Lectin-Activated CD4+CD45RA+ T-Lymphocytes have no Ability to Kill Monocytes

Monocytes are eliminated from cell culture by antigen or mitogen activated cytotoxic CD4⁺ T-lymphocytes. In this report we asked the question whether CD4⁺CD45RA⁺ and CD4⁺CD45RO⁺ subpopulations differ in the ability to kill monocytes in pokeweed mitogen (PWM)-activated cultures. Data are presented that although CD4⁺CD45RA⁺ vigorously proliferate in the presence of PWM, they do not kill monocytes or secrete IFNγ.     

Activation status of T and NK cells in the endometrium throughout menstrual cycle and normal and abnormal early pregnancy

Endometrial lymphocytes were studied at all stages throughout the menstrual cycle and early pregnancy by flow cytometry to examine different lymphocyte subpopulations and the expression of the T- and NK-cell activation markers. After pregnancy, CD8⁺CD3⁺ lymphocytes were decreased in the decidua. In both endometrium and decidua, more T cells expressed CD69, CD71, HLA-DR, and CD38 antigens than in peripheral blood. After pregnancy, CD71⁺CD3⁺ lymphocytes were further increased. CD25⁺CD3⁺ lymphocytes decreased significantly in the endometrium and decidua of ectopic pregnancies, but not in the decidua of normal pregnancies. These findings indicate that T cells are regionally activated in the first trimester, and it may be the result of the stimulation by fetal antigens. NK cells were the most abundant cell type in the decidua, which expressed the phenotype CD16⁻ CD56⁺, and CD57⁻CD56⁺. The proportion of activated decidual NK cells was increased in anembryonic pregnancies more than in normal pregnancies, although the total NK subpopulation was similar in both groups. This might result in increased NK cytotoxicity in anembryonic pregnancies. In conclusion, T cells are activated, but NK cytotoxicity is decreased in the decidua of early normal pregnancies. This might be important in the control of trophoblast growth and placental development.     

A study of CD45RO, CD45RA and CD29 antigen expression on human decidual T cells in an early stage of pregnancy

The decidua is the place where the fertilized egg is implanted and where the immunocompetent cells of the mother come into direct contact with genetically disparate cells of the conceptus. Although the T cells in the decidua are exposed to fetal antigens, the fetus is not rejected by maternal immunocompetent cells. In the present study, we examined surface markers to determine whether the T cells in the human decidua are naive T cells without or memory T cells with a history of antigen stimulation. Although few T cells were present in the decidua, as compared to the peripheral blood, CD45RO⁺, CD29⁺ and CD45RA⁻ CD4⁺ T cells as well as CD45RO⁻, CD29⁺ and CD45RA⁻ CD8⁺ T cells, which are considered to be memory T cells, were in the majority, with only small numbers of CD45RO⁻, CD29⁻ and CD45RA⁺ CD4⁺ and CD8⁺ cells, which are naive T cells, present. Also, the decidual mononuclear cells secreted IL-2 and IL-4. Since IL-4 is secreted only by memory T cells, it is suggested that in the decidua memory T cells increase in number and secrete cytokines, thereby in some way influencing the phenomenon of fertility.     

