乙型肝炎组织中淋巴细胞的表型变化及意义

目的：探讨肝脏淋巴细胞在乙型病毒性肝炎中的作用。方法；运用流式细胞仪检测23例乙型病毒性肝炎患者肝穿标本中淋巴细胞CD3／CD56的表达。结果：在乙型肝炎发展的不同阶段，CD3^ CD56^-,CD3^ CD56^ CD3^-CD56^ 三类细胞群的比例随着病情的发展而发生变化。在急性乙型肝炎期，淋巴细胞总数明显增高，且以CD56^ 细胞的NK作用为主（P＜0．05），此期中的γδT淋巴细胞（CD3^ CD56^ 淋巴细胞）表现尤为突出。在轻度慢性肝炎期，CD56^ 细胞与CD3^ CD56^-细胞在数量上无明显差异。在中、重度慢性肝炎期，则出现了以CD3^ CD56^-类细胞，即CD8^ T淋巴细胞的CTL作用为主（P＜0．05）的免疫学分布。结论：在乙型肝炎病程发展的不同阶段，肝脏淋巴细胞的作用侧重点存在显著差异。     

原发性胆汁性肝硬化患者外周血T淋巴细胞亚群检测的意义

目的 探讨原发性胆汁性肝硬化（PBC）患者外周血T淋巴细胞亚群及细胞因子的特点及意义。方法 以35倒健康人作为对照组,用流式细胞术分析了35例确诊的PBC患者外周血T淋巴细胞亚群及其分泌IL-4和IFN-γ情况。结果 PBC患者组CD^4＋T细胞升高,CD8^＋T细胞下降,CD4^＋T／CD8^＋T比值上升（P〈0．05）;PBC患者CD4^＋T细胞IFN-1表达显著升高（P〈0．05）,而IL-4水平与对照组比较无显著性差异（P〉0．05）。结论 在PBC发病机制中,CD4^＋T细胞和TH1亚群细胞超重要作用。     

骨髓增生异常综合征患者Th1/Th2细胞平衡改变的临床意义

目的：探讨骨髓增生异常综合征（MDS）患者外周血CD4^＋、CD8^＋细胞亚群平衡的改变，以及与活化T细胞之间关系的临床意义。方法：选择14例难治性贫血患者（RA）和1例难治性贫血伴原始细胞增多患者（RAEB）的外周血，在PMA、Ionomycin、Monensin存在的条件下体外培养，进行细胞膜表面抗原和细胞内因子染色，并用流式细胞仪计数外周血表达CD3^＋ CD8^-IFN-γ^＋细胞（Th1）、CD3^＋CD8^-IL-4^＋细胞（Th2）、CD3^＋CD8^＋IF-γ^＋细胞（Tcl）、CD3^＋CD8^＋IL-4^＋细胞（Tc2）的百分率。结果与活化T细胞之间关系做相关性分析。结果：MDS组患者Th1细胞高于正常对照组（P〈0．05），体内的Th细胞向Th1细胞漂移，产生过多的造血抑制因子如IFN-γ，TNF-β等。Th2，Tc1，Tc2细胞与正常对照组相比无显著差异（P〉0．05）；CD4^＋CD45RO^＋细胞的百分率和Th1细胞的百分率呈正相关，相关系数为r=0．573（P〈0．05），CD4^＋CIN5RA^＋细胞的百分率和Th1细胞的百分率呈负相关，相关系数为r=-0．509（P=0．05），而和Th2，Tc1，Tc2的百分率无明显的相关性（P〉0．05）。结论：MDS患者存在有Th1／Th2细胞因子网络的失衡以及过多的造血抑制因子，且这些因子主要来源于T淋巴细胞。     

化脓性结核病患者T淋巴细胞亚群分类变化特征

目的：检测化脓性结核病患者外周血和病灶处脓液T淋巴细胞亚群分类,探讨化脓性结核病患者免疫状态。方法：对30例结核病组患者外周血标本和病灶处脓液标本各30份应用流式细胞仪在单个核细胞水平检测T淋巴细胞亚群分类;选取20例健康体检者外周血标本20份作对照。结果：结核病组外周血中CD4^＋淋巴细胞百分含量低于对照组,CD8^＋淋巴细胞百分含量高于对照组,2组比较差异有统计学意义（P〈0．01）;结核病组病灶处CD4^＋淋巴细胞数高于对照组（P〈0．01）,CD8^＋淋巴细胞数与对照组比较差异无统计学意义（P〉0．05）。结论：化脓性结核病的发生与T淋巴细胞亚群失衡有关,表现为外周血中CD4^＋细胞数量减少,而局部病灶中CD4^＋细胞增加。     

