非小细胞肺癌不同胸腔积液严重程度及预后患者lncRNA MEG3表达及其与Th17/CD4^(+)T细胞的关系

目的:研究非小细胞肺癌(NSCLC)不同胸腔积液严重程度及预后患者lncRNA MEG3表达及其与Th17/CD4^(+)T细胞的关系。方法:选取2020年1月至2022年12月福建医科大学附属泉州第一医院收治的104例NSCLC恶性胸腔积液患者作为研究对象,根据胸腔积液量分为3组:少量胸腔积液组(35例)、中量胸腔积液组(42例)、大量胸腔积液组(27例)。根据患者疾病实际发展转归分为预后良好组(29例未出现复发和转移)和预后不良组(75例出现复发和转移)。另选取同期于福建医科大学附属泉州第一医院治疗的60例肺炎良性胸腔积液患者作为对照组。实时荧光定量PCR检测两组胸腔积液中MEG3表达。收集受试者外周静脉血,流式细胞术检测外周血Th17细胞、CD4^(+)T细胞比例,并计算Th17/CD4^(+)T。对比各组患者lncRNA MEG3及外周血Th17、CD4^(+)T细胞水平。Logistic回归分析NSCLC胸腔积液及预后的影响因素。结果:NSCLC组胸腔积液lncRNA MEG3表达及CD4^(+)T细胞百分比低于对照组,Th17细胞百分比、Th17/CD4^(+)T高于对照组(P<0.05)。大量胸腔积液组lncRNA MEG3表达及CD4^(+)T细胞百分比低于少量胸腔积液组、中量胸腔积液组,中量胸腔积液组lncRNA MEG3表达及CD4^(+)T细胞百分比低于少量胸腔积液组,大量胸腔积液组Th17细胞百分比、Th17/CD4^(+)T高于少量胸腔积液组、中量胸腔积液组,中量胸腔积液组Th17细胞百分比、Th17/CD4^(+)T高于少量胸腔积液组(P<0.05)。预后不良组lncRNA MEG3表达及CD4^(+)T百分比低于预后良好组,而Th17细胞百分比、Th17/CD4^(+)T高于预后良好组(P<0.05)。Logistic回归分析结果显示,lncRNA MEG3为NSCLC胸腔积液的保护因素,Th17/CD4^(+)T为危险因素(P<0.05);lncRNA MEG3为NSCLC预后的保护因素,Th17/CD4^(+)T为危险因素(P<0.05)。结论:NSCLC不同胸腔积液严重程度及预后患者lncRNA MEG3表达及Th17/CD4^(+)T不同,且lncRNA MEG3为NSCLC胸腔积液及预后的保护因素,Th17/CD4^(+)T为危险因素,可作为胸腔积液严重程度及预后诊断的有效生物标志物。

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.

CD4^(+)T细胞Th1/Th2及Th17/Treg平衡状态在IgA肾病间质纤维化中的意义

目的:探讨CD4^(+)T细胞亚群Th1、Th2、Th17、Treg及Th1/Th2、Th17/Treg比值与IgA肾病(IgAN)间质纤维化的关系。方法:收集2022年6月至2023年12月在济南威高肾科医院经肾活检确诊的86例IgAN患者,按照肾间质纤维化程度分为T0组(n=36)、T1组(n=28)和T2组(n=22)。选取同期的健康体检者43例为对照组。采用流式细胞术检测外周血中CD4^(+)T细胞亚群的比例,酶联免疫吸附试验检测外周血中CD4^(+)T淋巴细胞因子:γ干扰素(IFN-γ)、白细胞介素(IL)-4、IL-17和转化生长因子β(TGF-β)。结果:随着肾间质纤维化程度的增加,Th1、Treg及Th1/Th2逐渐降低,而Th2、Th17及Th17/Treg逐渐升高(P<0.05)。IL-4、IL-17和TGF-β水平随着IgAN肾间质纤维化程度的增加而升高(P<0.05),而IFN-γ水平在各组间的差异无统计学意义。Pearson相关性分析显示,Th1/Th2、Th17/Treg与IgAN患者的临床指标呈弱相关性(P<0.05)。结论:IgAN患者外周血中CD4^(+)T细胞亚群Th1/Th2、Th17/Treg比值失衡,可能参与了肾间质纤维化的发生发展。

