Increased frequency of IFN-gamma-producing peripheral CD8+ T cells with memory-phenotype in patients with chronic hepatitis C

To identify the capacity for cytokine production and the phenotypic characteristics of peripheral CD8(+) T cells in patients with chronic hepatitis C, 31 patients with chronic hepatitis C and 22 healthy controls were studied at the single cell level by three-color flow cytometry. Whole blood was stained with surface CD8, intracellular interferon-gamma (IFN-gamma), and interleukin-4 (IL-4), surface CD8, CD28, and intracellular IFN-gamma after stimulation with PMA plus ionomycin, and then surface CD8, CD45RA, and CD28. IFN-gamma-producing peripheral CD8(+) T cells were found frequently in patients than in controls (P < 0.05), whereas IL-4-producing peripheral CD8(+) T cells were not. Although the frequency of peripheral CD28(+)CD8(+) and CD28(-)CD8(+) T cells in patients was not different from that of controls, CD28(+)CD8(+) T cells exceeded CD28(-)CD8(+) T cells in the capacity for IFN-gamma-production after mitogenic stimulation (P < 0.01). In a more detailed analysis of the CD28(+)CD8(+) T cells, CD45RA(-)CD28(+)CD8(+) T cells, defined phenotypically as memory cells, were found frequently in patients than in controls (P < 0.05). There were no significant correlations between the frequency of IFN-gamma-producing peripheral CD8(+) T cells and hepatitis C virus RNA level or serum alanine aminotransferase level in patients. These data suggest that functionally T cytotoxic type 1 and memory CD8(+) T cells are predominant in the peripheral blood of chronic hepatitis C patients and that such activated CD8(+) T cells are associated with liver damage.     

Lower percentage of CD8+ T cells in peripheral blood of patients with sporotrichosis

PurposeTo characterize the peripheral immunity and immunity response of patients with sporotrichosis, in this study we determined the lymphocyte subsets in the peripheral blood of Chinese patients with sporotrichosis.MethodsIn this retrospective study, peripheral blood was collected from 69 sporotrichosis patients (37, fixed cutaneous form; 32 lymphocutaneous) and 66 healthy controls. Lymphocyte subsets were analyzed using flow cytometry.ResultsCompared to controls, the percentage of CD8+ T cells was lower in sporotrichosis patients. The percentage of CD8+ T cells in peripheral blood tended to become lower with disease duration and disease severity, although the difference was not statistically significant for either acute, subacute and chronic patients or fixed cutaneous and lymphocutaneous patients.ConclusionOur data indicate that the decrease of CD8+ T cells in peripheral blood of patients with sporotrichosis is associated with disease severity, although the difference was not statistically significant for either duration or clinical forms of the disease. Combining antifungal agents and immunomodulators in patients with long disease duration and lymphocutaneous may be more beneficial than antifungal monotherapy.     

Type 1 CD8^＋ T Cells are Superior to Type 2 CD8^＋ T Cells in Tumor Immunotherapy due to Their Efficient Cytotoxicity, Prolonged Survival and Type 1 Immune Modulation

CD8^＋ cytotoxic T （Tc） cells play a crucial role in host immune responses to cancer, and in this context, adoptive CD8^＋ Tc cell therapy has been studied in numerous animal tumor models. Its antitumor efficacy is, to a large extent, determined by the ability of Tc cells to survive and infiltrate tumors. In clinical trials, such in vitro-activated T cells often die within hours to days, and this greatly limits their therapeutic efficacy. CD8^＋ Tc cells fall into two subpopulations based upon their differential cytokine secretion. In this study, we in vitro generated that ovalbumin （OVA）-pulsed dendritic cell （DCovA）-activated CD8^＋ type 1 Tc （Tcl） cells secreting IFN-T, and CD8^＋ type 2 Tc （Tc2） cells secreting IL-4, IL-5 and IL-10, which were derived from OVA-specific T cell receptor （TCR） transgenic OT I mice. We then systemically investigated the in vitro and in vivo effector function and survival of Tcl and Tc2 cells, and then assessed their survival kinetics after adoptively transferred into C57BL/6 mice, respectively. We demonstrated that, when compared to CD8^＋ Tc2, Tcl cells were significantly more effective in perforin-mediated cytotoxicity to tumor cells, had a significantly higher capacity for in vivo survival after the adoptive T cell transfer, and had a significantly stronger therapeutic effect on eradication of well-established tumors expressing OVA in animal models. In addition, CD8^＋ Tcl and Tc2 cells skewed the phenotype of CD4^＋ T cells toward Thl and Th2 type, respectively. Therefore, the information regarding the differential effector function, survival and immune modulation of CD8^＋ Tcl and Tc2 cells may provide useful information when preparing in vitro DC-activated CD8^＋ T cells for adoptive T cell therapy of cancer.     

