CD4+T细胞的新亚型：Th17细胞及其生物效应

辅助性T细胞通常分为Th1型和Th2型.20余年来,该分类方法形成了理解CD4 T细胞免疫生物学、固有免疫和适应性免疫调节理论的框架.近来研究发现,机体存在一种新型的不同于1型和2型的CD4 效应T细胞——辅助性17细胞(Thelp 17,Th 17),该细胞是由天然T细胞前体分化而来,具有独立的分化和发育调节机制,并特异性地产生白介素17(interleukin 17,IL-17)效应因子,在自身免疫性疾病和感染性疾病中发挥重要调节作用.这将对深入研究机体免疫调节、免疫病理和机体防御反应机制具有重要意义.就这种新型的辅助性T细胞的产生、发育分化机制和免疫调节效应研究进展做一简要综述.     

Knockout of LASP1 in CXCR4 expressing CML cells promotes cell persistence,proliferation and TKI resistance

Chronic myeloid leukaemia (CML) is a clonal myeloproliferative stem cell disorder characterized by the constitutively active BCR-ABL tyrosine kinase. The LIM and SH3 domain protein 1 (LASP1) has recently been identified as a novel BCR-ABL substrate and is associated with proliferation, migration, tumorigenesis and chemoresistance in several cancers. Furthermore, LASP1 was shown to bind to the chemokine receptor 4 (CXCR4), thought to be involved in mechanisms of relapse. In order to identify potential LASP1-mediated pathways and related factors that may help to further eradicate minimal residual disease (MRD), the effect of LASP1 on processes involved in progression and maintenance of CML was investigated. The present data indicate that not only overexpression of CXCR4, but also knockout of LASP1 contributes to proliferation, reduced apoptosis and migration as well as increased adhesive potential of K562 CML cells. Furthermore, LASP1 depletion in K562 CML cells leads to decreased cytokine release and reduced NK cell-mediated cytotoxicity towards CML cells. Taken together, these results indicate that in CML, reduced levels of LASP1 alone and in combination with high CXCR4 expression may contribute to TKI resistance.     

肺炎支原体肺炎伴喘息儿童血清25(OH)D_3、Th17/Treg表达水平与肺功能的关系

目的:探讨肺炎支原体肺炎伴喘息儿童血清25羟基维生素D3[25(OH)D_3]、辅助性17细胞/调节性T细胞(Th17/Treg)表达水平与肺功能的关系。方法:将新疆医科大学第五附属医院收治的肺炎支原体肺炎伴喘息患儿26例作为肺炎伴喘息组,肺炎支原体肺炎不伴有喘息患儿54例作为肺炎不伴喘息组,另选取健康儿童30例作为对照组,比较各组血清25(OH)D_3、白细胞介素(IL)-10、IL-17、Th17细胞及Treg细胞占CD4+T细胞比例及肺功能,并分析其相关性。结果:肺炎伴喘息组血清25(OH)D_3、IL-10、Treg细胞占CD4+T细胞比例低于肺炎不伴喘息组、对照组,Th17细胞占CD4+T细胞比例、Th17/Treg、IL-17高于肺炎不伴喘息组、对照组(P0.05)。各组第一秒最大呼气量占用力肺活量百分比(FEV1/FVC)比较差异无统计学意义(P0.05),肺炎伴喘息组FEV1占预计值百分比(FEV1%pred)、峰值呼气流量(PEF)低于肺炎不伴喘息组、对照组(P0.05),肺炎不伴喘息组与对照组FEV1%pred、PEF比较无统计学意义(P0.05)。肺炎伴喘息组患儿血清25 (OH)D_3与Th17/Treg、IL-17呈负相关(P0.05),与IL-10、FEV1%pred、PEF呈正相关(P0.05),血清Th17/Treg与IL-10、FEV1%pred、PEF呈负相关(P0.05),与IL-17呈正相关(P0.05)。结论:肺炎支原体肺炎伴喘息儿童血清25(OH)D_3、Th17/Treg表达水平异常,肺功能下降,且25(OH)D_3、Th17/Treg表达水平与肺功能相关。     