CD3+、CD4+和CD8+ T细胞Vβ亚家族中PD-1免疫表型检测方法的建立

 目的：建立分析CD3⁺、CD4⁺和CD8⁺ T细胞Vβ亚家族中免疫抑制性受体程序性细胞死亡蛋白1（PD-1）分子免疫表型的方法,从而了解人体外周血T细胞谱系中PD-1表型细胞的分布频率。方法：收集健康个体（HI）外周血10例,利用抗CD45、CD3、CD4、CD8、PD-1等多色荧光流式单抗,以及T细胞受体（TCR）Vβ谱系试剂盒[IOTest^® Beta Mark TCR Vβ Repertoire Kit,共8管,每一管均为FITC和PE偶联的复合抗体（A~H）,每一管复合抗体包含3个TCR Vβ亚家族的抗体],检测T细胞亚群TCR Vβ亚家族中PD-1的分布情况。结果：在10例HI的检测中,CD3⁺、CD3⁺CD4⁺和CD3⁺CD8⁺ T细胞亚群中检测的24个TCR Vβ亚家族总和符合试剂盒所提供的Quick Reference Card数据,初步结果显示,HI中CD3⁺ T细胞主要优势TCR谱系是Vβ2、Vβ3、Vβ8、Vβ9、Vβ5.1、Vβ13.1和Vβ13.2;而在CD3⁺CD4⁺ T细胞中的优势利用主要集中在Vβ2、Vβ3、Vβ8、Vβ9、Vβ5.1和Vβ13.1;在CD3⁺CD8⁺ T细胞中则主要为Vβ1、Vβ2、Vβ3、Vβ9、Vβ13.1和Vβ13.2等亚家族。进一步分析显示PD-1⁺细胞在HI的CD3⁺、CD3⁺CD4⁺和CD3⁺CD8⁺ T细胞24个TCR Vβ亚家族中均可以检测到,PD-1⁺细胞在T细胞各亚群中分布频率有个体差异,在CD4⁺ T细胞中,以Vβ5.2⁺ T细胞的分布频率较高,而在CD8⁺ T细胞中,以Vβ11⁺和Vβ13.6⁺ T细胞的分布频率较高。结论：本项工作利用健康人样本成功建立了多色荧光流式细胞术检测T细胞亚群TCR Vβ谱系中免疫抑制性受体PD-1分子免疫表型的方法,为进一步应用于分析白血病等病人TCR Vβ谱系的免疫抑制特点提供了新方法。     

Dye Dilution Proliferation Assay: Application of the DDPA to Identify Tumor-Specific T Cell Precursor Frequencies in Clinical Trials

A better understanding of immune effector and regulatory pathways has led to innovative, and complex, immunotherapy strategies. CD8⁺ cytolytic T lymphocytes (CTL) provide one common pathway of tumor cell destruction. The peripheral blood CTL compartment typically comprises a minority of anti-tumor CD8⁺ lymphocytes and the determination of their number during clinical trials is the focus of various laboratory methods. We have monitored tumor specific CD8⁺ as well as CD4⁺ lymphocyte precursor frequencies in the peripheral blood using a Dye Dilution Proliferation Assay (DDPA). We summarize our experience applying DDPA in a multi-parameter, antigen-specific assay, detailing some of its complexities and advantages. We provide examples of our clinical trial results showing tumor-specific CD8⁺ and CD4⁺ precursor frequency (PF) data in patients being treated on novel immunotherapy trials.     

内分泌和化疗对乳腺肿瘤患者干细胞的影响

BACKGROUND:Tumor stem cells are the root of cancer recurrence and metastasis, so clinical researches should focus on the effects of different treatments on tumor stem cells. OBJECTIVE:To explore the effects of endocrine therapy and chemotherapy on stem cells in patients with breast cancer. METHODS:After recovery and cultivation of estrogen receptor-positive human breast cancer cell lines MCF-7, passage 3 cells in logarithmic phase were selected and divided into three groups containing control, estradiol and estradiol with tamoxifen groups. The estradiol group was divided into three subgroups: 10^-7, 10^-8 and 10^-9 mol/L estradiol was added into the medium, respectively; the estradiol with tamoxifen group was divided into three subgroups: 10^-7, 10^-8 and 10^-9 mol/L estradiol with 10^-6 mol/L tamoxifen were added into the medium, respectively. The same amount of absolute ethyl ethanol was added into the medium of control group. Fifteen female patients with late recurrence and metastasis of breast cancer received chemotherapy as recurrence and metastasis group. Another 15 healthy volunteers were selected as healthy control group. RESULTS AND CONCLUSION:The proportion of CD44⁺CD24^-/low cell subsets in the estradiol and estradiol with tamoxifen groups was significantly higher than that of the control group (P < 0.05), and the proportion of CD44⁺CD24^-/low cell subsets in the estradiol group was significantly higher than that of the estradiol with tamoxifen group at the same concentration (P < 0.05). The proportion of CD44+CD24-/low cell subsets had no significant differences among groups at 10 and 20 days of culture (P < 0.05). The proportion of CD44⁺CD24^-/low cell subsets significantly increased in MCF-7 cells after 24-hour intervention with different chemotherapy drugs. But only the proportion of CD44⁺CD24^-/low cell subsets in the paclitaxel and doxorubicin groups was significantly higher than that of the control group after 20-day intervention (P < 0.05). Besides, the proportion of CD44⁺CD24^-/low cell subsets in the peripheral blood of healthy volunteers was significantly lower than that of the recurrence and metastasis group (P < 0.05). Among 15 patients with late recurrence and metastatic of breast cancer, 9 had stable disease, 5 had partial remission, 1 had failed chemotherapy and cancer progression. Moreover, the proportion of CD45^-CD44⁺CD24^-/low cell subsets in the peripheral blood of patients sensitive for chemotherapy was significantly lower than that before treatment (P < 0.05). In conclusion, both endocrine therapy and chemotherapy exert a certain effect on the CD44⁺CD24^-/low cell subsets of breast cancer positive for estrogen receptor. Given that CD44⁺CD24^-/low cell subsets in MCF-7 cells resist chemotherapy drugs, the proportion of CD45^-CD44⁺CD24^-/low cells in the peripheral blood of patients sensitive for chemotherapy is decreased.     