Up-regulation of microRNA-210 induces immune dysfunction via targeting FOXP3 in CD4 T cells of psoriasis vulgaris

Psoriasis vulgaris (PV) is a chronic inflammatory and T cell-mediated autoimmune skin disease. An immune dysfunction that is manifested by abnormally activated T cells and defective regulatory T (Treg) cells may play an important role in the pathogenesis of PV. However, the precise mechanism of the immune dysfunction in PV patients still remains unclear. In this study, we found that miR-210 expression is increased significantly in CD4⁺ T cells from patients with PV and confirmed that FOXP3 is a target gene of miR-210. We also found that overexpression of miR-210 inhibits FOXP3 expression and impairs the immunosuppressive functions of Treg cells in CD4⁺ T cells from healthy controls. In contrast, inhibition of miR-210 increases FOXP3 expression and reverses the immune dysfunction in CD4⁺ T cells from patients with PV. Our data demonstrates that increased miR-210 induces immune dysfunction via by FOXP3 in CD4⁺ T cells from patients with PV.     

CD3+CD56+ NK T cells are significantly decreased in the peripheral blood of patients with psoriasis

Psoriasis is a chronic, inflammatory, hyperproliferative skin disease, in which autoimmunity plays a great role. Natural killer T cells (NK T cells), are suggested to be involved in the pathogenesis of different autoimmune diseases. To examine the involvement of CD3+CD56+ NK T cells in the pathogenesis of psoriasis, we investigated the lymphocyte subpopulations obtained from blood samples of psoriatic patients before and after treatment, and of healthy controls, using two-colour flow cytometry. We found no significant differences between total T cells, total B cells, T helper cells, T cytotoxic cells and NK cells in patients with psoriasis before and after treatment and in controls. Increased percentage of memory T cells and decreased percentage of naive T cells was detected in psoriatic patients compared to controls, but these changes were not statistically significant. The CD3+CD56+ cells of psoriatic patients were significantly decreased relative to controls. The percentage of CD3+CD56+ cells increased after different antipsoriatic therapies, but remained significantly lower than those found in controls. CD3+CD56+ cells of healthy controls were capable of rapid activation, while in psoriatic patients activated NK T cells were almost absent. The decrease in the number of CD3+CD56+ cells may represent an intrinsic characteristic feature of patients with psoriasis, which is supported by the fact that after treatment NK T cells do not reach the values found in controls. In conclusion our results suggest that CD3+CD56+ NK T cells could be actively involved in the development of Th1 mediated autoimmune diseases.     