CD4^(+)/CD8^(+)、Th17/Treg与老年慢性阻塞性肺疾病患者病情及近期预后的相关性

目的探讨CD4^(+)/CD8^(+)、辅助性T细胞17(helper T cells 17,Th17)/调节性T细胞(regulatory T,Treg)检测在老年慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者病情及预后评估中的价值。方法选择老年COPD患者288例(COPD组)和老年健康体检者70例(对照组)。比较COPD组和对照组CD4^(+)、CD8^(+)、Th17、Treg、CD4^(+)/CD8^(+)、Th17/Treg,比较COPD组不同病情患者CD4^(+)、CD8^(+)、Th17、Treg、CD4^(+)/CD8^(+)、Th17/Treg;COPD组患者随访6个月,比较不同预后患者临床资料。多因素Logistic回归分析老年COPD近期不良预后的影响因素,绘制受试者工作特征(receiver operating characteristic,ROC)曲线分析CD4^(+)/CD8^(+)、Th17/Treg预测老年COPD近期不良预后的临床价值。结果COPD组外周血CD4^(+)、CD4^(+)/CD8^(+)、Treg含量均低于对照组,外周血CD8^(+)、Th17含量、Th17/Treg高于对照组(P<0.05);Ⅲ级组外周血CD4^(+)、CD4^(+)/CD8^(+)、Treg含量低于Ⅰ级组、Ⅱ级组,外周血CD8^(+)、Th17含量、Th17/Treg高于Ⅰ级组、Ⅱ级组,Ⅱ级组外周血CD4^(+)、CD4^(+)/CD8^(+)、Treg含量低于Ⅰ级组,外周血CD8^(+)、Th17含量、Th17/Treg高于Ⅰ级组(P<0.05);Pearson相关性分析显示,COPD患者CD4^(+)/CD8^(+)与病情分级呈负相关,Th17/Treg与病情分级呈正相关(P<0.05);预后不良组外周血CD4^(+)、CD4^(+)/CD8^(+)、Treg含量低于预后良好组,外周血CD8^(+)、Th17含量、Th17/Treg、血清IL-6、CRP水平高于预后良好组(P<0.05);多因素Logistic回归分析显示,CD4^(+)/CD8^(+)、Th17/Treg、IL-6、CRP是老年COPD患者近期不良预后的影响因素;ROC曲线分析显示,CD4^(+)/CD8^(+)、Th17/Treg预测老年COPD患者近期不良预后曲线下面积分别为0.813(95%CI:0.730~0.896)、0.852(95%CI:0.730~0.896)。结论CD4^(+)/CD8^(+)、Th17/Treg在老年COPD患者病情及近期预后评估中具有较高的临床价值。

Study on the effect and mechanism of the dysfunction of CD4^+ T cells in the disease process of chronic cardiac failure

Objective: To study the effect and mechanism of the dysfunction of CD4+ T cells in the disease process of chronic cardiac failure (CHF).Methods:According to different group technologies, 100 CHF patients were divided into the following groups: ischemia group and non-ischemia group, heart function Ⅰ-Ⅱ group and heart function Ⅲ-Ⅳ group, event group and non-event group, and 50 healthy volunteers were included in the control group. Realtime PCR was used to detect transcription factors T-bet and GATA-3 of Th1 and Th2; flow cytometry was applied to determine the ratio of Th17 and Treg cells; ELISA was employed to test cytokines IFN-γ, IL-4, IL-17 and IL-10 of peripheral blood Th1, Th2, Th17 and Treg cells, respectively; ultrasonic cardiogram was used to exploit to LVEF and LVEDd; and electrochemilu minescene immunoassay was used to examine plasma BNP. The differences of all indexes of all groups were analyzed and the correlation between CD4 T cells and clinical indexes was analyzed by Pearson correlation analysis. Results: As compared to the control group, the transcription factors T-bet and GATA-3 of Th1 and Th2, the ratio of cytokines Th17 and IFN-γ, cytokines IL-17, T-bet/GATA-3, IFN-γ/IL-4, Th17 cells/Treg cells, IL-17/IL-10 of the ischemia group and non-ischemia group, heart functionⅠ-Ⅱgroup and heart function Ⅲ-Ⅳ group, event group and non-event group were all increased significantly, while their transcription factor GATA-3 of Th2, cytokines IL4, Treg cells ratio, cytokines IL10 were decreased obviously. The differences showed statistical significance (P < 0.05). The increase or decrease of the partial CD4+ T cells of the ischemia group, heart function Ⅲ-Ⅳ group and event group was more distinctly. The results of Pearson correlation analysis showed that IFN-γ and IL-17 were significantly positively correlated with LVEDd and BNP, IL-4 and IL-10 were also significantly positively correlated with LVEF, but correlated negatively with BNP, and IL-17 was negatively correlative with LVEF. Conclusions: There was a correlation between CHF and the dysfunction of CD4+ T cells showing immune activation phenomenons of deviations from the Th1/Th2 balance towards Th1 and from the Th17/Treg balance towards Th17, which was also related to the types, severity and prognosis of the disease.

