ITP患者骨髓间质干细胞体外对CD4(+)CD25(+)Treg细胞增殖的影响

目的:体外研究免疫性血小板减少症(ITP)患者骨髓间质干细胞(MSCs)对CD4(+)CD25(+)Treg细胞的影响。方法:采用Ficoll分离ITP患者及健康对照者骨髓单个核细胞,通过体外培养,扩增出MSCs。通过Ficoll分离法和尼龙棉柱法获取ITP患者外周血T淋巴细胞,ITP患者MSCs经丝裂霉素C处理后按不同数量(1×103、1×104、1×105个细胞/孔)(A1、A2、A3组)接种培养板作为基底层细胞;健康对照者MSCs经丝裂霉素C处理后按不同数量(1×103、1×104、1×105个细胞/孔)(B1、B2、B3组)接种培养板作为基底层细胞;然后分别以2×105个细胞/孔接种体外分离纯化的ITP患者T淋巴细胞,设立空白组C组(ITP患者T淋巴细胞单独培养)。培养第3天各自收集T淋巴细胞,用流式细胞术检测各组CD4(+)CD25(+)Treg细胞比例。结果:在血凝素作用下,实验组及对照组各浓度亚组CD4(+)CD25(+)Treg细胞数量均较空白组明显减少(P0.05);A2组较B2组CD4(+)CD25(+)Treg细胞数量显著减少(P0.05),A3组较A2、B3组CD4(+)CD25(+)Treg细胞数量明显减少(P0.05)。结论:ITP患者可能存在MSCs质或量的改变,导致免疫调节功能异常,对CD4(+)CD25(+)Treg细胞增殖抑制作用明显加强,CD4(+)CD25(+)Treg细胞的不足引起外周免疫耐受减弱,MSCs的免疫调节功能异常可能在ITP发病过程中起重要作用。     

Reconstitution of CMV pp65 and IE-1-specific IFN-γ CD8(+) and CD4(+) T-cell responses affording protection from CMV DNAemia following allogeneic hematopoietic SCT

Threshold levels of CMV-specific T-cell populations presumably affording protection from active CMV infection in allo-SCT recipients have been proposed, but lack extensive validation. We quantified CMV pp65 and immediate-early 1-specific IFN-γ CD8(+) and CD4(+) T cell responses at days +30, +60 and +90 after transplantation in 133 patients, and established cutoff cell levels protecting from CMV DNAemia within the first 120 days after transplantation. No patients showing IFN-γ CD8(+) or IFN-γ CD4(+) T-cell counts >1.0 and >1.2?cells/μL, respectively, developed a subsequent episode of CMV DNAemia. Initial or recurrent episodes of CMV DNAemia occurred in the face of IFN-γ T-cell levels below defined thresholds. Negative predictive values at day +30 for the IFN-γ CD8(+) and CD4(+) T-cell markers were 68.1 and 61.8%, respectively. Recipients of grafts from CMV seropositive, related or HLA-matched donors, or receiving non-myeloablative conditioning had nonsignificant tendencies to reach more frequently protective levels of both T-cell subsets at early and late (day +365) times after transplantation. The use of anti-thymocyte globulin and umbilical cord blood transplantation were associated with impaired CMV-specific T-cell reconstitution. CMV-specific IFN-γ CD8(+) and CD4(+) T-cell recovery occurred irrespective of detectable CMV DNAemia.     

Unique roles of Schistosoma japonicum protein Sj16 to induce IFN-γ and IL-10 producing CD4(+) CD25(+) regulatory T cells in vitro and in vivo

Various proteins are expressed during different stages of schistosome development that are essential for cercarial penetration of vertebrate skin and evasion of host immune response. CD4(+) CD25(+) regulatory T cells are important in modulating immune responses towards helminth infections. Schistosoma japonicum protein Sj16 present in the secretions of schistosomula has been shown to have anti-inflammatory effects; however, it is uncertain whether Sj16 can induce CD4(+) CD25(+) regulatory T cells to participate in the regulation of early infection. In this study, we demonstrate a relationship between recombinant Sj16 (rSj16) and the induction of CD4(+) CD25(+) Foxp3(+) regulatory T cells. An increase in CD4(+) CD25(+) T cells was observed both in splenic cells from mice injected with rSj16 and the cells pretreated with rSj16, respectively. The induced CD4(+) CD25(+) T cells suppressed CD4(+) CD25(-) T-cell proliferation; furthermore, IFN-γ and IL-10 released from rSj16-stimulated cells contribute to this suppression. Additionally, rSj16-treated bone marrow dendritic cells (BMDCs) demonstrate an immature phenotype and play a role in the conversion of CD4(+) CD25(-) T cells into suppressive CD4(+) CD25(+) regulatory T cells. Our study identified a new CD4(+) CD25(+) T-cell population that induced by rSj16 and suggests that an IFN-γ-biased microenvironment during early infection of schistosome may favour the establishment of infection.     