系统性红斑狼疮患者外周血CD4+和CD8+T细胞PD-1的表达和意义

目的 探讨程序性死亡分子1 (PD-1)在系统性红斑狼疮(SLE)患者外周血CD4+和CD8+T细胞上的表达及临床意义.方法 应用流式细胞仪检测51例SLE患者和38例健康对照者外周血T细胞亚群表面PD-1表达水平,比较SLE稳定组、活动组和健康对照组以及狼疮肾炎组和无狼疮肾炎组之间CD4+和CD8+T细胞表面PD-1表达的百分比,并分析其与临床表现及实验室检查数据的相关性.结果 SLE活动组CD4+T细胞PD-1表达水平高于健康对照组和不活动组,差异均有统计学意义(P＜0.05).SLE活动组、稳定组CD8+T细胞PD-1表达水平均高于健康对照组,差异有统计学意义(P＜0.05).狼疮肾炎患者CD4+PD-1+和CD8+PD-1+T细胞分别高于无狼疮肾炎患者(P＜0.01).SLE患者中抗dsDNA抗体、抗Sm抗体、抗核小体抗体阳性组外周血CD4+和CD8+T细胞PD-1表达水平均高于对应阴性组.SLE患者CD4+和CD8+T细胞PD-1表达百分率与SLE疾病活动度指数(SLEDAI)、尿蛋白定量呈正相关,与补体C3呈负相关.结论 SLE患者外周血CD4+和CD8+T细胞PD-1表达异常,与SLEDA1和自身抗体产生有明确的相关性.     

The frequency of CLA+ CD8+ T cells in the blood of psoriasis patients correlates closely with the severity of their disease.

Psoriasis is thought to be a T cell-mediated skin disease and the cutaneous lymphocyte antigen (CLA) is an important skin homing epitope for T cells. We have studied the relationship between disease severity (PASI) and phenotypic analysis of T cells in the blood of 36 patients with psoriasis focusing on the expression of CLA, VLA-4 and CD25 on CD4+ and CD8+ T cells. The patients had a higher frequency of circulating CLA+ CD8+ cells than healthy controls. Furthermore, a much stronger correlation was observed between PASI and the frequency of CLA+ CD8+ than CLA+ CD4+ T cells. The frequency of CLA+D8+ T cells correlated more strongly with redness, thickness and scaling of the skin lesions than the total affected body surface area. In contrast to CLA the T cell expression of VLA-4 did not demonstrate any such correlation. Finally, the expression of the activation marker CD25 on CD8+ T cells showed a strong correlation with disease severity in patients with moderate to severe psoriasis (PASI > 10) but such correlation was not observed for CD4+ T cells. These findings support the notion that circulating CLA+ CD8+ T cells may play an important role in the pathogenesis of psoriasis.     

慢性乙型肝炎患者外周血中CD4+CD25+调节性T细胞的变化和意义

目的 观察慢性乙型肝炎感染者外周血CD4 CD25 调节性T细胞(Treg)的频率,特征性表型及与临床指标的相关性.方法 采集44例慢性乙型肝炎,29例健康人外周血,多色流式分析Treg的频率,CD4 CD25 Foxp3 调节性T细胞的频率.所有病例均经酶联免疫吸附实验(ELISA)检测HBsAg、HBsAb、HBeAg、HBeAb和HBcAb,实时荧光定量RT-PCR检测血清HBV DNA载量及肝功能的检测.结果 慢性乙型肝炎组Treg的频率(5.02%±2.07%)显著高于正常对照组(3.35%±1.51%,P＜0.05);Foxp3在Treg上特异性表达,慢性乙型肝炎组CD4 CD25 Foxp3 调节性T细胞的频率(2.05%±1.03%)显著高于正常对照组(1.30%±0.68%,P＜0.05);慢性乙型肝炎组Treg的频率与病毒载量呈正相关.结论 Treg在慢性乙型肝炎感染者中明显增高,并与病毒载量相关,提示Treg在慢性乙型肝炎中担负着重要的免疫调节作用,可能是造成乙肝病毒感染慢性化的重要因素.     

Type 1 and type 2 cytokine-producing mouse CD4+ and CD8+ T cells in acute Schistosoma mansoni infection