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.

Analysis of CD4^+CD25^+ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

Depletion of CD4^+CD25^+ regulatory T cells can promote local immunity to suppress tumor growth in benzo[a]pyrene-induced forestomach carcinoma

AIM： To elucidate the distribution of CD4^＋CD25^＋ regulatory T cells （Tregs） in different lymphoid tissues and its local enhancement on tumor growth before and after depletion of CD4^＋CD25^＋ Tregs. METHODS： Female ICR mice were garaged with benzo[a]pyrene （BaP） to induce forestomach carcinoma. CD4^＋CD25^＋ Tregs were intraperitoneally depleted with monoclonal antibody PC61. These mice were divided into BaP-only, BaP ＋ IgG, BaP ＋ PC61, and control groups. The forestomach of mice was dissected for histological analysis, and tunnel test was performed for apoptosis of tumor cells. CD4^＋CD25^＋ Tregs were sorted from different lymphoid tissues and expression of Foxp3, IL-10, and chemokine receptors was analyzed by flow cytometry, semi-quantitative and veal-time polymerase chain reaction. RESULTS： The mice gavaged with only BaP showed increased forestomach papilloma and carcinoma at wk 16 and 32. The proportion of CD4^＋CD25^＋ Tregs was significantly higher in peri-stomach regional lymph nodes than in other lymphoid tissues. These CD4^＋CD25^＋ Tregs in regional lymph nodes expressed higher levels of Foxp3 and IL-10, enriched in the CD62L-subset, and CCR1 and CCR5 chemokine receptors. In mice gavaged with BaP ＋ PC61, the number of tumor nodules and tumor volume decreased significantly with massive infiltrating cells and apoptosis of tumor cells. In the draining regional lymph nodes, the number of CD4^＋CD25^＋ Tregs also decreased significantly. CONCLUSION： Inducible and activated CD4^＋CD25^＋ Tregs in the draining regional lymph nodes suppress host local immunity during tumor growth. Depletion of CD4^＋CD25^＋ Tregs can promote host local immunity to suppress tumor growth.

Super-resolution immunohistochemistry study on CD4 and CD8 cells and the relation to macrophages in human cochlea

Recently, the human cochlea has been shown to contain numerous resident macrophages under steady-state. The macrophages accumulate in the stria vascularis, among the auditory nerves, and are also spotted in the human organ of Corti. These macrophages may process antigens reaching the cochlea by invasion of pathogens and insertion of CI electrode. Thus, macrophages execute an innate, and possibly an adaptive immunity. Here, we describe the molecular markers CD4 and CD8 of T cells, macrophage markers MHCⅡ and CD11b, as well as the microglial markers TEME119 and P2Y12, in the human cochlea. Immunohistochemistry and the advantageous super-resolution structured illumination microscopy(SR-SIM) were used in the study. CD4^+ and CD8^+ cells were found in the human cochleae. They were seen in the modiolus in a substantial number adjacent to the vessels, in the peripheral region of the Rosenthal's canal, and occasionally in the spiral ligament. While there are a surprisingly large number of macrophages in the stria vascularis as well as between the auditory neurons,CD4^+ and CD8^+ cells are hardly seen in these areas, and neither are seen in the organ of Corti. In the modiolus,macrophages, CD4^+ and CD8^+ cells appeared often in clusters. Interaction between these different cells was easily observed with SR-SIM, showing closely placed cell bodies, and the processes from macrophages reaching out and touching the lymphocytes. Otherwise the CD4^+ and CD8^+ cells in human cochlear tissue are discretely scattered. The possible roles of these immune cells are speculated.