原发性胆汁性肝硬化患者调节性T细胞及其与肝功能损伤的研究

目的检测原发性胆汁性肝硬化（primary biliary cirrhosis，PBC）患者外周血CD4^＋ CD8^＋ T、CD4^＋ CD25^＋调节性T细胞亚群，探讨其与肝功能损伤、抗AMA—M2抗体产生的关系。方法采用流式细胞术检测北京协和医院住院和门诊PBC患者（n=27）外周血CD4^＋ CD8^＋ T细胞、CD4^＋ CD25^＋ T细胞群比例，以26例其他肝脏疾病患者作为疾病对照组，30例健康体检者作为正常对照，观察调节性T细胞亚群与患者的肝功能指标及自身抗体如AMA—M2、ANA的关系。结果PBC患者外周血CD4^＋ CD25^＋ T细胞比例与正常对照组比较结果差异无统计学意义（P〉0．05），但显著高于其他肝脏疾病组（P〈0．05）。CD4^＋ CD8^＋细胞群与正常人相比结果差异无统计学意义，其他肝病组明显高于正常对照组（P〈0．05）。PBC患者外周血NK细胞比例显著低于正常对照组（P〈0．05）。肝功严重损伤的PBC患者外周血CD4^＋ CD25^＋ T细胞比例明显高于肝功能轻度损伤者（P〈0．05）。PBC患者外周血CD4^＋ CD25^＋ T细胞数量与肝功能损伤指标ALB呈负相关（r=－0．338，P〈0．05），与TBIL、DBIL呈正相关（r分别为0．362，0．386，P〈0．05）。CD4^＋ CD25^＋ ／CD4^＋比例与GGT呈负相关（r=－0．335，P〈0．05），与TBIL、DBIL呈正相关（r分别为0．333，0．339，P〈0．05）。抗AMA—M2抗体阳性患者外周血CD4^＋ CD25^＋细胞比例明显高于AMA—M2阴性患者（P〈0．01）。未发现CD4^＋ CD25^＋ T、CD4^＋ CD8^＋T细胞比例在ANA^＋和ANA—PBC患者之间差异有统计学意义。结论PBC患者外周血CD4^＋ CD25^＋T细胞群比例与肝功能损害和抗AMA—M2抗体的产生密切相关，因此推测CD4^＋ CD25^＋T细胞的变化可能是导致病情发展以及肝脏损伤的关键环节之一。     

ALL病儿T淋巴细胞CD28表达及意义

目的探讨儿童B细胞型急性淋巴细胞白血病（ALL）治疗前后外周血T淋巴细胞表面CD28的表达及临床意义。方法采用流式细胞术,检测27例初发的B细胞型AI。L病儿化疗前后外周血T淋巴细胞表面CD28的表达,其中经化疗后达到完全缓解（cR）21例,未缓解（NR）6例。以同期23例健康体检儿童作为正常对照组。结果与正常对照组比较,治疗前初发ALL病儿外周血CD4^＋T淋巴细胞表面、CD8^＋T淋巴细胞表面CD28的表达均显著降低（t=21．56、44．02,P〈0．05）。化疗后CR组病）LPb周血CD4^＋T淋巴细胞表面、CD8^＋T淋巴细胞表面CD28较治疗前均显著升高（t=10．84、25．30,P〈0．05）,而NR组病儿CD4^＋T淋巴细胞表面、CD8^＋T淋巴细胞表面CD28与治疗前比较差异无显著性（P〉0．05）。结论CD28是参与AI。L发病的重要共刺激分子,其表达异常可能是ALL的发病因素之一;其表达水平可能是判断ALI。预后及化疗敏感性的因素之一。     

