Altered Expression of the Lymphocyte Activation Markers CD30 and CD27 in Patients with Pouchitis

Background: The mechanism underlying the development of ileal pouch inflammation in ulcerative colitis patients (pouchitis) after restorative proctocolectomy is unclear. Persistent systemic T cell activation or expansion of specific memory cell populations could predispose certain patients to develop local inflammation within the neo-rectum. Therefore, the aim was to study the expression of the lymphocyte activation markers CD27, CD30, CD25 and CD69 on the CD45RO+ memory cell subset of isolated peripheral blood mononuclear cells (PBMC), soluble CD30 levels and mucosal CD30 expression in patients with pouchitis and in controls. Methods: Flow cytometry was performed on PBMC isolated from patients with pouchitis (n = 9), without pouchitis (n = 10) and normal controls (n = 9). Serum CD30 was measured in patients with pouchitis (n = 25), without pouchitis (n = 26) and normal controls (n = 20) by ELISA. CD30 expression was quantified in pouchitis (n = 15) and normal pouch (n = 15) mucosa using a three-stage immunoperoxidase method. Results: Naïve CD45RO-CD27+ PBMC were significantly decreased in pouchitis (25.6%) compared to normal controls (34.4%), (P = 0.03). CD30, CD25 and CD69 subsets did not differ between the groups. Serum CD30 was increased in pouchitis patients 58 (1-380) U/ml compared to non-pouchitis 16.5 (1-290) U/ml, P = 0.007, and normal controls 11 (2-80) U/ml, P = 0.0005. In the mucosa, the numbers of CD30+ cells were increased in pouchitis compared to noninflamed pouches (P = 0.02). Conclusions: Increased sCD30 in pouchitis is associated with elevated mucosal expression. Of the activation markers studied, only the circulating naïve CD27+ population differed in pouchitis patients compared with controls. The observed decrease in this cell type may reflect antigen priming and subsequent loss of CD27 implying that antigen driven activation of specific T cell subsets may occur in pouchitis.     

Persistence of high CD40 and CD40L expression after restorative proctocolectomy for ulcerative colitis

AIM:To focus on the role of CD40 and CD40L in their pathogenesis. METHODS:We analyzed by immunohistochemistry the CD40 and CD40L expression in the pouch mucosa of 28 patients who had undergone RPC for UC, in the terminal ileum of 6 patients with UC and 11 healthy subjects. We also examined by flow cytometry the expression of CD40 by B lymphocytes and monocytes in the peripheral blood of 20 pouch patients, 15 UC patients and 11 healthy controls. RESULTS:Ileal pouch mucosa leukocytes presented a significantly higher expression of CD40 and CD40L as compared to controls. This alteration correlated with pouchitis, but was also present in the healthy pouch and in the terminal ileum of UC patients. CD40 expression of peripheral B lymphocytes was significantly higher in patients with UC and pouch, respect to controls. Increased CD40 levels in blood B cells of pouch patients correlated with the presence of spondyloarthropathy, but not with pouchitis, or inflammatory indices. CONCLUSION:High CD40 expression in the ileal pouch mucosa could be implied in the pathogenesis of pouchitis following proctocolectomy for UC, whereas its increased levels on peripheral blood B lymphocytes are associated with the presence of extraintestinal manifestations.     

原发性胆汁性肝硬化患者外周血单个核细胞Toll样受体表达与调节性T细胞相关性分析

目的 探讨原发性胆汁性肝硬化(PBC)患者外周血单个核细胞(PBMC)Toll样受体2、4、9(TLR2,TLR4,TLR9)表达及PBMC中CD4+ CD25+调节性T细胞(Tregs)特点和影响因素.方法 采用流式细胞术检测PBC患者(52例)和健康对照者(22例)外周血PBMC中TLR2、4、9阳性细胞及Tregs比例,比较其在PBC患者和健康对照者中的差别并分析TLR与Tregs相关性.结果 PBC患者Tregs比例低于健康对照者[(1.53+1.33)vs(4.42±1.43),P＜0.0001],TLR4阳性细胞比例高于健康对照者[(36.95±3.53)vs(32.84±8.06),P=0.003];PBC患者Tregs比例与TLR9阳性细胞比例呈负相关(R2 =0.115,P=0.016),健康对照者Tregs比例与TLR2,4和TLR2,4,9联合表达阳性细胞比例呈负相关(R2=0.326,P=0.007;R2 =0.226,P=0.034).结论 PBC患者外周血PBMC中Tregs比例降低,TLR4表达升高,可能受肝硬化失代偿期腹腔感染的影响.     