Naive and memory CD4+ T-cells in the cerebrospinal fluid of children with aseptic meningitis following measles-mumps-rubella vaccination and enteroviral meningitis

We investigated the distribution of memory (CD45RO⁺) and naive (CD45RA⁺CD62L⁺) CD4⁺ T-cells as well as CD8⁺ T-cells and total T-cells in the CSF of children with aseptic meningitis following measles-mumps-rubella (MMW) vaccination and those with enteroviral meningitis. Flow cytometric analysis of CSF cells was performed in 12 children with MMR vaccine-associated meningitis and 11 children with enteroviral meningitis. Percentages of total T-cells, CD4⁺ and CD8⁺ T-cells and monocytes in CSF of patients from the two groups were not significantly different. The majority of CD4⁺ T-cells in the CSF of both patient groups were of memory phenotype. Percentages of CSF naive CD4⁺ T-cells were increased in children with aseptic meningitis following MMR vaccination. Further studies focused on the more detailed immunophenotyping of CSF cells are needed to fully establish the usefulness of flow cytometry in the diagnostic workup of inflammatory CNS diseases in children.     

THE PERIPHERAL BLOOD LEUKOCYTE PHENOTYPE IN PATIENTS WITH BREAST CANCER: EFFECT OF DOXORUBICIN/PACLITAXEL COMBINATION CHEMOTHERAPY

Presence of functional immune system is critical for any attempt aimed at improving survival of breast cancer patients by strategies based on immune system manipulation. We evaluated by flow cytometry the phenotype of peripheral blood leukocyte of 43 breast cancer patients. In 11 patients, the phenotype was evaluated before and during the chemotherapy by combination of doxorubicin and paclitaxel (AT). Compared with controls breast cancer patients had significantly higher relative and absolute numbers of CD3^-HLADR⁺, CD3^-CD69⁺ and CD14⁺CD16^-, and significantly lower percentages of CD3^- and CD8^-CD28⁺ cells. After one cycle of AT, the absolute numbers of CD3⁺, CD3^-CD4^-, CD3⁺CD8⁺ and CD8^-CD28⁺ cells increased significantly. Present data show a presence of T-cell activation in breast cancer patients. Administration of AT may lead to an increase in functional T-cells in peripheral blood, indicating a potential for combining chemotherapy with immunotherapy in the treatment of breast cancer patients.     

