调节性及辅助性T细胞在人类IgA肾病中的表达及意义

目的 探讨CD4+CD25high调节性T细胞（Treg）及辅助性T细胞亚群（Th1、Th2）比例失衡在IgA肾病（IgAN）免疫发病机制中的作用。 方法 用流式细胞仪检测IgAN患者外周血Treg及Th1、Th2的比例。以胞内染色技术检测叉头框蛋白3（FOXP3）的表达。Treg及Th1、Th2的比例与IgAN各项临床病理指标的相关性分析采用Spearman或Pearson相关分析法。 结果 IgAN患者外周血中Treg比例明显高于健康人[（2.14±0.82）％比（1.59±0.53）％，P ＜ 0.05]，与血IgA水平呈正相关（r = 0.397，P ＜ 0.05），与eGFR呈负相关（r = -0.376，P ＜ 0.05）。IgAN患者外周血中Th2细胞比例显著高于健康对照组[（2.57±0.72）％比（1.81±1.10）％，P ＜ 0.05]，与血IgA水平呈正相关（r = 0.468，P ＜ 0.05）。IgAN患者Th1/Th2比值显著低于健康对照组（5.75±1.89比12.73±9.79，P ＜ 0.05），但与临床各指标间没有相关性。 结论 IgAN患者体内存在T细胞亚群表达紊乱。Treg在外周血中的增多以及以Th2为优势的Th1/Th2失衡可能在IgAN的发病中起重要作用。     

CD4+ CD25+ CD62+ T-Regulatory Cell Subset Has Optimal Suppressive and Proliferative Potential

CD4+ CD25+ regulatory T cells (Treg) are potent suppressors, and play important roles in autoimmunity and transplantation. Recent reports suggest that CD4+ CD25+ Treg are not a homogeneous cell population, but the differences in phenotype, function, and mechanisms among different subsets are unknown. Here, we demonstrate CD4+ CD25+ Treg cells can be divided into subsets according to cell-surface expression of CD62L. While both subsets express foxp3 and are anergic, the CD62L+ population is more potent on a per cell basis, and proliferates and maintains suppressive function far better than the CD62L- population and unseparated CD4+ CD25+ Treg. The CD62L+ population preferentially migrates to CCL19, MCP-1 and FTY720. Both CD62L+ and CD62L- subsets prevent the development of autoimmune gastritis and colitis induced by CD4+ CD25-CD45RBhigh cells in severe combined immunodeficiency (SCID) mice. Overall, these results suggest CD4+ CD25+ Treg are not a homogenous cell population, but can be divided into at least two subsets according to CD62L expression. The CD62L+ subset is a more potent suppressor than the CD62L- population or unfractionated CD4+ CD25+ Treg cells, can be expanded far more easily in culture, and is more responsive to chemokine-driven migration to secondary lymphoid organs. These properties may have significant implications for the clinical manipulation of the CD4+ CD25+ CD62L+ cells.     

CD4+CD25+FOXP3+T细胞在肝癌肝移植肿瘤复发中的作用

目的 探讨肝癌肝移植病人移植前后外周血和肿瘤组织中CD4+CD25+FOXP3+T细胞比例变化及其临床意义.方法 用流式细胞仪检测肝癌肝移植病人和其他肝移植病人术后外周血中CD4+CD25+FOXP3+T细胞的比例,并采用正常人作对照.用免疫组化法检测肝癌病人和非肝癌病人肿瘤组织中FoxP3的表达及CD8+T细胞浸润的比例.观察肝癌肝移植病人术后及肿瘤复发后调节性T细胞的变化及其对肿瘤复发的影响.结果 流式细胞检测显示肝癌肝移植、非肝癌肝移植的病人术后外周血中CD4+CD25+FOXP3+T细胞占CD4+T细胞的比例较正常人明显升高,分别为(10.15±1.00)%、(5.30±1.64)%和(3.20±1.18)%,P<0.05.肝癌肝移植肿瘤复发病人较未复发病人外周血CD4+CD25+FOXP3+T比例明显升高,分别为(15.15±1.50)%和(6.80±1.50)%,P<0.01.免疫组化检测显示肿瘤组织中FOXP3+T细胞增多,CD8+T细胞浸润明显减少.结论 肝癌肝移植肿瘤复发的病人外周血中CD4+CD25+FOXP3+T细胞比例升高.调节性T细胞可能通过减少CD8+T细胞浸润,加速肿瘤复发.     

