相比,AR^(-)和AR^(+)患者外周血Th1细胞中IL-18蛋白的表达增加,IL-18BPa蛋白的表达减少;AR^(+)患者外周血Th1细胞中IL-18Rα蛋白的表达增加;过敏原可增强HC Th1细胞IL-18Rα蛋白的表达,并诱导AR^(+)患者和HC CD4^(+)T细胞IL-18、IL-18BPa和IL-18RαmRNA的表达。结论过敏原极可能通过促进外周血CD4^(+)Th1细胞表达IL-18和IL-18Rα进而参与AR发病。

基于Notch信号通路探讨芪蛭皱肺颗粒对慢性阻塞性肺疾病大鼠Th1/Th2免疫平衡的调节机制

目的探讨果蝇双翅边缘缺刻同源基因(Notch)信号通路在慢性阻塞性肺疾病(COPD)辅助性T细胞1(Helper T cells 1,Th1)和辅助性T细胞2(Helper T cells 2,Th2)失衡中的作用及芪蛭皱肺颗粒的干预机制。方法70只Wistar大鼠随机挑选10只作为空白对照组,其余大鼠均采用香烟烟雾(CS)联合气管滴注脂多糖(Lipopolysaccharide,LPS)法建立COPD模型,空白对照组及造模组各随机挑选3只大鼠验证造模是否成功。造模结束进行灌胃给药干预,造模组大鼠随机分为模型对照组、阳性对照组(67.5μg·kg^(-1))及芪蛭皱肺颗粒高中低剂量组(3.24、1.62、0.81 g·kg^(-1)),分别给予生理盐水、醋酸地塞米松混悬液、芪蛭皱肺高、中、低剂量混悬液进行灌胃干预,空白对照组同模型对照组,灌胃等体积生理盐水。经28天造模及28天治疗后,采用动物肺功能测试系统检测吸气峰流速(Peak Inspiratory Flow,PIF)和呼气峰流速(Peak Expiratory Flow,PEF),处死大鼠提取肺脏、脾脏、血清及支气管肺泡灌洗液(BALF),苏木素-伊红(HE)染色评价肺组织病理变化,酶联免疫吸附实验法(ELISA)测定血清及BALF中肿瘤坏死因子-α(TNF-α)含量,流式细胞仪检测脾脏Th1/Th2细胞水平,免疫组织化学法(Immunohistochemistry,IHC)及蛋白免疫印迹法(Western blot)检测肺组织Notch1、Hes家族发状分裂相关增强子1(Hes1)、Hey家族发状分裂相关增强子1(Hey1)蛋白水平,实时荧光定量聚合酶链式反应(Real-time PCR,RT-PCR)检测肺组织Notch1、Hes1、Hey1基因表达水平。结果与空白对照组比较,模型对照组大鼠肺功能显著降低(P<0.05),肺组织出现炎性细胞浸润、支气管结构破坏等病变,血清及BALF中TNF-α含量显著升高(P<0.05),脾Th1细胞百分比显著降低(P<0.05),Th2细胞百分比显著升高(P<0.05),肺组织Notch1、Hes1、Hey1蛋白及mRNA表达显著升高(P<0.05),差异均具有统计学意义;与模型对照组比较,各给药组大鼠肺功能显著升高(P<0.05),肺组织病理损伤均有所减轻,血清及BALF中TNF-α含量显著降低(P<0.05),脾Th1细胞百分比显著升高(P<0.05),Th2细胞百分比显著降低(P<0.05),肺组织Notch1、Hes1、Hey1蛋白及mRNA表达显著降低(P<0.05),差异均具有统计学意义。结论芪蛭皱肺颗粒通过抑制Notch信号通路调节Th1/Th2平衡,从而改善COPD大鼠肺功能及病理损伤,影响其免疫功能。