系统性红斑狼疮患者外周血淋巴细胞亚群分析

目的探讨T淋巴细胞亚群、B淋巴细胞和NK细胞在系统性红斑狼疮fSLE忡的作用。方法利用流式细胞仪对29例活动期、24例非活动期SLE患者及50例健康对照者的外周血中的CD3＋、CD4＋、CD8＋、CD3-CD16＋CD56＋（NK）及CD19＋（B）淋巴细胞进行检测。结果活动期组与非活动期组及健康对照组比较：CD4＋T淋巴细胞、CD4＋／CD8＋比值及NK细胞明显降低（P〈0．05）,B淋巴细胞明显增高（P〈0．05）,CD3＋T和CD8＋T淋巴细胞无统计学意义（P〉0．05）。非活动期组与健康对照组比较．T淋巴细胞各亚群、NK细胞及B淋巴细胞均无统计学意义（P〉0．05）。结论检测外周血淋巴细胞亚群．对判断病情及指导临床治疗具有重要意义。     

多种炎性疾病患者外周血CD4＋和CD8＋T淋巴细胞的检测与临床意义分析

目的：探讨恶性肿瘤、肺结核、干燥综合征、葡萄膜炎等多种炎性疾病患者外周血CD4＋和CD8＋淋巴细胞的变化及其临床意义。方法应用BD FACSCanto II流式细胞仪检测上述疾病患者外周血CD4＋、CD8＋ T细胞并计算两者比值,同时检测健康者标本20例作为健康对照组,进行统计分析。结果与健康对照组比,恶性肿瘤组CD4＋ T淋巴细胞、CD4＋/CD8＋均明显降低,CD8＋ T淋巴细胞增高,差异有统计学意义（ P＜0．05）;全身炎性反应综合征组CD4＋ T淋巴细胞、CD4＋/CD8＋均明显降低,CD8＋ T 淋巴细胞增高,差异有统计学意义（ P＜0．05）;葡萄膜炎组CD8＋ T淋巴细胞增高,差异有统计学意义（P＜0．05）;肺结核组CD4＋ T淋巴细胞降低,差异有统计学意义（ P＜0．05）。结论患者的 CD4＋、CD8＋ T 淋巴细胞异常,表明机体细胞免疫功能异常。流式细胞仪检测CD4＋、CD8＋ T淋巴细胞可以作为患者临床免疫功能的初步诊断指标,辅助疾病治疗和病情观察。     

H7N9禽流感患者外周血T淋巴细胞亚群的分析及意义

目的探讨H7N9禽流感患者外周血T淋巴细胞亚群的变化情况。方法将18例H7N9禽流感患者（感染组）根据治疗预后分为治愈组13例和死亡组5例,以门诊健康献血者20名作为正常对照组。采用特异性引物聚合酶链反应（PCR）检测H7N9禽流感病毒,并用流式细胞术检测外周血T淋巴细胞亚群的变化。结果感染组CD3＋、CD8＋和CD4＋T细胞绝对数及CD4＋／CD8＋比值均低于正常对照组（P〈0．05）。治愈组CD3＋、CD8＋及CD4＋T细胞绝对数均低于正常对照组（P〈0．05）。与治疗前比较,治愈组治疗后CD3＋、CD8＋及CD4＋T细胞绝对数明显升高（P〈0．05）;但CD3＋、CD4＋T细胞绝对数及CD4＋／CD8＋比值仍低于正常对照组（P〈0．05）。死亡组治疗前、后CD3＋、CD8＋及CD4＋T细胞绝对数均低于治愈组和正常对照组（P均〈0．05）;且治疗前、后各项指标之间差异均无统计学意义（P〉0．05）。治愈组CD3＋、CD8＋及CD4＋T细胞绝对数均高于死亡组（P〈0．05）。结论H7N9禽流感患者存在比较严重的T淋巴细胞亚群失衡,死亡患者免疫失衡更为严重。T淋巴细胞亚群失衡可能与H7N9禽流感的发病有关。     