雷公藤内酯醇对系统性红斑狼疮病人B细胞表达CD86分子的影响

目的 了解系统性红斑狼疮(SLE)患者外周血B细胞表达CD86的情况，以及雷公藤内酯醇(Tripto1ide，TL)对其的影响。方法 用流式细胞仪检测外周血单个核细胞(PBMC)新鲜分离时以及与不同浓度TL在体外培养后的CB86阳性率。结果 SLE B细胞的CD86阳性率在新鲜分离时(P＜0．002)和培养48h后(P＜0．001)均较正常人B细胞高。25ng／m1的TL可明显降低SLE和正常人B细胞的CD86阳性率(P＜0．001)，2．5ng／ml则只对SLE B细胞起作用(P＜0．001)。结论 SLE病人B细胞CD86阳性率高于正常人；TL对SLE或正常人的CD86^ B细胞均有显著的下调作用，在某一浓度下仅抑制SLEB细胞的CD86表达。     

Quantitation of IL-2Rp75 (CD122) expression on mononuclear cells in rheumatic disease.

OBJECTIVE: To compare expression of the p75 chain of the interleukin-2 receptor (IL-2Rp75, CD122) on peripheral and synovial mononuclear cells in rheumatoid and non-rheumatoid inflammatory arthritis. METHODS: Peripheral blood (PBMC) and synovial (SFMC) mononuclear cells were isolated from subjects with rheumatoid arthritis (n = 16) and non-rheumatoid inflammatory arthritis (n = 12). PBMC were isolated from six healthy controls. Expression of CD122 was examined using indirect immunofluorescence and quantitative flow cytometry. RESULTS: There was no difference in IL-2Rp75 expression on PBMC from rheumatoid arthritis patients, non-rheumatoid arthritis patients, and controls. In subjects with rheumatoid arthritis there was no difference in IL-2Rp75 expression on PBMC and SFMC. However, in the non-rheumatoid arthritis group there was an increase in IL-2Rp75 expression on SFMC compared with PBMC (P = 0.0032). On SFMC there was a greater expression of IL-2Rp75 in non-rheumatoid arthritis than in rheumatoid arthritis (P = 0.0007). Expression was greater on CD8 positive cells and in subjects with shorter duration of disease. CONCLUSIONS: The p75 chain of the IL-2 receptor, an important T cell activation antigen, is not upregulated in synovial fluid. This appears to be a disease specific defect and provides further support for the concept of "frustrated" or incomplete T cell activation in this disease.     

Increased serum levels of soluble IL-2 receptor, CD30 and CD8 molecules, and gamma-interferon in angioimmunoblastic lymphadenopathy: possible pathogenetic role of immunoactivation mechanisms

Abnormal immunoreaction associated with increased cell activation phenomena might play a role in the pathogenetic events leading to the development of angioimmunoblastic lymphadenopathy (AILD). In the present study we investigated the serum levels of some soluble molecules related to cell activation in 24 patients with AILD at presentation. In particular, we measured by immunoenzymatic or immunoradiometric techniques the levels of the Tac peptide (sIL-2R), soluble CD30 (sCD30) and CD8 (sCD8) antigens, and gamma-IFN (gIFN). The results show that all the above molecules are increased as compared to normal controls, with a different pattern of increase for the different molecules. The sIL-2R levels were very high in all cases with no overlap between AILD and control samples (mean 6315 +/- 3374 U/ml, controls 271 +/- 112 U/ml, P less than 0.001). Very high values of the sCD30 antigen (722 +/- 895 U/ml) were detected in all cases but five, as opposed to the lack of detectable levels in controls. A significant increase of sCD8 (978 +/- 646 U/ml, controls 334 +/- 95 U/ml, P less than 0.01) and gIFN (329 +/- 236 U/ml, controls 97 +/- 43 U/ml, P less than 0.01) was also observed with some overlap between AILD samples and controls. The above findings further support the view that a condition of abnormally enhanced cell activation is likely to play a central role in the pathogenetic events leading to the composite clinicopathological picture of AILD.     

