Peripheral CD4^(+)CD8^(+) double positive T cells:A potential marker to evaluate renal impairment susceptibility during systemic lupus erythematosus

Lupus nephritis(LN) has a high incidence in systemic lupus erythematosus(SLE) patients, but there is a lack of sensitive predictive markers. The purpose of the study was to investigate the association between the CD4^(+)CD8^(+)double positive T(DPT) lymphocytes and LN. The study included patients with SLE without renal impairment(SLE-NRI), LN, nephritic syndrome(NS), or nephritis. Peripheral blood lymphocyte subsets were analyzed by flow cytometry. Biochemical measurements were performed with peripheral blood in accordance with the recommendations proposed by the National Center for Clinical Laboratories. The proportions of DPT cells in the LN group were significantly higher than that in the SLE-NRI group(t=4.012, P<0.001), NS group(t=3.240,P=0.001), and nephritis group(t=2.57, P=0.011). In the LN group, the risk of renal impairment increased significantly in a DPT cells proportion-dependent manner. The risk of LN was 5.136 times(95% confidence interval, 2.115–12.473) higher in cases with a high proportion of DPT cells than those whose proportion of DPT cells within the normal range. These findings indicated that the proportion of DPT cells could be a potential marker to evaluate LN susceptibility, and the interference of NS and nephritis could be effectively excluded when assessing the risk of renal impairment during SLE with DPT cell proportion.

Association between the level of CD4^+T lymphocyte microRNA-155 and coronary artery disease in patients with unstable angina pectoris

Objective To study the association between the expression of microRNA-155(miRNA-155)in peripheral blood CD4^+T lymphocytes and the level of semrn interferon-7(IFN-7)concentration and the severity of coronary artery disease (CAD).Methods After coronary angiography,252patients with suspected unstable angina pectoris (UAP)were divided into the UAP group (128patients with CAD confirmed by angiography)and the control group (124patients without CAD confirmed by angiography).Fresh peripheral blood was extracted 16-24h before coronary angiography,CD4^+T lymphocytes was tested using immunomagnetic beads,the expression ofmiRNA-155was tested using quantitative PCR and the expression of IFN-7was tested using enzyme-linked immunosorbent assay (ELISA).According to the results of angiography,Gensini score of coronary artery lesions was analyzed.Furthermore,we also analysis the association between the level of miRNA-155in peripheral blood CD4^+T lymphocytes,the level of serum IFN-γand Gensini score of coronary lesion.Results The levels ofmiRNA-155(0.49±0.08vs.0.23±0.09)and IFN-7(227.58±26.01vs.141.23±17.89)in the UAP group were significantly higher than that of the control group,the difference was statistically significant.The level of miRNA-155and IFN-γwere positively correlated with Gensini score of CAD (r =0.534,r =0.713,respectively,all P <0.05).The level of miRNA-155was positively correlated with the level of IFN-γ,(r =0.686,P <0.05).Conclusions The level of miRNA-155in peripheral blood CD4^+T lymphocytes and the level of IFN-γ are closely correlated with the severity of CAD.

外周血CD4^(+)PD-1^(+)Tcells及CD4^(+)T淋巴细胞ATP含量与复发性卵巢癌疗效的相关性分析

目的 探讨外周血CD4^(+)程序性细胞死亡受体-1(PD-1)^(+)T cells及CD4^(+)T淋巴细胞三磷酸腺苷(ATP)含量与复发性卵巢癌疗效的相关性。方法 选取30例复发性卵巢癌患者为复发组,30例未复发卵巢癌患者为非复发组;另选取30名同期体检健康者作为对照组。评估外周血CD4^(+)PD-1^(+)T cells及CD4^(+)T淋巴细胞ATP含量与复发性卵巢癌疗效的相关性。结果 复发组和非复发组的CD4^(+)PD-1^(+)T cells较对照组明显升高(P<0.05)。复发组和非复发组的CD4^(+)T淋巴细胞ATP含量较对照组明显降低(P<0.05)。复发组治疗后CD4^(+)PD-1^(+)Tcells显著低于治疗前(P<0.05),治疗后CD4^(+)T淋巴细胞ATP含量显著高于治疗前(P<0.05)。CD4^(+)PD-1^(+)T cells与复发性卵巢癌疗效成负相关(r=-0.393,P=0.039),CD4^(+)T淋巴细胞ATP含量与复发性卵巢癌疗效成正相关(r=0.449,P=0.031)。结论 复发性卵巢癌患者外周血CD4^(+)PD-1^(+)T cells及CD4^(+)T淋巴细胞ATP含量与疗效密切相关。