前列腺素E2对Th1／Th2和Tcl／Tc2淋巴细胞免疫平衡的调节

本研究探讨前列腺素E2（PGE2）对外周血T淋巴细胞体外增殖的影响,以及对Th1／Th2和Tc1／Tc2细胞的免疫平衡的调节作用。不同浓度PGE2与抗CD3和抗CD28单克隆抗体（mAb）和健康成人外周血单个核细胞（MNC）共同培养120小时,测定细胞增殖程度。ELISA方法测定24、48、72和120小时细胞培养上清液中IFN-γ和IL-4浓度变化。流式细胞仪测定CD4＋IL-4＋T细胞和CD4＋1FN-γT细胞以及CD8＋IL-4＋T细胞和CD8＋IFN-γ＋ T细胞比值。各实验均以不加PGE2为对照。结果表明：①随PGE：的浓度增加,T细胞体外增殖的抑制率明显增高（p=0．001）;T细胞增殖抑制率与PGE2浓度之间呈明显的正相关（r=0．889,P=0．000）。②实验组培养120小时的IFN-γ浓度与第72小时的IFN-γ浓度差异无显著性（P=0．917）,对照组细胞培养上清液中的IFN-γ浓度随时间持续增高（P=0．046）;实验组不同时间的IFN-γ浓度均明显低于对照组（P〈0．05）。实验组在不同时间产生IL-4浓度无明显变化（P=0．400）;对照组24小时细胞培养上清中IL-4浓度高于48、72和120小时（P值分别为0．007、0．003和0．002）;实验组细胞培养24小时时IL-4浓度明显低于对照组（P=0．037）;实验组细胞培养48、72和120小时与对照组IL-4浓度差异无显著性（P〉0．05）。⑧实验组与对照组CD4＋IFN-γT细胞的比例无明显变化（P=0．767）;实验组CD4＋IL-4＋T细胞比例略高于对照组（P=0．051）;实验组CD4 IL-4＋T细胞与CD4＋IFN-γT细胞的比值明显高于对照组（P=0．011）。实验组与对照组CD8＋IFN-γT细胞的比例无明显变化（P=O．441）;实验组CD8＋IL-4＋T细胞的比例明显高于对照组（P=0．015）;实验组CD8＋ IL4＋T细胞与CD8＋IFN-γT细胞的比值明显高于对照组（P=0．038）。结论：PGE2体外抑制外周血T细胞的增殖;PGE2作用24小时即可抑制IFN-γ和儿4的产生,并且明显影响T细胞1FN-γ的高峰出现,对IFN-γ具有持续性的抑制作用,对见-4的持续性影响并不明显;PGE2使CD4＋IL-4＋T细胞与CD4＋IFN-γT细胞的比值和CD8＋IL-4＋T细胞与CD8＋IFN-γT细胞的比值增加,调节T细胞的免疫反应向Th2／T02方向发展。     

Sensitivity of whole-blood T lymphocytes in individual patients to tacrolimus (FK 506): impact of interleukin-2 mRNA expression as surrogate measure of immunosuppressive effect

BACKGROUND: To optimize immunosuppressive treatment in individual transplant patients, functional measurements of the effects of tacrolimus (FK 506) are of clinical importance. Previous investigations have demonstrated the occurrence of tacrolimus-resistant production of interleukin-2 (IL-2) in vitro, which may explain in part why rejection episodes are still a frequent problem despite attainment of therapeutic blood concentrations and HLA matching. However, an adequate surrogate marker to define the tacrolimus response in individual patients has not been established. METHODS: We investigated the immunosuppressive effects of tacrolimus on anti-CD3/anti-CD28 T-cell costimulation in a human whole-blood assay, analyzing T-cell proliferation, activation marker expression (CD25, CD69), IL-2 protein expression, and cytokine mRNA expression in vitro (n = 11 healthy individuals). We also quantified IL-2 mRNA expression in patients undergoing tacrolimus (n = 4) or cyclosporin A (CsA; n = 4) monotherapy before ex vivo living-donor kidney transplantation. RESULTS: T-cell proliferation; CD25, CD69, and IL-2 concentrations; and IL-4 mRNA were significantly decreased in vitro. In contrast, cytokine mRNA profiles revealed variable tacrolimus sensitivity. Whole-blood samples from 3 of 11 healthy individuals demonstrated marked suppression of IL-2 mRNA expression (>50%) when tacrolimus was administered in vitro. When CsA was added to whole-blood cultures, the influence on IL-2 mRNA expression was comparable to that of tacrolimus in 9 of 11 individuals. Two individuals responded conversely, indicating that differences in the in vitro response to tacrolimus and CsA among individuals may be attributable to potential heterogeneity in the involvement of the CD28 pathway. Kinetic profiles of IL-2 mRNA expression also revealed individually distinct degrees of calcineurin inhibitor sensitivity in patients undergoing tacrolimus or CsA monotherapy before living-donor kidney transplantation. CONCLUSIONS: Our results suggest an individual degree of calcineurin inhibitor sensitivity of activated whole-blood lymphocytes based on IL-2 mRNA expression. Our approach is potentially valuable for identifying transplant patients in whom IL-2 mRNA expression is unaffected or even enhanced after initiation of immunosuppressive therapy. Such individuals may be less sensitive to the immunosuppressive agent and therefore at increased risk of transplant rejection. Prospective studies are necessary to determine the correlation of IL-2 mRNA expression with the clinical risk of transplant rejection.     