Downregulation of CD4+CD25+ regulatory T cells may underlie enhanced Th1 immunity caused by immunization with activated autologous T cells

Regulatory T cells （Treg） play important roles in immune system homeostasis, and may also be involved in tumor immunotolerance by suppressing Th1 immune response which is involved in anti-tumor immunity. We have previously reported that immunization with attenuated activated autologous T cells leads to enhanced anti-tumor immunity and upregulated Thl responses in vivo. However, the underlying molecular mechanisms are not well understood. Here we show that Treg function was significantly downregulated in mice that received immunization of attenuated activated autologous T cells. We found that Foxp3 expression decreased in CD4＋CD25＋ T cells from the immunized mice. Moreover, CD4＋CD25＋Foxp3＋ Treg obtained from immunized mice exhibited diminished immunosuppression ability compared to those from naive mice. Further analysis showed that the serum of immunized mice contains a high level ofanti-CD25 antibody （about 30 ng/ml, p〈0.01 vs controls）. Consistent with a role ofanti-CD25 response in the downregulation of Treg, adoptive transfer of serum from immunized mice to naive mice led to a significant decrease in Treg population and function in recipient mice. The triggering of anti-CD25 response in immunized mice can be explained by the fact that CD25 was induced to a high level in the ConA activated autologous T cells used for immunization. Our results demonstrate for the first time that immunization with attenuated activated autologous T cells evokes anti-CD25 antibody production, which leads to impeded CD4＋CD25＋Foxp3＋ Treg expansion and function in vivo. We suggest that dampened Treg function likely contributes to enhanced Thl response in immunized mice and is at least part of the mechanism underlying the boosted anti-tumor immunity.

Proliferation and Apoptosis of Bone Marrow CD4^+ T Cells in Patients with Aplastic Anemia and Impacts of the Secreted Cytokines on Hematopoietic Stem Cells from Umbilical Cord Blood

Recent studies indicate that immune-associated aplastic anemia(AA)resembles such autoimmune diseases as insulin-dependent diabetes and chronic autoimmune thyroiditis that belong to organ-specific autoimmune diseases.Many independent investigation groups have successfully isolated the pathopoiesis-associated T cell clone causing hematopoiesis failure with a CD4 phenotype from peripheral blood and bone marrow(BM)in AA patients.In the current study,BM CD4+ T cells were isolated from AA patients and healthy con...

CD4^+CXCR5^+ T cells activate CD27^+IgG^+ B cells via IL-21 in patients with hepatitis C virus infection

BACKGROUND： Chronic hepatitis C virus （HCV） infection causes the skewing and activation of B cell subsets, but the characteristics of IgG＋ B cells in patients with chronic hepa- titis C （CHC） infection have not been thoroughly elucidated. CD4＋CXCR5＋ follicular helper T （Tfh） cells, via interleukin （IL）-21 secretion, activate B cells. However, the role of CD4＋CXCR5＋ T cells in the activation ofIgG＋ B cells in CHC patients is not clear. METHODS： The frequency of IgG＋ B cells, including CD27-IgG＋ B and CD27＋IgG＋ B cells, the expression of the activation markers （CD86 and CD95） in IgG＋ B cells, and the percentage of circu- lating CD4＋CXCR5＋ T cells were detected by flow cytometry in CHC patients （n=70） and healthy controls （n=25）. The con- centrations of serum IL-21 were analyzed using ELISA. The role of CD4＋CXCR5＋ T cells in the activation of IgG＋ B cells was investigated using a co-culture system. RESULTS： A significantly lower proportion of CD27＋IgG＋ B cells with increased expression of CD86 and CD95 was observed in CHC patients. The expression of CD95 was negatively correlated with the percentage of CD27＋IgG＋ B cells, and it contributed to CD27＋IgG＋ B cell apoptosis. Circulating CD4＋CXCR5＋ T cells and serum IL-21 were significantly increased in CHC patients. Moreover, circulating CD4＋CXCR5＋ T cells from CHC patients induced higher expressions of CD86 and CD95 in CD27＋IgG＋ B cells in a co-culture system; the blockade of the IL-21 decreased the expression levels of CD86 and CD95 in CD27＋IgG＋ B cells.CONCLUSIONS： HCV infection increased the frequency of CD4＋CXCR5＋ T cells and decreased the frequency of CD27＋IgG＋ B cells. CD4＋CXCR5＋ T cells activated CD27＋IgG＋ B cells via the secretion of IL-21.