肝移植后他克莫司血药浓度对外周血单个核细胞内HBV DNA的影响

背景：肝移植后他克莫司等免疫抑制剂的长期应用导致机体细胞免疫功能降低,并有可能影响机体对乙肝病毒的清除。 目的：分析乙肝相关肝移植患者后不同浓度他克莫司对外周血单个核细胞中的HBV DNA含量的影响。 方法：纳入乙肝相关终末期肝病肝移植受者23例,根据移植后12周清晨空腹他克莫司血药浓度,分为高浓度组(≥ 10 μg/L) 9例和低浓度组(< 10 μg/L)14例,同时用荧光标记单克隆抗体结合流式细胞技术检测外周血T细胞亚群的百分比,用实时荧光定量PCR检测外周血单个核细胞内的HBV DNA。 结果与结论：用多元线性回归分析外周血单个核细胞内的HBV DNA含量与CD8⁺CD152⁺呈正相关,与CD8⁺CD28⁺呈负相关。高血药浓度他克莫司的患者外周血单个核细胞内的HBV DNA高于低浓度组,其改变与反映细胞免疫功能的指标CD8⁺CD152⁺和CD8⁺CD28⁺的变化有关。     

老年初诊急性髓细胞白血病患者CD4+CD25+调节性T细胞的表达

背景：研究证实,很多恶性肿瘤患者体内CD4⁺CD25⁺调节性T细胞存在高表达,近期也有研究发现,急性髓细胞白血病患者外周血CD4⁺CD25⁺调节性T细胞同样表现出高比例表达。 目的：分析老年初诊急性髓细胞白血病患者CD4⁺CD25⁺调节性T细胞的表达特点。 方法：纳入初诊急性髓细胞白血病患者92例,将年龄在60岁以下者设为中青年组(n=22),年龄在60岁以上者设为老年观察组(n=70)。在老年观察组中,32例经规范化疗后完全缓解,设为完全缓解组;将余下38例设为老年组,依据FAB分型标准,分为M2 6例、M3 19例、M4 7例、M5 6例。另选择同期体检健康人群42名作为正常对照组。抽取受试者外周静脉血,检测CD4⁺CD25⁺调节性T细胞表达情况。 结果与结论：老年组、完全缓解组CD4⁺CD25^highFOXP3⁺调节性T细胞比例高于正常对照组(P < 0.01),并且老年组CD4⁺CD25^high FOXP3⁺调节性T细胞比例高于完全缓解组(P < 0.01)。老年组、完全缓解组CD4⁺FOXP3⁺T细胞比例高于正常对照组(P < 0.01),并且老年组CD4⁺ FOXP3⁺T细胞比例高于完全缓解组(P < 0.01)。老年组CD4⁺CD25^high FOXP3⁺调节性T细胞与CD4⁺ FOXP3⁺T细胞比例高于中青年组(P < 0.01)。老年组不同分型间CD4⁺CD25^high FOXP3⁺调节性T细胞和CD4⁺ FOXP3⁺T细胞比例比较差异均无显著性意义(P > 0.05)。Pearson相关性检验结果显示,老年初诊急性髓细胞白血病患者外周血CD4⁺CD25^high FOXP3⁺调节性T细胞比例和CD4⁺ FOXP3⁺T细胞比例呈正相关(r=0.87,P=0.019)。表明老年初诊急性髓细胞白血病患者CD4⁺CD25⁺调节性T细胞比例高于健康人群和中青年急性髓细胞白血病患者。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;组织工程全文链接：     

