蕈样肉芽肿患者外周血单一核细胞CD45RA及CD45RO表达的研究

目的探讨蕈样肉芽肿（Mycosis fungoides,MF）患者外周血单个核细胞CD45RA及CD45RO的表达及其与MF发病的关系。方法应用双荧光抗体标记、流式细胞仪检测15例MF患者外周血单个核细胞CD45RA及CD45RO的表达。结果（1）MF患者外周血CD3^＋、CD3^＋CD8^＋细胞与正常对照比较差异不显著（P〉0.05）。（2）MF患者外周血CD4^＋细胞低于正常对照,差异非常显著（P〈0.001）。（3）MF患者外周血T细胞CD3^＋CD4^＋/CD3^＋CD8^＋比值低于正常对照,差异显著（P〈0.001）。（4）MF患者外周血CD45RA^＋细胞低于正常对照,差异非常显著（P〈0.001）,CD45RO^＋细胞高于正常对照,差异非常显著（P〈0.001）。（5）MF患者外周血CD45RO^＋/CD45RA^＋比值高于正常对照,差异非常显著（P〈0.001）。（6）MF患者外周血CD4^＋CD45RA^＋细胞低于正常对照,差异非常显著（P〈0.001）。（7）MF患者外周血CD4^＋CD45RO^＋细胞及CD8^＋CD45RO^＋细胞均高于正常对照,差异非常显著（均P〈0.001）。（8）MF患者外周血CD4^＋CD45RO^＋/CD4^＋CD45RA^＋比值及CD8^＋CD45RO^＋/CD8^＋CD45RA^＋比值均高于正常对照,差异非常显著（P〈0.01及P〈0.001）。结论MF患者外周血中,不仅存在CD4^＋亚群失调和CD4^＋/CD8^＋比值降低,而且在CD4^＋和CD8^＋亚群中也存在CD45RA^＋、CD45RO^＋亚群失调和CD45RO^＋/CD45RA^＋比值升高,从而导致的机体免疫功能紊乱,可能与MF的发病或病情加剧有关。     

人胎盘间充质干细胞CD200+亚群细胞对同种异基因移植排斥的调节效应

文题释义： CD200⁺亚群细胞：从人胎盘间充质干细胞中分选获得的高表达CD200抗原的一群细胞。属于免疫球蛋白超级家族的膜糖蛋白CD200在调节免疫反应方面很重要,在维持免疫稳态方面起着关键作用。 移植排斥：在同种异基因组织、器官移植中,受者的免疫系统会对移植物产生排斥反应,这是一个涉及多种免疫反应的免疫学现象。排斥反应的轻重取决于供者与受者之间人类主要组织相容抗原的差异程度。 背景：免疫排斥反应仍是皮肤异体移植面临的重要难题,实验室前期研究发现高表达CD200的人胎盘间充质干细胞亚群细胞具备较强的免疫调节能力。 目的：进一步研究人胎盘间充质干细胞CD200⁺亚群细胞对同种异基因移植排斥的调节作用。 方法：构建同种异基因小鼠皮肤移植模型,经尾静脉分别将PBS(对照组)、人胎盘间充质干细胞(PMSCs组)、人胎盘间充质干细胞CD200⁺亚群细胞(CD200⁺-PMSCs组)输注C57BL/6小鼠体内。观察移植物的开始坏死时间、存活时间、皮片状态;细胞治疗7 d,采集小鼠外周血进行白细胞计数,采用Q-PCR、ELISA方法检测小鼠脾脏与外周血中白细胞介素10、干扰素γ及肿瘤坏死因子α的表达。 结果与结论：①与对照组相比,PMSCs组与CD200⁺-PMSCs组移植物状态良好,存活时间明显延长(P < 0.001);CD200⁺-PMSCs组移植物状态及存活时间均优于PMSCs组(P < 0.01);②细胞治疗7 d,PMSCs组与CD200⁺-PMSCs组白细胞数量明显少于对照组(P < 0.01);③与对照组相比,CD200⁺-PMSCs组脾脏中白细胞介素10 mRNA表达明显升高(P < 0.05),PMSCs组、CD200⁺-PMSCs组干扰素γ及肿瘤坏死因子α mRNA表达量则明显下调(P < 0.05,P < 0.01),同时CD200⁺-PMSCs组干扰素γ及肿瘤坏死因子α mRNA表达量明显低于PMSCs组(P < 0.01,P < 0.05);④与对照组相比,PMSCs组、CD200⁺-PMSCs组血液中白细胞介素10水平明显升高(P < 0.05,P < 0.01),而干扰素γ及肿瘤坏死因子α水平则明显下调(P < 0.05,P < 0.001;P < 0.01,P < 0.001),同时CD200⁺-PMSCs组干扰素γ及肿瘤坏死因子α水平明显低于PMSCs组(P < 0.05);⑤结果表明,人胎盘间充质干细胞对同种异基因皮肤移植排斥具有调节作用;CD200⁺亚群细胞抑制免疫排斥反应能力更强,其机制可能是CD200通过调节白细胞介素10、干扰素γ及肿瘤坏死因子α等免疫相关因子参与免疫反应。 ORCID: 0000-0002-9945-9094(刘婷) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