CD8+T细胞和调节性T细胞在食管癌中表达及与预后的关系

目的 探讨食管癌组织中CD8+T细胞、Foxp3阳性调节性T细胞(regulatory T cells,Treg)表达及对预后的影响.方法 应用免疫组织化学SP法检测CD8、Foxp3在90例食管癌组织间质、癌巢中的表达,计数阳性细胞,分析阳性细胞数目与预后的关系.结果 间质中CD8+T细胞的数量与浸润深度、分化程度呈负相关(P<0.05);Foxp3+Treg细胞数量与淋巴结转移、病变长度呈正相关(P<0.05).本组病例3年总生存率66.67%(60/90).单因素生存分析显示,无论癌巢或间质中,CD8+T细胞计数高者的总体生存曲线优于计数低者,差异有统计学意义(P<0.05);roxp3+Treg细胞计数高者的累计生存情况较计数低者差(P<0.05).多因素分析显示,间质浸润的CD8+T、Foxp3+Treg细胞数量和病变长度是影响生存期的独立因素(P<0.05).结论 间质中浸润的CD8+T细胞数量、Foxp3+Treg细胞数量是影响食管癌患者预后的独立因素,Foxp3+Treg细胞数量增多预后不良,CD8+T细胞数量增多则预后良好.     

Regulatory effect of FK506 on CD152 and PD-1 in the liver allorecipients

AIM: We dynamically observed the expression of CD152 and PD-1 on T-cell surfaces in peripheral blood of liver allorecipients to explore the regulatory effect of FK506 on negative costimulatory molecules. METHODS: We evaluated 20 liver allorecipients, 19 end-stage liver disease patients, and 20 healthy volunteers for FK506 concentrations measured by an enzyme-multiplied immunoassay technique and flow cytometry to determine T-cell subsets as well as CD152 and PD-1 expression. RESULTS: After liver transplantation, the frequency of CD4+ T cells gradually decreased to significantly lower level than those in the disease controls (P < .05). CD8+ T cells in each treatment group were obviously higher than those in the disease controls (P < .05). The expression of CD152 on CD4+ and CD8+ T cells was greater than those in healthy controls (P < .05); and at 2 and 4 weeks, higher than those in disease controls (P < .05). The expression of PD-1 on CD4+ T cells from the 2 weeks after treatment was significantly greater than that in healthy controls (P < .05), and that on CD8+ T cells increased obviously from the 4 weeks compared with disease controls (P < .05). CONCLUSION: FK506 up-regulated the expression of CD152 and PD-1 on the T-cell surface inhibiting proliferation and activation of effector T cells, favoring the survival of allorecipients.     

Further Analysis of the T-Cell Subsets and Pathways of Murine Cardiac Allograft Rejection

The present study examined the role of CD4+ and CD8+ T cells in cardiac allograft rejection when either the direct or indirect pathway was eliminated for the CD4+ portion of the response. To study the pathways in vivo, we used genetically altered mouse strains that lack class II antigens as either the donors or recipients for cardiac transplantation. In contrast to earlier published studies, which used different strain combinations, we found that either CD4- or CD8-depletion prolonged cardiac allograft survival moderately, but not indefinitely, in an MHC-mismatched, minor-matched combination. When the CD4+ indirect pathway was eliminated, rapid graft rejection occurred when both T-cell subsets were present and when either CD4+ or CD8+ T cells were depleted. When the CD4+ direct pathway was eliminated, rapid graft rejection occurred when both T-cell subsets were present, there was slow rejection when CD4+ T cells were eliminated, and no rejection was seen for more than 100 days when CD8+ T cells were eliminated. However, the long-surviving allografts on the recipients with only CD4+ cells and an indirect pathway did show evidence of chronic vasculopathy. Thus, either CD4+ or CD8+ T cells can mediate acute cardiac allograft rejection in these experiments when both pathways are available. In addition, CD4+ T cells can provide help for acute rejection through either the direct or indirect pathway. Finally, recipients who have only CD4+ cells and an indirect pathway do not demonstrate acute rejection, but do show evidence of chronic rejection.     

Acute rejection after liver transplantation: Is there a specific immunological pattern?