骨髓间充质干细胞调节心脏移植大鼠的免疫功能

BACKGROUND:Heart transplantation is an effective method for treatment of end-stage heart failure, but immune rejection that seriously impact therapeutic effacicy is easy to occur after transplantation. OBJECTIVE:To investigate the regulatory effect of bone marrow mesenchymal stem cells on the immune function of rats undergoiong heart transplantation. METHODS:Twenty Lewis rats were enrolled as donors, and 20 Wistar rats as recipients. Heart transplantation models were established in the Wistar rats. These 20 model rats were randomized into cell transplantation and control group with 10 rats in each group. Forty-eight hours after heart transplantation, rats in the cell transplantation group were given bone marrow mesenchymal stem cell suspension (1 mL, 2×10⁸ cells/L) via the tail vein, while rats in the control group were given normal saline in the same dose. Then, the expression levels of serum interleukin-2, interleukin-10 and percentage of CD4⁺, CD8⁺, CD4⁺/CD8⁺, CD4⁺CD25^high, CD4⁺CD25^high Foxp3⁺ T cells in the venous blood were detected in the two groups at 7 days after cell transplantation. Additionally, rat myocardial tissues were taken and observed pathologically. RESULTS AND CONCLUSION:The survival time of the cell transplantation group was significantly longer than that of the control group (P < 0.05). The expression level of interleukin-2 showed no significant difference between the two groups (P > 0.05), but the level of interleukin-10 in the cell transplantation group was significantly higher than that in the control group (P < 0.05). Compared with the control group, the percentage of CD4⁺/CD8⁺, CD4⁺CD25^high, CD4⁺CD25^high Foxp3⁺ and CD4⁺ T cells was significantly higher, and the percentage of CD8⁺ T cells was significantly lower in the cell transplantation group (P < 0.05). Histopathological findings showed that there were a small amount of infiltrated lymphocytes in the cell transplantation group with the presence of slight bleeding and edema, and these inflammatory reactions were milder than those in the control group. These findings indicate that bone marrow mesenchymal stem cell transplantation can effectively reduce the rejection in rats undergoing heart transplantation.     

血红素加氧酶1在间充质干细胞上调哮喘患者外周血调节性T细胞中的作用

背景：上调CD4⁺CD25⁺CD127^low/-调节性T细胞是目前治疗哮喘的新靶点。骨髓间充质干细胞在体外可上调正常人外周血的调节性T细胞,但机制尚未明确。 目的：观察血红素加氧酶1对间充质干细胞上调哮喘患者外周血CD4⁺CD25⁺CD127^low/-调节性T细胞的影响。 方法：分别加入0,15,30,45,60 μmol/L氯化高铁血红素和0,5,10,15,20 μmol/L锌-原卟啉对骨髓间充质干细胞进行预处理,荧光定量PCR检测各组间充质干细胞中血红素加氧酶1 mRNA的表达量。密度梯度离心法分离哮喘急性发作期患者和健康对照者的外周血单个核细胞,分别与经氯化高铁血红素预处理、经锌-原卟啉预处理、不经预处理的间充质干细胞共孵育,加入植物血凝素反应72 h,流式细胞仪检测各组CD4⁺CD25⁺CD127^low/-调节性T细胞占CD4⁺T细胞的比例。 结果与结论：人骨髓间充质干细胞中有血红素加氧酶1表达,其表达在体外可被诱导和抑制。间充质干细胞中血红素加氧酶1 mRNA的表达量随氯化高铁血红素浓度增加而增加(P < 0.05),随锌-原卟啉浓度增加而减少(P < 0.05),具有剂量依赖性。间充质干细胞在体外可提高哮喘患者和健康对照者CD4⁺CD25⁺CD127^low/-调节性T细胞的水平(P < 0.01),诱导间充质干细胞中血红素加氧酶1的表达可使共孵育后CD4⁺CD25⁺CD127^low/-调节性T细胞水平提高(P < 0.01),抑制间充质干细胞中血红素加氧酶1的表达可使共孵育后CD4⁺CD25⁺CD127^low/-调节性T细胞的水平降低(P < 0.01),但仍高于对照组(P < 0.01)。提示骨髓间充质干细胞部分通过血红素加氧酶1上调哮喘患者外周血CD4⁺CD25⁺CD127^low/-调节性T细胞。     

Characterization of CD28CD4 T cells in living kidney transplant patients with long-term allograft acceptance

CD28⁻CD4⁺ T-cell subpopulation is expanded in kidney allograft patients with long graft survival. To seek for the roles of CD28⁻CD4⁺ T cells in the long-term acceptance of kidney allografts, we characterized this population by analyzing cell surface molecules, TCR V_β repertoire, mixed lymphocyte reaction (MLR), and cytokine production. The number of CD28⁻CD4⁺ T cells increased correlatively with time after transplantation in this group of patients. The CD28⁻CD4⁺ T cells did not express detectable levels of CD25, CD69, V24, or CTLA-4 but expressed heterogeneous amounts of CD45 RA on the surface. Freshly sorted CD28⁻CD4⁺ T cells revealed a restricted V_β repertoire, whereas the V_β usage of CD28⁺CD4⁺ T cells from the same patients was much diversified. Expression levels of TGF-β and IFNγ gene were significantly higher in the CD28⁻ CD4⁺ T cells than in the CD28⁺CD4⁺ T cells from the kidney allograft patients. These findings suggest that an oligoclonal CD28⁻ CD4⁺ T-cell population is continuously activated in patients with long allograft survival, which may be linked with the long-term acceptance.     