哮喘患者辅助T细胞活化与白细胞介素5释放

目的 了解过敏状态和哮喘状态下辅助（ＣＤ４＾＋）Ｔ细胞活化及白细胞介素５（ＩＬ－５）释放的原因和作用。方法 对过敏性哮喘组１２例（ＡＡ）、非过敏性哮喘组１０例（ＮＡＡ）、过敏性非哮喘组９例（ＡＮ）及正常对照组１０名（Ｎ）在有无抗原刺激下进行支气管肺泡灌洗液（ＢＡＬＦ）细胞及周围血单个核细胞（ＰＢＭＣ）培养，比较组内和组间ＣＤ４＾＋Ｔ细胞活化（标志物ＣＤ２５＾＋）与ＩＬ－５释放水平。结果 ＰＢＭＣ在     

系统性红斑狼疮患者外周血CD4+CD25highFoxp3+调节性T细胞数量和Foxp3mRNA的表达水平与疾病活动性的相关性

目的 研究系统性红斑狼疮(SEE)患者CD4+CD25highFoxp3+调节性T细胞的数量及其功能基因Foxp3 mRNA的表达水平与SLE疾病活动性和肾脏损伤的相关性.方法 采用四色流式细胞术以Foxp3-异硫氰酸荧光素(FITC )/CD25-藻红蛋白/CD4-多甲藻叶绿素蛋白(PerCP)/CD3-藻蓝蛋白7抗体组合检测40名健康对照者及42例SLE患者外周血CD4+CD25highFoxp3+调节性T细胞的数量,实时荧光定量聚合酶链反应(PCR)检测特异性转录因子Foxp3 mRNA的表达水平,并分析其与SLE患者疾病活动指数(SLEDAI)、补体C3及血清抗双链DNA(dsDNA)抗体的关系.统计学方法采用t检验和Spearman相关分析.结果 活动期SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量显著低于健康对照组[(4±3)％与(7±4)％,P＜0.05],稳定期与健康对照组差异无统计学意义(P＞0.05);活动期SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量及CD4+CD25highFoxp3+调节性T细胞/CD4+比值显著低于稳定期患者[(4±3)％,(9±6)％与(5±4)％,(10±6)％,P均＜0.05];活动期SLE患者外周血Foxp3 mRNA的表达水平明显低于稳定期和对照组(P＜0.01,P＜0.05);SLE患者并发肾病组外周血CD4+CD25highFoxp3+调节性T细胞数量及CD4+CD25highFoxp3+调节性T细胞/CD4+比值显著低于SLE非肾病组(P＜0.05).相关分析显示,SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量与SLEDAI呈负相关(r=-0.5782,P＜0.05);CD4+CD25highFoxp3+调节性T细胞/CD4+比值与SLEDAI呈负相关(r=-0.4913,P＜0.05),与补体C3呈正相关(r=0.3687,P＜0.05);SLE患者外周血CD4+CD25highFoxp3+调节性T细胞数量与Foxp3 mRNA的表达水平呈正相关(r=0.6142,P＜0.0l).结论 SLE患者外周血CD4+CD25highFoxp3+调节性T细胞和Foxp3 mRNA的变化可能是导致SLE疾病发生和发展的关键因素之一,与疾病的活动性有密切关系.     