The aim of the study was to assess various T-cell subsets and cytokine secretion patterns both in liver tissue and in the peripheral blood of 24 liver transplant patients to assess possible specific immunological involvement in early acute rejection episodes after liver transplantation. Particularly, we studied CD4+ CD7+, CD8+ CD38+, and CD4+ CD25+ T cells by flow cytometry, as well as contemporaneously, interleukin (IL)-2 and IL-10 secretion by ELISpot to determine possible Th1-like immune responses and the immunomodulation expressed by Treg cells in acute liver rejection, respectively. As a control group we included patients transplanted without acute rejection. Early acute rejection within the first 4 weeks was proven histologically in 42% of patients. It was associated with a greater expression of CD4+ CD7+ and CD8+ CD38+ T cells in the liver than in the blood (P < .001). A contemporaneous reduced expansion of liver Treg cells was evident in patients with acute rejection (P < .001). Our data suggested that a preferential Th1-like immune mechanism operated in local fashion as characterized by a decreased presence in the liver and blood of Treg cells.     

抗原特异性CD4+CD25+T细胞对同种异体胰岛移植影响及作用机制研究

目的:探讨抗原特异性CD4+CD25+Treg细胞免疫对同种异体胰岛急性移植排斥反应的影响和机制。方法:用MACS分选供体抗原特异性CD4+CD25+Treg细胞免疫糖尿病BALB/cByJ受体小鼠，以ICR小鼠胰岛为供体行同种异体胰岛移植。观察移植后小鼠的存活时间、移植前后外周血CD4+和CD8+T细胞亚群的变化和移植物中Th1/Th2细胞因子mRNA表达水平的变化。结果:抗原特异性Treg细胞联合胰岛移植组(C组)胰岛移植物平均生存期为(34.57±17.15)d，显著长于单纯胰岛移植组(B组)的(10.6±1.82) d (P<0.01)；移植后第3天，C组外周血CD4+/CD8+的值显著低于B组(P<0.01)；C组移植物中IL-10,TGF-β mRNA表达比B组显著增强。B组移植物中IL-1β，IL-2及IFN-γ mRNA表达明显强于C组。结论:抗原特异性CD4+CD25+ Treg细胞可通过调节Th2/Th1之间的反应平衡而延长同种异体胰岛移植物的存活时间。     

Patterns of CD4/CD8 T-cell ratio in dialysis effluents predict the long-term outcome of peritonitis in patients undergoing peritoneal dialysis.

BACKGROUND: The peritoneal immune compartment is a microenvironment with a particular T-cell repertoire and susceptible to local inflammation. To clarify the role of T lymphocytes in peritoneal immunity, the changes in T-cell subpopulations in peritoneal dialysis effluents (PDEs), and their influence on the response to the treatment of peritonitis and on its prognosis were studied in patients undergoing long-term, continuous ambulatory peritoneal dialysis (CAPD). METHODS: A cohort of 36 patients treated with CAPD and who had histories of peritonitis were divided into a group with rapid and a group with delayed response to antibiotics, and were followed for 3 years. CD4/CD8 T-cell ratios, T-cell cytokine mRNA expression patterns and transforming growth factor-beta1 (TGF-beta1) concentrations were examined in PDE during bouts of peritonitis. The change in 4 h D/P creatinine during the peritoneal equilibration test (PET) between year 0 and year 3 was expressed as deltaD/P creatinine. RESULTS: The serial changes in T-cell subsets in PDE during peritonitis showed two patterns: (i) pattern 1, manifest as a progressive increase in the CD4/CD8 ratio, and associated with a rapid response to treatment; and (ii) pattern 2, manifest as a progressive decrease in the CD4/CD8 ratio, and associated with a delayed response to treatment. The major T-cell phenotypes in PDE during peritonitis were Th1-CD4(+) and Tc2-CD8(+), determined by cloning techniques, RT-PCR and double immunofluorescence staining. TGF-beta1 in the effluent was undetectable in pattern 1 after 7-8 days, but remained detectable at 2 weeks in pattern 2. Pattern 2 patients had a significantly greater decrease (deltaD/P creatinine: -0.198+/-0.086) in solute transport than pattern 1 patients (deltaD/P creatinine: -0.036+/-0.077, P<0.05). CONCLUSIONS: These results suggest that a progressive decrease of the CD4/CD8 ratio in PDE correlates with a persistent expression of TGF-beta1, and plays a pathogenetic role in the evolution of peritonitis, PET deterioration and peritoneal fibrosis. Therefore, patterns of CD4/CD8 T-cell ratio in PDE may predict clinical outcomes of peritonitis in CAPD patients.     