巨型艾美耳球虫Th1类细胞因子刺激性分子EmARM-β对鸡PBMC和T细胞亚群免疫功能的影响

[目的]本文旨在探讨巨型艾美耳球虫Th1类细胞因子刺激性分子Armadillo/β-catenin样重复序列蛋白(EmARM-β)对鸡免疫细胞功能的影响。[方法]利用密度梯度离心法和免疫磁珠分选法分别分离出鸡外周血单个核细胞(peripheral blood mononuclear cell,PBMC)、CD8^(+)T细胞和CD4^(+)CD25^(-)T细胞,之后将EmARM-β重组蛋白(rEmARM-β)分别与上述细胞进行体外共孵育6 h,利用CCK-8试剂和qPCR法分别检测并分析其对上述细胞增殖能力及相关细胞因子分泌水平的影响。[结果]经免疫磁珠分选后,流式细胞术检测鸡CD8^(+)T细胞和CD4^(+)CD25^(-)T细胞的纯度分别为93.01%和88.88%。与对照组相比,rEmARM-β显著促进鸡PBMC的增殖能力,显著上调Th1类细胞因子(ifn-γ和il-2)和促炎性细胞因子(il-1β、il-6和il-17a)mRNA水平;显著促进CD8^(+)T细胞的增殖能力,显著上调其ifn-γ、il-2和tnf-α mRNA水平,显著上调穿孔素、fasl(Fas配体)和fas mRNA水平;能显著促进CD4^(+)CD25^(-)T细胞的增殖能力,显著上调其ifn-γ和il-2 mRNA水平,下调il-4和il-10 mRNA水平。[结论]EmARM-β可以有效促进鸡PBMC和T细胞亚群(CD8^(+)T细胞和CD4^(+)CD25^(-)T细胞)的增殖能力,提升其Th1类细胞因子和促炎性细胞因子的分泌水平,提示EmARM-β具有研发鸡球虫病新型疫苗候选抗原的潜力。

Rapamycin ameliorates experimental autoimmune uveoretinitis by inhibiting Th1/Th2/Th17 cells and upregulating CD4+CD25+ Foxp3 regulatory T cells

· AIM: To determine the effects of rapamycin on experimental autoimmune uveoretinitis(EAU) and investigate of role of rapamycin on T cell subsets in the disease.·METHODS: EAU was induced in rats using peptides1169 to 1191 of the interphotoreceptor binding protein(IRBP). Rapamycin(0.2 mg/kg/d) was administrated by intraperitoneal injection for a consecutive 7d after immunization. Th1/Th2/Th17 cytokines, TGF-β1, and IL-6produced by lymphocyteswere measured by ELISA, while Th17 cells and CD4 +CD25 + regulatory T cells(Tregs)from rat spleen were detected by flow cytometry.·RESULTS: Intraperitoneal treatment immediately after immunization dramatically ameliorated the clinical course of EAU. Clinical responses were associated with reduced retinal inflammatory cell infiltration and tissue destruction. Rapamycin induced suppression of Th1/Th2/Th17 cytokines, including IFN-γ, IL-2, IL-17, IL-4, and IL-10 release from T lymphocytes of EAU rats, in vitro.Rapamycin also significantly increased TGF-β1production but had no effect on IL-6 productionof T lymphocytes from EAU rats in vitro. Furthermore,rapamycin decreased the ratio of Th17 cells/CD4 +T cells and upregulated Tregs in EAU, as detected by flow cytometry.·CONCLUSION: Rapamycin effectively interferes with T cell mediated autoimmune uveitis by inhibiting antigen-specific T cell functions and enhancing Tregs in EAU.Rapamycin is a promising new alternative as an adjunct corticosteroid-sparing agent for treating uveitis.

Keratinocytes costimulate naive human T cells via CD2: a potential target to prevent the development of proinflammatory Th1 cells in the skin

The interplay between keratinocytes and immune cells,especially T cells,plays an important role in the pathogenesis of chronic inflammatory skin diseases.During psoriasis,keratinocytes attract T cells by releasing chemokines,while skin-infiltrating selfreactive T cells secrete proinflammatory cytokines,e.g.,IFN γand IL-17A,that cause epidermal hyperplasia.Similarly,in chronic graftversus-host disease,allogenic IFN γ-producing Th1/Tc1 and IL-17-producing Th17/Tc17 cells are recruited by keratinocyte-derived chemokines and accumulate in the skin.However,whether keratinocytes act as nonprofessional antigen-presenting cells to directly activate naive human T cells in the epidermis remains unknown.Here,we demonstrate that under proinflammatory conditions,primary human keratinocytes indeed activate naive human T cells.This activation required cell contact and costimulatory signaling via CD58/CD2 and CD54/LFA-1.Naive T cells costimulated by keratinocytes selectively differentiated into Th1 and Th17 cells.In particular,keratinocyte-initiated Th1 differentiation was dependent on costimulation through CD58/CD2.The latter molecule initiated STAT1 signaling and IFN γproduction in T cells.Costimulation of T cells by keratinocytes resulting in Th1 and Th17 differentiation represents a new explanation for the local enrichment of Th1 and Th17 cells in the skin of patients with a chronic inflammatory skin disease.Consequently,local interference with T cell–keratinocyte interactions may represent a novel strategy for the treatment of Th1 and Th17 cell-driven skin diseases.