Effect of Mg^(2+)level on the functions of CD8^(+)T lymphocytes and NK cells in patients with COVID-19

Objective:To investigate the changes of Mg^(2+) levels in serum and peripheral blood mononuclear cells(PBMCs)of patients with COVID-19 and its effects on the functions of CD8^(+)T lymphocytes and NK cells.Methods:A total of 165 COVID-19 patients hospitalized in Ezhou Central Hospital from January 20 to February 20,2020 were divided into mild/common group(98 cases)and severe/critical group(67 cases).At the same time,34 healthy persons were selected as the control group.Peripheral blood was collected and PBMCs were isolated,the level of Mg^(2+) in serum and PBMCs was detected.The subsets of CD8^(+)T lymphocytes and NK cell and the expression levels of their surface inhibitory molecular PD-1 and activator molecular NKG2D were detected by flow cytometry.The correlation between Mg^(2+) concentration and the expression levels of PD-1 and NKG2D was also analyzed.Results:Compared with the control group,the concentration of Mg^(2+) in serum and PBMCs,the counts of CD8^(+)T lymphocytes and NK cell in patients with mild/common and severe/critical groups were significantly reduced(P<0.05),while the expression level of surface inhibitory molecular PD-1 were significantly increased(P<0.05),while the expression level of the activation molecule NKG2D were significantly decreased(P<0.05).However,the changes of the above indicators in patients with severe/critical group were greater than those in the mild/common group(P<0.05).In addition,the Mg^(2+) concentration in COVID-19 patients was negatively correlated with the expression level of PD-1 on CD8^(+)T lymphocytes and NK cells(P<0.05),and positively correlated with the expression levels of NKG2D(P<0.05).Conclusion:The concentration of Mg^(2+) in the serum and PBMCs of COVID-19 patients is significantly reduced,which may cause the function of CD8^(+)T lymphocytes and NK cells to be inhibited.

MBD2 promotes Th2 differentiation in ovalbumin-induced CD4+T cells

Introduction:Allergen-specific CD4+T cells play a central role in autoimmune disorders,allergies and asthma,with Th2-type immunity being the typical functional response of CD4+T cells.This study aimed to investigate the role of MBD2 in regulating Th2 cell differentiation.Methods:Splenic mononuclear cells were extracted from C57BL/6 mice,and CD4+T cells were isolated using magnetic beads and confirmed through flow cytometry.Lentivirus was employed to construct MBD2-silenced CD4+T cells.In vitro experiments were performed to treat splenogenic mononuclear cells and CD4+T cells with Ovalbumin(OVA),and Th2 cell ratios and IL-4 levels were assessed using flow cytometry and ELISA.Results:The purity of the isolated CD4+T cells was 95.73%,confirming successful isolation of primary CD4+T cells.Compared to the control group,the Th2 cell ratio exhibited an increase in the Th2-induced group.Treatment with 5-Aza(concentrations,1-100μM)promoted Th2 cell differentiation and increased IL-4 levels.Notably,when combined with Th2 induction and 10μM 5-Aza treatment,silencing MBD2 further amplified Th2 cell ratios and elevated IL-4 levels in cell supernatants.Furthermore,OVA(concentration,200μg/mL)induced the differentiation of CD4+T cells into Th2 cells and increased IL-4 secretion.Interestingly,silencing MBD2 significantly increased the Th2 cell ratio and IL-4 levels in OVA-treated CD4+T cells.Conclusion:In summary,OVA promoted CD4+T cell differentiation into Th2 cells and enhanced IL-4 levels.MBD2 was identified as a mediator of Th2 cell differentiation in splenic-derived CD4+T cells,influenced by OVA or 5-Aza treatment.