CD4⁺ and CD8⁺ T cells can be divided based on the cytokines that they secrete into type 1 (Th1, Tc1) and type 2 (Th2, Tc2) subsets. Schistosoma mansoni infection in mice is characterized by a type 2-dominated response. We have used intracellular cytokine staining to demonstrate dramatic changes in the relative numbers of Tc1 and Th2 cells in the spleens of mice during acute schistosome infection. In infected mice prior to egg laying a generalized type 1 response dominated, and was associated with an expansion in the frequency of Tc1 and Th1 cells. By week 7 after infection the cytokine response was of type 2, with an increase in the numbers of Th2 cells and a dramatic reduction in the frequency of Tc1 cells. Following the onset of egg laying there was apoptosis of cells in the spleens of mice, with CD4⁺ and in particular CD8⁺ T cells undergoing apoptosis. The loss of CD8⁺ T cells may in part be attributable to the development of a type 2 environment, following egg laying, with type 2 responses mediating the apoptosis of Tc1 cells. Schistosome regulation of Tc1 during egg laying may be required to prevent type 1 inflammatory responses from exacerbating egg-induced pathology.     

Circulating CD137+CD8+ T cells accumulate along with increased functional regulatory T cells and thoracic tumour burden in lung cancer patients

CD137 is a promising target for immunostimulation strategies against cancer. Previous studies showed that CD137⁺CD8⁺ T cells are enriched in antitumour effector T cells in both preclinical tumour models and cancer patients, but to date, such T cells in the blood of lung cancer patients have not been sufficiently investigated. In this study, circulating antigen‐activated CD8⁺ T cell subsets, identified as CD137⁺CD8⁺ or PD‐1⁺ (programmed cell death protein 1) CD8⁺, and regulatory T cells (Treg), identified as CD4⁺CD25⁺CD127^low/?, in 40 untreated lung cancer patients and in 49 age‐ and sex‐matched healthy controls (HCs) were assessed by flow cytometry. Results were evaluated for associations with lung cancer patient clinical characteristics. Correlations between antigen‐activated CD8⁺ T cells and effector Treg (CTLA‐4⁺ [cytotoxic T‐lymphocyte antigen 4] CD4⁺CD25⁺CD127^low/?) were also investigated. Higher percentages of PD‐1⁺, CD137⁺ and PD‐1⁺CD137⁺ amongst CD8⁺ T cells were observed in lung cancer patients compared with HCs. The percentages of CD137⁺CD8⁺ and PD‐1⁺CD137⁺CD8⁺ T cell subsets amongst CD8⁺ T cells were positively correlated with thoracic tumour burden and were strongly positively correlated with the percentage of effector Treg subset. Smoking patients harboured higher percentages of the PD‐1⁺CD8⁺ T cell subset compared with non‐smoking patients. This study demonstrated that circulating antigen‐activated CD8⁺ T cells accumulated in lung cancer patients along with increased effector Treg and thoracic tumour burden. These findings aid a better understanding of immune‐host interactions in lung cancer patients using peripheral blood, and further support immunotherapeutic intervention strategies using combination therapy for differential control of Treg and activation of tumour‐specific effector T cells.     

Upregulation of CCR4 in activated CD8+ T cells indicates enhanced lung homing in patients with severe acute SARS-CoV-2 infection

COVID-19 is a life-threatening disease leading to bilateral pneumonia and respiratory failure. The underlying reasons why a smaller percentage of patients present with severe pulmonary symptoms whereas the majority is only mildly affected are to date not well understood. Comparing the immunological phenotype in healthy donors and patients with mild versus severe COVID-19 shows that in COVID-19 patients, NK-/B-cell activation and proliferation are enhanced independent of severity. As an important precondition for effective antibody responses, T-follicular helper cells and antibody secreting cells are increased both in patients with mild and severe SARS-CoV-2 infection. Beyond this, T cells in COVID-19 patients exhibit a stronger activation profile with differentiation toward effector cell phenotypes. Importantly, when looking at the rates of pulmonary complications in COVID-19 patients, the chemokine receptor CCR4 is higher expressed by both CD4 and CD8 T cells of patients with severe COVID-19. This raises the hypothesis that CCR4 upregulation on T cells in the pathogenesis of COVID-19 promotes stronger T-cell attraction to the lungs leading to increased immune activation with presumably higher pulmonary toxicity. Our study contributes significantly to the understanding of the immunological changes during COVID-19, as new therapeutic agents, preferentially targeting the immune system, are highly warranted.     