In vitro-derived insulin-producing cells modulate Th1 immune responses and induce IL-10 in streptozotocin-induced mouse model of pancreatic insulitis

Background: Insulitis is defined by the presence of immune cells infiltrating in the pancreatic islets that might progress into the complete β-cell loss. The immunomodulatory properties of bone marrow-derived mesenchymal stem cells(BM-MSCs) have attracted much attention. This study aimed to evaluate the possible immunomodulatory effects of rat BM-MSCs and MSCs-derived insulin-producing cells(IPCs) in a mouse model of pancreatic insulitis. Methods: Insulitis was induced in BALB/c mice using five consecuti ve doses of streptozotocin. MSCs or IPCs were directly injected into the pancreas of mice and their effects on the expression of Th subsetsrelated genes were evaluated. Results: Both BM-MSCs and IPCs significantly reduced the expression of pancreatic Th1-related IFN-γ( P < 0.001 and P < 0.05, respectively) and T-bet genes(both P < 0.001). Moreover, the expression of IL-10 gene was significantly increased in IPC-treated compared to BM-MSC-or PBS-treated mice( P < 0.001 both comparisons). Conclusions: BM-MSCs and IPCs could successfully suppress pathologic Th1 immune responses in the mouse model of insulitis. However, the marked increase in IL-10 gene expression by IPCs compared to BM-MSCs suggests that their simultaneous use at the initial phase of autoimmune diabetes might be a better option to reduce inflammation but these results need to be verified by further experiments.

外周血Th1/Th2细胞及部分细胞因子对活动性肺结核患者治疗效果的评估价值

目的探讨外周血辅助性T细胞(Th)1/Th2及部分细胞因子对活动性肺结核(APTB)患者治疗效果的评估价值。方法选取初诊APTB患者和健康体检者各100例作为研究组和对照组,收集2组研究对象的一般临床资料,研究组患者予以6个月标准化抗结核治疗,并于治疗开始后第2、6月随访评估治疗效果;抽取对照组体检者体检日和研究组患者治疗前及治疗后第2、6个月的清晨空腹静脉血,采用流式细胞仪检测外周血Th1、Th2细胞,采用酶联免疫法检测血清干扰素γ(IFN-γ)、白细胞介素2(IL-2)、IL-4及IL-10水平;采用受试者工作(ROC)曲线分析研究组患者Th1/Th2细胞及其细胞因子对APTB患者治疗有效的预测价值。结果治疗前研究组患者外周血Th1、Th1/Th2和血清IFN-γ、IL-2低于对照组(P<0.05),外周血Th2和血清IL-4、IL-10水平高于对照组(P<0.05);与治疗前相比,研究组患者治疗后第2、6月时外周血Th1、Th1/Th2和血清IFN-γ、IL-2升高(P<0.05),外周血Th2和血清IL-4、IL-10水平下降(P<0.05);与无效组比较,有效组APTB患者治疗前外周血Th1、Th1/Th2和血清IFN-γ、IL-2升高(P<0.05),Th2占比和血清IL-4、IL-10水平降低(P<0.05);ROC曲线显示,研究组患者外周血Th1/Th2评估APTB治疗价值的特异度最高(68.37%)、血清IL-4敏感度最高(78.53%),Th1、Th2、Th1/Th2、IFN-γ、IL-2、IL-4及IL-10联合检测预测APTB患者抗结核治疗有效曲线下面积(AUC)为0.913(95%CI为0.546~0.910),敏感度和特异度(86.23%、76.28%)较单项指标检测提高。结论APTB患者机体存在Th1/Th2细胞调节失衡,抗结核治疗有助于Th1/Th2平衡调节,监测外周血Th1/Th2细胞及部分细胞因子可评估APTB临床治疗效果。