Phenotypical Characterization of Cells in the Thoracic Duct of Patients with and without Systemic Inflammatory Response Syndrome and Multiple Organ Failure

The subset composition and recirculation properties of the migrating lymphocyte pool in humans is largely unknown. The present study was conducted in order to phenotypically characterize cells in human thoracic duct lymph of patients under non-inflammatory and inflammatory conditions. These data were compared with data from peripheral blood, with special emphasis on those cells homing to the gut. Thoracic duct lymph and peripheral blood contained comparable proportions of B and T lymphocytes and CD8⁺ cells. Thoracic duct lymph contained proportionally more CD4⁺ cells, more CD4⁺CD45RO⁺ that express α₄β₇ cells and more CD8⁺CD45RO⁺ that express α₄β₇, as compared to peripheral blood. These data suggest an equal recirculation rate of B and T lymphocytes; a more active recirculation of CD4⁺ cells compared to CD8⁺ cells; and a more active recirculation of memory cells to the gut as compared to other extra-lymphoid sites in patients under non-inflammatory conditions. Data were also obtained in patients with the system inflammatory response syndrome and multiple organ failure. Although it is generally assumed that granulocytes and monocytes do not recirculate, lymph of multiple organ failure patients contained significantly more granulocytes than monocytes, indicating that in severe generalized inflammatory states these cells re-enter the circulation through the thoracic duct. Furthermore, no increased activation of cells homing to the gut was found in these patients.     

供体凋亡淋巴细胞预输注对猪肝移植受体CD4+T、CD8+T、CD4+CD25+调节性T细胞的影响

背景：凋亡细胞能够主动调节机体的免疫功能,并能通过调节机体细胞免疫和体液免疫的途径诱导免疫耐受,但这些结果只在大鼠肝脏移植模型中证实。 目的：探讨通过⁶⁰Co γ射线体外处理后的供体淋巴细胞预输注诱导猪肝移植特异性免疫耐受的作用中,对淋巴细胞亚群的影响。 方法：建立非转流小型猪原位肝移植模型。将受体猪随机摸球法均分为2组：空白对照组,受体猪无特殊处理,行肝移植;淋巴细胞组：受体猪在肝移植前7 d经耳静脉注射⁶⁰Co γ射线处理过的5×10⁸个供体淋巴细胞。观察两组受体猪移植后的存活时间,移植后T淋巴细胞亚型CD4⁺T、CD8⁺T、CD4⁺CD25⁺Tr变化及病理。 结果与结论：移植后3 d,两组病理活检均呈急性中、重度排斥反应;移植后6 d,两组均呈急性重度排斥反应。移植后1,3,6 d CD4⁺T、CD8⁺T、CD4⁺CD25⁺Tr升降趋势,两组间差异无显著意义(P > 0.05)。提示,⁶⁰Co γ射线体外处理过的淋巴细胞预输注未能够诱导猪同种异体肝移植特异性免疫耐受,未能引起T淋巴细胞亚群变化有关。     