Do CD8(+)CD25(+) cells predict immune reconstitution syndrome in HIV-positive patients who begin HAART?

Although undesirable, immune reconstitution syndrome (IRS) indicates a favourable effect of HAART and it should be differentiated from an opportunistic infection because of their distinct implications and management. Our group analysed different immunological parameters with the aim of identifying IRS predictors in patients who begin HAART. We found that CD8(+)CD25(+) cell pretreatment values in patients who developed IRS were four times higher than in those patients who did not develop IRS.     

Sodium selenite ameliorates dextran sulfate sodiuminduced chronic colitis in mice by decreasing Th1, Th17, and γδT and increasing CD4(+)CD25(+) regulatory T-cell responses

AIM To assess the effect of sodium selenite on the severity of dextran sulfate sodium(DSS)-induced colitis in C57BL/6 mice.METHODS Mice were randomly divided into four groups(n = 10/group): normal group, selenium(Se) group, chronic colitis group, and Se + chronic colitis group. The mice were sacrificed on day 26. Survival rates, clinical symptoms, colon length, and histological changes were determined. The percentages and absolute numbers of immune system cells in the lamina propria lymphocytes(LPL) of the colon, the expression of m RNA in colon tissue, and the concentrations of Th1, Th17, and Treg cytokines in LPL from the large intestine, were measured.RESULTS Se significantly ameliorated the symptoms of colitis and histological injury(P 0.05 each), increasing the proportions of neutrophils and CD4+ CD25+ T cells(P 0.05 each) and decreasing the proportions of γδT cells, CD4+, CD4+CD44+, and CD4+ CD69+ T cells in LPL(P 0.05 each). Moreover, Se reduced the expression of IL-6, IFN-γ, IL-17 A, IL-21, T-bet, and RORγt(P 0.05 each), but enhanced the expression of IL-10 and Foxp3(P 0.05 each). CONCLUSION These results suggest that Se protects against DSSinduced chronic colitis perhaps by increasing the number of CD4(+)CD25(+) Tregs that suppress the secretion of proinflammatory cytokines and populations of Th1, Th17, and γδT cells.     

Homing and clonogenic outgrowth of CD34(+) peripheral blood stem cells: a role for L-selectin?

OBJECTIVE: After transplantation of hematopoietic stem cells, adhesion molecules play a major role in the multistep process of engraftment in which L-selectin is suggested to be of relevance. A positive correlation previously was found between the number of reinfused L-selectin(+) stem cells and platelet recovery. In the present study, we determined the role of L-selectin in different engraftment steps, i.e., adhesion to endothelial cells, migration, and clonogenic outgrowth by in vitro assays that closely mimic the in vivo situation. MATERIALS AND METHODS: Flow adhesion and migration experiments were performed using the human bone marrow endothelial cell line 4LHBMEC and isolated peripheral CD34(+) cells with or without blocking of L-selectin-ligand interaction. Various clonogenic assays, including serum-free colony-forming unit-megakaryocytes (CFU-MK) and burst-forming unit-megakaryocytes (BFU-MK), were performed with sorted L-selectin(+)L-selectin(-) cells or in the presence of antibodies. RESULTS: Blocking of L-selectin on CD34(+) cells did not significantly affect rolling over and firm adhesion to 4LHBMEC. In addition, no role for L-selectin was found in transendothelial migration experiments. Finally, in clonogenic outgrowth of sorted or anti-L-selectin monoclonal antibody-incubated CD34(+) cells, no key role for L-selectin expression could be defined in BFU-MK and CFU-MK assays. CONCLUSION: Using in vitro assays for CD34(+) stem cell adhesion, migration, and clonogenic capacity, we were not able to define a major role for L-selectin.     