原发免疫性血小板减少症患者外周血淋巴细胞亚群、NKT细胞及Th1/Th2细胞因子水平分析

目的分析原发免疫性血小板减少症患者外周血淋巴细胞亚群、自然杀伤T细胞(natural killer T-cell,NKT)及Th1/Th2细胞因子水平的变化及临床意义。方法纳入2021年6月—2022年6月麻城市人民医院收治的60例原发免疫性血小板减少症患者为观察组,均施予流式细胞术检测,将血小板(platelet,PLT)<20×109/L患者分为观察1组(30例),将PLT≥20×109/L患者分为观察2组(30例);纳入同期30例健康体检者为对照组,对上述受试者的外周血淋巴细胞亚群、NKT细胞及Th1/Th2细胞因子水平进行测定。结果3组CD3^(+)、CD3^(+)CD4^(+)、CD3^(+)CD8^(+)、CD3-CD19^(+)、CD3-CD16^(+)CD56^(+)、CD4^(+)/CD8^(+)、NKT比较,差异有统计学意义(F_(CD3+)=16.806,F_(CD3+CD4+)=24.271,F_(CD3+CD8+)=4.880,F_(CD3-CD19+)=34.955,F_(CD3-CD16+CD56+)=11.051,F_(CD4+/CD8)=13.014,F_(NKT)=38.576,P<0.001);原发免疫性血小板减少症患者的CD3^(+)、CD3^(+)CD4^(+)、CD3-CD16^(+)CD56^(+)、CD4^(+)/CD8^(+)、NKT与PLT计数为正相关关系(P<0.001);CD3^(+)CD8^(+)、CD3-CD19^(+)、白细胞介素2、白细胞介素4、白细胞介素6、白细胞介素10、肿瘤坏死因素、干扰素-γ与PLT计数为负相关关系(P<0.001)。结论原发免疫性血小板减少症患者体内的淋巴细胞亚群、NKT细胞、Th1/Th2细胞因子紊乱,且病情进展与T、B淋巴细胞亚群与NKT细胞为正相关关系。

消痈散结汤对浆细胞性乳腺炎模型大鼠Th1/Th2细胞因子平衡及上皮连接蛋白表达的影响

目的观察消痈散结汤对浆细胞性乳腺炎模型大鼠Th1/Th2细胞因子平衡及上皮连接蛋白表达的影响,探讨消痈散结汤的可能作用机制。方法将36只SPF级SD雌性大鼠随机分为正常对照组、模型组、消痈散结汤组,每组各12只。模型组、消痈散结汤组大鼠乳腺组织注射临床PCM患者浆乳组织混悬液,建立浆细胞性乳腺炎大鼠模型。造模成功后,消痈散结汤组给予消痈散结汤0.3 mg/(kg·d)灌胃,正常组和模型组等体积生理盐水灌胃,干预2周。干预结束后,酶联免疫吸附法(Enzyme-linked immuno sorbent assay,Elisa)检测大鼠乳腺匀浆组织血清白细胞介素4(Interleukin-4,IL-4)、白细胞介素-6(Interleukin-6,IL-6)及干扰素-γ(Interferon-γ,IFN-γ)含量,免疫组化法检测大鼠乳腺组织CD138阳性浆细胞比例,HE检测大鼠乳腺组织形态学改变,免疫组化法检测大鼠乳腺组织中上皮连接蛋白E-cadherin、ZO-1及Claudin3的表达水平。结果与正常组比较,模型组大鼠乳头内陷、乳房肿块,部分可见乳头溢液、皮肤破溃,血清IL-4水平减少,IL-6、IFN-γ增加,乳腺组织导管上皮E-cadherin、ZO-1及Claudin3的蛋白表达降低,浆细胞标记CD138表达水平升高,差异均有统计学意义(P<0.05)。与模型组比较,消痈散结汤组大鼠乳头内陷及乳头溢液程度减轻,肿块体积变小。血清IL-6、IFN-γ减少,乳腺导管上皮E-cadherin、ZO-1及Claudin3的表达增加,CD138表达减少,差异均有统计学意义(P<0.05)。结论消痈散结汤能够减轻浆细胞性乳腺炎模型大鼠病理损伤,可能与平衡Th1/Th2细胞因子,增加上皮连接蛋白表达有关。