Upregulated adenosine 2A receptor accelerates post-infectious irritable bowel syndrome by promoting CD4+T cells’T helper 17 polarization

BACKGROUND Post-infectious irritable bowel syndrome(PI-IBS)is generally regarded as a functional disease.Several recent studies have reported the involvement of lowgrade inflammation and immunological dysfunction in PI-IBS.T helper 17(Th17)polarization occurs in IBS.Adenosine and its receptors participate in intestinal inflammation and immune regulation.AIM To investigate the role of Th17 polarization of CD4+T cells regulated by adenosine 2A receptor(A2AR)in PI-IBS.METHODS A PI-IBS model was established by infecting mice with Trichinella spiralis.The intestinal A2AR and CD4+T lymphocytes were detected by immunohistochemistry,and the inflammatory cytokines were detected by enzyme-linked immunoassay.CD4+T lymphocytes present in the animal’s spleen were separated and cultured with or without A2AR agonist and antagonist.Western blotting and real-time quantitative polymerase chain reaction were performed to determine the effect of A2AR on the cells and intestinal tissue.Cytokine production was determined.The protein and mRNA levels of A2AR associated signaling pathway molecules were also evaluated.Furthermore,A2AR agonist and antagonist were injected into the mouse model and the clinical features were observed.RESULTS The PI-IBS mouse model showed increased expression of ATP and A2AR(P<0.05),and inhibition of A2AR improved the clinical features in PI-IBS,including the abdominal withdrawal reflex and colon transportation test(P<0.05).The number of intestinal CD4+T cells and interleukin-17(IL-17)protein levels increased during PI-IBS,which was reversed by administration of the A2AR antagonist(P<0.05).CD4+T cells expressed A2AR and produced IL-17 in vitro,which was regulated by the A2AR agonist and antagonist.The A2AR antagonist increased the production of IL-17 by CD4+T cells via the Janus kinase-signal transducer and activator of transcriptionreceptor-related orphan receptorγsignaling pathway.CONCLUSION The results of the present study suggested that the upregulation of A2AR increases PI-IBS by promoting the Th17 polarization of CD4+T cells.

Influence of Danshen Injection on airway inflammation and CD4^+ CD25^+ regulatory T cells of asthmatic rats

Objective： To investigate the influence of Danshen Injection on airway inflammation and CD4^＋CD25^＋ regulatory T cells（CD4^＋CD25^＋ Tr） of asthmatic rats, and elucidate the possible mechanism of Danshen Injection in treatment of asthma. Methods： 30 Wister rats were randomly divided into control group, asthma group and Danshen Injection treated group. Bronchoalveolar lavage fluids （BALF） were collected, and cytology studies were conducted. Lung tissues were obtained and pathologic analyses were done with hematoxylin and eosin stain （HE）. Flow cytometry was used to detect the CD4^＋CD25^＋ Tr ratio in peripheral blood mononuclear cells （PBMCs）. Results： Total cell, the percentage of lymphocytes, neutrophils and eosinophils （Eos） in BALF of Danshen Injection-treated group were lower than that in asthma group （P〈0.05, P〈0.01）. Compared with asthma group, less infiltration of inflammatory cells in lung tissues was observed in Danshen Injection-treated group. CD4^＋CD25^＋ Tr of asthma group was lower than that of control and Danshen Injection treated group （P〈0.05）. Conclusion： Danshen Injection can suppress airway inflammation of asthmatic rats, probably by increasing the number of CD4^＋CD25^＋ Tr.