罕见CD8(+)蕈样肉芽肿伴体液免疫缺失1例

正1病例资料患者,男,27岁,因"皮疹1年半,加重伴面部浮肿3个月"于2015年8月19日入院。2013年12月患者因日晒后出现头部红疹伴脱屑至当地医院就诊,经治疗无明显好转。红疹进一步扩散,进展至颈部、前胸部皮肤。2014年12月,患者前胸部皮肤皮疹逐渐增多,呈片状红疹样改变,伴红肿痒痛,出现水疱,同时伴有发热,就诊于武汉市第一医院皮肤科,经抗感染及静脉丙球冲击治疗,患者体温正常,水疱消退,但皮疹仍存在,并扩展至背部。后     

Allogeneic transplantation of CD34(+) selected cells from peripheral blood from human leukocyte antigen-identical siblings: detrimental effect of a high number of donor CD34(+) cells?

Clinical results after T-cell-depleted allografts might be improved by modifying the graft content of progenitor and accessory cells. Although the association of the number of donor T cells with the clinical outcome has been studied extensively, the optimum number of progenitor cells that should be administered to patients is unknown. The characteristics of 84 consecutive human leukocyte antigen (HLA)-identical sibling transplants of granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood progenitor cells depleted of T cells by CD34(+) positive selection (allo-PBT/CD34(+)) were analyzed for their effect on clinical outcome. After a median follow-up of 24 months (range, 1-70 months), 50 patients remain alive (59.5%) and 34 have died (21 [25%] as a result of the transplant and 13 [15.5%] due to disease relapse). The median number of CD34(+) cells administered to the patients was 3.9 x 10(6)/kg (range, 1.2-14.3 x 10(6)/kg). A number of CD34(+) cells in the inoculum of 1 x 10(6)/kg to 3 x 10(6)/kg was associated with increased survival: 21 of 28 (75%) patients are alive, as compared with 29 of 56 (52%) patients receiving more than 3 x 10(6)/kg (actuarial probability 75% vs. 42%, respectively; P =.01). In the multivariate analysis, the independent prognostic variables for survival were CD34(+) cell dose 1 x 10(6)/kg to 3 x 10(6)/kg (RR = 4.8; P =.0008), sex-pairing match (RR = 3.2; P =.002), and early stage of disease (RR = 2.8; P =.007). From these results it appears that, in allo-PBT/CD34(+) from HLA-identical siblings, a number of CD34(+) cells in the inoculum between 1 x 10(6)/kg to 3 x 10(6)/kg is an important factor for better survival, and that higher CD34(+) cell doses might be associated with a poorer outcome.     

Differential antigen specificity of hepatitis C virus-specific interleukin 17- and interferon γ-producing CD8(+) T cells during chronic infection

A subset of CD8(+) T cells can secrete interleukin 17 (IL-17). However, very little information is currently available about their antigen specificity, tissue distribution, and biological relevance in chronic human viral infection. To address these issues, we comprehensively analyzed peripheral and intrahepatic CD8(+) T-cell responses in a cohort of patients with chronic hepatitis C virus (HCV) infection for the antigen-specific production of IL-17 and interferon (IFN) γ. We found that HCV-specific IL-17-producing and retinoic acid receptor related orphan receptorγt-expressing CD8(+) T cells are detectable in blood and liver and target different epitopes, compared with IFN-γ-producing CD8(+) T cells. Their highest frequency was found in patients with low inflammatory activity, suggesting a protective role in chronic HCV infection.     

Vasorelaxant effects of cicletanine and its (+)- and (−)-enantiomers in isolated human pulmonary arteries