过敏性鼻炎患者外周血CD4^(+)2型辅助T(Th2)细胞中IL-18的表达升高

目的 探讨过敏性鼻炎(AR)患者外周血2型辅助T(Th2)细胞白细胞介素18(IL-18)、 IL-18结合蛋白亚型a(IL-18BPa)和IL-18受体α(IL-18Rα)的表达水平及过敏原对其表达的影响。方法 采用大籽蒿花粉过敏原粗提液(ASWE)和梧桐花粉过敏原粗提液(PPAE)、室尘螨过敏原粗提液(HDME)激发外周血单个核细胞和免疫磁珠分选后的CD4^(+)T细胞,流式细胞术检测上述标本CD4^(+)Th2细胞中IL-18、 IL-18BPa和IL-18Rα的表达,BioPlex检测血浆IL-4的水平,并分析其与IL-18+Th2细胞比例的相关性。结果 与正常对照相比,AR患者外周血CD4^(+)Th2细胞中IL-18+细胞的比例增加,IL-18的平均荧光强度(MFI)增强;IL-18Rα MFI降低。过敏原可直接诱导分选后的HC CD4^(+)Th2细胞表达IL-18和IL-18Rα, AR患者CD4^(+)Th2细胞表达IL-18Rα。AR患者血浆IL-4水平升高,并与IL-18+Th2细胞的比例相关。结论 过敏原可能通过诱导外周血CD4^(+)Th2细胞表达IL-18参与AR发病。

H3N2流感病毒HA保守Th表位对CD4^(+)T细胞活化及分化的影响

目的探讨H3N2病毒HA保守Th表位对CD4^(+)T细胞活化及分化的影响,为后续研究通用流感疫苗奠定基础。方法利用NetMHCⅡpan-4.0软件预测H3N2疫苗株A/Kansas/14/2017 HA的Th表位,以IC 50<500 nM为标准筛选阳性预测表位;采用SWISS-MODLE对A/Kansas/14/2017 HA进行同源建模,在3D模型上定位保守Th表位,根据表位的亲和力、保守性及核心序列的位置筛选高保守表位;合成6个非重叠的高保守Th表位,用保守Th表位体外刺激健康人外周血单核细胞,流式细胞术分析CD4^(+)T细胞表面分子CD69及细胞因子IFN-γ、IL-4的表达。结果在A/Kansas/14/2017 HA中共筛选到48个阳性预测Th表位,其中33个在H3-HA中是保守的;成功构建A/Kansas/14/2017 HA 3D模型,27个保守Th表位位于HA头部,其余6个位于杆部;人外周血PBMC经保守Th表位刺激后,CD69分子表达增加,细胞因子IFN-γ和IL-4合成增加。结论筛选到的6个H3N2 HA高保守Th表位具有免疫原性,可刺激CD4^(+)T细胞活化及分化。

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.

Changes of CD4^+CD25^+ Regulatory T Cells in Patients with Acute Coronary Syndrome and the Effects of Atorvastatin