老年特应性皮炎患者外周血Th1、Th2、Th17细胞与鳞状上皮细胞癌抗原水平的变化及意义

目的探究外周血辅助性T细胞Th1、Th2、Th17和鳞状细胞癌抗原(SCCA)水平在老年特应性皮炎患者中的变化及其意义。方法以2022年3月—2022年7月于四川省人民医院皮肤科就诊的25例老年特应性皮炎患者(老年特应性皮炎组)及同期33例健康体检者(健康对照组)为研究对象。采取流式细胞术测定外周血Th1、Th2和Th17细胞占CD4+T细胞的比例;检测老年特应性皮炎患者的外周血嗜酸性粒细胞(EOS)计数、总免疫球蛋白E(IgE)及血清SCCA水平;采用Spearman法进行相关性分析。结果老年特应性皮炎患者皮损严重且分布广泛,主要表现为四肢伸侧及背部苔藓样湿疹,部分患者有1种或多种系统性疾病。老年特应性皮炎组的外周血EOS计数较健康对照组明显增加(P<0.05)。老年特应性皮炎组外周血中Th2和Th17细胞比例均高于健康对照组(P<0.05),而Th1细胞比例与健康对照组比较,差异无统计学意义(P>0.05)。老年特应性皮炎组外周血EOS计数与Th2细胞比例、SCCA水平有相关性(rs=-0.399和0.607,均P<0.05)。老年特应性皮炎患者的SCCA水平与SCORAD评分呈正相关(rs=0.416,P<0.05)。结论老年特应性皮炎患者具有特有的临床特征,以Th2和Th17型炎症为主。老年特应性皮炎患者血清SCCA水平可在一定程度上反映其疾病的严重程度。

Activated rat hepatic stellate cells influence Th1/Th2 profile in vitro

AIM: To investigate the effects of activated rat hepatic stellate cells(HSCs) on rat Th1/Th2 profile in vitro.METHODS: Growth and survival of activated HSCs and CD4+ T lymphocytes cultured alone or together was assessed after 24 or 48 h. CD4+ T lymphocytes were then cultured with or without activated HSCs for 24 or 48 h and the proportion of Th1 [interferon(IFN)-γ+] and Th2 [interleukin(IL)-4+] cells was assessed by flow cytometry. Th1 and Th2 cell apoptosis was assessed after 24 h of co-culture using a caspase-3 staining procedure. Differentiation rates of Th1 and Th2 cells from CD4+ T lymphocytes that were positive for CD25 but did not express IFN-γ or IL-4 were also assessed after 48 h of co-culture with activated HSCs. Galectin-9 expression in HSCs was determined by immunofluorescence and Western blotting. ELISA was performed to assess galectin-9 secretion from activated HSCs.RESULTS: Co-culture of CD4+ T lymphocytes with activated rat HSCs for 48 h significantly reduced the proportion of Th1 cells compared to culture-alone conditions(-1.73% ± 0.71%; P < 0.05), whereas the proportion of Th2 cells was not altered; the Th1/Th2 ratio was significantly decreased(-0.44 ± 0.13; P < 0.05). In addition, the level of IFN-γ in Th1 cells wasdecreased(-65.71 ± 9.67; P < 0.01), whereas the level of IL-4 in Th2 cells was increased(82.79 ± 25.12; P < 0.05) by co-culturing, as measured by mean fluorescence intensity by flow cytometry. Apoptosis rates in Th1(12.27% ± 0.99%; P < 0.01) and Th2(1.71% ± 0.185%; P < 0.01) cells were increased 24 h after co-culturing with activated HSCs; the Th1 cell apoptosis rate was significantly higher than in Th2 cells(P < 0.01). Galectin-9 protein expression was significantly decreased in HSCs only 24 h after coculturing(P < 0.05) but not after 48 h. Co-culture for 48 h significantly increased the differentiation of Th1 and Th2 cells; however, the increase in the proportion of Th2 cells was significantly higher than that of Th1 cells(1.85% ± 0.48%; P < 0.05).CONCLUSION: Activated rat HSCs lower the Th1/Th2 profile, inhibiting the Th1 response and enhancing the Th2 response, and this may be a novel pathway for liver fibrogenesis.

