Th17细胞和调节性T细胞在系统性红斑狼疮患者中的研究

目的 研究系统性红斑狼疮(SLE)患者外周血Th17细胞、调节性T细胞及联系两者的细胞因子白细胞介素(IL)-6的变化及其与病情活动的相关性,探讨Th17细胞、调节性T细胞在SLE发病机制中的作用.方法 收集103例SLE患者及28名健康者,SLE活动性的判断采用SLE疾病活动指数(SLEDAI)评分方法,其中非活动组(SLEDAI≤9分)患者37例,活动组(SLEDAI＞9分)患者66例.用四色分选流式细胞仪检测外周血Th17细胞、调节性T细胞百分数,用流式细胞仪微球捕获芯片技术(CBA)检测血清IL-6浓度,并分析其与病情活动的相关性.统计学处理采用t检验、秩和检验和Spearman相关分析.结果 ①SLE患者组Th17细胞[(1.2±1.1)%]、IL-6[(35±92)pg/ml]显著高于健康对照组[(0.6±0.4)%、(6±3)pg/ml],而调节性T细胞[(1.6±1.2)%]显著低于健康对照组[(2.6±1.8)%].②SLE活动组Th17细胞[(1.6±1.7)%]显著高于SLE非活动组[(1.0±0.7)%]与健康对照组(P均＜0.05),SLE活动组调节性T细胞[(1.5±1.3)%]显著低于SLE非活动组[(2.1±2.0)%]与健康对照组(P均＜0.05),SLE活动组Th17/调节性T细胞(0.68±0.34)显著高于SLE非活动组(0.24±0.20)与健康对照组(0.13±0.09,P均＜0.05),SLE活动组IL-6[(41±22) pg/ml]显著高于SLE非活动组[(32±28) pg/ml]与健康对照组(P均＜0.05);SLE非活动组与健康对照组之间Th17细胞、调节性T细胞、Th17/调节性T细胞、IL-6差异均无统计学意义(P＞0.05).③Th17细胞百分数与SLEDAI呈正相关,调节性T细胞百分数与SLEDAI呈负相关,Th17/调节性T细胞与SLEDAI呈正相关,IL-6浓度与SLEDAI评分之间呈正相关,Th17细胞与调节性T细胞呈负相关.结论 SLE患者外周血Th17细胞、调节性T细胞及IL-6浓度较健康对照组显著增高,并与疾病活动性密切相关,说明Th17细胞、调节性T细胞可能在SLE的发生、发展中起重要作用.

Abstract：

Objective To study the changes of Th17 cell, regulatory T cell (Treg) and interleukin (IL)-6 in the peripheral blood of patients with systemic lupus erythematosus (SLE) and their relationship with disease activity. Methods Percentage of Th17 and Treg in the peripheral blood of 103 patients with SLE and 28 healthy volunteers were detected by flow cytometry. The concentration of IL-6 in SLE patients and healthy volunteers was detected by cytometric bead array (CBA). The disease activity of SLE was measured by SLEDAI. SLE patients were divided into two groups: stable SLE (SLEDAI≤ 9, n=37) and active SLE (SLEDAI＞9, n= 66). The change of Th17, Treg, IL -6 and their relationship with disease activity were analyzed. Nonparamentric tests, t -test and spearman correlation were used for statistical analysis. Results The percentage of Th17 cells and the concentration of IL-6 in the peripheral blood in patients with SLE was higher than that in normal controls [respectively for (1.2±1.1)%, (35±92) pg/ml and (0.6±0.4)%, (6±3) pg/ml, P＜0.05]. However, the percentage of Treg in patients with SLE was lower than that in normal controls [respectively for (1.6±1.2)%,(2.6±1.8)%, P＜0.05]. The percentage of Th17, Th17/Treg IL-6 level in active SLE patients was higher than those in inactive SLE and those in normal controls (P＜0.05). However, the percentage of Treg in active SLE was lower than that in stable SLE patients and that in normal controls (P＜ 0.05). The percentage of Th17, Th17/Treg and concentration of IL-6 was positively correlated to disease activity(P＜0.05). But the percentage of Treg had negative correlation with the percentage of Th17 and disease activity (P＜0.05). Conclusion Th17, Treg and serum IL-6 in SLE patients are abnormal and they maybe contribute to the pathogenesis of SLE.     

系统性红斑狼疮患者外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的变化及糖皮质激素对其的影响

目的 研究系统性红斑狼疮(SLE)患者外周血CD4+CXCR5+T细胞占CD4+T细胞百分率以及糖皮质激素对其的影响,探讨其在SLE发病机制中的作用.方法 采用流式细胞术检测45例活动期、20例缓解期SLE患者及20名健康对照外周血中CD4+CXCR5+T细胞占CD4+T细胞的百分率,比较其在各组中的差异及糖皮质激素治疗对其的影响,同时检测各组中CD19+B细胞上CXCR5的表达.2组间比较用独立样本t检验,3组间比较采用多变量方差分析,与临床指标之间的相关性分析采用非参数的Spearman相关分析,治疗前后的差异用重复测量的方差分析.结果 ①SLE患者外周血CD4+CXCR5+T细胞占CD4+T细胞的比例高于健康对照组[(16±7)%与(12±3)%,P＜0.01],其中活动组[(18±7)%]高于健康对照组(P＜0.05),而缓解组[(11±4)%]和健康对照组之间差异无统计学意义(P＜0.05);狼疮肾炎组高于非狼疮肾炎组,但差异无统计学意义[(18±7)%与(14±7)%,P=0.05].②CD4+CXCR5+T细胞百分率与SLE疾病活动指数(SLEDAI)、抗核抗体滴度和红细胞沉降率(ESR)呈正相关,与补体C3呈负相关(P均＜0.05),与C反应蛋白、病程、免疫球蛋白无相关性(P＞0.05).抗双链DNA抗体升高组与正常组之间、抗sm抗体、抗SSMSSB抗体阴性组和阳性组之间差异无统计学意义(P均＞0.05).③活动期SLE患者CD19+B细胞上CXCR5的表达比例低于健康对照组[(85±11)%与(94±3)%,P＜0.05].④10例初发、未接受治疗的活动期患者在接受地塞米松(20 mg/d)治疗后第1、3、7天外周血中CD4+CXCR5+T细胞百分率均低于治疗前(P均＜0.05).治疗前后CD19+CXCR5+B细胞的百分率无变化(P均＞0.05).结论 外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的异常町能参与SLE的发病.