The purpose of the study was to investigate, in isolated human pulmonary artery, the ability of cicletanine and its (−) and (+)-enantiomers to attenuate the endothelin-1 (Et-1) induced vasoconstriction, and to potentiate vasorelaxation (relative to plateau of the effect of Et-1) by sodium nitroprusside (SNP) and human atrial natriuretic peptide (ANP). In pulmonary artery rings, Et-1 induced a concentration-dependent vasoconstriction with median effective concentration (EC₅₀ = 26 ± 2.8 nmol/L. Pretreatment of the vessels with 100 μmol/L (±)-cicletanine reduced the effect of Et-1 (EC₅₀ = 36 ± 3.5 nmol/L; P < .01). (−)-enantiomer displayed greater capacity to antagonize the vasoconstrictor action of Et-1 (EC₅₀ = 47 ± 4.2 nmol/L) v (+)-enantiomer (EC₅₀ = 29.9 ± 6.5 nmol/L; P < .01). In arterial rings, precontracted with 10 nmol/L Et-1, ANP caused vasorelaxation (EC₅₀ = 9.7 ± 1.9 nmol/L). The relaxant effect of ANP was potentiated by 100 μmol/L of (−)-(EC₅₀ = 4.2 ± 0.6 nmol/L; P < .01), but not (+)-cicletanine (EC₅₀ = 7.6 ± 0.7 nmol/L). Sodium nitroprusside relaxed pulmonary artery rings precontracted with 10 nmol/L Et-1 (EC₅₀ = 41 ± 11 nmol/L). The effect of SNP was potentiated by 10 μmol/L (±)-cicletanine (EC₅₀ = 9.0 ± 0.7 nmol/L; P < .05). The potentiating effect of 10 μmol/L (+)-cicletanine was weaker (EC₅₀ = 7.9 ± 1.8 nmol/L) than that of (−)-enantiomer (EC₅₀ = 3.3 ± 0.54 nmol/L; P < .05). The relaxant effect of SNP was not further potentiated by 100 μmol/L (±)-cicletanine. The present results demonstrate that, cicletanine antagonizes Et-1 induced vasoconstriction in an isolated human pulmonary artery and potentiates vasorelaxation by two guanylate cyclase activators, ANP and SNP. (−)-Cicletanine displays greater vasorelaxant activity v (+)-enantiomer.     

庆阳市新发现HIV/AIDS病例首次CD4^(+)T淋巴细胞检测结果分析

目的 分析新发现HIV/AIDS病例首次CD4+T淋巴细胞检测值分布及其影响因素,为制定随访、检测及治疗等干预措施提供参考依据。方法 从中国艾滋病综合防治信息系统中导出2012—2020年甘肃省庆阳市报告的HIV/AIDS病例首次CD4+T淋巴细胞检测数据,录入EXCEL 2007中,按研究变量分类整理,两组计量资料用非参数MannWhitney u检验,多组计量资料比较采用多个独立样本Kruskal-Wallis检验,线性关系采用非参数Spearman检验。结果2012—2020年庆阳市报告新发现HIV/AIDS病例542例,首次检测CD4+T淋巴细胞且符合入组的病例538例,不同性别(Z=-0.562)、地区(Z=-0.040)、民族(Z=-0.182)和性伴数(Z=-1.357)病例的首次CD4+T淋巴细胞检测绝对值,差异无统计学意义(P>0.05);不同疾病状态(Z=-16.640)、样本来源(Z=-6.756)、感染途径(Z=-3.094)以及有无性病史(Z=-2.315)病例的首次CD4+T淋巴细胞检测绝对值,差异均有统计学意义(P<0.05);不同年龄(χ^(2)=38.875)、职业(χ^(2)=23.739)、文化程度(χ^(2)=25.071)、婚姻状态(χ^(2)=13.057)和报告机构(χ^(2)=27.803)的新发现HIV/AIDS病例首次CD4+T淋巴细胞检测绝对值,差异均有统计学意义(P<0.05);进一步对其两两比较,≤24岁组高于50岁~组(Z=-5.922)、学生高于农民工(Z=-3.314)、大专高于小学(Z=-4.184)、未婚高于离异/丧偶(Z=-3.614)、血液检测机构高于医疗机构(Z=-3.244),差异均有统计学意义(P<0.05);年龄与首次CD4+T淋巴细胞检测绝对值呈负相关(r=-0.294,P<0.05)。结论 年龄、职业、文化程度、婚姻状态、报告机构、疾病状态、感染途径、是否主动检测和有无性病是影响庆阳市新发现HIV/AIDS病例首次CD4+T淋巴细胞检测绝对值的因素;应分类施策,提高HIV抗体检测率和及时检测CD4+T淋巴细胞,为扩大抗病毒治疗覆盖率提供支撑。     

运德素治疗HBV DNA(+)、HBeAb(+)肝硬化的临床观察

拉米夫定耐药或者HBeAg变异者,须谨慎选用干扰素。我们试用国产α-1b干扰素-运德素治疗慢性乙型肝炎后代偿期肝硬化且拉米夫定耐药HBVDNA（＋）、HBeAg（-）、HBeAb（＋）的患者32例,并与同期用冰莲健肝灵的32例进行了对照。现将临床治疗、观察结果报告如下。     

Characterization of CD4 and CD8 T cells producing IFN-γ in human latent and active tuberculosis