Ribavirin and IFN-α combination therapy induces CD4+ T-cell proliferation and Th1 cytokine secretion in patients with chronic hepatitis B

AIM: To investigate the anti-viral mechanism of combination therapy of interferon (IFN)-α and ribavirin in patients with chronic hepatitis B. METHODS: Twenty patients were assigned to receive either IFN-α plus ribavirin (group A,n = 14) or no treatment as a control (group B,n = 6). Patients were analyzed for T-cell proliferative responses specific for hepatitis B virus (HBV)-antigen and cytokine production by peripheral blood mononuclear cells (PBMCs). RESULTS: Combination therapy induced HBV-antigen specific CD4+ T-cell proliferative responses in four patients (28.6%). Production of high levels of HBV-specific IFN-γ,tumor necrosis factor (TNF)-α,interleukin (IL)-12 by PBMCs was found in five patients (35.7%),who showed significantly lower HBV DNA levels in serum at 12 mo after treatment ended (P = 0.038) and at 24 mo of follow-up (P = 0.004) than those without high levels of cytokine production. CONCLUSION: HBV-antigen specific CD4+ T cells may directly control HBV replication and secretion of anti-viral T helper 1 (Th1) cytokines by PBMCs during combination therapy of chronic hepatitis B with ribavirin and IFN-α.

All-transRetinoic Acid Regulates Th1/Th2 Balance in CD4+T cells When GATA-3 is Deficient

The essential effect of vitamin A on immune function occurs through various mechanisms including direct effect on ThloTh2 balance modulation. However, it is unclear whether or not vitamin A can regulate Thl-Th2 balance under a strong Thl-polarizing condition. Therefore, the purpose of our study was to examine the effect of vitamin A metabolite allotrans retinoic acid （ATRA） on ThloTh2 differentiation in CD4~ T cells under GATA-3 deficiency, which can induce Thl-polarizing condition. In the present study, GATA-3 deficiency T cells were induced by siRNA and checked by real-time quantitative PCR and western blot. GATA-3 deficiency CD4＋ T cells and normal CD4＋ T were treated for 48 h with or without ATRA.

不同类型人乳头状瘤病毒感染者CD4+T细胞、CD8+T细胞、Th1/Th2因子水平分析

目的:探讨不同类型人乳头状瘤病毒(HPV)感染与外周血CD4^(+)T细胞、CD8^(+)T细胞、Th1/Th2因子表达。方法:收集2021年7月-2022年6月就诊于本院的HPV感染者172例临床资料,均行HPV分型检测,并完成检测外周血CD4^(+)T细胞、CD8^(+)T细胞、Th细胞因子,及白细胞介素-2(IL-2)、干扰素-γ(IFN-γ)IL-4和IL-10。比较高危HPV感染与低危HPV感染者以及多重感染与单一感染外周血炎症因子差异。结果:172例HPV感染者中高危型HPV感染80.2%,低危型HPV感染19.8%;单一感染64.5%,多重感染35.5%。高危HPV感染以HPV 16最多,其次为HPV52、HPV58,低危HPV感染以HPV61最多。高危型HPV感染者外周血CD4^(+)T细胞、CD4^(+)/CD8^(+)比值和IL-2、IFN-γ水平均低于低危HPV感染者,CD8^(+)T细胞和血清IL-4、IL-10水平显著高于低危HPV感染者;多重感染者外周血CD4^(+)T细胞、CD4^(+)/CD8^(+)比值和IL-2、IFN-γ水平均低于单一感染者,CD8^(+)T细胞和血清IL-4、IL-10水平显著高于单一感染者(均P<0.05)。结论:不同类型HPV感染者CD4^(+)T细胞、CD8^(+)T细胞的表达存在差异,高危HPV感染者有着较低的CD4^(+)T细胞、CD4^(+)/CD8^(+)、Th1细胞因子和较高的CD8^(+)T细胞、Th 2细胞因子表达,且多重感染者更为明显。