CD4+CXCR5+ T cells activate CD27+IgG+ B cells via IL-21 in patients with hepatitis C virus infection

BACKGROUND: Chronic hepatitis C virus (HCV) infection causes the skewing and activation of B cell subsets, but the characteristics of IgG+ B cells in patients with chronic hepa-titis C (CHC) infection have not been thoroughly elucidated. CD4+CXCR5+ follicularhelperT(Tfh)cells,viainterleukin (IL)-21 secretion, activate B cells. However, the role of CD4+CXCR5+T cellsintheactivationof IgG+ BcellsinCHCpatientsis not clear.
METHODS: The frequency of IgG+ B cells, including CD27?IgG+B and CD27+IgG+ B cells,the expression of the activation markers (CD86 and CD95) in IgG+ B cells, and the percentage of circu-lating CD4+CXCR5+ T cells were detected by flow cytometry in CHC patients (n=70) and healthy controls (n=25). The con-centrations of serum IL-21 were analyzed using ELISA. The role of CD4+CXCR5+ T cells in the activation of IgG+ B cells was investigated using a co-culture system.
RESULTS: A significantly lower proportion of CD27+IgG+ B cells with increased expression of CD86 and CD95 was observed in CHC patients.The expression of CD95 was negatively correlated with the percentage of CD27+IgG+ B cells, and it contributed to CD27+IgG+ B cell apoptosis. Circulating CD4+CXCR5+ T cells and serum IL-21 were significantly increased in CHC patients. Moreover, circulating CD4+CXCR5+ T cells from CHC patients induced higher expressions of CD86 and CD95 in CD27+IgG+B cells in a co-culture system; the blockade of the IL-21 decreased the expression levels of CD86 and CD95 in CD27+IgG+ B cells.
CONCLUSIONS: HCV infection increased the frequency of CD4+CXCR5+ T cells and decreased the frequency of CD27+IgG+B cells. CD4+CXCR5+ T cells activated CD27+IgG+ B cells via the secretion of IL-21.     

Increased state of activation of CD4 positive T cells and elevated interferon γ production in pouchitis

Background—Immunoregulatory abnormalities of Tcells might be of importance in the pathogenesis of pouchitis afterileoanal pouch anastomosis (IAP).
Aims—To characterise T cell subsets, their stateof activation, and production of cytokines in inflamed and non-inflamedpouches in patients with ulcerative colitis (UC) and familialadenomatous polyposis (FAP). The influence of T cell activation onmucosal transformation was also studied.
Patients—Mucosal biopsy specimens were taken from42 patients with IAP (33 with UC and nine with FAP).
Methods—Mononuclear cells were isolated bystandard techniques and characterised by three colour flow cytometry.Interferon γ (IFN-γ) production was studied using the ELISPOT technique.
Results—In patients with UC with pouchitis therewas a significant increase in the CD4:CD8 ratio, expression ofactivation markers on CD3+ cells, and number of IFNγ producingmononuclear cells compared with patients with UC without pouchitis(CD4:CD8 ratio 1.3 (range 0.7-2.7) versus 0.6 (0.1-1.0), p=0.012). Inaddition, a positive correlation between increased crypt depth and thenumber of CD4+ cells (r=0.57) was shown.
Conclusion—The observed increase in activatedmucosal CD4+ T cells and IFN-γ production might lead to mucosaldestruction and crypt hyperplasia as seen in pouchitis.

Keywords:pouchitis; T cell activation; mucosaltransformation

     