Role of LAP^+CD4^+ T cells in the tumor microenvironment of colorectal cancer

AIM To investigate the abundance and potential functions of LAP^+CD4^+ T cells in colorectal cancer(CRC). METHODS Proportions of LAP^+CD4^+ T cells were examined in peripheral blood and tumor/paratumor tissues of CRC patients and healthy controls using flow cytometry. Expression of phenotypic markers such as forkhead box(Fox)p3, cytotoxic T-lymphocyte-associated protein(CTLA)-4, chemokine CC receptor (CCR)4 and CCR5 was measured using flow cytometry. LAP^-CD4^+ and LAP^+CD4^+ T cells were isolated using a magnetic cellsorting system and cell purity was analyzed by flow cytometry. Real-time quantitative polymerase chain reaction was used to measure expression of cytokines interleukin (IL)-10 and transforming growth factor(TGF)-β.RESULTS The proportion of LAP^+CD4^+ T cells was significantly higher in peripheral blood from patients (9.44% ± 3.18%) than healthy controls (1.49% ± 1.00%, P < 0.001). Among patients, the proportion of LAP^+CD4^+ T cells was significantly higher in tumor tissues(11.76% ± 3.74%) compared with paratumor tissues (3.87% ± 1.64%, P < 0.001). We also observed positive correlations between the proportion of LAP^+CD4^+ T cells and TNM stage(P < 0.001), distant metastasis(P < 0.001) and serum level of carcinoembryonic antigen(P < 0.05). Magnetic-activated cell sorting gave an overall enrichment of LAP^+CD4^+ T cells (95.02% ± 2.87%), which was similar for LAP^-CD4^+ T cells(94.75% ± 2.76%). In contrast to LAP^-CD4^+ T cells, LAP^+CD4^+ T cells showed lower Foxp3 expression but significantly higher levels of CTLA-4, CCR4 and CCR5(P < 0.01). LAP^+CD4^+ T cells expressed significantly larger amounts of IL-10 and TGF-β but lower levels of IL-2, IL-4, IL-17 and interferon-γ, compared with LAPCD4+ T cells.CONCLUSION LAP^+CD4^+ T cells accumulated in the tumor microenvironment of CRC patients and were involved in immune evasion mediated by IL-10 and TGF-β.

再生障碍性贫血外周血Th17和CD4^+ CD25^+ Treg细胞表达情况研究

本研究旨在探讨成人再生障碍性贫血(aplastic anemia,AA)外周血Th17和CD4+CD25+调节性T细胞(Treg)表达情况。45例初发AA患者分为轻型AA(n=25)和重型AA(n=25)两组,15例正常人作为对照。应用流式细胞术检测AA患者外周血中Th17和CD4+CD25+Treg的比例,ELISA检测血清及刺激后外周血单个核细胞(PBMNC)上清液中的白介素(IL-17)、γ干扰素(IFN-γ)及肿瘤坏死因子-α(TNF-α)水平,并与正常对照组比较。结果表明:重型AA(SAA)组外周血Th17细胞比例,血清中IL-17及IFN-γ表达高于轻型AA(MAA)和正常人组,而重型AA组外周血CD4+CD25+Foxp3+Treg细胞比例降低。与轻型AA和正常对照相比,重型AA患者单个核细胞培养后上清液中IL-17及IFN-γ表达水平显著增加。AA患者血红蛋白计数与Th17细胞及血清IL-17表达呈负相关,与CD4+CD25+Treg表达成正相关。结论:重型AA患者外周血Th17细胞应答增强,而CD4+CD25+Treg细胞缺乏,贫血的严重程度与外周血Th17/Treg免疫应答失衡密切相关。

Th17细胞与CD4^+CD25^+调节性T细胞在儿童过敏性紫癜发病机制中的作用

目的探讨Th17细胞与CD4+CD25+调节性T细胞在儿童过敏性紫癜(HSP)免疫发病机制中的作用。方法过敏性紫癜急性期患儿48例,采用流式细胞术检测外周血CD4+CD25+调节性T细胞(CD4+CD25+Treg)的比例,采用荧光定量PCR(real-time PCR)检测CD4+ T细胞IL-17A、IL-17F、转录因子ROR-γt、Foxp3及细胞因子IL-6、TGF-β等mRNA表达;同期40例同龄健康儿童作为对照。结果过敏性紫癜急性期患儿CD4+ T细胞高表达IL-17A及IL-17F(P<0.01)。Th17细胞转录因子ROR-γt及前炎症细胞因子IL-6 mRNA表达明显增高(P<0.01)。CD4+CD25+调节性T细胞比例明显低于正常对照组(P<0.01),其转录因子Foxp3表达亦明显降低(P<0.01)。结论Th17等促炎性T细胞高表达和CD4+CD25+调节性T细胞数量减少导致的免疫抑制效应不足,是导致过敏性紫癜免疫失衡的重要原因,IL-6高表达与此密切相关。