Abstract：

Objective To investigate the frequencies of CD4+CXCR5+T cells in the CD4+T cells of peripheral blood of patients with systemic lupus erythematosus (SLE) and the effect of glucocorticoid on it.Methods Frequencies of CD4+CXCR5+T cell were analyzed by flow cytometry in 45 active,20 inactive SLE patients and 20 healthy controls.Differences between groups and the effect of glucocorticoid were analyzed.Meanwhile, the expression of CXCR5 on CDI9+B cells was analyzed. Independent sample t test was used for statistical analysis between twogroups, ANOVA was applied for data analysis between 3 groups,,nonparameterical Spearman's analysis was used for correlation analysis and repeated measurement ANOVA were used to compare the parameters before and after treatment. Results The percentage of CD4+CXCR5+ in CD4+T cells was increased in patients with SLE compared with healthy controls[(16±7)% vs (12±3)%, P＜0.01].It was increased in patients with active SLE [(18±7)%] compared with healthy controls (P＜0.05) but there was no significant difference between inactive SLE[(11±4)%] and healthy controls(P＞0.05). The percentage in patients with LN was higher than that in patients without LN, but without significant difference[(18±7)%vs (14±7)%, P=0.05 ]. The percentage of CD4+CXCR5+T cells was positively correlated with SLEDAI,the titer of ANA and level of ESR but negatively correlated with the level of C3 (P<0.05 for each).No correlation was found between duration and the levels of CRP and immunoglobulin.. The percentage in patients with high anti-dsDNA group was also higher than that of the low group, but no differences were found between anti-Sm antibody positive and negative groups neither between anti-SSA/SSB antibody positive and negative groups(P＞0.05 for each).The expression level of CXCR5 on CD19+B cells in active SLE patients was lower than that of healthy controls[(85±11)% vs (94±3)%, P＜0.05 ]. The percentages of CD4+CXCR5+T cells in 10 untreated active SLE patients were decreased at day 1,day 3 and day 7 after being treated with dexamethasone (20mg/d) when compared with those before the treatment (P＜0.05 for each), but the percentages of CD19+CXCR5+B cells had no significant change (P＞0.05 for each).Conclusion These results demonstrate that the abnormality of CD4+CXCR5+T cells may play an important role in the pathogenesis of SLE.     

系统性红斑狼疮患儿外周血辅助性T17细胞表达的初步研究

目的 观察系统性红斑狼疮(SLE)患儿外周血辅助性T17(Th17)细胞表达变化,探讨Th17细胞及其相关的细胞因子在SLE患儿发病中的作用.方法 以15名健康儿童为健康对照组,流式细胞术(FCM)检测25例SLE患儿(SLE组)外周血Th17细胞百分率,用酶联免疫吸附试验(ELISA)法检测外周血血浆白细胞介素(IL)-17、IL-21水平,分析Th17细胞与SLE活动度之间的关系.统计学处理采用t检验和Spearman相关性检验.结果 与健康对照组相比,SLE患儿外周血中CD3+CD8-IL-17+T细胞的频率[(1.24±0.64)%,(0.59±0.21)%],CD3+CD8-IL-21+T细胞的频率[(1.5±0.6)%,(0.8±0.4)%]明显增高,P均＜0.01;血浆中IL-17,IL-21水平明显升高(P＜0.01).CD3+CD8-IL-17+T细胞频率与疾病活动度相关(r=0.732,P＜0.01),CD3+CD8-IL-21+T细胞频率与疾病活动度无相关性(r=-0.002,P＞0.05).结论 Th17细胞及其相关细胞因子参与儿童SLE发病及病情发展.

Abstract：

Objective To investigate the Th17 cell expression in peripheral blood of childron with systemic lupus erythematosus (SLE) and explore the role of Th17 cells and the cytokines in the pathogenesis of SLE. Methods Twenty-five children with SLE were enrolled and 15 healthy children were controls. Flow cytometry (FCM) was employed to detect the expression of Th17 cells in peripheral blood of SLE children (SLE group, n=25), and IL-17, IL-21 levels in plasma were detected by ELISA. Two-independent sample t-test and Spearmen's test were used for correlation analysis. Results Compared with that of the control, the frequencies of CD3+CD8-IL-17+T[(1.24±0.64)% vs (0.59±0.21)%], CD3+CD8-IL-21+T cells[(1.5±0.6)%vs (0.8±0.4)% ] increased significantly in SLE patients (P＜0.01) and the plasma concentrations of IL-17, IL-21 were significantly higher (P＜0.01). The SLE activity was positively correlated with the frequencies of CD3+CD8-IL-17+T cells, but not with CD3+CD8-IL-21+T cells. Conclusion Th17 cells and the related cytokinesplay an important role in the pathogenesis of childhood SLE.     

系统性红斑狼疮患者外周血CD4+CXCR5+滤泡辅助性T细胞样细胞的变化及糖皮质激素对其的影响

Objective To investigate the frequencies of CD4+CXCR5+T cells in the CD4+T cells of peripheral blood of patients with systemic lupus erythematosus (SLE) and the effect of glucocorticoid on it.Methods Frequencies of CD4+CXCR5+T cell were analyzed by flow cytometry in 45 active,20 inactive SLE patients and 20 healthy controls.Differences between groups and the effect of glucocorticoid were analyzed.Meanwhile, the expression of CXCR5 on CDI9+B cells was analyzed. Independent sample t test was used for statistical analysis between twogroups, ANOVA was applied for data analysis between 3 groups,,nonparameterical Spearman's analysis was used for correlation analysis and repeated measurement ANOVA were used to compare the parameters before and after treatment. Results The percentage of CD4+CXCR5+ in CD4+T cells was increased in patients with SLE compared with healthy controls[(16±7)% vs (12±3)%, P＜0.01].It was increased in patients with active SLE [(18±7)%] compared with healthy controls (P＜0.05) but there was no significant difference between inactive SLE[(11±4)%] and healthy controls(P＞0.05). The percentage in patients with LN was higher than that in patients without LN, but without significant difference[(18±7)%vs (14±7)%, P=0.05 ]. The percentage of CD4+CXCR5+T cells was positively correlated with SLEDAI,the titer of ANA and level of ESR but negatively correlated with the level of C3 (P<0.05 for each).No correlation was found between duration and the levels of CRP and immunoglobulin.. The percentage in patients with high anti-dsDNA group was also higher than that of the low group, but no differences were found between anti-Sm antibody positive and negative groups neither between anti-SSA/SSB antibody positive and negative groups(P＞0.05 for each).The expression level of CXCR5 on CD19+B cells in active SLE patients was lower than that of healthy controls[(85±11)% vs (94±3)%, P＜0.05 ]. The percentages of CD4+CXCR5+T cells in 10 untreated active SLE patients were decreased at day 1,day 3 and day 7 after being treated with dexamethasone (20mg/d) when compared with those before the treatment (P＜0.05 for each), but the percentages of CD19+CXCR5+B cells had no significant change (P＞0.05 for each).Conclusion These results demonstrate that the abnormality of CD4+CXCR5+T cells may play an important role in the pathogenesis of SLE.     