自身免疫甲状腺炎患者外周血miR-145表达及其与Th1/Th2比值的关系

目的探讨miR-145在自身免疫甲状腺炎(AIT)患者外周血中表达情况,并分析其与Th1/Th2比值的关系。方法回顾性选取2020年1月至2021年12月武汉市中医医院收治的245例AIT患者病例资料,作为研究对象(AIT组);选取同时期本院体检健康者250名为对照组。通过实时荧光定量PCR法检测所有受试者血浆中miR-145的表达情况,以流式细胞术对外周血中Th1细胞比例、Th2细胞比例进行检测,并计算Th1/Th2比值;采用酶联免疫吸附分析检测所有受试者血浆中细胞因子γ干扰素(IFN-γ)、白细胞介素-4(IL-4)水平。Pearson相关性分析miR-145与Th1、Th2、Th1/Th2比值及IL-4水平的相关性。结果AIT组患者血浆miR-145表达水平为0.33±0.11,低于对照组(1.02±0.36),差异有统计学意义(P<0.05)。AIT组患者外周血Th1细胞比例、Th1/Th2比值分别为(7.78±1.88)%、2.69±0.84,均高于对照组[(3.64±0.96)%、0.88±0.26],Th2细胞比例为(3.17±1.89)%,低于对照组[(5.48±2.04)%],差异均有统计学意义(P<0.05)。AIT组患者血浆中IFN-γ水平为(35.73±10.82)pg/mL,高于对照组[(22.47±6.15)pg/mL],IL-4水平为(3.67±1.06)ng/mL,低于对照组[(6.74±2.04)ng/mL],差异均有统计学意义(P<0.05)。Pearson分析显示,miR-145表达水平与Th1细胞比例、Th1/Th2比值、IFN-γ、IL-4负相关,与Th2细胞比例、IL-4正相关(P<0.05)。结论AIT患者血浆中miR-145低表达,Th1/Th2比值升高,miR-145可能经对Th1/Th2比值产生影响参与AIT发生。

Novel heat shock protein Hsp70L1 activates dendritic cells and acts as a Th1 polarizing adjuvant

Heat shock proteins (HSPs) are reported to act as effective adjuvants to elicit anti-tumor and anti-infection immunity. Here, we report that Hsp70-like protein 1 (Hsp70L1), a novel HSP derived from human dendritic cells (DCs), has potent adjuvant effects that polarize responses toward Th1. With a calculated molecular weight of 54.8 kDa, Hsp70L1 is smaller in size than Hsp70 but resembles it both structurally and functionally. Hsp70L1 shares common receptors on DCs with Hsp70 and can interact with DCs, promoting DC maturation and stimulating secretion of the proinflammatory cytokines interleukin 12p70 (IL-12p70), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), and the chemokines IP-10, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and normal T cell expressed and secreted (RANTES). The induction of interferon-gamma-inducible protein 10 (IP-10) secretion by Hsp70L1 is not shared by Hsp70, and other functional differences include more potent stimulation of DC IL-12p70, CC-chemokine, and CCR7 and CXCR4 expression by Hsp70L1. Immunization of mice with the hybrid peptide Hsp70L1-ovalbumin(OVA)(257-264) induces an OVA(257-264)-specific Th1 response and cytotoxic T lymphocyte (CTL) that results in significant inhibition of E.G7-OVA tumor growth. The ability of Hsp70L1 to activate DCs indicates its potential as a novel adjuvant for use with peptide immunizations; the Hsp70L1 antigen peptide hybrid may serve as a more effective vaccine for the control of cancer and infectious diseases.

Effects of the Overall Alkaloid of a Traditional Chinese Medicine “Tongbiling” on the Cytokine Expression in Th1 and Th2 Cells of Rheumatoid Arthritis and Their Signaling Pathway