Human alpha defensin in HIV-exposed but uninfected individuals

Human alpha defensins 1, 2, and 3 are produced by CD8 T cells of HIV-infected long-term nonprogressors and have an antiviral activity. alpha Defensins were examined in peripheral blood mononuclear cells (PBMCs), cervical-vaginal mononuclear cells (CVMCs), and cervical biopsies of 9 HIV-1-exposed but uninfected women (ESNs), 10 HIV-infected patients (HIV), and 13 low-risk healthy controls (HCs). Results showed that, whereas alpha defensin production and alpha defensin-expressing CD8 lymphocytes were comparable in ESNs and HIV patients, constitutive alpha defensin production by peripheral CD8 and CVMCs was augmented in ESNs compared with HCs (P = 0.001 and P = 0.058, respectively); alpha defensin mRNA was increased in PBMCs of ESNs; unstimulated, alpha defensin-expressing peripheral and mucosal CD8 lymphocytes were 10-fold higher in ESNs compared with HCs (P = 0.003 and P = 0.01, respectively); and alpha defensin mRNA and alpha defensin-expressing cells were augmented in cervical biopsies of ESN compared with HCs (mRNA:P = 0.03). The differences were reduced upon in vitro mitogen stimulation. A robust constitutive production of alpha defensin is seen in HIV-exposed uninfected individuals; these peptides could have a role in the potentially protective immune response that characterizes ESNs.     

Increased Microchimeric CD4 T Lymphocytes in Peripheral Blood from Women with Systemic Sclerosis

Recent studies have demonstrated the presence of microchimeric cells in peripheral blood and skin lesions from patients with systemic sclerosis (SSc). In a previous study we found that some peripheral blood CD3⁺ cells from female patients with SSc contained male DNA. Here, peripheral blood samples from 47 patients with SSc (30 with diffuse cutaneous SSc and 17 with limited cutaneous SSc) and 22 healthy controls were sorted for CD4⁺ and CD8⁺ T cells. Both positively and negatively selected populations were analyzed for male DNA by quantitative PCR. Analysis of Y chromosome sequences in the sorted cells demonstrated the presence of microchimerism in 82.9% of SSc patients compared to 63.6% of controls. The numbers of CD4⁺ and CD8⁺ T cells were found to be significantly higher in the SSc patients than in controls. Furthermore, patients with dcSSc were observed to have significantly more CD4⁺ microchimeric T cells than the controls. In the CD8⁺ T-cell population, there was a trend toward more microchimeric cells in the patients but this did not reach significance. These results support the hypothesis that microchimeric CD4⁺ T cells may be involved in the pathogenesis of SSc.     

Expression of activation, adhesion molecules and intracellular cytokines in acute pancreatitis.