系统性红斑狼疮患儿外周血辅助性T17细胞表达的初步研究

Objective To investigate the Th17 cell expression in peripheral blood of childron with systemic lupus erythematosus (SLE) and explore the role of Th17 cells and the cytokines in the pathogenesis of SLE. Methods Twenty-five children with SLE were enrolled and 15 healthy children were controls. Flow cytometry (FCM) was employed to detect the expression of Th17 cells in peripheral blood of SLE children (SLE group, n=25), and IL-17, IL-21 levels in plasma were detected by ELISA. Two-independent sample t-test and Spearmen's test were used for correlation analysis. Results Compared with that of the control, the frequencies of CD3+CD8-IL-17+T[(1.24±0.64)% vs (0.59±0.21)%], CD3+CD8-IL-21+T cells[(1.5±0.6)%vs (0.8±0.4)% ] increased significantly in SLE patients (P＜0.01) and the plasma concentrations of IL-17, IL-21 were significantly higher (P＜0.01). The SLE activity was positively correlated with the frequencies of CD3+CD8-IL-17+T cells, but not with CD3+CD8-IL-21+T cells. Conclusion Th17 cells and the related cytokinesplay an important role in the pathogenesis of childhood SLE.     

Effects of splenectomy in patients with cirrhosis undergoing hepatic resection for hepatocellular carcinoma

AIM: To study the effects of splenectomy in patients with cirrhosis undergoing hepatic resection for hepatocellular circinoma.METHODS: Twenty-six patients with HCC associated with cirrhosis were divided into hepatectomy with splenectomy group (splenectomy group, n=11) and hepatectomy without splenectomy group (non-splenectomy group, n=15). T lymphocyte subsets such as CD4, CD8, CD4/CD8, helper T (Th) lymphocyte cytokines such as interferon γ(IFN-γ),interleukin 2(IL-2), interleukin 10(IL-10) and white blood cell (WBC), platelet (PLT), total bilirubin (T-Bil) were measured and used as parameters to evaluate the effects of splenectomy.RESULTS: There was no significant difference in CD4, CD8,CD4/CD8, IL2, IFN-γ, IL10, WBC, PLT, T-Bil levels between two groups before surgery. Two months after operation,the levels of CD4 (41.2 %±4.2 % vs 34.7 %±3.8 %), CD4/CD8 (1.7±0.2 vs 1.0±0.2), IFN-γ (102.3±15.9 pg/ml vs 86.5±14.8 pg/ml), IL-2(97.2±15.6 pg/ml vS77.6±14.5 pg/ml) were increased and those of CD8 (25.6±3.9 vs32.8 %±4.1%),IL-10 (56.9±10.4 pg/ml vs 72.6±15.3 pg/ml) were decreased in splenectomy groups as compared with those in nonsplenectomy group (P<0.05). WBC and PLT counts in the splenectomy group were 8.9±1.6×109 and 310±32×109,respectively, which were significantly higher than those in non-splenectomy group (3.7±1.4×109 and 104±41×109) respectively on the 14th post-operative day. T-Bil concentration in the splenectomy group (24±7 μmol/L) was significantly lower than that in the non-splenectomy group (37±13 μmol/L) on the 7th post-operative day (P<0.05).CONCLUSION: Splenectomy combined with hepatectomy for HCC associated with cirrhosis is helpful for the recovery of T-lymphocyte subsets and the maintenance of Th:L/Th2 cytokine balance.     

HBV特异性抗原对HBV携带者T淋巴细胞免疫功能的影响

目的 通过分析不同类型HBV携带者外周血单个核细胞(PBMCs)的细胞免疫功能,分析HBV抗原对其的影响,探索HBV慢性感染的机制并寻求可能的免疫治疗方法.方法 用不同的抗原和(或)细胞因子刺激培养的无症状HBV携带者PBMCs,酶联免疫吸附法检测细胞培养上清液中不同细胞因子的水平;流式细胞术检测PBMCs的细胞表型.对数据进行t检验分析和相关性分析.结果 HBsAg刺激无症状HBV携带者PBMCs后产生的干扰素(IFN)γ为(48.3±19.8)Pg/ml,较健康对照人群低[(196.2±104.3)Pg/ml(t=3.023,P＜0.05)].HBsAg和HBcAg刺激HBeAg阳性患者PBMCs分泌的IFN y水平分别为(50.4±51.6)Pg/ml和(63.2±36.9)pg/ml,明显低于HBeAg阴性组[(86.2±42.3)Pg/ml和(101.4±32.5)pg/ml],t值分别为2.468和3.184,P值均＜0.05;HBeAg阳性患者组分泌白细胞介素(IL)-12 P70明显低于HBeAg阴性组(P＜0.05);补偿外源性IL-12可明显促进HBV携带者PBMCs分泌IFN γ(P＜0.01),IL-12协同HBV抗原可激活CD8+CD45RA+CCR7及CD8+CD45RA CD62L+细胞.结论 HBeAg阳性患者PBMCs分泌IL-12减少,这可能是HBV携带者持续感染的重要原因;外源性IL-12可促进HBV携带者PBMCs中的中枢记忆性T淋巴细胞的免疫功能.

Abstract：

Objective To investigate the effect of HBV antigens and pathological mechanism of chronic HBV infection by analyzing the cellular immune function of peripheral blood mononuclear cells (PBMCs) from HBsAg carriers. Methods PBMCs were prepared from individuals with chronic asymptomatic HBV infection and cultured in the presence of different antigens and/or cytokines. The levels of cytokines in culture supernatants were detected by ELISA method. The phenotype of the cells was detected by FACS.Results The levels of IFN γ secreted by PBMCs from HBsAg carriers were (48.3 ± 19.8) pg/ml, significantly lower than that from healthy controls (t = 3.023, P ＜ 0.05＝; The IFN γ produced by PBMCs from HBeAg positive patients due to HBsAg and HBcAg stimulation were (50.4±51.6) pg/ml and (63.2 ± 36.9)pg/ml, significantly lower than that of HBeAg negative patients (t = 2.468 and 3.184, P ＜ 0.05, respectively＝.The IL-12p70 secreted by PBMCs from HBeAg positive patients was also significantly lower than that of HBeAg negative patients (P ＜ 0.05＝; Exogenous IL-12 promoted significantly PBMCs to secrete IFN γ (P ＜0.01＝ and IL-12 combined with HBV antigens activated CD8+CD45RA+CCR7+ and CD8+CD45RA-CD62L+cells. Conclusion IL-12 secreted by PBMCs decreased in HBeAg positive patients, which may be the crucial reason of viral persistence in chronic HBV carriers. Exogenous IL-12 combined with specific HBV antigen could promote the central memory CD8+ T cells to produce IFN γ.     