羟氯喹抑制紫外线诱导的系统性红斑狼疮CD4+T细胞白细胞介素-10和干扰素-γ的表达

目的 研究紫外线对系统性红斑狼疮(SLE)CD4+T细胞因子的影响和羟氯喹的抑制作用.方法 选择SLE 30例,健康对照10名.磁珠分选SLE患者和健康人的CD4+T细胞,紫外线311 nm窄谱中波紫外线暴露,加入羟氯喹共培养,酶联免疫吸附试验(ELISA)检测培养上清白细胞介素(IL)-10和干扰素-γ的表达水平.采用t检验进行统计学分析.结果 SLE患者CD4+T细胞IL-10表达高于健康对照[(27±4)和(18±3) pg/ml,P=0.011];经45、100 mJ/cm2紫外线暴露后,SLE活动患者CD4+T细胞IL-10表达升高[(27±4)和(77±42) pg/ml,(40±18)和(77±42) pg/ml,P=0.022,P=0.048],经100 mJ/cm2紫外线暴露后,活动患者CD4+T细胞IL-10表达高于稳定患者[(77±42)和(24±4)pg/ml,P=0.029];羟氯喹降低SLE活动患者CD4+T细胞IL-10和干扰素-γ表达[(2.6±4.0)和(17.9±2.3)pg/ml,P=0.018,P=-0.017)];羟氯喹降低经45,100 mJ/cm2紫外线暴露后SLE活动患者T细胞IL-10表达[(40±18)和(22±6)pg/ml,(77±42)和(21±5) pg/ml,P=0.037,P=0.04];羟氯喹降低经100 mJ/cm2紫外线暴露的SLE活动和稳定患者T细胞干扰素-γ表达[(18±3)和(13±14) pg/ml,(19±7)和(12±5) pg/ml,P=0.013,P=0.049].结论 紫外线加重SLE患者体内Th1/Th2细胞因子的比例失衡;羟氯喹抑制了紫外线诱发SLE患者干扰素-γ和IL-10的表达.

Abstract：

Objective To explore the role of hydroxychloroquine (HCQ) in ultraviolet B (UVB)- induced expression of interleukin (IL)-10 and interferon (IFN)-γ from CD4+T cells in patients with systemic lupus erythematosus (SLE). Methods Thirty patients with SLE and 10 healthy controls were enrolled in the study. CD4+ T cells were isolated using magnetic beads from SLE patients and healthy controls. HCQ was added in culture media before and after irradiation with UVB 311 nm narrow band ultraviolet B (NB-UVB). The levels of IL-10 and IFN-γ in the supernatant were detected with enzyme-linked immunosorbent (ELISA). Comparisons between groups were performed by t-test. Results The level of IL-10 was higher in SLE patients [(27±4) pg/ml] than that in healthy controls [(18±3) pg/ml, P=0.011]. After exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages, the level of IL-10 was increased significantly in patients with active disease (P=0.022, P=0.048). After exposure of CD4+T cells to UVB in 100 mJ/cm2 dosages, the levels of IL-10 was higher in patients with active disease [(77±42) pg/ml] than patients with stable disease [(24± 4) pg/ml, P=0.029]. When CD4+ T cell were cultured with HCQ, IL-10 and IFN-γ levels in patients with active disease [(2.6±4.0), (17.5±2.3) pg/ml] were decreased significantly (P=0.018, P=0.017). HCQ reversed UVB-induced IL-10 expression in active SLE patients after exposure of CD4+T cells to UVB in 45 or 100 mJ/cm2 dosages (P=0.037, P=0.04). HCQ also reversed UVB-induced IFN-7 expression in active SLE patients and stable SLE patients after exposure to CD4+T cells with UVB in 100 mJ/cm2 dosages (P=0.013, P= 0.049). Conclusion UVB can aggravate the imbalance of Th1 and Th2 cytokines. HCQ inhibits UVB-induced IL-10 and IFN-7 expression of CD4+T cells in patients with SLE, especially in patients with active disease.     

Depletion in blood CD11c-positive dendritic cells from HIV-infected patients.

OBJECTIVES: To quantify blood dendritic cells from HIV-positive patients and to study the expression of functional molecules, in relation to HIV viral load, CD4 cell counts and antiretroviral treatment. DESIGN AND METHODS: Three-colour flow cytometry analysis was used to quantify blood dendritic cells without previous isolation from whole blood and to study the expression of functional molecules (MHC class II, CD11c, CD83, CD86) by dendritic cells from 30 HIV-positive patients, 15 of whom were treated with combined antiretroviral therapy (viral loads from undetectable to 5.4 log copies/ml, CD4 cell counts 1-1895 cells/mm3) and 11 non-infected controls. RESULTS: The median proportion of blood dendritic cells from HIV-positive patients was significantly decreased when the plasma viral load was above 200 copies/ml: 0.2% (0.1-1.1, n = 19) compared with 0.4% (0.2-0.8, n = 11) in patients with undetectable viral load whether they were treated or not, and to 0.4% (0.2-1.3, n = 11) in controls (P = 0.02). A major decrease of the CD11c positive dendritic cells was observed in all HIV-positive samples, with only 18% (mean; range: 0.3-80%, median 4.2%) compared with 44% (11-70%, median 42%) of control dendritic cells (P = 0.0006). In contrast, the proportion of dendritic cells expressing CD86, was slightly higher in HIV-positive patients than in controls (P = 0.03). CONCLUSIONS: The decreased proportion of blood dendritic cells correlated with virus replication and the lack of dendritic cells expressing CD11c are the first evidence of strong dendritic cell alterations in HIV-positive patients. Although the proportion of blood dendritic cells are in the normal range in treated HIV-positive patients with undetectable viral load, the CD11c alterations persist indicating that antiretroviral therapy might only partly correct the alterations of the circulating dendritic cells.     