Pacientes com lúpus eritematoso sistêmico e síndrome antifosfolípide secundária possuem números reduzidos de células B CD4+ CD25+ Foxp3+ (células Treg) e células B CD3– CD19+ circulantes

IntroductionCD4+CD25+Foxp3+ regulatory T (Treg) cell depletion has been reported in systemic lupus erythematosus (SLE) and, recently, in primary antiphospholipid syndrome (APS); the issue has not been studied in SLE patients with secondary APS (SLE/APS) so far.ObjectiveTo quantify total lymphocytes, Treg cells, CD3+CD19– T cells and CD3–CD19+ B cells in SLE/APS patients and healthy controls.MethodsCell subtypes underwent immunophenotyping using specific monoclonal antibodies (anti–CD3 CY5, anti–CD4 FITC, anti–CD25, anti–Foxp3, anti–CD19 PE) and flow cytometry.ResultsTwenty–five patients with SLE/APS (mean age 43.5 years, 96% females, 96% caucasians, mean duration of disease 9.87 years, mean SLEDAI 10 ± 5.77) and 25 age and sexmatched controls entered the study. It was realized that the numbers of Treg and CD3– CD19+ B cells were significantly lower in SLE/APS patients than in controls (all p < 0.05). Treg and CD3–CD19+ B cells remained numerically low after controlling (ANCOVA) for percentage of total lymphocytes (p < 0.05). Decreasing levels of circulating Treg and CD3–CD19+ B cells correlated to higher scores of lupus activity (rs = –0.75, p < 0.0001; rs = –0.46, p = 0.021, respectively). Number of Treg cells and CD3–CD19+ B lymphocytes did not significantly differ in users or nonusers of chloroquine, azathioprine and corticosteroids (all p > 0.05).ConclusionsIn this preliminary study, patients with SLE and secondary APS showed depletion of Treg and CD3–CD19+ B cells; decreasing numbers of both subtypes correlated to a higher SLEDAI. Treg cells depletion might contribute to the autoimmune lesion seen in patients with SLE/APS. The reduced number of CD3–CD19+ B cells seen in these patients deserves more studies in order to get further elucidation.     

Differences in Type 1 and Type 2 intracytoplasmic cytokines, detected by flow cytometry, according to immunosuppression (cyclosporine A vs. tacrolimus) in stable renal allograft recipients

Recent multicenter, randomized clinical trials have shown that in renal transplant patients tacrolimus (FK506) was more efficient than cyclosporine A (CsA) at preventing acute rejection. In order to try and evaluate whether this difference was related to a different in vivo T-cell suppression we assessed, in a prospective study, the frequencies of interleukin (IL)-2-, IL-4-, IL-5-, IL-6-, IL-10-, interferon-gamma (IFN-gamma)- and double-positive IL-2/IFN-gamma-producing whole T cells, CD4 + and CD8 + T-cell subsets by means of cytokine flow cytometry. This was performed after in vitro stimulation of peripheral blood mononuclear cells (PBMCs) with phorbol myristate acetate (PMA) and ionomycin, in the presence of monensin, in 14 healthy volunteers (controls) and in 14 renal transplant patients. The immunosuppression of the latter was based either on CsA (n = 7) or on FK506 (n = 7). Cytokine-expressing T-cell frequencies were assessed immediately pretransplantation (DO), and subsequently 3 months (M3) and 6 months (M6) afterwards in fasting patients prior to the morning intake of the immunosuppressive drug. We found that at DO the frequencies of IL-2-(22 +/- 2% vs. 22.2 +/- 2%), IFN-gamma-(26 +/- 3% vs. 29 + 3.4%) and IL-4-(0.8 +/- 0.2% vs. 1.4 +/- 0.2%)-expressing T lymphocytes were not significantly different between the controls and the patients, respectively. Conversely, the frequency of IL-2/IFN-gamma double positive cells was higher in the latter (9.3 +/- 1.6%) than in the controls (5.6 +/- 0.8); p = 0.06. Finally, on D0 the frequencies of IL-5-, IL-6-, and IL-10-producing T lymphocytes were lower than 1%, in both groups, as well as after grafting, i.e. on M3 and M6. As compared to baseline (DO): (a) chronic immunosuppression significantly decreased the frequencies of IL-2-, IL-4- and IL-2/IFN-gamma-expressing T cells, whereas those of IFN-gamma, IL-5, IL-6, and IL-10 were not significantly affected; (b) the frequencies of cytokine-expressing T cells were not statistically different between M3 and M6; (c) the decrease in the frequencies of IL-2- and IL-2/IFN-gamma-expressing T cells affected CD4 + and CD8 + cells equally; (d) there was a marginal decrease in the frequency of IFN-gamma-expressing cells only in the CD4 + subset but not in the CD8 population; and (e) for CsA, but not for FK506, the frequency of the IL-2-expressing T cells was negatively correlated with the whole blood trough levels. When we compared the frequencies of cytokine-expressing cells in FK506- and CsA-treated patients, we found that the frequency of IL-2-expressing T cells was significantly lower with FK506 (10.9+/-1.61%) than with CsA (16.3 +/- 1.8%; p = 0.03), whereas the frequencies of the other cytokine-expressing cells were not statistically different between the two groups. In conclusion, our study clearly demonstrated that studied ex vivo, FK506 and CsA decrease the frequencies of cells expressing IL-2, IL-4 and IL-2/IFN-gamma in vivo but do not affect those expressing IFN-gamma. Meanwhile, the frequency of IL-2-producing T cells was more affected with FK506 than with CsA and was negatively correlated with the CsA trough level. Finally, our results regarding IL-2 might explain to some extent the higher efficiency of FK506 in vivo than CsA.     