The function of CD4＋CD25＋ regulatory T lymphocytes （Treg） in patients with acute coronary syndrome （ACS） and the effects of atorvastatin were investigated. Forty-eight patients with ACS were randomly divided into two groups： group C receiving conventional therapy （n=24）, and group C＋A receiving conventional therapy＋atorvastatin （10 mg/day, n=24）. T lymphocytes from ACS patients （before and 2 weeks after the treatment） or 18 healthy subjects were separated and the flow cytometry was used to measure the percentage of Treg. The inhibitory ability of Treg on effector T cells was determined by mixed lymphocyte reaction （MLR）. ELISA was used to measure the serum levels of cytokines （IL-10, TGF-β1 and IFN-γ） before and after treatment. The results showed that as compared with normal control group, Treg percentage was decreased significantly （P〈0.01）, the inhibitory ability of Treg on the T lymphocytes proliferation was reduced （P〈0.01）, IFN-γ levels were increased and IL-10 and TGF-β1 levels were lowered in ACS patients. After treatment with atorvastatin, Treg percentage and the inhibitory ability of Treg on T lymphocytes proliferation were significantly increased in ACS patients. Serum IFN-γ was decreased significantly, while IL-10 and TGF-β1 were elevated significantly as compared with the non-atorvastatin group. The number of Treg was positively correlated with serum TGF-β1, but negatively with serum IFN-γ and CRP. It was concluded that ACS was associated with decreased number and defected function of Treg, which may play an important role in initiating immune-inflammatory response in ACS. The inhibitory effects of atorvastatin on inflammation in ACS may be due to its beneficial effects on Treg and restoration of immune homeostasis.

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.

钩吻素子对小鼠CD4^+T淋巴细胞Th1类及Th2类细胞因子分泌的影响

目的研究钩吻素子(Koumine,Kou)对小鼠CD4+T淋巴细胞Th1类及Th2类细胞因子分泌的影响。方法分离纯化小鼠脾脏CD4+T淋巴细胞,经刀豆球蛋白A(ConcanavalinA,ConA)刺激诱生细胞因子,然后加入不同浓度的Kou,继续培养24小时后离心收集细胞培养上清。用酶联免疫法(ELISA法)检测细胞上清中Th1类细胞因子IL2、IFNγ以及Th2类细胞因子IL4、IL10的含量变化情况,观察药物对CD4+T淋巴细胞因子表达的影响。结果对于Th1型细胞因子,各浓度组Kou作用24h后均可显著降低IL2、IFNγ的水平,并呈剂量依赖性;对于Th2型细胞因子,Kou对IL4表达影响不大,但中浓度组Kou(100μg/ml)可显著升高IL10的水平。结论对CD4+T细胞Th细胞因子分泌的调节可能是Kou治疗银屑病等T细胞活化异常炎症性疾病的免疫药理机制之一。

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。

Detection of microbial antigenic components of circulating immune complexes in HIV patients:Involvement in CD4^+ T lymphocyte count depletion

Objective:To investigate the prevalence of microbial antigenic components of circulating immune complexes amongst grades of CD4 T lymphocyte counts in HIV sero positive and seronegative participants.Methods:Polyethelene glycol(PEG-600) and buffering methods of precipitation and dissociation of immune complexes was used to generate immune solution from sera of 100 HIV sero-positive and 100 HIV sero-negative participants.These were categorized into 3 grades based on CD4 count:】 500 cell/mm,200-499 cell/mm3 and 【200 cell/mm3.The immune solutions were assayed using membrane based immunoassay and antibody titration, along side its unprocessed serum for detection of various microbial antigens and or antibodies. CD4 T cell counts were estimated using Patec Cyflow SL-3 Germany.Results:Antigenic component of immune complexes of various infectious agents was detected in 99 and 70 HIV seropositive and HIV sero-negative participants,respectively.In group A,there were 10 HIV positive participants,including 4(40.0%) had circulating immune complexes(CICs) due to Salmonella species only:1(10.0%) due to Salmonella-Plasmodium falciparum(P.falciparum),SalmonellaP. falciparum-HCV and P.falciparum antigens,respectively.In group B,45(45.4%) HIV seropositive participants with CICs had CD4 T lymphocyte count between 200-499 cells/mm^3.Out of these,20(44.4%) had CICs due to Salmonella species only:9(20%) due to Salmonella-P. falciparum.In group C,there were 44(44.4%) HIV sero-positive participants,including 3(6.8%) due to Salmonella species only:24(54.4%) due to Salmonella-P.falciparum:2(4.5%) due to P. falciparum only.Conclusions:In HIV sero-positive participants,presence of heterogeneity of Salmonella species-P.falciparum antigens was highly incriminated in CD4 count depletion but not homogeneity of malaria parasites antigens.Malaria parasites antigens only were incriminated in CD4^+ count depletion amongst HIV sero-negative participants.Before taking any decision on the management of HIV-1-positive individuals,their malaria and Salmonella paratyphi status should be assessed,but not malaria status alone.