Effects of UVB radiation on cytokine generation, cell adhesion molecules, and cell activation markers in T-lymphocytes and peripheral blood HPCs

BACKGROUND: Immunomodulatory effects of UV light have increasingly become a focus in transfusion medicine, BMT and transplantation immunology. In the transplant setting, the use of UVB radiation may reduce or abolish T-cell activation without compromising either bone marrow (BM) engraftment or graft-versus-leukemia effect. In this study, BM and apheresis-derived peripheral blood HPCs were used to investigate the effects of UVB on colony-forming ability, CD34+ cell viability, and growth potential, as well as on the secretion of MNC cytokines and the expression of cell surface markers and adhesion molecules. STUDY DESIGN AND METHODS: After UVB radiation, enriched populations of T cells and antigen-presenting cells (APCs) were treated with PHA, and the MNC response was measured, as was colony-forming ability. CD34+ cells were quantified and their growth potential was determined in culture. Next, T-cell activation status, cell adhesion molecule and cell surface activation marker expression, and cytokine profiles were evaluated, and cytokine mRNA was quantitated. Parallel studies were done in unirradiated control cell populations. RESULTS: Low-dose (10 mJ/cm(2)) UVB mitigates MNC proliferative responses by 94 percent while maintaining 60 and 80 percent of colony-forming ability in peripheral blood HPC and BM preparations, respectively, and >50 percent of colony-forming ability in CD34+ cell-enriched samples. Low-dose UVB radiation also significantly reduces T-cell production of TNFalpha, TNFalpha mRNA, TNFbeta, IL-2, and IL-6 and downregulates T-cell expression of CD28, CD25, CD69, and intercellular adhesion molecule 1. CONCLUSION: These findings have shown that a "window" of low-dose UVB radiation (10 mJ/cm(2)) exists, at which BM- and peripheral blood-derived MNC proliferation is inactivated, while the HPCs are relatively spared. UVB light selectively affects T cells, while APCs are resistant to low doses of UVB. UVB radiation also alters the expression of some cell surface markers and cytokines that are important in T-cell activation pathways. Reduction of T-cell activation without cytocidal effect may allow UVB radiation to become an immunomodulating agent in BM or HPC transplantation.     

Increased number of suppressor T-cells and impaired IL-2 mediated T-cell function in peripheral blood of gastric cancer patients

The present study was designed to investigate the mechanisms responsible for suppressed T-cell immunity in gastric cancer patients. The peripheral blood T-cell functions in gastric cancer patients were evaluated by measuring responses to PHA and IL-2, and T-cell subpopulations were assessed by flow cytometry. Both the PHA and IL-2 responses decreased in patients with gastric cancer, although the proportions of CD3+ and CD25+ cells were the same for gastric cancer patients and healthy volunteers. The PHA response, but not that of IL-2, was lower in patients with liver metastasis or peritoneal dissemination than in patients without these conditions. Single regression analysis showed that with lymph node involvement in the disease the IL-2 response was more strongly suppressed than PHA response. The serum level of IAP did not correlate with the response to either PHA or IL-2. The proportion of CD4+ cells was not affected by any factor related to gastric cancer, although CD4+/CD8+ and CD8+11b-/CD8+11b+ ratios did decrease in patients with distant lymph node involvement. These changes may be due to a comparative increase in suppressor cells. These results suggest that gastric cancer patients exhibit impaired IL-2 mediated T-cell function and that the number of suppressor cells may increase with progressive lymph node involvement. These two serial effects may be indeed responsible for the impaired blood T-cell function in patients with gastric cancer.     