Human CD4+CD25+ Regulatory T Cells Suppress Anti-Porcine Xenogeneic Responses

Due to the shortage of human organs, xenotransplantation is being explored as an alternative to allotransplantation, but immune rejection remains a major hurdle to its implementation. We tested the ability of human CD4+CD25+ T cells (Treg cells) to suppress CD4+ T cell-mediated anti-porcine xenoresponses usingin vitroassays. Human Treg cells were hyporesponsive to porcine cell stimulation and suppressed the proliferative response of CD4+CD25- T cells in a dose-dependent manner, and comparison of the allo- and xenoresponses indicated that more Treg cells might be required to suppress the xenogeneic response than the allogeneic response. Stimulation of CD4+CD25- T cells with porcine cells resulted in secretion of IFN-gamma, TNF-alpha, IL-10, IL-6 and IL-2, and Treg cells suppressed the secretion of these cytokines, as well as the CD4+CD25- T-cell cytolytic response against porcine cells. These results suggest a potential role for Treg cells in promoting xenograft survival.     

宫颈上皮内病变患者人乳头状瘤病毒感染与Th17细胞、Treg细胞及相关细胞因子表达的相关性

目的探讨宫颈上皮内病变（CIN）患者人乳头状瘤病毒（HPV）感染与Th17细胞、Treg细胞及相关细胞因子表达的相关性。 方法选择2017年2月至2018年9月于新疆自治区人民医院就诊的CIN患者共140例进行回顾性分析,根据是否感染HPV分为感染组（86例）和非感染组（54例）,采用免疫组织化学方法比较两组患者T细胞CD3、CD4及CD8水平以及Treg和Th17的表达水平,采用流式细胞技术比较两组患者的外周血T细胞亚群含量（CD3⁺ T细胞、CD4⁺ T细胞、CD8⁺ T细胞的百分比）、Treg细胞和Th17细胞百分含量（CD4⁺CD25⁺ Treg、CD8⁺CD25⁺CD127^-Treg、CD4⁺Foxp3⁺ Treg、CD8⁺CD25⁺Foxp3⁺ Treg和CD4⁺IL17⁺Th17以及Th17/Treg比值）以及相关细胞因子[肿瘤坏死因子（TGF）-β、白细胞介素（IL）-10、IL-17和干扰素（IFN）-γ]含量的差异,应用多因素Logistic回归模型分析CIN患者感染HPV的危险因素。 结果免疫组织化学结果显示,两组患者CIN组织中T细胞CD3、CD4和CD8的表达水平差异无统计学意义,感染组患者CIN组织中T细胞抗-FOX3（Treg）和Th17表达水平显著高于非感染组。流式细胞术检测结果显示两组患者CD3⁺ T细胞、CD4⁺ T细胞、CD8⁺ T细胞百分比差异无统计学意义（t = 0.870、1.509、0.236;P = 0.385、0.133、0.813）,感染组患者CD4⁺CD25⁺ Treg、CD8⁺CD25⁺CD127^—Treg、CD4⁺Foxp3⁺ Treg、CD8⁺CD25⁺Foxp3⁺ Treg、CD4⁺IL17⁺ Th17以及Th17/Treg比值均显著高于非感染组（P均< 0.05）。感染组患者外周血TGF-β、IL-10和L-17均高于非感染组（t =－11.601、－42.251、－40.31,P均< 0.001）,而IFN-γ低于非感染组（t = 10.316、P < 0.001）。多因素Logistic回归分析显示,CD4⁺CD25⁺ Treg（OR = 3.673、P = 0.031）、CD4⁺IL17⁺Th17（OR = 1.974、P = 0.021）和Th17/Treg（OR = 3.585、P = 0.024）均为CIN患者感染HPV的危险因素,IFN-γ（OR = 0.612、P = 0.028）为CIN患者感染HPV的保护因素。 结论CIN患者HPV感染与Treg细胞、Th17细胞及IFN-γ因子存在相关性,HPV感染的CIN患者Treg和Th17细胞增高,IFN-γ降低。     