To investigate the effects of overall alkali of a traditional Chinese medicine “Tongbiling” (brucine and strychnine alkaloids in main) on the cytokines expression in Th1 and Th2 cells in the synovial fluid of patients with rheumatism arthritis and their signal pathway, the mononuclear cells in the synovial fluid (SFMC) of patients were isolated by Ficoll-Hypaque gradient centrifugation, and the CD3 + CD69 + and CD3 + HLA-DR antigen were analyzed by flow cytometry in comparison with those of the peripheral blood. The rest of cells were cultured after resuspension with RPMI 1640 culture medium. Phorbol 12,13-dibutyrate (PDB) and ionomycin were added successively into the culture with various concentration of overall alkali Tongbiling (TBL). After 4 h of cultivation, the expression of IFN-γ and IL-4 in CD3 + cells were analyzed by flow cytometry. The influence of overall alkali TBL (100?mg/L) on the intracellular calcium was investigated after Fluo-3/AM labeling and stimulation with PDB and ionomycin at 1, 2, 4 and 10?min, and the influence of TBL on the expression of CD3 + CD69 + cells were determined with stimulation of PDB for 24?h in the whole blood lymphocytes culture. It was found that the percentage of T cells bearing CD69 was significantly up-regulated (77%), while that of T cells bearing HLA-DR was 44% in the synovial mononucleated cells. After PDB and ionomycin stimulation, the expression of IFN-γ in CD3 + cells were up-regulated, but there was no change on the expression of IL-4 in CD3 + cells, indicating that ratio of Th1/Th2 was significantly increased and Th cells differentiate to Th1 cells in mainly. Four concentrations of overall alkaloid of TBL (200?mg/L, 100?mg/L, 50?mg/L, 25?mg/L) could down-regulated the expression of IFN-γ in CD3 + cells and the Th1/Th2 ratio obviously, but all the concentrations of the overall alkaloids had no effect on the expression of IL-4 in CD3 + cells. 100?mg/L concentration of the overall alkaloid did not down-regulate the intracellular calcium level. Each concentration of the overall alkaloid could down-regulated the expression CD69 obviously on the PDB-activated mouse T cells. It concluded from the above observations that the overall alkaloid of TBL could relieve the inflammatory and immune damages by suppressing the expression of Th1 type cytokines and Th1 cell differentiation, regulating the imbalance of Th1/Th2 cells and inhibiting the early activation of the T lymphocytes bearing CD69. There was no remarkable influence on the intracellular calcium signaling transduction pathway. The inhibitory effected on T cells to express IFN-γ might be due to the suppression of PKC-MAPK signaling pathway. From the standpoint of traditional Chinese medicine, this might be due to the regulation of “Yin” and “Yang” imbalance of joints to modify the pathological status in rheumatoid arthritis. This study provided an experimental basis for the application of overall alkaloids of TBL in the treatment of rheumatoid arthritis.

不同血糖水平的冠心病患者外周血中Th1、Th2细胞比例变化分析

目的探讨不同血糖水平的冠心病患者外周血中1型辅助性T细胞(Th1)、2型辅助性T细胞(Th2)的细胞比例变化.方法选取冠心病患者86例,根据血糖检测结果分为冠心病血糖正常组34例,冠心病合并糖耐量异常组20例,冠心病合并糖尿病组32例;另收集同期健康志愿者25名为健康对照组.采集空腹外周静脉血,检测空腹血糖、糖化血红蛋白、γ干扰素、白细胞介素4、白细胞介素5、白细胞介素10水平.分离外周血单个核细胞,流式细胞仪检测Th1、Th2细胞比例.分离并培养人脐静脉内皮细胞,并使用免疫磁珠清除外周血单个核细胞中CD4+T细胞,进行人脐静脉内皮细胞毒性试验.出院后对冠心病患者进行1 a的随访,记录主要不良心血管事件发生情况.结果冠心病合并糖尿病组患者空腹血糖、糖化血红蛋白水平明显高于健康对照组、冠心病血糖正常组、冠心病合并糖耐量异常组(P<0.05).冠心病合并糖尿病组Th1细胞比例明显高于冠心病合并糖耐量异常组、冠心病血糖正常组、健康对照组,Th2细胞比例明显低于冠心病合并糖耐量异常组、冠心病血糖正常组、健康对照组(P<0.05).冠心病合并糖尿病组γ干扰素水平明显高于冠心病合并糖耐量异常组、冠心病血糖正常组、健康对照组,白细胞介素4、白细胞介素5、白细胞介素10水平明显低于冠心病合并糖耐量异常组、冠心病血糖正常组、健康对照组(P<0.05).冠心病合并糖尿病组患者外周血单个核细胞水平明显低于冠心病合并糖耐量异常组、冠心病血糖正常组、健康对照组(P<0.05);清除CD4+T细胞的外周血单个核细胞在各组间比较差异无统计学意义(P>0.05).空腹血糖、糖化血红蛋白、Th1细胞比例、Th2细胞比例对冠心病患者预后均具有较高的预测价值(P<0.05);其中各指标联合检测的预测价值最高(AUC=0.942),灵敏度、阳性预测值均明显高于各指标单独检测.结论高糖状态下冠心病患者外周血中Th1细胞比例升高,Th2细胞比例降低,能够加速血管内皮细胞损伤,与空腹血糖和糖化血红蛋白联合检测可用于患者预后评估.