Patients with pulmonary tuberculosis (PTB) frequently have reduced IFN-γ production in response to mycobacterial antigens, compared to individuals with latent Mycobacterium tuberculosis infection (LTBi). However, it is not clear whether this reduced responsiveness is restricted to a particular T cell subset. Herein, PBMCs from 26 PTB patients, 30 household contacts (HHCs) of PTB, and 30 tuberculin positive (TST+) healthy subjects not recently exposed to PTB, were stained with CFSE and stimulated non-specific (PPD) for 120?h, and specific (CFP-10/ESAT-6) and latency (HSpX) mycobacterial antigens for 144?h and the percentage of CD4(+) and CD8(+)IFN-γ(+) T cells responding determined by flow cytometry, in addition to their memory phenotype by the CD45RO and CD27 expression. PTB had decreased frequency of both CD4(+) and CD8(+) precursor cells, as well as decreased number of CD4(+)IFN-γ(+) cells in response to all antigens, whereas CD8(+)IFN-γ(+) cells were decreased in response to PPD and ESAT-6, but not to CFP-10 and HSpX. HHCs exhibited the highest precursor frequencies and IFN-γ responses, irrespective of the antigen employed. The CD4(+)/CD8(+) cell ratios showed that in response to PPD CD4(+) precursor and IFN-γ-producer cells are more frequent than their CD8(+) counterparts, and that PTB have a decreased CD4(+)IFN-γ(+)/CD8(+)IFN-γ(+) ratio in response to PPD, CFP-10, and ESAT-6. CD4(+)IFN-γ(+) and CD8(+)IFN-γ(+) cells exhibited a central memory phenotype (CD45RO(+)CD27(+)), irrespective of the group of subjects and the antigen used for stimulation. In conclusion, PTB patients had a decreased percentage of CD4(+) and CD8(+) precursor cells and CD4(+)IFN-γ(+). HHCs exhibited the highest frequency of CD4(+) and CD8(+) precursors and CD4(+)IFN-γ(+)-producing cells.     

Increased CD4+CD16+ and CD4+CD56+ T cell subsets in Behçet's disease

Behçet's disease is a systemic vasculitis of unknown etiology. Various immune abnormalities have previously been shown in Behçet's disease. We investigated T lymphocyte subsets associated with cytotoxic activity and natural killer (NK) cells by flow cytometry in 37 patients with Behçet's disease, 38 healthy controls, and 17 diseased control patients. Compared to the healthy controls, CD4⁺CD16⁺ and CD4⁺CD56⁺ subsets were found to be higher in the Behçet's disease group as well as in the disease control group (CD4⁺CD16⁺: BD=5?±?3, DC=14?±?14, HC= 3?±?2, P=0.001; CD4⁺CD56⁺: BD=11?±?5, DC= 18?±?17, HC=8?±?6, P=0.01). CD8⁺CD16⁺ and CD8⁺CD56⁺ T cell subsets were at normal levels in Behçet's disease but found to be elevated in disease controls. Similarly, NK cells (CD16⁺CD56⁺) were high only in the disease control group. Significant increases in CD4⁺CD16⁺ and CD4⁺CD56⁺ cell subsets in Behçet's patients and disease controls suggest that T cell activation patterns of these subsets in Behçet's disease are similar to those in other inflammatory disorders.     

Replenishing Peripheral CD4(+) Regulatory T Cells: A Possible Immune-Intervention Strategy in Type 1 Diabetes?

Controlling the diabetogenic activity of peripheral islet antigen-specific T cells is essential to halt the progression of autoimmunity that leads to the development of type 1 diabetes mellitus (T1DM). Over the past years, evidence has been gathered to suggest that the dysfunction of CD4(+)CD25(+) regulatory T (Treg) cells, and the interleukin-10 (IL10) -secreting type 1 regulatory T (Tr1) cells are associated with disease onset in diabetic patients. Although CD4(+)CD25(+) Treg cells develop as a distinct lineage of T cells in the thymus, results from recent studies have shown that they can also arise independently from the peripheral pool of conventional CD4(+) lymphocytes. These observations have led to the development of various methods to convert peripheral CD4(+) T cells into CD4(+)CD25(+) Treg and Tr1 cells in vitro or to induce the development and expansion of Treg cell subsets in vivo. This article reviews the progress made in Treg cell recruitment in vivo that involves the potential for the prevention or even reversal of T1DM.