Upregulated voltage-gated potassium channel Kv1.3 on CD4+CD28null T lymphocytes from patients with acute coronary syndrome

Objective The purpose of our study is to observe the voltage-gated potassium channel Kv1.3 expressed on CD4+ CD28null T cells from the peripheral blood of acute coronary syndrome (ACS) patients by the patch clamp technique. Methods Kvl.3 potassium channels expression from 17 patients with ACS and 11 healthy age-match controls was detected in single cell(CD4+CD28null T cells and CD4+CD28+T cells) by fluorescence microscopy and patch clamp. Results The percentage of CD4+CD28null T cells was higher in the ACS patients [(6.97±2.05)%] than that in the controls [(1.38±0.84)%, P<0.05]. The concentration of hsCRP was directly correlated with the number of the CD4+CD28null T cells in the ACS patients (r=0.52, P<0.05). The conductance (6.89±1.17ns vs 3.36±0.66ns), dens (1.95±0.80 μm2 vs 1.13±0.57 μm2) and numbers (574.5±97.6 n/cell vs. 280.3±55.3 n/cell) of the Kvl.3 channels on the CD4+CD28null T cells were significantly higher than those on the CD4+CD28+T cells (all P<0.0l) in ACS patients, but were similar on CD4+CD28+T between ACS patients and controls. Conclusion The CD4+CD28null T cells and the numbers of Kv1.3 channels on the CD4+CD28null T cells from patients with ACS are significantly upregulated and might contribute to the pathogenesis of ACS.     

慢性乙型肝炎患者应用拉米夫定前后T淋巴细胞亚群、CD4+CD25+调节性T淋巴细胞及白细胞介素-10、干扰素-γ的变化

目的 观察慢性乙型肝炎(CHB)患者应用拉米夫定抗病毒治疗前后外周血T淋巴细胞亚群、CD4+CD25+调节性T淋巴细胞(CD4+CD25+Treg)及IL-10、IFN-γ水平的变化.方法 选择CHB患者90例,在应用拉米夫定抗病毒治疗前及治疗后52周时,用流式细胞仪检测患者外周血T淋巴细胞亚群;在治疗前及治疗后12、24、36、52周时,用流式细胞仪检测外周血CD4+CD25+Treg频率,用双抗体夹心ELISA法检测IL-10、IFN-γ水平.组间及组内总体均数比较采用方差齐性的单因素方差分析或Dunnett's检验,组内治疗前后两时段均数比较采用配对t检验.结果 在90例CHB患者中,完全应答的32例患者的CD4+T淋巴细胞、CD8+T淋巴细胞及CD4+/CD8+较治疗前升高(t=4.055、3.267、2.328,均P＜0.05),外周血CD4+CD25+Treg频率在0、12、24、36、52周时分别为(5.40±0.60)%、(4.99±0.59)%、(4.54±0.72)%、(3.86±0.95)%、(3.44±0.76)%;IFN-γ水平、IFN-γ/IL-10逐步上升,IL-10水平逐步下降.部分应答的43例患者的CD4+T淋巴细胞及CD4+/CD8+较治疗前升高(t=3.484、2.018,均P＜0.05),CD8+T淋巴细胞较治疗前无明显差异,外周血CD4+CD25+Treg频率在0、12、24、36、52周时分别为(5.65±0.60)%、(5.23±0.63)%、(4.65±0.98)%、(4.42±0.97)%、(4.32±0.82)%,IFN-γ水平、IFN-γ/IL-10升高,IL-10水平降低.无应答的15例患者的外周血CD4+T淋巴细胞、CD8+T淋巴细胞及CD4+/CD8+、外周血CD4+CD25+Treg频率及IFN-y水平、IFN-γ/IL-10、IL-10水平较治疗前无明显变化.结论 应用拉米夫定治疗过程中,获得满意应答的CHB患者的外周血CD4+CD25+Treg频率下降,CD4+/CD8+、IFN-γ/IL-10的比例升高.

Abstract：

Objective To explore the correlation between the efficacy of lamivudine (LAM)therapy and changes of T lymphocyte subsets,CD4+ CD25+ regulatory T lymphocytes (Treg),and levels of interleukin-10 (IL-10)and interferon-gamma (IFN-γ)in the peripheral blood of patients with chronic hepatitis B (CHB).Methods Ninety CHB patients were enrolled in this study.T lymphocyte subsets in the peripheral blood were detected by flow cytometry at baseline and week 52 of LAM therapy.The frequencies of CD4+ CD25+ Treg in the peripheral blood were detected by flow cytometry and levels of IL-10 and IFN-γ were detected by enzyme-linked immunosorbent assay (ELISA)at baseline,week 12,24,36 and 52.The comparisons of overall means between groups and within groups were done by analysis of variance or Dunnett's test.The comparison of means before and after LAM therapy was done by paired t test.Results In 32 complete-responders of 90 CHB patients,the proportions of CD4+ T lymphocytes,CD8+ T lymphocytes and CD4+/CD8+ ratio were increased significantly after LAM therapy (t=4.055、3.267、2.328,all P＜0.05); the frequencies of CD4+CD25+ Treg at baseline,week 12,24,36 and 52 were (5.40±0.60)%,(4.99±0.59)%,(4.54± 0.72)%,(3.86±0.95)% and (3.44±0.76)%,respectively; the levels of IFN-γ,IFN-γ/IL-10 ratio were increased,while the IL-10 level was decreased after LAM therapy.In 43 partial-responders,the proportion of CD4+T lymphocytes and ratio of CD4+/CD8+ were increased after LAM therapy (t= 3.484,2.018,both P＜0.05); the proportion of CD8+ T lymphocytes was not changed significantly after therapy; the frequencies of CD4+ CD25+ Treg at baseline,week 12,24,36 and 52 were (5.65±0.60)%,(5.23±0.63)%,(4.65±0.98)%,(4.42±0.97)% and (4.32±0.82)%,respectively;IFN-γ level,IFN-γ/IL-10 ratio were increased,while IL-10 level was decreased.In 15 non-responders,the proportion of T lymphocyte subsets,the frequency of CD4+ CD25+ Treg,the levels of IFN-γ and IL-10 were not changed significantly after LAM treatment.Conclusions In CHB patients who have achieved response after LAM therapy,the frequency of CD4+ CD25+ Treg in the peripheral blood is decreased,while ratios of CD4+/CD8+ and IFN-γ/IL-10 in the peripheral blood are increased.     

克罗恩病患者血清可溶性细胞毒T淋巴细胞相关抗原4水平高表达及其临床意义

目的 研究可溶性细胞毒T淋巴细胞相关抗原4(sCTLA4)在克罗恩病(CD)患者血清中的表达状况,分析sCTLA4水平与红细胞沉降率(ESR)和C反应蛋白(CRP)的关系及其临床意义,探讨CTLA4基因-1661A/G和-1722T/C多态性与CD的相关性.方法 收集126例CD患者和300名健康对照者,酶联免疫吸附实验测定血清sCTLA4水平.全自动红细胞沉降系统分析仪测定ESR水平.速率散射比浊法测定CRP水平.DNA测序技术检测CTLA4基因-1661A/G和-1722T/C多态性.结果 CD患者血清中sCTLA4水平为(18.70±3.72)ng/ml,明显高于健康对照者[(1.72±0.32)ng/ml,P＜0.01].活动期CD患者血清sCTLA4水平为(19.83±4.35)ng/ml,比缓解期CD患者[(18.02±3.14)ng/ml,P=0.015]增高.CD患者血清sCTLA4水平与ESR和CRP水平呈正相关(r=0.267,P=0.003;r=0.524,P＜0.01).疾病行为影响CD患者血清sCTLA4水平,狭窄型CD患者血清sCTLA4水平高于非狭窄非穿透型和穿透型CD患者,差异均有统计学意义(P值分别=0.021和0.015).CD组和健康对照组中CTLA4基因-1661A/G,-1722T/C等位基因及基因型频率差异无统计学意义(P＞0.05).结论 CD患者血清sCTLA4水平增高,增高的sCTLA4水平不仅与疾病活动状态及狭窄型疾病行为相关,且与CRP正相关,提示sCTLA4分子在CD免疫学发病中可能起重要作用.