Adhesion and activation molecules as well as cytokines play an important role in an immune scenario. In acute pancreatitis, we have studied some of these in order to evaluate dysregulation. For this we took peripheral blood mononuclear cells and pancreatitis tissue cells. We analysed activation markers like CD69, CD25 and HLA-DR and found a marked elevation of CD69 as well as CD25 in both peripheral blood cells and tissue mononuclear cells when compared to controls. In PBMC-CD69: P<0.01 and CD25: P<0.01; in tissue-CD69: P<0.001 and CD25: P<0.001. The HLA-DR levels, however, were reduced in the disease state (in acute pancreatitis patient blood (P<0.01) and tissue cells (P<0.001)). The adhesion molecules showed unanimous rise in the blood and the tissue samples. In blood samples, CD11a: P<0.05 and CD11b: P<0.05 and tissue samples CD11a: P<0.01 and CD11b: P<0.01and CD54 in peripheral blood (P<0.05) and tissue (P<0.01) of AP was high as compared to controls. By simultaneous flowcytometric analysis, we determined the co-expression of a surface marker (CD4/CD8/CD14) and intracellular cytokine (TNF-alpha and IFN-gamma) in individual cells. The IFN-gamma producing CD8+T cells were elevated in pancreatic tissue (P<0.01). TNF-alpha producing cell numbers were significantly higher in tissue cells than in blood and also in CD8+ T cells (P<0.001). We conclude that monocyte function is affected in AP as shown by reduced HLA-DR numbers and lowered TNF-alpha producing cells. Moreover, the CD8+T cells appear to play an important role in cytokine synthesis at the effector site.     

IL-27诱导原发性胆汁性肝硬化患者CD4+T细胞增殖分化作用机制研究

目的 探讨IL-27对原发性胆汁性肝硬化(primary biliary cirrhosis,PBC)患者外周血CD4+T细胞的增殖分化作用及其相关免疫学机制.方法 收集PBC患者、慢性乙肝患者(choronic hepatitis B,CHB)、健康体检者(health controls,HCs)外周血,磁珠分离CD4+T细胞.IL-27体外作用后,CCK-8测定细胞增殖情况,ELISA法检测细胞因子,定量PCR分析T-bet和GATA3基因表达情况,免疫印迹测定p-STAT-1和p-STAT-3的表达.结果 IL-27作用后,PBC组、CHB组和HCs组CD4+T细胞增殖能力均显著增强,PBC组CD4+T细胞增殖能力强于CHB和HCs组,差异有统计学意义(P＜0.001),同时PBC组细胞培养液中IL-2和IFN-γ在IL-27作用后较CHB和正常对照组均显著增高(P＜0.001),IL-10表达无明显变化.未经IL-27诱导情况下,PBC组T-bet表达高于CHB组(P=0.007),IL-27诱导后PBC组CD4+T淋巴细胞T-bet基因表达显著增加,同时对GATA3具有抑制作用,作用前后差异有统计学意义(P＜0.001),CHB组作用前后无显著变化(P=0.3).免疫印迹测定p-STAT-1、p-STAT-3发现,正常情况下各研究组均不表达p-STAT-1、p-STAT-3,IL-27作用后,表达明显升高,其中PBC组升高尤为明显.结论 IL-27可以诱导PBC患者CD4+T细胞增殖,并通过活化p-STAT-1、p-STAT-3信号通路,诱导CD4+T细胞向Th1分化,同时分泌相关细胞因子,可能在PBC早期免疫炎症反应过程中具有重要作用.     

Stage-dependent detection of CD14+ and CD16+ cells in the human heart after myocardial infarction

Monocytes are critically involved in cardiovascular wound healing processes. Human monocytes can be classified into two subsets based on the expression of CD14 and CD16. Here, we examined the temporal and spatial distribution of CD14⁺ and CD16⁺ cells after myocardial infarction (MI) in human heart and spleen tissue and correlated it with markers of cardiac repair. Heart samples obtained at autopsy were histologically classified into acute (AMI; n?=?11), subacute (SAMI; n?=?10) and old (OMI; n?=?16) MI, or control myocardium (CONTR; n?=?8). Histochemical analyses revealed marked fibrosis in OMI (p?<?0.001 vs. CONTR). The adhesion molecule CD56 was also strongly expressed in OMI (p?<?0.01 vs. CONTR) and found to correlate with fibrosis (p?<?0.001). The number of capillaries was reduced in OMI (p?<?0.01 vs. CONTR; p?<?0.05 vs. AMI), whereas the hypoxia indicator carbonic anhydrase IX was predominantly expressed in AMI (p?<?0.01 vs. OMI and CONTR) and SAMI (p?<?0.05 vs. OMI and CONTR). The monocyte chemoattractrant osteopontin was also more highly expressed in hearts of SAMI patients (p?<?0.01 vs. CONTR). Numbers of CD14⁺ monocytes were found to correlate with CD16⁺ cells (p?<?0.05) and inversely with fibrosis (p?<?0.05). Regarding a MI-associated release of monocytes from spleen reservoirs, a non-significant reduction of splenic CD14⁺ and CD16⁺ cells was detected in subjects with AMI. In conclusion, disease stage-specific alterations in CD14⁺ and CD16⁺ cells in human heart may contribute to cardiac repair processes following MI.     