EV71感染手足口病患儿T细胞亚群表达

目的分析EV71感染手足口病患儿T细胞亚群的表达。 方法选取2015年10月至2017年10月于惠州市中心人民医院接受治疗的手足口病患儿共302例,其中EV71感染患儿110例,依据EV71感染患儿中枢神经有无被累及分成轻症组（69例）与重症组（41例）,选取同期于本院体检的健康儿童110例作为对照组;流式细胞仪对T淋巴细胞亚群CD3⁺、CD3⁺CD8^－、CD3⁺CD8⁺、CD3⁺CD8^－/CD3⁺CD8⁺、Th1、Th2、Th1/Th2、Tc1、Tc2、Tc1/Tc2、Th17、Tc17、调节性T淋巴细胞（Treg）及Th17/Treg所占比率进行检测,ELISA法检测各组对象血清内转化生长因子-β1（TGF-β1)、白细胞介素4（IL-4）、白细胞介素17A（IL-17A）及γ干扰素（IFN-γ）含量变化。 结果轻症组患儿血清Th1、Th1/Th2、Tc1和Tc1/Tc2所占比率为（9.59 ± 2.15）%、（8.67 ± 2.19）%、（28.82 ± 5.49）%和（56.39 ± 10.48）%,重症组分别为（13.58 ± 2.74）%、（9.45 ± 2.29）%、（40.46 ± 6.37）%和（54.87 ± 9.61）%,均显著高于对照组患儿,差异均有统计学意义（F= 12.159、11.470、13.925、10.542,P = 0.016、0.013、0.008、0.035）;轻症组患儿血清IFN-γ和IL-17A水平分别为（11.32 ± 2.76）pg/ml和（6.38 ± 2.10）pg/ml,重症组分别为（11.38 ± 2.65）pg/ml和（12.59 ± 3.58）pg/ml,均显著高于对照组患儿,差异均有统计学意义（F= 12.590、11.664,P = 0.024、0.019）。 结论EV71感染后手足口患儿机体内Th17/Treg与Th1/Th2比例失衡。     

Dynamic changes in Th1, Th17, and FoxP3+ T cells in patients with acute cellular rejection after cardiac transplantation

Previously, studies suggest that CD4(+) effector T-cell subsets participate in allograft rejection. However, the dynamic changes and relative roles of these CD4(+) effector T-cell subsets, especially Th17 cells, have not been systemically examined in patients with acute rejection after cardiac transplantation. In this study, we have studied and compared these CD4(+) T-cell subsets in peripheral blood and endomyocardial biopsies (EMB) in patients with stable-graft and acute cellular rejection. We observed that the gene expressions including T-bet, IFN-γ, RORγt, IL-17, IL-23, and FoxP3, the functional marker of Th1, Th17, and FoxP3(+) CD4(+) T cells, were elevated in EMB samples from patients with acute graft rejection. Accordingly, the percentages of circulating Th1, Th17, and FoxP3(+) CD4(+) T cells were also significantly increased. The data suggest that Th1, Th17, and FoxP3(+) CD4(+) T cells are associated with acute graft rejection in patients with cardiac transplantation.     