Abstract：

Objective To investigate the expression of serum soluble cytotoxic T lymphocyte associated antigen 4 (sCTLA4), the association of sCTLA4 level with erythrocyte sedimentation rate (ESR) and C reactive protein (CRP), as well as its role in patients with Crohn's disease (CD). The relationship-1661A/G and -1722T/C polymorphisms of CTLA4 gene and between disease susceptibility and phenotype of CD was analyzed. Methods A total of 126 CD patients and 300 healthy controls were enrolled in the study. Serum sCTLA4 level was determined by enzyme-linked immunosorbent assay. The concentrations of ESR and CRP were analyzed by automatic ESR Analyzer SRS 100/Ⅱ and rate nephelometry, respectively. The polymorphisms of CTLA4-1661A/G and -1722 T/C were genotyped by DNA sequencing. Results Serum sCTLA4 level was higher in CD patients than in healthy controls [(18. 70±3. 72) ng/ml vs (1.72±0. 32) ng/ml, P＜0. 01)]. Among CD patients, sCTLA4 level was higher in patients with active disease when compared to those with inactive disease [(19.83±4.35) ng/ml vs (18. 02±3.14) ng/ml, P=0. 015)]. sCTLA4 level was positively correlated with ESR and CRP levels (r=0. 267, P=0. 003; r=0. 524 P ＜0.01, respectively). In CD patients, serum sCTLA4 level was significantly higher in those with stricturing disease behavior than that in those without stricturing and penetrating or with penetrating disease behavior (P= 0.021; P=0. 015, respectively). Detection of CTLA4 -1661A/G and -1722T/C polymorphisms showed no significant difference between CD patients and healthy controls. Conclusions The high level of serum sCTLA4 in CD patients is correlated with disease activity, CRP levels and disease behavior. It suggests that sCTLA4 may play an important role in pathogenesis of CD.     

中试生产的结核DNA疫苗免疫效能研究

目的 以300 L发酵罐中试规模生产的冻干Mtb Ag85A质粒DNA疫苗为基础免疫小鼠,对该疫苗的免疫效能进行研究。 方法 将4~6周龄的BALB/c小鼠20只通过数字表法随机分为2组,一组免疫Mtb Ag85A质粒DNA疫苗（结核DNA疫苗免疫组）,另一组免疫PBS作为阴性对照（PBS对照组）;于免疫前采血,肌内注射免疫3次。在第3次免疫后3周摘眼球处死小鼠,收集血清,进行血清特异性抗体检测和细胞因子(IFN-γ、IL-4)检测;同时对每只小鼠取脾细胞进行特异性γ干扰素 (IFN-γ) 分泌检测和脾细胞CD4⁺、CD8⁺百分率检测。 结果 （1）免疫前后免疫组与对照组抗体水平吸光度A值均小于0.2。（2）结核DNA疫苗免疫组血清细胞因子IL-4水平［（146.2±34.3）pg/ml］低于PBS对照组［（177.7±28.1）pg/ml］,差异有统计学意义（t=2.244,P=0.038）,而IFN-γ水平［（129.6±159.0）pg/ml］虽然高于PBS对照组［（76.5±21.5）pg/ml］,但差异无统计学意义（t=－1.047,P=0.309）。（3）酶联免疫斑点检测结果表明结核DNA疫苗免疫组脾细胞特异性IFN-γ分泌水平（分泌IFN-γ细胞个数）（103.60±112.14）高于PBS对照组（5.78±5.83）,差异有统计学意义（t=36.538,P=0.018）。（4）结核DNA疫苗免疫组CD4⁺细胞百分率均值（21.57%）高于PBS对照组（12.17%）,差异有统计学意义（t=3.043,P=0.038）;CD8⁺细胞百分率均值（13.70%）与PBS对照组（10.57%）相比,差异无统计学意义（t=0.847,P=0.445）。 结论 Mtb Ag85A质粒DNA疫苗主要刺激机体Th1型细胞免疫,而对诱导机体产生Th2型细胞免疫应答没有显著的促进作用。     

骨髓间充质干细胞移植对MRL/Ipr鼠B细胞活化因子及B细胞活化的影响

目的 探讨正常鼠骨髓问充质干细胞(BM-MSCs)移植对MRL/Ipr狼疮鼠B细胞活化因子(BAFF)表达及B细胞活化的影响.方法 18只雌性MRL/Ipr鼠随机分为MSCs治疗组和对照组,18周龄时治疗组经尾静脉移植MSCs 1×10~6/只;5只同周龄雌性BAL B/C小鼠作为健康阴性对照.酶联免疫吸附试验(ELISA)法检测血清BAFF、干扰素(IFN)-γ、白细胞介素(IL)-2、IL-10水平,流式细胞术检测脾脏中边缘区、T1期、T2期B细胞百分率及细胞数.结果 ①MSCs治疗8周后,治疗组血清BAFF水平[(32±14)ng/ml]显著低于对照组[(47±13)ng/ml](P＜0.05);血清IL-10[(19±7)pg/ml]显著低于对照组[(40±13)pg/ml](P＜0.01);血清IFN-γ、IL-2低于对照组[(26±20)pg/ml与(38±25)pg/ml、(73±10)pg/ml与(80±14)pg/ml],但差异无统计学意义.②MSCs治疗8周后,可降低治疗组脾脏边缘区B细胞百分率(15±4)%,对照组为(21±5)%,但差异无统计学意义,并能显著降低边缘区B细胞数[(9±6)×10~6与(19±10)×10~6,P＜0.05].③MSCs治疗8周后,可降低治疗组脾脏T1期、T2期B细胞百分率[(3.4±2.1)%与(7.3±4.0)%]、[(2.6±1.4)%与(4.8±2.7)%],但差异无统计学意义,并能显著降低T1期B细胞绝对数[(2.7±1.7)×10~6与(5.1±2.0)×10~6,P＜0.05]、T2期B细胞绝对数[(2.0±1.2)×10~6与(3.7±1.7)×10~6,P＜0.05].结论 BM-MSCs移植可能通过抑制体内BAFF的过量表达,进而抑制狼疮鼠B细胞的过度活化.     