Inability to mediate prolonged reduction of regulatory T Cells after transfer of autologous CD25-depleted PBMC and interleukin-2 after lymphodepleting chemotherapy

CD25CD4 regulatory T cells (Treg) regulate peripheral self-tolerance and possess the ability to suppress antitumor responses, which may explain the poor clinical response of cancer patients undergoing active immunization protocols, and provides the rationale for neutralizing Treg cells in vivo to strengthen local antitumor immune responses. Because interleukin-2 (IL-2) mediates tumor regression in about 15% of treated patients but simultaneously increases Treg cells, we hypothesized that transient elimination of Treg cells will enhance the clinical effectiveness of IL-2 therapy. In the current study, 5 patients with metastatic melanoma who were refractory to prior IL-2 received a lymphodepleting preparative regimen followed by the adoptive transfer of autologous lymphocytes depleted of CD25 Treg cells and high-dose IL-2 administration. CD25 cells were eliminated from patient leukapheresis samples using a clinical-grade, large-scale immunomagnetic system, leaving CD8 and CD25CD4 T cells intact. In the early aftermath of CD25 Treg cell-depleted cell infusion, CD25FOXP3+ CD4 Treg cells rapidly repopulated the peripheral blood of treated patients with 18% to 63% of CD4 T cells expressing FOXP3. Recovering CD25CD4 T cells exhibited suppressive activity against CD25CD4 effector T-cell proliferation in vitro. No patient experienced objective tumor regression or autoimmunity. Our results indicate that in vivo transfer of autologous CD25-depleted mononuclear populations to lymphopenic patients in combination with high-dose IL-2 is not sufficient to mediate prolonged reduction of Treg cells after IL-2 administration.     

Delayed cytokine mRNA expression kinetics after T-lymphocyte costimulation: a quantitative measure of the efficacy of cyclosporin A-based immunosuppression

BACKGROUND: Because cyclosporin A (CsA) and glucocorticoids inhibit the production of interleukin-2 (IL-2) and other cytokines, quantitative analysis of cytokine mRNA might constitute a pharmacodynamic measure for immunosuppressive drug effects. We investigated whether immunosuppressive drugs influence cytokine mRNA expression kinetics during T-cell costimulation. METHODS: We used a human whole blood assay to determine basal (unstimulated) IL-2, IL-4, and tumor necrosis factor-alpha (TNF-alpha) mRNA concentrations and expression kinetics after anti-CD3/anti-CD28 monoclonal antibody costimulation in kidney transplant recipients undergoing CsA-based immunosuppressive triple therapy and in healthy controls (ex vivo study I). The effect of CsA on IL-2 mRNA expression kinetics was also determined ex vivo in patients undergoing CsA monotherapy (ex vivo study II) and after in vitro addition of CsA. RESULTS: In ex vivo study I, basal TNF-alpha mRNA but not IL-2 and IL-4 mRNA was decreased in kidney transplant patients. We observed shifts in peak IL-2 and IL-4 (from 8 to 24 h) and TNF-alpha (from 4 to 8 h of costimulation) mRNA expression in kidney transplant patients after T-cell costimulation. In patients undergoing CsA monotherapy (ex vivo study II), the inhibitory effect of CsA was detectable as an individually delayed increase in IL-2 mRNA during costimulation. In vitro addition of CsA also induced a dose-independent displacement of IL-2 mRNA expression kinetics (i.e., a delay). CONCLUSIONS: A delayed increase in cytokine mRNA expression during T-cell costimulation may represent a sensitive effect of immunosuppression. The single analysis of one absolute or peak mRNA value could be misleading. For prospective studies involving measurement of cytokine mRNA, we therefore suggest the parameter "area of cytokine mRNA expression over time", which should include absolute cytokine mRNA values at two different time points of mRNA kinetics.     