不同致敏模式支气管哮喘患儿外周血T淋巴细胞亚群比较及临床意义

目的：分析比较不同过敏原致敏模式下支气管哮喘患儿外周血T淋巴细胞亚群,为进一步明确不同变应原致敏哮喘患儿机体的免疫功能状态差异提供更多依据。方法：入组374例在北京儿童医院过敏反应科确诊的哮喘患儿,行过敏原皮肤点刺试验确定患儿致敏模式,流式细胞术测定外周血T细胞（CD3⁺CD19⁺）、CD4⁺T细胞（CD3⁺CD4⁺）、CD8⁺T细胞（CD3⁺CD8⁺）、Tregs（CD4⁺CD25⁺FOXP3⁺）、Th1（CD4⁺IFN-γ⁺T）、Th2（CD4⁺IL-4⁺T）以及Th17细胞亚群（CD4⁺IL-17A⁺T）,横断面比较上述免疫学指标在不同致敏模式哮喘患儿组间是否存在差异。结果：根据SPT检出阳性过敏原数量,将哮喘患儿分为未致敏哮喘组、单一致敏哮喘组及多重致敏哮喘三组。与健康对照组儿童相比,未致敏哮喘组、单一致敏哮喘组以及多重致敏哮喘三组哮喘患儿的外周血T细胞、CD4⁺T细胞、CD8⁺T细胞、CD4/CD8比值、Th1细胞、Th2细胞、Th1/Th2比值、Tregs、Th17细胞、Tregs/Th17比值等指标的组间差异均具有统计学意义。进一步对三组患儿进行两两比较发现,Tregs、Th17细胞以及Tregs/Th17比值在三组间差异具有统计学意义（均P<0.05）。根据SPT阳性检出过敏原种类,将入组哮喘患儿致敏谱主要分为尘螨组、霉菌组、动物皮屑组、花粉组及其他组。与健康对照组儿童相比,尘螨组、霉菌组、动物皮屑组、花粉组以及其他组过敏原致敏患儿的外周血T细胞、CD4⁺T细胞、CD8⁺T细胞、CD4/CD8比值、Th1细胞、Th2细胞、Th1/Th2比值、Tregs、Th17细胞、Tregs/Th17比值等指标的组间差异均具有统计学意义（均P<0.05）。同样地,行两两比较发现,五组患儿的外周血Th1细胞、Th2细胞、Th1/Th2比值、Tregs、Th17细胞以及Tregs/Th17比值在三组间差异具有统计学意义（均P<0.05）。结论：不同致敏模式的哮喘患儿外周血T淋巴细胞亚群分布状态存在差异,因此应注意不同致敏模式的哮喘患儿免疫应答状态差异。     

Differentiation of T Lymphocytes in the Human Adenoid as Measured by the Expression of CD45 Isoforms

Encounter of antigen by T lymphocyte on antigen-presenting cells results in changes in the expression of several cell surface molecules, including the abundant cell surface glycoprotein CD45. We have characterized the expression of the CD45 isoforms CD45RA and CD45RO in CD4⁺ and CD8⁺ T lymphocytes in the adenoids and peripheral blood of young children. We found that the relative proportions of CD45RA⁻,CD45RO⁺ antigen-experienced T cells was higher in the adenoids than in peripheral blood, and that the proportion of naive or resting CD45RA⁺,CD45RO⁻ T cells was lower in the adenoids than in peripheral blood. The frequency of bright double-positive CD45RA⁺,CD45RO⁺ T cells, which represent cells in transition from the CD45RA⁺ to CD45RO⁺ phenotype, was higher in the adenoids than in peripheral blood. The frequency of another double-positive cell population, but with unknown ontogeny, expressing both CD45RA and CD45RO at a low level, was higher in peripheral blood than in adenoidal T cells. It was found that the frequency of adenoidal antigen-experienced CD45RA⁻,CD45RO⁺ T lymphocytes increased with increasing age of the child. These results are consistent with the model that the adenoids serve as a site for conversion of CD45RA⁺ to CD45RO⁺ T lymphocytes, and that the maturation of the immune system in young children is associated with phenotypic changes in T lymphocytes residing in secondary lymphoid organs.     