前列腺癌患者外周血中CD4~+CD25~(high)调节性T细胞及调节因子TGF-β1和COX-2的表达

目的:通过检测前列腺癌患者以及健康志愿者外周血中CD4+CD25high调节性T细胞、TGF-β1及COX-2的表达,初步探讨CD4+CD25high调节性T细胞在前列腺癌发病机制中的作用及其与TGF-β1和COX-2的相关性。方法:应用流式细胞术检测30例前列腺癌患者治疗前后(其中前列腺癌局限组11例,非局限组19例)及20例健康志愿者外周血单个核细胞(PBMC)中CD4+CD25high调节性T细胞占CD4+T细胞的比例;应用酶联免疫吸附试验(ELISA)检测其外周血清中TGF-β1和COX-2的表达。对前列腺癌患者上述指标进行术前术后对比分析,另对CD4+CD25high调节性T细胞与TGF-β1及COX-2的相关性进行分析;并探讨上述指标在前列腺癌患者局限组和非局限组间是否存在差异性。结果:流式细胞术检测显示,前列腺癌患者治疗前PBMC中CD4+CD25high调节性T细胞占CD4+T细胞的比例为(18.32±7.49)%,高于健康志愿者对照组(7.77±1.86)%(P<0.05)。前列腺癌患者治疗后其比值为(17.34±5.87)%,较治疗前稍减低,但两者相比无显著差异(P>0.05)。ELISA检测外周血清中TGF-β1和COX-2显示,前列腺癌组分别为(215.97±55.16)ng/ml和(6.88±5.14)ng/ml,对照组分别为(149.75±47.11)ng/ml和(5.65±2.69)ng/ml;前列腺癌患者外周血清中TGF-β1的表达水平高于健康志愿者对照组(P<0.05),COX-2的表达水平与对照组无显著差异(P>0.05)。通过多重线性回归分析表明,前列腺癌患者PBMC中CD4+CD25high调节性T细胞的表达与血清中TGF-β1和COX-2的表达无显著相关。前列腺癌局限组和非局限组外周血中CD4+CD25high调节性T细胞、TGF-β1及COX-2的表达均无显著性差异(P>0.05)。结论:前列腺癌患者PBMC中CD4+CD25high调节性T细胞可能参与前列腺癌的发生,其增殖机制与血清中TGF-β1和COX-2的表达无关,可能与肿瘤本身及肿瘤局部微环境相关。     

T-helper and T-cytotoxic cell subsets monitoring during active cytomegalovirus infection in liver transplantation

OBJECTIVE: The objectives of this study was to analyze the peripheral blood T-lymphocyte subsets of the Th1-related versus Th2-related cytokines of CD4+ cells, and the Tc1 versus Tc2 cytokines of CD8+ cells liver transplant recipients with versus without active cytomegalovirus (CMV) infection. METHODS: Isolated peripheral blood mononuclear cells (PBMC) were stimulated using PMA/Ionomycin/Monensin. Interleukin (IL)-4 and interferon gamma (IFN-gamma) production by CD4+ and CD8+T cells were determined using fluorescence-activated cell sorter (FACS) analysis. RESULTS: The ratios of CD4/CD8 were significantly lower among active CMV-infected patients. The levels of Th2 and Tc2 cytokines (IL-4) were similar between CMV-infected and uninfected patients. However, the levels of Th1-type and Tc1-type cytokines (IFN-r) were significantly lower among active CMV-infected patients. CONCLUSIONS: Low levels of Th1-type cytokines seem to correlate with active CMV infection in liver transplant recipients.     

In vitro cytokine responses in liver transplant recipients treated with cyclosporine A and tacrolimus