Comparison of common gamma-chain cytokines, interleukin-2, interleukin-7, and interleukin-15 for the in vitro generation of human tumor-reactive T lymphocytes for adoptive cell transfer therapy

The adoptive transfer of human tumor-reactive T lymphocytes into autologous patients can mediate the regression of metastatic melanoma. Here, the in vitro generation of melanoma-reactive T lymphocytes was compared using 3 common gamma-chain cytokines, interleukin (IL)-2, IL-7, and IL-15, alone or in combination. The proliferation, function, and phenotype were evaluated for tumor-reactive T cells derived from peripheral blood mononuclear cells (PBMCs) from patients previously immunized with the melanoma-associated peptide gp100:209-217(210M) and PBMCs transduced with a retrovirus encoding the alpha and beta chains of a gp100-reactive T-cell receptor (TCR). IL-7 alone did not induce significant proliferation of any tumor-reactive T-cell population, whereas IL-2 and IL-15 induced significant proliferation of tumor-reactive T lymphocytes from both sources. Cells cultured in the presence of IL-2 or IL-15 secreted comparable amounts of interferon-gamma and IL-2 in response to melanoma cells in vitro and were phenotypically similar in terms of costimulatory molecules (CD27 and CD28), cytokine receptors (CD25, CD122, and CD127), and a lymphoid homing molecule (CD62L). In addition, the proliferation, function, and phenotype of T cells cultured with combinations of IL-2, IL-7, and IL-15 were similar to those grown with IL-2 alone. The effects of these cytokines on TCR stimulation of CD45RA+ naive cells derived from adult patients and from human umbilical cord blood were also compared. Similar to the data with activated tumor-reactive T lymphocytes, IL-7 alone did not support significant proliferation of naive T cells after TCR stimulation with anti-CD3, although IL-2 and IL-15 induced comparable proliferation of T lymphocytes with similar phenotypic attributes.     

环孢素A联合供者骨髓细胞输注延长同种大鼠移植心脏存活时间

背景:同种异体器官移植后需长期使用免疫抑制剂控制机体免疫状态以免发生排斥反应,但免疫抑制治疗使受者的感染发生率增高,诱导移植免疫耐受是解决这一问题的理想方法.在啮齿类动物,已能诱导出长期免疫耐受,其中供者骨髓细胞输注在诱导免疫耐受中具有重要作用.目的:观察环孢素A联合供者骨髓细胞输注对同种大鼠移植心脏存活时间的影响.设计:随机对照动物实验.单位:浙江大学医学院附属第一医院肾脏病中心.材料:实验于2002-03/2004-12在浙江大学医学院实验动物中心完成.选用SPF级近交系雄性Lewis大鼠40只和雄性BN大鼠60只,实验方法符合动物伦理学要求.方法:①制作Lewis→BN大鼠的异位(腹部)心脏移植模型(n=40),将40组动物模型按随机数字表法分为4组(n=10):对照组不进行特别处理;环孢素A组术后连续7 d给予环孢素A:联合处理组分别于术中及术后第6天输注供者骨髓细胞,术后连续7 d给予环孢素A;供者骨髓细胞组分别于术中及术后第6天输注供者骨髓细胞.另以BN大鼠间的异位(腹部)心脏移植作为BN对照组(n=10).②观察各组受体大鼠移植心脏的存活时间,检测术后第6天血清白细胞介素2含量以及移植心脏组织中肿瘤坏死因子mRNA表达水平,应用流式细胞仪检测术后第6,12,18天时受体外周血有核细胞中的供体来源细胞、CD3 CD25 细胞、CD4 CD25 细胞的百分比以及共刺激分子CD86表达水平、CD4 CD45RC /CD4 CD45RC-比率等.主要观察指标:各组受体大鼠移植心脏存活时间、血清白细胞介素2含量及心肌组织中肿瘤坏死因子α mRNA水平、排斥反应分级、外周血有核细胞中的供者来源细胞、CD3 CD25 细胞、CD4 CD25 细胞的百分比及共刺激分子CD86表达、CD4 CD45RC /CD4 CD45RC-比率等.结果:Lewis大鼠40只及雄性BN大鼠60只全部进入结果分析.①联合处理组大鼠移植心脏存活时间长于对照组和供者骨髓细胞组(P＜0.05).②联合处理组大鼠血清白细胞介素2含量及心肌组织中肿瘤坏死因子α mRNA表达水平均低于对照组和供者骨髓细胞组(P＜0.05).③联合处理组大鼠排斥反应程度轻于其他3个移植组.④联合处理组大鼠外周血有核细胞上CD86表达受到明显抑制:术后6.12d联合处理组的CD4 CD45RC /C134 CD45RC比率及D3 cD25 细胞百分比均低于对照组和供者骨髓细胞组:接受供者骨髓细胞输注大鼠外周血中供者来源的有核细胞多于未输注大鼠.结论:短疗程环孢素A治疗联合供者骨髓细胞输注能减轻大鼠心脏移植急性排斥反应程度,延长移植心脏存活时间.     