Th17及CD4^+CD25^+调节性T细胞在结肠癌患者外周血中的表达

目的检测结肠癌患者外周血Th17及CD4+CD25+调节性T细胞水平及功能状态,探讨其在结肠癌发病过程中的免疫机制。方法应用流式细胞仪测定结肠癌患者及对照组外周血中Th17和CD4+CD25+调节性T细胞占CD4+T细胞的百分比,采用ELISA法测定两组患者外周血IL-17、IL-23、TGF-β1和IL-6的水平。结果结肠癌患者外周血Th17、CD4+CD25+调节性T细胞占CD4+T细胞的百分比及各相关的细胞因子较对照组升高,差异均有统计学意义(P<0.05);中晚期结肠癌组外周血Th17、CD4+CD25+调节性T细胞占CD4+T细胞的百分比及各相关的细胞因子较早期结肠癌组升高,差异均有统计学意义(P<0.05)。结论 Th17、CD4+CD25+调节性T细胞分化失衡与结肠癌发生、发展密切相关,为结肠癌免疫治疗提供新思路。

三种不同方法体外诱导CD4^+T细胞向Th17细胞分化与增殖的效率

背景:研究认为Th17细胞参与了许多炎症性疾病的发生,但其体外诱导分化效率尚不明确。目的:比较采用3种不同方法在体外诱导CD4+T细胞向Th17细胞分化与增殖的效率。方法:分别采取以下3种方法体外诱导Th17细胞:①给予CD3及CD28抗体刺激,并在培养体系中加入白细胞介素6及转化生长因子β,培养3d。②在方法①的基础上加入白细胞介素1β及肿瘤坏死因子α,培养3d。③在方法②的基础上第3天洗去细胞因子,培养2d;再次给予CD3及CD28抗体刺激,并在培养体系中加入白细胞介素23,培养3d。结果与结论:3种方法培养后CD4+T细胞中Th17细胞的比例分别为(8.5±2.8)%,(26.9±4.3)%,(44.3±5.5)%,三组之间差异有显著性意义(P<0.01)。提示在培养体系中加入白细胞介素1β、肿瘤坏死因子α及白细胞介素23,更有利于增加CD4+T细胞向Th17细胞分化的比例。

原发性肝癌患者外周血CD4^(+)T、CD8^(+)T、Tc17、Th17和Treg淋巴细胞的变化及其意义

目的了解原发性肝癌(PLC)患者外周血CD4^(+)T、CD8^(+)T、Tc17、Th17和Treg淋巴细胞的变化。方法2018年6月~2019年12月我院诊治的PLC患者83例(巴塞罗那临床肝癌分期A期25例,B期23例,C期18例,D期17例)和健康人35例,采用流式细胞技术检测外周血CD4^(+)T、CD8^(+)T及Tc17(CD8^(+)IL-17)、Th17(CD4^(+)IL-17)和CD4^(+)CD25^(+)CD45RA^(+)Treg淋巴细胞百分比。结果PLC患者外周血CD4^(+)T淋巴细胞百分比为(32.8±8.5)%,显著低于健康人[(43.3±7.4)%,P<0.05],CD4^(+)/CD8^(+)细胞比值为(0.9±0.3),显著低于健康人[(1.2±0.1),P<0.05];PLC患者外周血Treg细胞[(6.4±0.9)%对(3.0±0.1)%]、Th17细胞[(5.0±1.1)%对(3.1±1.5)%]及Tc17细胞[(2.3±0.4)%对(1.0±0.2)%],均较健康人显著升高(P<0.05);BCLC C/D期患者外周血CD4^(+)CD25^(+)CD45RA^(+)Treg细胞百分比较A/B期患者显著升高[(7.6±0.4)%对(5.3±0.5)%,P<0.001],Th17(CD4^(+)IL-17)和Tc17(CD8^(+)IL-17)细胞百分比较A/B期亦显著升高[分别为(6.9±1.1)%对(5.4±0.6)%,P<0.01和(2.8±0.6)%对(1.6±0.4)%,P<0.01]。结论PLC患者外周血淋巴细胞亚群出现异常改变,可能与肿瘤的分期密切相关,为从免疫学角度开展调节T淋巴细胞失衡的免疫治疗提供了理论依据。