A profile of TNFR2+ regulatory T cells and CD103+ dendritic cells in the peripheral blood of patients with asthma

The interaction of tolerogenic CD103⁺ dendritic cells (DCs) with regulatory T (Tregs) cells modulates immune responses by inducing immune tolerance. Hence, we determined the proportion of these cells in the peripheral blood mononuclear cells (PBMC) of asthmatic patients. We observed lower trends of CD11b^-CD103⁺ DCs and CD86 within CD11b^-CD103⁺ DCs, while increased levels of Foxp3 expressing CD25^+/-TNFR2⁺ cells in asthmatics. There was a positive correlation in the expression of Foxp3 within CD3⁺CD4⁺CD25⁺TNFR2⁺ Tregs and CD11b^-CD103⁺ as well as the expression of CD86 within HLA-DR⁺CD11c⁺CD11b^-CD103⁺ DCs. In conclusion, we suggest that the increased levels of Tregs in blood could continuously suppress the T helper 2 (Th2) cells activation in the circulation which is also supported by the increase of anti-inflammatory cytokines IL-10 and TNF. Overall, functional immunoregulation of the regulatory cells, particularly Tregs, exhibit immune suppression and induce immune tolerance linked with the immune activation by the antigen presenting cells (APC).     

CD30 expression on circulating memory CD4+ T cells as a Th2-dominated situation in patients with atopic dermatitis

Th2 clones have been reported to express CD30 preferentially, but whether T cells producing Th2-type cytokines may favor CD30 expression in the in vivo state remains unknown. We investigated the expression of CD30 on circulating T cells in atopic dermatitis (AD) as a Th2-dominated disorder. Peripheral blood mononuclear cells were prepared from 51 AD patients and 14 nonatopic controls, and their phenotypes were analyzed with flow cytometry. Cytokine production by stimulated CD4+ T cells was also assessed by the single-cell-staining method. Flow cytometric analysis clearly revealed that CD30+ T cells were identifiable in the blood of AD patients with greater frequency compared to controls. The important finding was that CD30 expression was restricted to a small but substantial population of memory (CD45RO+) CD4+ T cells, but not CD8+ ones. In AD patients, it was demonstrated that the percentages of CD30+ cells within CD45RO+ CD4+ T cells correlated well with the disease severity, serum IgE levels, peripheral eosinophil counts, and tendency toward Th2-dominant cytokine pattern as determined by the ratio of interleukin-4 to interferon-gamma production. This study suggests that CD30 expression in circulating T cells might serve as an in vivo marker for the Th2-dominated condition.     

The frequency of QAP2.1 is increased in psoriasis vulgaris patients

The heritage of psoriasis has a polygenic mode, the most essential antigens being Cw6,DR7,DQA1^*0201 in many ethnic groups. We have found a strong association between the high risk HLA haplotypes carrying DQA1^*0201 and psoriasis. Thus, the aim of this study was to further examine if the flanking promotor genes, URRs of DQ (QAP and QBP) are involved in the susceptibility to this disease. The series consisted of 62 patients and 50 control subjects and the PCR-SSO method was used. The frequency of the promoter gene QAP2.1 was significantly increased in the psoriatics (P_c=1.5×10⁻², RR=4.6) and the frequency of QAP4.1 was decreased in the patients group (P_c=3.9×10⁻²). The psoriasis risk allele DQA1^*0201 was associated always with QAP2.1, except once with QAP3.1.

: The combination of QAP2.1 and DQA1^*0201 is associated to psoriasis. Promotor region could exert its influence by gene expression or functionally through different inducibility by cytokines.     

类风湿性关节炎患者外周血TH1/TH2细胞的研究

目的探讨CD4+TH1/TH2细胞在类风湿性关节炎(RA)发生发展过程中的作用.方法采用酶联免疫斑点法(ELISPOT)对15例RA患者和30例健康人外周血中T淋巴细胞亚群及CD4+TH1/TH2功能亚型进行检测.结果RA患者外周血中TH1细胞的百分率较正常对照组升高(P＜0.05).结论 TH1细胞介导的细胞免疫可能与RA的发生发展有关.     