Immunophenotypic analyses of CD34(+) cell subsets in bone marrow from HIV-infected patients during highly- active antiretroviral therapy

BACKGROUND: Because increased bone marrow lymphopoiesis might contribute to immunologic reconstitution during highly-active antiretroviral therapy (HAART), we examined the effect of HAART on CD34(+) cell subsets in bone marrow from HIV-infected patients. MATERIALS AND METHODS: In 12 HIV-infected patients, bone marrow and peripheral blood were collected before then 4 and 26 weeks after initiating HAART. Bone marrow in 28 HIV-seronegative controls was also examined. Immunophenotypic analyses of CD34(+) cell subsets in bone marrow were performed by flow cytometry. RESULTS: Our main findings in bone marrow were: (i) HIV-infected patients had increased proportions of CD34(+)cells expressing T- and B-cell markers before initiating HAART; (ii) in contrast, these patients had decreased proportions of CD34(+) cells expressing myeloid-associated markers; (iii) although HAART induced an increase in peripheral T-cell counts, the percentage of CD34(+)cells expressing T-cell markers tended to decrease during such therapy; (iv) HAART induced a decrease in serum IgG accompanied by a slight decrease in the proportion of CD34(+)cells expressing B-cell markers; (v) in contrast, HAART induced a significant increase in peripheral granulocyte counts, accompanied by a slightly increased proportion of CD34(+) cells expressing myeloid-associated molecules. CONCLUSION: Our findings are compatible with an HIV-related block in T-cell differentiation, leading to accumulation of T-cell progenitors in bone marrow, and such a block may be removed by HAART.     

Immunomodulatory effects of three macrolides, midecamycin acetate, josamycin, and clarithromycin, on human T-lymphocyte function in vitro.

The effect of three macrolide antibiotics, midecamycin acetate, josamycin, and clarithromycin, on human T-cell function was investigated in vitro. Midecamycin acetate and josamycin suppressed the proliferative response of peripheral blood mononuclear cells stimulated by polyclonal T-cell mitogens at concentrations between 1.6 and 8 micrograms/ml. At higher concentrations (40 to 200 micrograms/ml), all these drugs showed a marked inhibitory effect. At concentrations of 1.6 to 40 micrograms/ml, these drugs suppressed interleukin-2 (IL-2) production induced by mitogen-stimulated T cells, but not the expression of IL-2 receptor (CD25), in a dose-dependent manner. Therefore, the suppressive action on T-lymphocyte proliferation seems to be based on the ability of these drugs to inhibit IL-2 production by T cells. The drug also inhibited mixed lymphocyte reaction at the same concentrations. Combined treatment with these macrolides and the known immunosuppressants such as FK506 and cyclosporin A resulted in an increased inhibition of T-cell proliferation. The immunomodulatory properties of the antibiotics may have clinical relevance for modulation of the immune response in transplant patients and in patients with inflammatory diseases.     

CD4~＋ CD25~＋细胞和CD8~＋ CD28~-调节性T细胞在原发性肝癌患者外周血中的水平变化

目的研究原发性肝癌（HCC）患者外周血中CD4＋ CD25＋细胞和CD8＋ CD28-调节性T细胞的表达及意义。方法采用流式细胞仪测定93例HCC患者和24名健康人外周血中的CD4＋ CD25＋细胞和CD8＋ CD28-调节性T细胞水平。结果 HCC患者外周血中的CD4＋ CD25＋细胞[（14.39±4.58）%]和CD8＋ CD28-调节性T细胞[（24.05±5.64）%]表达均显著高于对照组[（8.37±1.73）%、（15.93±4.84）%,P〈0.05];CD8＋CD28-调节性T细胞与HCC的分期呈正相关（r=0.463,P〈0.001）。结论 CD4＋ CD25＋细胞和CD8＋ CD28-调节性T细胞在HCC患者外周血中呈高表达,且CD8＋ CD28-调节性T细胞与HCC的病情及预后相关。