血清CCL17、CXCR4与重度子痫前期患者Th17细胞的相关性分析及对母婴结局的影响

摘要 目的：探讨血清C-C基序趋化因子配体17（CCL17）、CXC趋化因子受体4（CXCR4）与重度子痫前期（SPE）患者辅助性T细胞（Th）17细胞的相关性分析及对母婴结局的影响。方法：选择2018年3月至2021年3月苏州大学附属第二医院妇产科收治的169例SPE患者（SPE组）和77例健康孕产妇（对照组）。检测血清CCL 17、CXCR4水平和外周血Th17细胞及其细胞因子。Pearson分析血清CCL 17、CXCR4水平与外周血Th17细胞及其细胞因子的关系,多因素Logistic回归分析SPE母婴结局不良的相关因素。结果：SPE组血清CCL17、CXCR4水平低于对照组（P＜0.05）,外周血Th17细胞占比、血清白细胞介素（IL）-17水平高于对照组（P＜0.05）。血清CCL17、CXCR4水平与外周血Th17细胞占比、血清IL-17水平呈负相关（P＜0.05）。母婴结局不良53例,母婴结局良好116例,母婴结局不良组血清CCL 17、CXCR4水平低于母婴结局良好组（P＜0.05）。高Th17细胞占比、年龄大、低水平CCL17、CXCR4是SPE患者母婴结局不良的危险因素（P＜0.05）。结论：SPE患者血清CCL17、CXCR4水平降低,且与外周血Th17细胞占比增加,血清IL-17水平增高有关,低水平CCL17、CXCR4是SPE患者母婴结局不良的危险因素。     

IL-17 and IL-22 elicited by a DNA vaccine encoding ROP13 associated with protection against Toxoplasma gondii in BALB/c mice

Toxoplasma gondii, an intracellular parasitic protozoan, is capable of infecting man and all warm-blooded animals. Cell-mediated immunity is vital in mounting protective responses against T. gondii infection. Recent studies have shown that T-helper (Th) 17 responses may play a key role in parasite control. In this current study, we constructed a DNA vaccine encoding T. gondii ROP13 in a pcDNA vector. Groups of BALB/c mice were immunized intramuscularly with pcROP13 or controls and challenged with the RH strain of T. gondii. The results showed that immunization with pcROP13 could elicit an antibody response against T. gondii. The expression of the canonical Th17 cytokines, interleukin (IL)-17 and IL-22, were significantly increased after immunization with pcROP13 compared with control groups ( p < 0.05). Furthermore, vaccination resulted in a significant decrease in parasite load ( p < 0.05). The induction of Th17 related cytokines, using a ROP13 DNA vaccine, against T. gondii should be considered as a potential vaccine approach for the control of toxoplasmosis.     

Diverse signaling pathways through the SDF-1/CXCR4 chemokine axis in prostate cancer cell lines leads to altered patterns of cytokine secretion and angiogenesis

The establishment of metastatic bone lesions in prostate cancer (CaP) is a process partially dependent on angiogenesis. Previously we demonstrated that the stromal-derived factor-1 (SDF-1 or CXCL12)/CXCR4 chemokine axis is critical for CaP cell metastasis. In this investigation, cell lines were established in which CXCR4 expression was knocked down using siRNA technology. When CaP cells were co-transplanted with human vascular endothelial cells into SCID mice, significantly fewer human blood vessels were observed paralleling the reductions in CXCR4 levels. Likewise, the invasive behaviors of the CaP cells were inhibited in vitro. From these functional observations we explored angiogenic and signaling mechanisms generated following SDF-1 binding to CXCR4. Differential activation of the MEK/ERK and PI3K/AKT pathways that result in differential secretion IL-6, IL-8, TIMP-2 and VEGF were seen contingent on the cell type examined; VEGF and TIMP-2 expression in PC3 cells are dependent on AKT activation and ERK activation in LNCaP and LNCaP C4-2B cells leads to IL-6 or IL-8 secretion. At the same time, expression of angiostatin levels were inversely related to CXCR4 levels, and inhibited by SDF-1 stimulation. These data link the SDF-1/CXCR4 pathway to changes in angiogenic cytokines by different signaling mechanisms and, suggest that the delicate equilibrium between proangiogenic and antiangiogenic factors may be achieved by different signal transduction pathways to regulate the angiogenic phenotype of prostate cancers. Taken together, our results provide new information regarding expression of functional CXCR4 receptor-an essential role and potential mechanism of angiogenesis upon SDF-1 stimulation.     