系统性红斑狼疮患者外周血单个核细胞CD28 mRNA的表达

目的　探讨CD28在系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中的表达水平及其意义。方法　应用反转录-聚合酶链反应(RT-PCR)检测了34例活动期SLE患者和30名正常人PBMC中CD28mRNA的表达水平。结果　活动期SLE患者CD28的阳性表达率为20.6%,明显低于正常人对照组(70.0%),差异非常显著(P＜0.001);活动期SLE组CD28的平均表达水平(0.19±0.21)亦明显低于正常对照组(0.43±0.11),差异显著(P＜0.05)。结论　CD28的异常表达可能在SLE发病机制中起作用,CD28mRNA的低水平表达可能与外周血CD28＋T细胞凋亡增加或迁移到炎症部位有关。     

急性冠状动脉综合征患者的CD4+CD28null T淋巴细胞电压门控性Kv1.3钾通道数量增加

目的 运用膜片钳技术检测急性冠状动脉综合征(ACS)患者的CD4+CD28nullT淋巴细胞Kv1.3钾通道的数量是否增加,探讨CD4+CD28nullT淋巴细胞上Kv1.3钾通道的表达.方法 选择17例ACS患者,11例年龄、性别匹配的正常体检者作为对照.运用荧光染色标记及膜片钳技术检测单个T淋巴细胞(CD4+CD28nullT淋巴细胞和CD4+CD28+T淋巴细胞)上Kv1.3钾通道的数量,比较Kv1.3通道在两种细胞上的差别.结果 ACS患者的CD4+CD28nullT淋巴细胞比例为(6.97±2.05)%,明显高于对照组的(1.38±0.84)%(P<0.05).ACS患者的CD4+CD28nullT淋巴细胞数量与hsCRP浓度呈正相关(r=0.52,P<0.05).ACS患者CD4+CD28nullT淋巴细胞Kv1.3通道的电导、密度、数量明显高于对照组CD4+CD28+T淋巴细胞(P<0.01).而两组CD4+CD28+T淋巴细胞间Kv1.3通道的电导、密度、数量差异无统计学意义(P>0.05).结论 ACS患者CD4+CD28nullT淋巴细胞明显增多.ACS患者CD4+CD28nullT淋巴细胞上Kv1.3钾通道数量比自身和对照的CD4+CD28+T淋巴细胞明显增多,CD4+CD28nullT淋巴细胞的功能可能与Kv1.3钾通道增多相关.     

Immune modulatory effects of cyclooxygenase type 2 inhibitors in HIV patients on combination antiretroviral treatment