SOCS1和SOCS3低甲基化对川崎病Th1/Th2细胞失衡的影响

目的 探讨SOCS1和SOCS3低甲基化对川崎病Th1/Th2细胞失衡的影响.方法 急性期川崎病患儿36例,健康同年龄对照组16名.酶联免疫吸附试验(ELISA)检测血浆白细胞介素(IL)-6蛋白浓度;实时荧光定量聚合酶链反应(PCR)检测CD4+T细胞SOCS1、SOCS3、T-bet、干扰素(IFN)-γ、GATA3、IL-4基因mRNA表达;流式细胞术检测外周血Th1/Th2细胞比例和CD4+T细胞磷酸化STAT3(pSTAT3)蛋白平均荧光强度(MFI);甲基化特异性定量PCR(MethySYBR PCR)检测CD4+T细胞SOCSl基因外显子2、SOCS3基因5'端非翻译区(5'-UTR)3个可能的STAT3结合位点CpG岛甲基化水平.采用t检验进行统计分析.结果 ①急性期川崎病患儿血浆IL-6浓度[分别为(51.8±16.3)pg/ml和(8.6±2.0)pg/ml]、CD4+T细胞pSTAT3 MH水平[分别为(52±14)和(10±4)]显著上调(P＜0.05).其中川崎病合并冠状动脉损伤组(川崎病-CAL+)IL-6和pSTAT3 MFI水平均明显高于无冠状动脉损伤组[IL-6为(87.2±27.4)pg/ml与(36.2±12.8)pg/ml,P＜0.05;pSTAT3 MFI为(75±15)和(42±11),P＜0.05].②急性期川崎病患儿Th1、Th2细胞比例及相关因子(T-bet、IFN-γ、GATA3和IL-4)表达明显增高(P＜0.05),Th1/Th2比值低于健康对照组(P＜0.05).其中川崎病-CAL+组Th1、Th2细胞比例及相关因子表达水平高于川崎病-GAL-组(P＜0.05),而Th1/Th2比值则略低于后者(P＜0.05).③急性期川崎病患儿CD4+T细胞SOCS1和SOCS3 mRNA水平显著高于同年龄对照组(P＜0.05),其中川崎病-CAL+组 SOCS1和SOCS3 mRNA表达低于川崎病-CAL-组(P＜0.05);健康对照组SOCS1基因外显子2、SOCS3基因5'-UTR区第3个STAT3结合位点的CpG岛完全去甲基化,急性期川崎病患儿呈低甲基化状态(P＜0.05),其中川崎病-CAL+组去甲基化水平明显低于川崎病-CAL组(P＜0.05);各组SOCS3基因5'-UTR区第1、2个STAT3结合位点CpG岛均处于完全去甲基化状态(P＞0.05).结论 SOCS1和SOCS3基因低甲基化所致表达相对不足可能是川崎病Th1/Th2细胞失衡的因素之一.     

CD4+白细胞介素-17+辅助性T细胞对香烟暴露小鼠肺部炎症及肺气肿的作用

目的 观察香烟烟雾暴露小鼠肺实质中CD4+白细胞介素(IL)-17+辅助性T细胞(Th17)数量及活性的表达,探讨其在香烟暴露小鼠肺部CD4+ γ-干扰素+(Th1)炎症及肺气肿中的作用及相关机制.方法 将40只雄性Balb/c小鼠按随机数字表法分为4组:对照12周(C12)组、对照24周(C24)组、烟雾暴露12周(S12)组、烟雾暴露24周(S24)组,每组10只.香烟烟雾暴露法建立小鼠肺气肿模型.HE染色观察小鼠肺气肿的改变,计算平均内衬间隔和肺泡破坏指数(DI);流式细胞术检测小鼠肺实质中CD4+IL-17+T(Th17)细胞、CD4+γ-干扰素+T(Th1)细胞、CD4+IL-17+γ-干扰素+T(Th17/Th1)细胞、CD8+γ-干扰素+T(Tc1)细胞、CD8+IL-21R+细胞及CD4+IL-17+IL-21+细胞比例;荧光定量PCR法检测小鼠肺实质中维甲酸相关孤独受体(RORγt)和IL-17的mRNA表达,并分析这些指标的相互关系.结果 S12组和S24组的平均内衬间隔[(39±4)μm和(47±7)μm]和DI(39.1±1.6和45.2±3.1)明显高于C12组[(32±4)μm和28.2±1.6]和C24组[(33±3)μm和28.9±2.1],且以S24组的增高更为明显,差异均有统计学意义(t值为4.378～15.188,均P＜0.05);S12组和S24组Th17细胞比例[(3.3±1.1)%和(7.2±2.2)%]均明显高于C12组和C24组[(1.8±0.8)%和(2.0±0.6)%];S12组和S24组RORγtmRNA表达量[(25±4)和(35±3)]及IL-17的mRNA表达量[(26±3)和(36±3)]亦明显高于C12组[(10±5)和(13±5)]和C24组[(11±7)和(8±6)],以S24组增高更为明显,差异均有统计学意义(P＜0.05);S12组和S24组Th1细胞比例[(10.0±3.7)%和(26.2 ±6.0)%]、Th17/Th1细胞比例[(0.61±0.30)%和(1.82±0.52)%]及Tc1细胞比例[(17.0±4.5)%和(26.8±8.5)%]均明显高于C12组[(3.8±1.7)%、(0.27±0.17)%和(4.8±1.9)%]和C24组[(4.2±1.3)%、(0.28±0.11)%和(5.2±1.0)%],以S24组增高更为明显,差异均有统计学意义(P＜0.05);S12组和S24组小鼠Th17细胞与Th1、Tc1细胞比例、平均内衬间隔、DI值均呈显著正相关(r值为0.519～0.797,均P＜0.01);Th17/Th1细胞比例与平均内衬间隔、DI值呈显著正相关(r值分别为0.742和0.802,均P＜0.01);S12组和S24组CD4+IL-17+IL-21+细胞比例[(0.19±0.04)和(0.55±0.24)]明显高于C12组和C24组[(0.07±0.03)和(0.08±0.03)],S24组增高更为明显,差异均有统计学意义(P＜0.05).S12组和S24组的CD8+IL-21R+细胞比例[(2.94±1.26)和(4.12±2.26)]高于C12组和C24组[(1.22±0.31)和(1.34±0.18)](P＞0.05);S12组及S24组小鼠CD4+IL-17+IL-21+细胞比例与Th1、Tc1细胞比例、平均内衬间隔和DI值均呈显著正相关(r值为0.694～0.754,均P＜0.05);S12及S24组小鼠CD8+IL-21R+细胞比例与平均内衬间隔和DI呈显著正相关(r值分别为0.516和0.725均P＜0.05).结论 香烟暴露导致肺气肿小鼠肺内Th17细胞数量及活性上调,并随烟雾暴露时间延长而增强;Th17细胞通过IL-21及IL-21R在肺部Th1/Tc1炎症中起重要促进作用;这对探讨COPD肺部炎症和肺气肿发生机制以及新的治疗靶点具有重要意义.