Characterization of bronchoalveolar lavage T cell subsets in sarcoidosis on the basis of CD57, CD4 and CD8

T cells expressing CD57 (a natural killer cell marker) with interferon-gamma (IFN-gamma) producing capacity increase under various conditions. CD57+ T cells are also present in the bronchoalveolar lavage fluid (BALF) of sarcoidosis, and several phenotypical and functional analyses of these cells have been reported. In the present study, BALF T cells obtained from 52 patients with sarcoidosis were classified further into CD4+CD57+ T cells, CD4+CD57- T cells, CD8+CD57+ T cells and CD8+CD57- T cells and their phenotypes and functional characteristics were assessed. Substantial proportions of these T cell subsets expressed natural killer cell markers CD161 and CD122. The biased expansion of Vbeta2 T cells was observed in both CD4+CD57+ T cells and CD4+CD57- T cells in BALF from most patients, while the expansion of other Vbeta T cells was also observed in some patients. Unexpectedly, the biased expansion of certain Vbeta T cells was also seen in either CD8+CD57+ T cells or CD8+CD57- T cells, while the expanded Vbeta T cells in CD8+ T cells differed substantially among individuals. BALF T cells showed a remarkably lower T cell receptor (TCR) intensity than that of peripheral blood T cells. Both CD8+ T cell subsets in BALF of sarcoidosis expressed the intracellular perforin/granzyme B, while all four subsets expressed intracellular IFN-gamma after in vitro activation, and CD4+ T cells, especially CD4+CD57+ T cells, expressed tumour necrosis factor-alpha. These findings indicate that CD57+ T cells as well as CD57- T cells in the BALF are phenotypically and functionally different from peripheral blood T cells and may play an important role in the Th1 dominant state and inflammation in pulmonary sarcoidosis.     

Distinct immune profiles characterize patients with diffuse or limited systemic sclerosis

Mitogen-stimulated IL-2, IFN-gamma, TNF-alpha (type 1 cytokines), and IL-10 (type 2 cytokine) production by peripheral blood mononuclear cells, as well as expression of surface markers on immune cells, was evaluated in systemic sclerosis (SSc) patients. Fifty-four SSc patients with either diffuse (dSSc) or limited (lSSc) disease and 20 age- and sex-matched healthy controls (HCs) were examined. Fourteen patients were treated with prednisone and 9 patients with prednisone and cyclophosphamide pulses. Results showed that (1) IL-2 production is significantly decreased, whereas IL-10 is higher in untreated patients compared to HCs; IL-10, IFN-gamma, and TNF-alpha production is higher in lSSc compared to dSSc patients; (2) CD4+25+ (IL-2R), CD8+, and CD8+45RA-28+57- (memory) lymphocytes are reduced in patients compared to HCs; (3) CD95-expressing CD4+ and CD8+ cells are significantly higher in dSSc patients; and (4) steroids are more effective alone than in combination with cyclophosphamide in reducing IL-10 and IFN-gamma production in these patients. These results confirm that a complex imbalance in cytokine production is present in SSc patients and suggest that peculiar phenotypic populations are underrepresented in these patients. Overexpression of Fas in dSSc could results from the attempt of the immune system to induce apoptosis of autoreactive T-cell clones.     