OBJECTIVES: To examine the immune modulating effects of cyclooxygenase type 2 (COX-2) inhibitors (COX-2i) in HIV-infected patients on combination antiretroviral treatment (CART). DESIGN: In-depth substudy from an approved, open, controlled, randomized study comparing the immune modulating effects of CART in combination with COX-2i after 12 weeks. METHODS: Patients (n = 38) on long-term CART with stable viral load (VL) < 50,000 copies/ml and CD4+ T-cell counts > 100/microl were randomized to CART and rofecoxib 25 mg bid (n = 12) or celecoxib 400 mg bid (n = 12), or CART only without placebo (n = 14). Routine clinical chemistry, CD4+ and CD8+ counts and VL were safety parameters. Immunological parameters included C-reactive protein, beta2-microglobulin, Ig isotypes and IgG subclasses as well as several T-lymphocyte subsets. Non-parametric analyses were used throughout. RESULTS: Prestudy experiments showed higher median intracellular expression of COX-2 in CD4+ (P = 0.048) and possibly CD8+ (P = 0.09) T cells from patients on CART compared with uninfected controls. In the clinical study, increased CD4+ T-cell counts were observed only in patients on COX-2i with VL < 50 copies/ml (P = 0.02). Decreased expression of CD38+ on CD8+ T cells and subsets as well as reductions in IgA and IgM (P < 0.03) were most pronounced in patients on COX-2i who had detectable VL (n = 6). COX-2i treatment enhanced the perforin content particularly in the differentiated CD27-/CD8+ T-cell subsets compared with controls (P = 0.05). CONCLUSIONS: COX-2i together with CART improved markers for persistent immune activation, particularly in patients with viraemia, as well as enhanced perforin expression, and thereby strengthened COX-2 as a potential therapeutic target in HIV infection.     

CD8+/DR+/CD25--T-lymphocytes associated with marrow graft failure

Phenotypic and functional characteristics of peripheral blood mononuclear cells (PBMC) were studied in eight patients with poor graft function following HLA-identical T cell-depleted marrow transplantation. Similar patients with good graft function and normal individuals were used as controls. Freshly isolated PBMC from patients with failing grafts contained more CD3+ and CD8+ cells than PBMC from well engrafted patients. The CD8+ cells appeared activated insofar as they expressed DR antigens, but they did not express the low affinity IL-2 receptor recognized by Tac antibody (CD25) and they did not have increased cytolytic activities. After culture with phytohemagglutinin (PHA) and IL-2, PBMC from patients with poor graft function contained fewer CD2+ and CD4+ cells than cultured PBMC from patients with good graft function. Cultured cells from patients with poor graft function acquired lymphokine activated killer (LAK) activity against NK-sensitive and NK-insensitive targets, but still did not express CD25. Host-mediated anti-donor cytotoxic activity could be demonstrated in one patient only after presensitization with donor cells and culture with IL-2 and PHA. The abnormalities in T cell activation observed in patients with poor graft function did not correlate with the donor or host origin of lymphoid cells. These data indicate that some cases of graft failure may be associated with defective T cell maturation. These abnormalities may simply represent a consequence of marrow failure or they may actually contribute to failure by not providing critical hematopoietic accessory functions.     

急性冠状动脉综合征患者外周血CD4+T细胞及CD28null/CD28+亚型活化后Kv1.3钾通道的表达和意义

目的 研究急性冠状动脉综合征(ACS)患者外周血中CD4+T细胞及CD28null/CD28+亚型活化前后Kv1.3钾通道数目的 变化以及Kv1.3钾通道阻滞剂对CD4+T细胞活化表达的影响,探讨Kv1.3钾通道在不稳定斑块中的意义.方法 用免疫磁珠法分离出27例ACS患者外周血中的CD4+T细胞,其中12例进一步分出亚型CD4+CD28null和CD4+CD28+T细胞,采用全细胞膜片钳技术记录细胞活化前及经CD3抗体活化72 h后的Kv1.3钾电流.CD4+T细胞活化时分别加入终浓度为0.1、1、10 nmol/L特异性Kv1.3钾通道阻滞剂rMargatoxin(rMgTX),共同培养72 h后用反转录-PER法检测干扰素-γ、肿瘤坏死因子(TNF)-α及颗粒酶B mRNA的表达.结果 活化后CD4+、CD4+CD28null、CD4+CD28+T细胞的Kv1.3钾通道的峰电流均明显增加,细胞平均通道数分别增加约90%、60%、80%[活化前后每个细胞的通道数分别为(402±88)个比(752±275)个、(553±328)个比(874±400)个、(392±133)个比(716±251)个,均P<0.05].活化前CD4+CD28nullT细胞Kv1.3钾通道的平均数目比CD4+CD28+T细胞多约40%(P<0.05),活化后两者差异无统计学意义(P=0.102).不同浓度的rMgTX均下调CD4+T细胞活化后干扰素-γ、TNF-α、颗粒酶B mRNA的表达,各浓度组间干扰素-γ、TNF-α、颗粒酶B mRNA的表达差异均有统计学意义(均P<0.01),浓度越高,各mRNA表达越低.结论 ACS患者外周血CD4+T细胞及CD28null/CD28+亚型活化后Kv1.3钾通道表达增加,特异性Kv1.3通道阻滞剂rMgTX呈浓度依赖性地抑制CD4+T细胞活化时干扰素-γ、TNF-α及颗粒酶B mRNA的表达,提示CD4+T细胞特别是CD4+CD28nullT细胞的Kv1.3钾通道可作为预防动脉粥样斑块不稳定的潜在治疗靶点.     