Abstract：

Objective To evaluate the expression and the role of Th 17 in cigarette smoke-induced lung inflammation and emphysema in mice.Methods Forty male BALB/c mice were randomly divided into 4 groups, including a control group C12, a control group C24, a smoke-exposure 12 week group (S12) and a smoke-exposure 24 week group S24 (n = 10 each).Morphological changes were evaluated by mean linear intercepts and destructive index (DI).The proportion of CD4+ IL-17 + Th17, CD4+ IFN-γ+ Th1, CD4+ IL-17 +IFN-γ+ T( Th17/ Th1 ), CD8+ IFN-γ+ Tc1, CD8+ IL-21R + and CD4+ IL-17 + IL-21 + T cells in lungs of mice was determined by flow cytometry.The mRNA expressions of RORγt and IL-17 were evaluated by real-time PCR.Results Mean linear intercepts and DI were significantly higher in S12 and S24 groups [(39 ± 4)μm, (47 ±7) μm], (39.1 ± 1.6, 45.2 ±3.1 ) as compared to C12[(32 ±4) μm,28.2 ± 1.6] and C24groups [(33 ± 3 ) μm ,28.9 ± 2.1], all P ＜ O.05.The percentage of Th17 of S12 and S24 groups [(3.3 ±1.1 )%, (7.2 ±2.2)%] was significantly increased as compared with that of C12 and C24 groups [( 1.8± 0.8) %, (2.0 ± 0.6) %], all P ＜ 0.05.The mRNA levels of RORγt [( 25 ± 4), ( 35 ± 3 )] and IL-17 [(26 ± 3), (36 ± 3 )] in S12 and S24 groups were higher than in C12 [(10 ± 5 ), (13 ± 5 )] and C24 groups [( 11 ± 7 ), (8 ± 6)], all P ＜ 0.05.The percentage of Th 1, Th17/Th1 and Tc1 cells of S12 and S24 groups [(10.0 ±3.7)%, (26.2 ±6.0)%], [(0.61 ±0.30)%, (1.82 ±0.52)%], [(17.0±4.5 ) %, ( 26.8 ± 8.5 ) %] was significantly increased as compared with that of C12 [( 3.8 ± 1.7 ) %,(0.27±0.17)%, (4.8 ±1.9)%] and C24 groups [(4.2±1.3)%, (0.28±0.11)%, (5.2±1.0)%], all P＜0.05.Moreover, the frequency of Th17 cells had a positive correlation with Th1, Tc1 cells and emphysematous lesions ( r =0.519 - 0.797, all P ＜ 0.01 ).In addition, a positive correlation between Th17/Th1 cells and emphysematous lesions was also found (r =0.742, 0.802, all P ＜0.01 ).The percentage of CD4+ IL-17+ IL-21 +T cells was significantly increased in S12 and S24 groups [(0.19 ±0.04) %, (0.55 ± 0.24) %] compared to controls [(0.07 ± 0.03 ) %, (0.08 ± 0.03 ) %], all P ＜ 0.05.Meanwhile, as compared with that of the controls [( 1.22 ± 0.31 ), ( 1.34 ± 0.18 )], the percentage of CD8+ IL-21 R + T cells was also increased in SI 2 and S24 groups [( 2.94 ± 1.26 ), (4.12 ± 2.26 )], but there were no differences among smoke-exposure groups ( P ＞0.05 ).The frequency of CD4+ IL-17 + IL-21 + T cells had a positive correlation with Th 1, Tc1 cells and emphysematous lesions (r = 0.694 -0.754, all P ＜0.05).And the frequency of CD8+ IL-21R+ T cells also had a positive correlation with emphysematous lesions ( r = 0.516, 0.725, all P ＜ 0.05).Conclusions Cigarette smoke increased the expression and the activity of Th17 in mice.Th17 may play a potential (active) role in the development of lung inflammation through IL-21/IL-21R pathway.     

初发系统性红斑狼疮患者外周血CD4+调节性T细胞CD73表达

目的 研究CD73在初发活动性系统性红斑狼疮(SLE)患者外周血CD4+调节性T细胞的表达情况,探讨其在SLE发病中的作用.方法 采用流式细胞术检测29例初发未经治疗的活动期SLE患者(SLE组)和22例健康人(健康对照组)外周血CD4+CD25+CD73+T细胞百分率及CD4+CD73+、CD4+CDhi25、CD4+CD25+T细胞中叉头状转录因子3(FOXP3)蛋白表达,同时对CD73表达水平与SLE活动指标进行相关性分析.结果 SLE组患者外周血CD4+ CD25+ CD73+T细胞百分率低于健康对照组[(1.25±1. 32)%vs(2.35±1.09)%,P＜0.01].SLE组和健康正常对照组,CD73在C4+ CDhi25T细胞的表达水平[(29.05±12.53)%、(43.35±10.09)%]高于CD4+ CD25+T细胞[(17.48±6.92)%、(29.98±10.39)%,P＜0.001];FOXP3蛋白在CD4+ CD73+ T细胞[(65.36±14.40)%、(63.80±14.05)%]、CD4+ CDhi25 T细胞的表达水平[(67.30±13.04)%、(56.30±9.21)%]明显高于CD4+ CD25+ T细胞[(45.70±12.74)%、(43.98±5.17)%,P＜0.001],在CD4+ CD73-T细胞几乎不表达,而在CD4+ CD73+ T细胞、CD4+ CDhi25 T细胞中的表达差异无统计学意义(P值均大于0.05).CD73在CD4+ CD25+ T细胞的表达水平与SLE疾病活动指数、ESR、C反应蛋白、抗补体C1q、抗核小体抗体均无相关性(P＞0.05).结论 CD73可作为调节性T细胞新的表面标记,其在调节性T细胞中的异常表达可能参与SLE的发病机制.     

胰岛素抵抗的慢性丙型肝炎患者外周血CD4+CD25+调节性T细胞的数量及其功能

目的 了解胰岛素抵抗的慢性丙型肝炎患者外周血CD4 +CD25+调节性T细胞(Treg)数量和功能的变化.方法 筛选40例HLA-A2+慢性丙型肝炎患者(其中20例合并胰岛素抵抗),流式细胞仪检测患者CD4+CD25+Treg细胞占外周血中CD4+T细胞的频率,液闪计数仪检测对HCV特异性CD8+T细胞增殖的抑制作用,ELISA法检测IFN-y水平.统计学处理采用t检验.结果 胰岛素抵抗的慢性丙型肝炎患者外周血CD4 +CD25+ Treg细胞占CD4+T细胞的(9.5±1.9)％,明显低于慢性丙型肝炎患者的(11.2±2.2)％(t=2.615,P＜0.05).胰岛素抵抗指数(HOMA-IR)≥4患者的CD4+CD25+ Treg细胞比例为(9.0±1.8)％,明显低于HOMA-IR＜4患者的(10.8±2.3)％(t=2.413,P＜0.05).胰岛素抵抗的慢性丙型肝炎患者CD4+CD25+ Treg细胞和去除Treg的外周血单个核细胞(PBMC)共培养上清液中IFN-y为(4 050±580) pg/mL,明显高于慢性丙型肝炎患者的(2 005±330)pg/mL(t=13.705,P＜0.01).HOMA-IR≥4患者IFN-y为(5 682±986)pg/mL,明显高于HOMA-IR＜4患者的(2 819±660) pg/mL(t=7.630,P＜0.01).结论 随着胰岛素抵抗程度加重,慢性丙型肝炎患者外周血CD4+ CD25+ Treg细胞频率减低,对HCV特异性CD8+T细胞增殖的抑制作用减弱.     