Regulation of VEGF, MMP-9 and metastasis by CXCR4 in a prostate cancer cell line

To investigate the mechanisms involved in PCa (prostate cancer) metastasis and CXCR4 (CXC chemokine receptor-4)-mediated VEGF (vascular endothelial growth factor) and MMP-9 (matrix metalloproteinase-9) expression, we used lentivirus-mediated RNAi (RNA interference) to reduce the expression of CXCR4 in a PCa cell line. We found that the silencing of CXCR4 led to a significant down-regulation of VEGF and MMP-9 at both the mRNA and protein levels compared with the control in vitro. Using an animal model, we confirmed that CXCR4 silencing via subcutaneous injection could reduce tumour growth as well as inhibit metastasis, particularly bone metastasis, of PCa. Using in vivo immunohistochemistry, we also found that the expression of VEGF and MMP-9 were reduced by the knockdown of CXCR4 in the primary tumours of mice. Collectively, our results indicate that CXCR4 plays an important role in PCa metastasis through the up-regulation of VEGF and MMP-9. These findings may aid future intervention strategies.     

枯草杆菌二联活菌肠溶胶囊联合泮托拉唑对UC患者炎性因子、肠黏膜功能及外周血Th17、CD4+CD25+Treg细胞表达的影响

摘要 目的：观察枯草杆菌二联活菌肠溶胶囊联合泮托拉唑对溃疡性结肠炎（UC）患者炎性因子、肠黏膜功能及外周血Th17、CD4⁺CD25⁺Treg细胞表达的影响。方法：研究对象选择2014年7月~2018年9月期间来我院香山路门诊部接受诊治的80例UC患者,随机分为联合组（枯草杆菌二联活菌肠溶胶囊联合泮托拉唑治疗）、对照组（泮托拉唑治疗）,各40例。对比两组的疗效、炎性因子、肠黏膜功能及外周血中Th17及CD4⁺CD25⁺Treg细胞表达 。记录两组治疗期间不良反应发生情况。结果：联合组的临床总有效率为92.50%（37/40）,对照组为70.00%（28/40）,两组比较差异有统计学意义（P<0.05）。两组不良反应发生率对比无差异（P>0.05）。联合组治疗6个月后血清白介素-8（IL-8）、肿瘤坏死因子-α（TNF-α）、C反应蛋白（CRP）水平均明显比对照组低（P<0.05）。联合组治疗6个月后血清D-乳酸含量、二胺氧化酶（DAO）水平均明显比对照组低（P<0.05）。联合组治疗6个月后外周血中Th17细胞表达比对照组低,CD4⁺CD25⁺Treg细胞表达比对照组高（P<0.05）。结论：枯草杆菌二联活菌肠溶胶囊联合泮托拉唑治疗UC患者,可有效改善肠道环境,使外周血中的 Th17 细胞表达降低,CD4⁺CD25⁺Treg细胞表达增加,并缓解炎症状态,临床效果满意且安全性好。     

槐杞黄颗粒治疗原发免疫性血小板减少症患儿疗效及对血小板参数、外周血Treg/Th17和CD4/CD8的影响

摘要 目的：探讨槐杞黄颗粒治疗原发免疫性血小板减少症（ITP）患儿疗效及对血小板参数、外周血Treg/Th17和CD4/CD8的影响。方法：2019年8月到2021年5月选择在本院诊治的免疫性血小板减少症患儿60例作为研究对象,根据1:1简单分配原则把患儿分为槐杞黄颗粒组与对照组各30例。对照组给予常规药物-注射用人免疫球蛋白治疗,槐杞黄颗粒组以对照组为基础,给予槐杞黄颗粒治疗,两组都治疗观察3个月,观察患儿治疗疗效与血小板参数、外周血Treg/Th17和CD4/CD8的变化情况。结果：治疗后槐杞黄颗粒组的总有效率为96.7 %,明显高于对照组的80.0 %（P<0.05）。槐杞黄颗粒组的起效时间少于对照组,疗效维持时间多于对照组,两两对比有明显差异（P<0.05）。两组治疗后的血小板计数、血小板参数都明显高于治疗前,槐杞黄颗粒组也明显高于对照组（P<0.05）。两组治疗后的外周血Treg/Th17和CD4/CD8比值都明显高于治疗前,槐杞黄颗粒组明显高于对照组（P<0.05）。结论：槐杞黄颗粒治疗原发免疫性血小板减少症患儿能有效提高治疗效果,可缩短起效时间,延长疗效持续时间,还能有效改善患儿的血小板计数、血小板参数,提高外周血Treg/Th17和CD4/CD8水平。     

Inhibition of chemokine (CXC motif) ligand 12/chemokine (CXC motif) receptor 4 axis (CXCL12/CXCR4)-mediated cell migration by targeting mammalian target of rapamycin (mTOR) pathway in human gastric carcinoma cells