HIV-1感染者CD73~+CD8~+ T细胞减少与T细胞异常活化的相关性研究

目的探讨HIV-1感染者外周血CD8~+T细胞上CD73的表达特点及其与T细胞异常活化和疾病进展的关系。方法研究入选65例HIV-1感染者和27例健康对照。通过流式细胞术检测研究对象外周血CD73~+CD8~+T细胞的频率和绝对计数,并将患者CD73~+CD8~+T细胞绝对计数和频率与其CD4~+T细胞计数、HIV-1载量以及CD38~+CD8~+T细胞频率进行相关性分析。结果与健康对照相比,HIV-1感染者外周血CD73~+CD8~+T细胞绝对计数和频率均明显降低(P均0.05);HIV-1感染者外周血CD73~+CD8~+T细胞绝对计数和频率与CD4~+T细胞计数呈正相关(r=0.555,P=0.001;r=0.342,P=0.005),与CD38~+CD8~+T细胞频率呈负相关(r=-0.384,P=0.002;r=-0.387,P=0.001);HIV-1感染者CD73~+CD8~+T细胞的绝对计数与HIV-1载量呈弱负相关(r=-0.261,P=0.035)。结论 HIV-1感染者外周血CD73~+CD8~+T细胞的减少不但与患者T细胞的活化程度呈显著负相关,而且与AIDS疾病进展相关。     

Combined carriership of TLR9-1237C and CD14-260T alleles enhances the risk of developing chronic relapsing pouchitis

AIM: To investigate the single nucleotide polymorphisms (SNPs) in genes involved in bacterial recognition and the susceptibility to pouchitis or pouchitis severity. METHODS: Analyses of CD14 -260C>T, CARD15/ NOD2 3020insC, Toll-like receptor (TLR)4 +896A>G, TLR9 -1237T>C, TLR9+2848G>A, and IRAKM + 22148G>A SNPs were performed in 157 Meal-pouch anal anastomosis (IPAA) patients (79 patients who did not develop pouchitis, 43 infrequent pouchitis patients, 35 chronic relapsing pouchitis patients) and 224 Italian Caucasian healthy controls. RESULTS: No significant differences were found in SNP frequencies between controls and IPAA patients. However, a significant difference in carriership frequency of the TLR9-1237C allele was found between the infrequent pouchitis and chronic relapsing pouchitis groups [P = 0.028, odd's ratio (OR) = 3.2, 95%d = 1.2-8.6]. This allele uniquely represented a 4-locus TLR9 haplotype comprising both studied TLR9 SNPs in Caucasians. Carrier trait analysis revealed an enhanced combined carriership of the alleles TLR9 -1237C and CD14 -260T in the chronic relapsing pouchitis and infrequent pouchitis group (P = 0.018, OR = 4.1, 95%CI = 1.4 -12.3). CONCLUSION: There is no evidence that the SNPs predispose to the need for IPAA surgery. The significant increase of the combined carriership of the CD14 -260T and TLR9 -1237C alleles in the chronic relapsing pouchitis group suggests that these markers identify a subgroup of IPAA patients with a risk of developing chronic or refractory pouchitis.