犀角地黄汤对免疫性血小板减少症模型大鼠外周血CD4^+ CD25^+调节性T细胞及Th17细胞的影响

目的探讨犀角地黄汤对免疫性血小板减少症(ITP)模型大鼠外周血CD4^+ CD25^+调节性T细胞(Treg)、Th17细胞及相关细胞因子的影响。方法将48只大鼠随机分为正常组、模型组、醋酸泼尼松组及犀角地黄汤低、中、高剂量组,每组8只。除正常组外,其余组均采用腹腔注射兔抗大鼠血小板血清(APS)建立ITP模型,于造模开始后第7天开始,各给药组灌胃给予相应剂量药物,连续14 d。采用血细胞计数仪检测造模前、灌胃前、灌胃结束后各组大鼠外周血中血小板数目;灌胃结束后,采用流式细胞术检测各组大鼠Treg、Th17细胞比例,采用实时荧光定量PCR法检测淋巴细胞Foxp3 mRNA相对表达量,采用酶联免疫吸附法检测血清白细胞介素-17(IL-17)、IL-35水平。结果灌胃结束后,与模型组比较,犀角地黄汤各剂量组大鼠血小板数目明显上升(P均<0.05),外周血淋巴细胞中Treg所占比例、Foxp3 mRNA相对表达量及IL-35水平均明显上升(P均<0.05),而Th17细胞所占比例及IL-17水平均明显下降(P均<0.05),且具有剂量依赖性;犀角地黄汤高剂量组各指标与醋酸泼尼松组比较差异均无统计学意义(P均>0.05)。结论犀角地黄汤可能通过上调ITP大鼠外周血中Treg细胞表达,下调Th17细胞表达,维持Treg/Th17平衡,进而调节相关细胞因子IL-17、IL-35的表达发挥抗血小板减少作用。

白三烯受体拮抗剂对哮喘气道重塑及Th17细胞／CD4^＋CD25^＋调节性T细胞表达的影响

支气管哮喘的气道重塑是气道炎症反复作用的结果,白三烯是气道重塑中的重要炎症介质之一。影响气道重塑的因素较多,近年来Th17细胞和CD4+CD25+调节性T细胞(CD4+CD25+treg细胞)在气道重塑中的作用日益受到重视。白三烯受体拮抗剂是治疗哮喘的有效药物,能在一定程度上抑制气道重塑,但其作用机制及对Th17细胞/CD4+CD25+treg细胞表达的影响机制尚不十分清楚。因此,阐明Th17细胞/CD4+CD25+treg细胞平衡在气道重塑中的表达变化、白三烯受体拮抗剂干预气道重塑的具体作用途径和生物效应及对Th17细胞/CD4+CD25+treg细胞表达的影响,将为以后哮喘患儿的预防和治疗提供新的靶点。