CXCL12/CXCR4 plays an important role in metastasis of gastric carcinoma. Rapamycin has been reported to inhibit migration of gastric cancer cells. However, the role of mTOR pathway in CXCL12/CXCR4-mediated cell migration and the potential of drugs targeting PI3K/mTOR pathway remains unelucidated. We found that CXCL12 activated PI3K/Akt/mTOR pathway in MKN-45 cells. Stimulating CHO-K1 cells expressing pEGFP-C1-Grp1-PH fusion protein with CXCL12 resulted in generation of phosphatidylinositol (3,4,5)-triphosphate, which provided direct evidence of activating PI3K by CXCL12. Down-regulation of p110β by siRNA but not p110α blocked phosphorylation of Akt and S6K1 induced by CXCL12. Consistently, p110β-specific inhibitor blocked the CXCL12-activated PI3K/Akt/mTOR pathway. Moreover, CXCR4 immunoprecipitated by anti-p110β antibody increased after CXCL12 stimulation and G(i) protein inhibitor pertussis toxin abrogated CXCL12-induced activation of PI3K. Further studies demonstrated that inhibitors targeting the PI3K/mTOR pathway significantly blocked the chemotactic responses of MKN-45 cells triggered by CXCL12, which might be attributed primarily to inhibition of mTORC1 and related to prevention of F-actin reorganization as well as down-regulation of active RhoA, Rac1, and Cdc42. Furthermore, rapamycin inhibited the secretion of CXCL12 and the expression of CXCR4, which might form a positive feedback loop to further abolish upstream signaling leading to cell migration. Finally, we found cells expressing high levels of cxcl12 were sensitive to rapamycin in its activity inhibiting migration as well as proliferation. In summary, we found that the mTOR pathway played an important role in CXCL12/CXCR4-mediated cell migration and proposed that drugs targeting the mTOR pathway may be used for the therapy of metastatic gastric cancer expressing high levels of cxcl12.     

不明原因复发性流产再次妊娠患者血清1,25(OH)2D3、sTim-3与Th17/Treg免疫失衡和妊娠结局的关系

摘要 目的：探讨不明原因复发性流产（URSA）再次妊娠患者血清1,25-二羟维生素D3[1,25（OH)）₂D₃]、可溶性T细胞免疫球蛋白黏蛋白分子3（sTim-3）与辅助性T细胞17（Th17）/调节性T细胞（Treg）免疫失衡和妊娠结局的关系。方法：选择于湖南省妇幼保健院2020年1月~2022年1月就诊的62例URSA再次妊娠患者作为研究组,另选择同期进行孕检的正常早孕妇女30例作为对照组。比较两组孕早期血清1,25(OH) ₂D₃、sTim-3及外周血Th17细胞、Treg细胞水平、Th17/Treg比值。Pearson法分析URSA再次妊娠患者血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值平的相关性。根据URSA再次妊娠患者妊娠结局的不同分为妊娠成功分娩组和妊娠再次流产组,比较两组孕早期血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值。受试者工作特征（ROC）曲线分析血清1,25(OH) ₂D₃、sTim-3与外周血Th17细胞、Treg细胞水平、Th17/Treg比值对妊娠结局的预测价值。结果：研究组血清sTim-3、外周血Th17细胞水平、Th17/Treg比值高于对照组,血清1,25(OH) ₂D₃、外周血Treg细胞水平低于对照组（P<0.05）。Pearson相关分析显示,URSA再次妊娠患者血清1,25(OH) ₂D₃与血清sTim-3、外周血Th17细胞水平、Th17/Treg比值呈负相关,与Treg细胞水平呈正相关（P<0.05）;血清sTim-3与外周血Treg细胞水平呈负相关,与Th17细胞水平、Th17/Treg比值呈正相关（P<0.05）。妊娠再次流产组血清sTim-3、外周血Th17细胞水平、Th17/Treg比值高于妊娠成功分娩组,血清1,25(OH) ₂D₃、外周血Treg细胞水平低于妊娠成功分娩组（P<0.05）。ROC曲线分析显示,血清1,25(OH) ₂D₃、sTim-3及外周血Th17细胞、Treg细胞水平及Th17/Treg比值均可预测URSA再次妊娠患者妊娠再次流产的发生风险,且上述指标联合检测的预测效能更高。结论：血清1,25(OH) ₂D₃水平异常降低、sTim-3水平异常升高可导致Th17/Treg免疫失衡,导致URSA再次妊娠患者再次发生流产。上述指标联合检测对URSA再次妊娠患者妊娠再次流产的预测效能更高。