Distribution of the natural killer-related receptor for HLA-C during highly active antiretroviral therapy for human immunodeficiency virus infection

Receptors interacting with Major Histocompatibility Complex class I molecules have been initially found on the surface of human natural killer (NK) cells, where they deliver inhibitory signals to the lysis, being thus defined killer inhibitory receptors (KIR). Subsequently, they were detected also on the surface of T-CD8⁺ lymphocytes and are particularly expanded during human immunodeficiency virus (HIV) infection, where they downregulate HIV-specific cytolysis. The expression of KIR recognizing human leukocyte antigen-C alleles was assessed in HIV-infected patients, undergoing highly active antiretroviral therapy (HAART). To this end, the combined expression of CD16/CD56, of CD3 and CD8 as well as of KIR (CD158a and CD158b) surface molecules was analyzed on peripheral blood mononuclear cells by monoclonal antibodies, and flow cytometry. An increase of CD3⁺CD8⁺CD158b⁺ cells was found after 6 months of HAART. This finding may have implications for the regulation of T-cell mediated cytolysis during HAART.     

Costimulatory strength influences the differential effects of transforming growth factor beta1 for the generation of CD8+ regulatory T cells

Transforming growth factor β1 (TGFβ1) is a pleiotropic cytokine, capable of exerting diverse biologic effects. Despite its central role in multiple immune activities, the molecular signals responsible for shaping TGFβ1's immunologic properties remain poorly elucidated. We report that costimulatory strength acts as a molecular switch, which influences the differential effects of TGFβ1 on the effector and regulatory development of naïve CD8⁺ lymphocytes. At low costimulation, TGFβ1 inhibits proliferation of CD8⁺ lymphocytes and cytokine secretion, but at high costimulation the response to TGFβ1 is quite different. High costimulation combined with TGFβ1 generates CD8⁺CD25⁺ T lymphocytes which maintain robust proliferative and survival capacity in the presence of low IL-2 concentrations. Furthermore, under these conditions, a subpopulation of CD8⁺ T lymphocytes is generated that express Foxp3, secrete IL-10, and inhibit naïve T lymphocyte proliferation via a contact-dependent mechanism. The adoptive transfer of these CD8⁺CD25⁺Foxp3⁺ T cells into mice inoculated intravenously with B16F10 melanoma appears to accelerate tumor progression as reflected by an increase in the number of pulmonary metastatic tumor foci. These findings indicate that costimulatory strength may act as a molecular switch in the generation of CD8⁺ T cells which possess a regulatory phenotype and the capacity to reduce antitumor immune responses within tumor-bearing mice.     

使用活化诱导标记法评价HIV-1感染者特异性CD4+T细胞免疫应答的研究

目的:建立一种检测HIV-1特异性CD4⁺T细胞亚群功能的活化诱导标记法(activation-induced markers,AIM),从而更有效地评价HIV-1抗原特异性CD4⁺T细胞免疫应答水平。方法:选取12例未经抗病毒治疗的HIV-1慢性感染者及6例未感染HIV-1的健康人,分别以基于多色流式细胞术的AIM法和细胞内因子染色法(intracellular cytokine staining,ICS)检测抗原特异性T淋巴细胞功能,并探讨两种方法用于评价HIV-1感染者抗原特异性T细胞免疫应答的能力。结果:AIM法检测HIV-1慢性感染者中HIV-1抗原特异性PD-1⁺CD25⁺CD4⁺T、CD69⁺CD200⁺CD4⁺T、CD69⁺ICOS⁺CD4⁺T细胞阳性的比例为11/12、8/12和7/12,检测CD69⁺ICOS⁺CD8⁺T、CD137⁺CD69⁺CD8⁺T、PD-1⁺CD25⁺CD8⁺、OX40⁺PD-1⁺CD8⁺T细胞阳性的比例为8/12、8/12、7/12、7/12。ICS法检测HIV-1抗原特异性IL-2⁺CD4⁺T、IFN-γ⁺CD4⁺T、TNF-α⁺CD4⁺T细胞阳性的占比为2/12、2/12、0;IFN-γ⁺CD8⁺T、TNF-α⁺CD8⁺T、IL-2⁺CD8⁺T细胞阳性的占比为12/12、10/12、5/12。结论:AIM法在评价CD4⁺T细胞功能方面更为敏感,可作为ICS法的补充,两种方法联合使用可更全面地评估抗原特异性T淋巴细胞反应。