CD4+CD25+调节性T细胞与系统性红斑狼疮病情活动性关系的研究

目的：检测系统性红斑狼疮患者外周血CD4^＋CD25^＋、CD4^＋CD8^＋调节性T细胞亚群，探讨其与疾病活动性、肾脏损伤、血清抗ds-DNA抗体及免疫球蛋白和补体C3含量的关系。方法：采用流式细胞术检测北京协和医院住院和门诊SLE患者（n=37）外周血CD4^＋CD25^＋T、CD4^＋CD8^＋T细胞群比例，以15例RA和15例SS组成自身免疫性疾病对照，30例健康体检者作为正常对照，观察调节性T细胞亚群与SLE患者疾病活动性指标SLEDAI、IgG、C3及血清抗ds-DNA抗体的关系。结果：①疾病活动期SLE患者外周血CD4^＋CD25^＋调节性T细胞群比例显著低于正常对照组（P〈0.01），疾病稳定期和风湿性疾病对照组与正常对照组结果差异无统计学意义。疾病活动期和稳定期SLE患者CD4＋CD8＋T细胞群比例都略高于正常对照组，但未发现结果差异有统计学意义（P〉0.05）。②疾病活动期SLE患者外周血CD4^＋CD25^＋T细胞比例及CD4^＋CD25^＋/CD4^＋值显著低于稳定期患者（P〈0.01）。SLE患者外周血CD4^＋CD25^＋/CD4^＋值与SLEDAI、补体C3呈低度相关（r分别为-0.491、0.368，P〈0.05），CD4^＋CD25^＋T细胞数量与SLEDAI呈负相关（r=-0.578，P〈0.05）。③SLE并发肾病组外周血CD4^＋CD25^＋T细胞群比例及CD4^＋CD25^＋/CD4^＋值显著低于非肾病组（P〈0.01；P〈0.05）。同一SLE患者治疗前后CD3^＋CD4^-CD8^-细胞和NK细胞降低，CD4^＋CD25^＋细胞、CD4^＋CD25^＋/CD4^＋值及CD8^＋T细胞增加，但未发现这些结果差异有统计学意义。本次研究未发现NK细胞、CD4^＋CD8＋T细胞、CD4^＋CD25^＋T细胞群比例在ds-DNA＋组与ds-DNA-组之间结果差异有统计学意义。结论：SLE患者外周血CD4^＋CD25^＋T细胞群比例与SLEDAI成负相关，与肾脏的损害也有密切关系，但与血清抗ds-DNA抗体产生的关系不明显。活动期SLE患者外周血CD4^＋CD25^＋T细胞减少，稳定期CD4^＋CD25^＋T细胞比例回升，因此推测CD4^＋CD25^＋T细胞的变化可能是导致疾病发生和病情发展及相关器官（如肾脏）损伤的关键环节之一。     

Increased proportion of CD3+CD4-CD8- double-negative T cells in peripheral blood of children with Behcet's disease

INTRODUCTION: Behcet's disease (BD) is a multi-system inflammatory disorder of poorly understood pathogenesis, which is characterized by oral aphtosis, genital ulcers and uveitis. OBJECTIVE: To assess the role of CD3+CD4-CD8- double negative (DN) T cells in pathogenesis of Behcet's disease. PATIENTS: Ten BD patients (age 12.2+/-2.2 years, 7 in remission, 3 in exacerbation state) treated at the Pediatric Rheumatology unit of Soroka University Medical Center and 3 age-matched controls participated in the study. METHODS: Peripheral blood lymphocytes of study subjects were isolated and stained with fluorescein-labeled anti-CD45, CD3, CD4, CD8 antibodies and analyzed by FACS assay. RESULTS: Proportion of CD4-CD8- DN T cells was significantly increased in BD patients (n=10) as compared to healthy controls (6.2+/-3.4% vs. 3.2+/-1.1% of total CD3+ cells, p<0.05), this cell group was additionally enhanced in BD exacerbation, compared to patients in remission (10+/-4.1% vs. 4.7+/-1.2%, p<0.05, respectively). DN T cells were significantly increased in BD patients in remission, compared to healthy controls (4.7+1.2% vs. 3.2+1.1% of total CD3+ cells, p<0.05, respectively). CONCLUSIONS: Behcet's disease is characterized by increased proportion of CD3+CD4-CD8- double negative T cells in peripheral blood. Further studies, that include additional immunophenotyping and analysis of gene expression, aimed at characterization of these cells are currently underway.     