Selective depletion of Vbeta2+CD8+ T cells in peripheral blood from rheumatic heart disease patients

Acute rheumatic fever (ARF) and its chronic valvular sequelae are the delayed consequence of a pharyngeal infection with group A Streptococcus (GAS). Several GAS proteins have been shown to be superantigens, raising the possibility that the expansion or deletion of T cells expressing specific Vbeta regions might play a role in the pathogenesis of ARF or chronic rheumatic heart disease (RHD). We therefore analyzed by four-color flow cytometry, the Vbeta repertoire on CD3, CD4 and CD8 T cells from four ARF patients, 10 RHD patients and also nine healthy controls. A selective depletion of Vbeta2+ T cells was found only in the CD8 subset of chronic RHD patients. This is of interest since a number of GAS superantigens exert their effects on Vbeta2+ cells and because only CD8+ T cells from ARF and RHD patients undergo anergy in response to GAS superantigens. Our results suggests that an ongoing immune process is present in RHD patients and that CD8+ T cells may have an important immunoregulatory role in the pathogenesis of the disease.     

替比夫定对慢性乙型肝炎患者外周血CD4~+ CD25~+ CD127~(low) T和CD8~+ CD25~+ T细胞频率的影响及临床意义

目的探讨核苷类药物替比夫定对慢性乙肝患者(CHB)外周血中CD4+CD25+CD127lowT细胞和CD8+CD25+T细胞比例的影响,并结合临床指标分析其临床意义。方法替比夫定抗病毒治疗22例CHB患者,在治疗前及治疗后3,6个月时,分别以流式细胞仪检测外周血中CD4+CD25+CD127lowT细胞和CD8+CD25+T细胞比例,实时荧光定量RT-PCR检测Foxp3 mRNA的表达水平,荧光定量PCR检测血清HBV DNA水平,酶联免疫吸附法检测HBV标志物,全自动生化分析仪检测ALT水平。结果 CHB患者外周血中CD4+CD25+CD127lowT细胞和CD8+CD25+T细胞比例显著高于对照组。替比夫定治疗3个月时,这两群细胞比例显著下降,Foxp3 mRNA的表达也显著下降;HBV DNA水平降至检测水平以下的CHB患者,其CD4+CD25+CD127lowT细胞也降至正常水平。治疗3、6个月时,HBeAg阴转率分别为9.1%和18.2%,发生HBeAg血清学转换者的CD4+CD25+CD127lowT细胞和CD8+CD25+T细胞比例均降至正常水平。结论替比夫定能快速有效抑制CHB患者的病毒复制...     

乙型肝炎病毒感染者外周血CD4＋T细胞CD25-CD127-表达的检测及临床意义

目的 检测乙型肝炎病毒（ HBV）感染者外周血CD4＋T细胞表面CD25 - CD127 -的表达情况及临床意义.方法 用流式细胞术比较分析53例慢性乙肝患者、53例HBV携带者和26例正常对照人群CD4＋T细胞表面CD25 - CD127 -的表达情况,并对20例用干扰素治疗的HBV-DNA阳性慢性乙肝患者随访.结果 ①与正常对照组比较,慢性乙肝患者、HBV携带者CD4＋ CD25 -CD127 -T细胞均显著升高,两者比较有统计学差异（Q =4.559,P＜0.05;Q=6.230,P＜0.05）;②HBV- DNA阳性患者（n=77） CD4＋ CD25 - CD127 -T细胞显著低于HBV- DNA阴性患者（n=29）,两者比较有统计学意义（t =2.290,P=0.024）;③与治疗前比,慢性乙肝患者干扰素治疗12周后CD4＋CD25- CD127 -T细胞显著降低,两者比较差异有统计学意义（t =2.469,P=0.024）.结论 乙型肝炎病毒感染者外周血CD4＋ CD25 - CD127 -T细胞与病毒的感染和清除相关,外源性干扰素可降低CD4＋ CD25 - CD127 -T细胞.