The inhibitory effect of Cyclosporine A (CsA) and Tacrolimus (Tacr) on interleukin 2 (IL-2) are well known, and the importance of Th1-type (IL-2, interferon gamma), and Th2-type (IL-4, IL-6, and IL-10) cytokine secretion in preventing allograft rejection is a controversial issue. The immunological mechanisms involved in long-term liver transplant recipients under CsA and Tacr were not studied precisely.
This study was designed to evaluate the effects of CsA and Tacr on the immune response of 62 long-term survivors following liver transplantation.
T-cell and B-cell subpopulations, the T helper (Th) cell activity, T-cell cytokine production, Staphylococcus aureus Cowan strain I (SAC-I)-stimulated B-cell responses and PWM-stimulated B-cell responses were examined.
CsA and Tacr decreased whole T-cell populations as well as CD4+T-cell IL-2 responses (p < 0.005, Tacr and p = 0.02, CsA), impaired CD4+ cell Th activity (p < 0.01, Tacr and p = 0.02, CsA) and reduced SAC-I-stimulated B-cell responses (immunoglobulin-secreting cells [ISC]: p = 0.001, Tacr and p < 0.05, CsA). A significantly impaired T-cell IL-10 secretion (p = 0.0001) and decreased Th function of whole T cells was found in Tacr-treated patients only, whereas unstimulated Th1 responses and SAC-I-stimulated B-cell IL-6 responses were reduced in CsA-treated patients.
Our data show that Tacr suppresses T-, CD4+-, and B-cell responses more effectively than CsA which may be relevant in the maintenance of long-term stable liver graft function.     

The Changed Balance of Regulatory and Naive T Cells Promotes Tolerance after TLI and Anti‐T‐Cell Antibody Conditioning

The goal of the study was to determine how the changed balance of host naïve and regulatory T cells observed after conditioning with total lymphoid irradiation (TLI) and antithymocyte serum (ATS) promotes tolerance to combined organ and bone marrow transplants. Although previous studies showed that tolerance was dependent on host natural killer T (NKT) cells, this study shows that there is an additional dependence on host CD4⁺CD25⁺ Treg cells. Depletion of the latter cells before conditioning resulted in rapid rejection of bone marrow and organ allografts. The balance of T‐cell subsets changed after TLI and ATS with TLI favoring mainly NKT cells and ATS favoring mainly Treg cells. Combined modalities reduced the conventional naïve CD4⁺ T cells 2800‐fold. The host type Treg cells that persisted in the stable chimeras had the capacity to suppress alloreactivity to both donor and third party cells in the mixed leukocyte reaction. In conclusion, tolerance induction after conditioning in this model depends upon the ability of naturally occurring regulatory NKT and Treg cells to suppress the residual alloreactive T cells that are capable of rejecting grafts.     

CD8+ T cell subsets TC1 and TC2 cause different histopathologic forms of murine cardiac allograft rejection

BACKGROUND: CD4+ T cell effector function is sufficient to mediate allograft rejection, and it is suggested that CD8+ T cell-mediated effects are dependent on CD4+ T cell help. CD8+ T cells can be classified into at least two functional subsets: Tc1, producing high amounts of interferon (IFN)-gamma and Tc2, producing interleukin (IL)-4, -5, -10, and -13 and low levels of IFN-gamma. Because these subsets express different chemokine receptors, they may have different capabilities of migrating into grafts. Once in the graft, each subset may perform different effector functions dependent on the cytokines it produces. We asked whether allospecific CD8+ T cells, in the absence of CD4+ T cells, are capable of mediating rejection of a primarily vascularized allograft, and if Tcl and Tc2 cells differ in their ability to mediate rejection. METHODS: Hearts from H-2d mice were transplanted into H-2b RAG 1-/- recipients. Without manipulation, these fully mismatched allografts would survive indefinitely due to the absence of mature T and B cells. We adoptively transferred allo-(H-2d)-reactive Tcl or Tc2 cells from H-2b mice into each recipient. Grafts were harvested and analyzed on predefined timepoints, rejection was graded on a modified ISHLT scale. RESULTS: On day 7, grafts from Tc1- or Tc2-injected animals showed grade 1-2 parenchymal rejection with stable phenotype and comparable distribution of graft infiltrating CD8+ T cells. Adoptive transfer of IFN-gammahigh Tc1, but not of IFN-gammalow Tc2 cells was followed by the development of graft vasculitis, as well as graft arteriopathy. Adoptive transfer of IL-4high IL-5high Tc2, but not of IL-4low IL-5low Tc1 cells lead to extensive infiltration of eosinophils and formation of giant cells. CONCLUSIONS: Both Tc1 and Tc2 cells can mediate murine cardiac allograft rejection in the absence of CD4+ T cell help, although each subset elicits a different type of inflammatory response. In this model, cytokine secretion of either functional CD8+ T effector cell subset is an important effector mechanism in the process of allograft rejection: IFN-gammahigh Tc1 cells are important in early graft vasculitis, although IL-4high IL-5high Tc2 cells promote recruitment of secondary effectors like eosinophils.