人外周血CD8+T细胞CD28、CD56及CD57表达水平的老年性改变

目的:通过对健康老年人与青年人外周血CD8⁺T细胞CD28、CD56及CD57表达水平的比较性研究,探讨免疫衰老的细胞学机制。方法:采用三色免疫荧光标记流式细胞术分析青年组(20-35岁)与老年组(60-75岁)外周血CD8⁺CD28⁺、CD8⁺CD56⁺及CD8⁺CD57⁺T细胞水平。结果:老年组外周血CD8⁺CD28⁺T细胞明显低于青年组,阳性百分率分别为34.07±5.29和49.84±7.43(P＜0.05);而老年组CD8⁺CD56⁺T细胞及CD8⁺CD57⁺T细胞均明显高于青年组,前者阳性百分率分别为6.60±2.40和2.10±0.35,后者阳性百分率分别为41.82±6.01和22.89±2.80(P＜0.05)。结论:老年人CD8⁺T细胞CD28、CD56及CD57的表达率均随年龄增长有明显改变;CD28表达下降可能是引起免疫系统功能降低的重要原因,而CD56、CD57表达水平的增加则可能是机体对细胞免疫功能下降的一种代偿性适应。     

SLE患者外周血中Foxp3+CD4+CD25+调节性T细胞的分析

目的分析SLE患者外周血中Foxp3 CD4 CD25 调节性T细胞和T细胞亚群上GITR的表达,以初步阐述其在SLE患者免疫稳态调节中的作用和意义。方法以流式细胞术检测SLE患者外周血中Foxp3 CD4 CD25 调节性T细胞和T细胞亚群上GITR的表达。结果稳定期及活动期SLE患者外周血中Foxp3 CD4 CD25 调节性T细胞比例显著低于健康对照(P<0.05),且活动期SLE患者Foxp3 CD4 CD25 调节性T细胞比例高于稳定期SLE患者(P>0.05)。SLE患者外周血CD3 CD4 T细胞和CD3 CD8 T细胞上GITR表达显著增加(P<0.05);随着疾病活动性增加,CD3 CD4 T细胞上GITR表达降低(P>0.05),CD3 CD8 T细胞上GITR表达增加(P>0.05)。结论SLE患者外周血中Foxp3 CD4 CD25 调节性T细胞表达降低,而患者T细胞亚群上GITR表达增加,其共同作用在诱导SLE患者外周耐受障碍中具有重要意义。     

CD4+CD25+ regulatory T cells in systemic sclerosis and other rheumatic diseases

Systemic sclerosis (SSc) is a generalized connective tissue disorder, characterized by a wide spectrum of microvascular and immunological abnormalities, leading to a progressive thickening and fibrosis of the skin and other organs, such as the lungs, GI tract, heart and kidneys. SSc is thought to be an autoimmune disease owing to the presence of high affinity antibodies and possible clinical overlap with other autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis. Autoimmune diseases arise because of a breakdown in immunological self tolerance. Self tolerance is maintained via multiple regulatory mechanisms within the immune system, including the thymic deletion of self-reactive T cells and mechanisms of peripheral tolerance. In recent years, the presence of CD4(+)CD25(+)FOXP3(+) Tregs has been identified as a major mechanism of peripheral tolerance, and accumulating evidence indicates that alterations in Treg frequencies and/or function may contribute to autoimmune diseases. Here, we will review recent data on the percentage, function and phenotype of CD4(+)CD25(+) Tregs in rheumatic disease, and discuss how recent developments may guide research in this area in SSc.     