半乳糖凝集素-1抑制过敏性哮喘小鼠Th2型炎症反应的研究

目的探讨半乳糖凝集素-1(galectin-1)抑制过敏性哮喘小鼠模型Th2型炎症反应的作用机制。方法54只BALB/c雌性小鼠随机均分为健康对照组、卵清蛋白(OVA)组和治疗组。在第0、3、7天,OVA组和治疗组小鼠分别皮下注射致敏液[含100μg OVA混合相同体积10%Al(OH)3佐剂],健康对照组注射等量生理盐水。最后1次注射后7 d,OVA组和治疗组小鼠分别进行滴鼻激发(50μg OVA),健康对照组使用50μl生理盐水,每天激发1次。治疗组在激发2 h后,给予galectin-1(1μg/ml)进行滴鼻治疗,健康对照组和OVA组使用生理盐水(10μl),连续7 d。第22天,检测3组小鼠气道高反应;收集肺泡灌洗液(BALF),吉氏染色后,对炎症细胞进行分类并计数;肺组织切片用HE染色后,镜下观察肺组织炎症病理变化;取眼球血,ELISA检测血清中过敏原特异性IgE和Th2型炎症细胞因子白细胞介素-4(IL-4)、IL-5、IL-13,以及γ干扰素(IFN-γ)、IL-10、转化生长因子-β(TGF-β)的表达水平。取脾淋巴细胞,流式细胞术检测调节性T细胞(Treg)的比例。结果气道高反应性结果显示,治疗组小鼠的气道高反应性(Penh值为2.08±0.17)较OVA组(Penh值为5.77±0.64)降低(P<0.05)。治疗组小鼠BALF中的嗜酸粒细胞、巨噬细胞和中性粒细胞数量分别为(1.33±0.52)×10^4/ml、(1.16±0.41)×10^4/ml、(1.33±0.52)×10^4/ml,与OVA组的[(7.00±1.41)×10^4/ml、(5.00±0.63)×10^4/ml、(5.50±0.84)×10^4/ml]比较,差异有统计学意义(P<0.05)。肺组织HE染色切片镜检显示,OVA组小鼠支气管周围出现炎症细胞浸润和气道壁增厚;治疗组小鼠的肺部炎症较OVA组改善,且与健康对照组相似。血清ELISA检测结果显示,治疗组血清中过敏原特异性IgE为0.24±0.06,IL-4、IL-5、IL-13的表达水平分别为(104.49±20.76)pg/ml、(82.82±7.71)pg/ml和(31.59±9.78)pg/ml,均较OVA组的[0.87±0.10,(442.72±14.97)、(445.18±35.60)和(434.67±9.78)pg/ml]降低(P<0.05)。治疗组血清中的IFN-γ、IL-10、TGF-β的表达水平分别为(120.80±9.71)、(63.05±6.05)、(67.89±6.64)pg/ml,均较OVA组的[(47.28±5.01)、(23.89±2.98)、(15.49±3.75)pg/ml]提高(P<0.05)。流式细胞术检测结果显示,治疗组的Treg比例为(9.64±0.41)%,较OVA组的(1.81±0.48)%增加(P<0.05)。结论Galectin-1通过促进机体调节性T细胞和调节性细胞因子IFN-γ、IL-10、TGF-β的生成抑制小鼠过敏性哮喘Th2型炎症反应。     

系统性红斑狼疮患者外周血Th17细胞与调节性T细胞的变化

目的 探讨外周血辅助性T细胞17(Th17)与调节性T细胞在系统性红斑狼疮(SLE)发生、发展中的变化.方法 选取32例SLE活动期患者作为SLE活动组、30例SLE稳定期患者作为SLE稳定组及25名健康者作为对照组,采用流式细胞术及定量聚合酶链反应(PCR)的方法,分别从蛋白质水平与mRNA水平检测其外周血CD4+T细胞IL-17及FoxP3的表达.采用单因素方差分析进行统计学处理.结果 SLE活动组外周血CD4+T细胞IL-17蛋白质(1.01±0.22)％及mRNA(2.04±0.63)表达水平显著高于SLE稳定组(0.48±0.16)％、(1.12±0.34)及健康对照组(0.41±0.12)％、1(P＜0.01),但SLE稳定组与健康者对照组差异无统计学意义(P＞0.05).SLE活动组外周血CD4+T细胞FoxP3蛋白质(2.36±0.54)％及mRNA(0.42±0.16)表达水平显著低于SLE稳定组(4.34±0.95)％、(0.87±0.28)及健康对照组(5.09±1.17)％、 1(P＜0.01),SLE稳定组又低于健康对照组(P＜0.05).结论 SLE患者外周血中可能存在Th17/调节性T细胞的失衡,且失衡程度可能与病情活动性相关.     

肝细胞癌患者免疫功能的研究

目的探讨肝细胞癌免疫功能的临床意义。方法对54例HCC患者、64例慢性乙型肝炎患者和23位正常献血员采用双抗体夹心法,一抗为生物素标记的抗IFN-γ、TNF-α、IL-2、IL-4、IL-10、IL-12P70抗体,根据样本的OD值及曲线的斜率计算出细胞因子的含量。采用免疫组织化学方法检测外周血CD3＋、CD4＋和CD8＋T淋巴细胞的百分率。银染法检测T淋巴细胞核仁形成区嗜银蛋白,流式细胞仪测定肝细胞癌患者外周血黏附单个核细胞CD83及HLA-DR表达。结果 HCC患者血清IFN-γ含量为（4.92±2.15）pg/mL,低于健康献血员（19.16±2.34）pg/mL和慢性乙型肝炎患者（11.42±6.86）pg/mL（P〈0.05）。HCC患者IL-2、IL-12P70含量降低,而TNF-α、TGF-β1、IL-4、IL-10含量增高。根据肝癌体积大小对患者分组比较血清IFN-γ水平,随着肿瘤体增大,血清IFN-γ水平下降,调节IFN-γ分泌的IL-12P70水平与IFN-γ的变化趋势一致,但HCC患者血液中都存在IFN-γ。根据甲胎蛋白的水平将未经治疗的HCC患者分成2组,结果提示甲胎蛋白含量高时血清IFN-γ水平下降,调节IFN-γ分泌的IL-12P70水平与IFN-γ的变化趋势一致。与慢性乙型肝炎患者和健康献血员相比,HCC患者CD4＋T淋巴细胞、CD4＋/CD8＋比值降低,反映T淋巴细胞增殖活性的AgNORs降低。黏附细胞悬液CD83和HLA-DR表达降低。结论 HCC患者的免疫功能低下,与肿瘤细胞的免疫逃逸有关。