脓毒症患者CD4^＋CD25^＋调节性T细胞和Thl7细胞检测的临床意义

目的探讨脓毒症患者外周血辅助性T细胞（THelper，Th）17及CD4^＋CD25^＋调节性T细胞（Treg细胞）表达的变化和检测的临床意义。方法将2008—01—2010—11在我医院ICU住院的脓毒症患者40例按照疾病严重程度分为三组：脓毒症组14例、严重脓毒症组15例和脓毒症休克组11例。再将脓毒症患者根据28d预后分为死亡组（D，n＝11）和存活组（S．n=29）。所有人选的40例脓毒症患者在入选当天行流式细胞术检测血中Thl7细胞及CD4^＋CD25^＋调节性T细胞的比例。比较脓毒症组、严重脓毒症组和脓毒症休克组患者血中Thl7细胞及CD4^＋CD25^＋调节性T细胞的比例变化，以及生存组和死亡组上述指标的差异。分析CD4^＋CD25^＋调节性T细胞和Thl7细胞免疫平衡紊乱与病情严重程度的关系。同时选取20例健康人为对照组。结果健康对照组Thl7细胞表达率为（0．91±0．38）％，CD4^＋CD25^＋调节性T细胞表达率为（0．39±0．23）％；脓毒症患者较正常对照组这两项指标明显升高，差异有统计学意义（P〈0．05）。其中脓毒症组分别为（2．09±0．53）％和（1．72±0．59）％；严重脓毒症组为（3．90±0．80）％和（2．72±0．22）％；脓毒性休克组为（1．85±0．35）％和（3.55±0．51）％。Thl7表达率严重脓毒症组最高，与其他两组比较差异有统计学意义（P〈0．05）。而脓毒症休克组与脓毒症组比较差异无统计学意义（P〉0．05）。CD4^＋CD25^＋调节性T细胞表达率脓毒症休克组最高，严重脓毒症组次之，脓毒症组最低，三组间两两比较差异有统计学意义（P〈0．05）。死亡组血中CD4^＋CD25^＋调节性T细胞表达率明显高于存活组[（3．03±0．91）％和（2．43±0．79）％，P〈0．05]，Thl7细胞表达率明显低于存活组[（2．01±0．66）％和（2．97±1．15）％，P〈0．05]。结论Thl7细胞和CD4^＋CD25^＋调节性T细胞在脓毒症的免疫发病机制中可能起着重要作用。检测脓毒症患者外周血．thl7细胞和CD4^＋CD25^＋调节性T细胞能够反映机体的细胞免疫状态，对评估患者预后有重要临床价值。

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。

Th9、Th17、CD4^+CD25^+FoxP3^+调节性T细胞在慢性淋巴细胞白血病患者外周血中的变化

目的探讨慢性淋巴细胞白血病患者外周血中Th9、Thl7和CD4^+CD25^+FoxP3^+调节性T细胞及相关细胞因子的表达水平及意义。方法采用流式细胞仪检测36例CLL患者和45例健康对照组外周血Th9、Th17和Treg细胞的表达率,采用酶联免疫吸附方法(ELISA)检测外周血中细胞因子IL-9、IL-17、TGF-β的表达水平。结果 1)CLL组患者外周血Th9、Th17细胞比例及细胞因子IL-9、IL-17浓度均明显高于健康对照组[(1.29±0.35)%vs(0.74±0.23)%,P<0.05;(2.15±0.46)%vs(1.08±0.37)%,P<0.05;(24.25±7.05)%vs(16.35±5.42)%,P<0.05;(11.47±3.99)%vs(7.36±2.85%),P<0.05]。CD4^+CD25^+FoxP3^+细胞比例及细胞因子TGF-β浓度明显低于对照组[(4.76±0.89)%vs(7.26±1.95)%,P<0.05;(3.82±1.39)%vs(6.82±2.01)%,P<0.05]。2)相关性分析:患者Th9、Th17细胞比例及IL-9、IL-17浓度与淋巴细胞计数负相关(γ_s=-0.374,P=0.034;γ_s=-0.382,P=0.032;γ_s=-0.445,P=0.011;γ_s=-0.413,P=0.024);患者Treg细胞比例及TGF-β浓度与淋巴细胞计数正相关(γ_s=0.416,P=0.023;γ_s=0.403,P=0.028)。结论慢性淋巴细胞白血病患者Th9、Th17与CD4^+CD25^+FoxP3^+调节性T细胞比例失衡,血清IL-9、IL-17和TGF-β水平变化,这些原因可能参与慢性淋巴细胞白血病的免疫发病过程。

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.