Contribution of CD8+ T cells to inflammatory cytokine production in systemic sclerosis (SSc)

Only limited attention has been paid to the role of CD8?+?T cells in the etiopathogenesis and progression of systemic sclerosis (SSc). CD8?+?T cells may have autoantigen-specific and pro-inflammatory but also immunomodulatory properties. To investigate the differentiation of CD8?+?T cells, staining of cell surface factors and of chemokine receptors were performed. In addition, the cytokine-producing ability of circulating CD8?+?T cells and their sensitivity to suppression by regulatory T cells (Tregs) were compared between patients with diffuse (dcSSc) or limited cutaneous SSc (lcSSc) and healthy individuals. We identified CD8?+?T cells as producers of pro-inflammatory type-2 cytokines with a significant contribution of memory CD8?+?T cells. Memory CD8?+?T cells of SSc patients stayed unaltered after suppression with autologous Tregs. Expression of chemokine receptors was significantly correlated with intracellular cytokine production in CD8?+?T cells with a clear dichotomy of type 1 and type 2 cytokines. High levels of intracellular cytokines, such as interleukin-(IL)-4, IL-13 and tumor-necrosis-factor-alpha (TNFalpha) were positively associated with the presence of Scl-70 or anti-centromere antibodies and negatively with the administration of glucocorticoids. Administration of glucocorticoids was positively associated with higher IFNgamma production. Lack of anti-centromere antibodies and therapy with methotrexate were positively associated with higher intracellular IL-10 production. CD8?+?T cells may significantly contribute to inflammation in SSc. Our findings suggest to not only focus on T helper cells in the development of therapeutic strategies but also to consider the role of CD8?+?T cells in the etiopathogenesis and perpetuation of SSc.     

Positive selection of self‐antigen‐specific CD8+ T cells by hematopoietic cells

In contrast to thymic epithelial cells, which induce the positive selection of conventional CD8⁺ T cells, hematopoietic cells (HCs) select innate CD8⁺ T cells whose Ag specificity is not fully understood. Here we show that CD8⁺ T cells expressing an H‐Y Ag‐specific Tg TCR were able to develop in mice in which only HCs expressed MHC class I, when HCs also expressed the H‐Y Ag. These HC‐selected self‐specific CD8⁺ T cells resemble innate CD8⁺ T cells in WT mice in terms of the expression of memory markers and effector functions, but are phenotypically distinct from the thymus‐independent CD8⁺ T‐cell population. The peripheral maintenance of H‐Y‐specific CD8⁺ T cells required presentation of the self‐Ag and IL‐15 on HCs. HC‐selected CD8⁺ T cells in mice lacking the Tg TCR also showed these features. Furthermore, by using MHC class I tetramers with a male Ag peptide, we found that self‐Ag‐specific CD8⁺ T cells in TCR non‐Tg mice could develop via HC‐induced positive selection, supporting results obtained from H‐Y TCR Tg mice. These findings indicate the presence of self‐specific CD8⁺ T cells that are positively selected by HCs in the peripheral T‐cell repertoire.     

成人隐匿性自身免疫性糖尿病患者CD4+CD25+T细胞的变化研究

目的：通过观察成人隐匿性自身免疫性糖尿病（LADA）患者CD4^＋CD25^＋T细胞的变化并与速发性1型糖尿病（T1 DM）比较，了解成人隐匿性自身免疫性糖尿病患者T细胞免疫功能的变化及与1型糖尿病的异同点。方法：LADA组24例，速发性T1DM18例，对照组20例，应用流式细胞技术测定3组人选者T细胞表面分子CD4、CD8、CD25、CD4^＋CD25^＋、CD8^＋CD25^＋、CD4^＋CD25^＋CD62L^＋，以百分比表示各表面分子阳性T细胞占外周血淋巴细胞的比例。结果：LADA与T1DM组CD4^＋T细胞、CD4／CD8比值明显高于健康对照组（P〈0．01），LADA与T1DM对比无差异。LADA组CD25^＋、CD4^＋CD25^＋、CD4^＋CD25^＋CD62L^＋T细胞高于健康对照组（P〈0．05），明显高于T1 DM组（P〈0．01）。T1 DM组CD25^＋、CD4^＋CD25^＋、CD4^＋CD25^＋CD62L^＋T细胞略低于健康对照组，但无统计学意义（P〉0．05）。结论：LADA患者外周血中诱导免疫耐受的CD4^＋CD25^＋、CD4^＋CD25^＋CD62L^＋T细胞较对照组升高并明显高于T1 DM患者。LADA患者胰岛B细胞功能下降较速发性T1 DM患者相对缓慢可能与CD4^＋CD25^＋T细胞的免疫保护有关。     

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.