PGE_2调节CD4~+T细胞分化的研究进展

前列腺素(prostaglandin,PG)是花生四烯酸的小分子衍生物,在内分泌系统、心血管系统、生殖系统和神经系统中发挥多种生物学效应。PGE_2作为PG比较有代表性的一种亚型,是人体内重要的炎症介质,参与调节CD4~+T细胞的分化及相关细胞因子的分泌,从而影响CD4~+T细胞所介导的疾病的发生发展。本文就PGE_2及其对CD4~+T细胞亚群分化的调节作用进行综述。     

Notch信号通路相关微小RNA在胶原蛋白诱导的关节炎小鼠免疫细胞中的表达

目的研究胶原蛋白诱发关节炎(CIA)小鼠T细胞中Notch信号通路相关微小RNA(miRNA)的表达情况。方法用牛Ⅱ型胶原蛋白和佐剂免疫DBA/1J小鼠建立CIA小鼠模型,分离外周血单个核细胞(PBMC),采用流式细胞术分选CIA小鼠和正常对照小鼠外周血、脾脏、淋巴结CD4~+T细胞和CD8~+T细胞。实时荧光定量PCR检测PBMC及T细胞中Notch信号通路相关miRNA分子的表达水平。结果与正常对照相比,CIA小鼠PBMC中miR-200b-3p、miR-30c-5p、miR-30b-5p及miR-23b-3p的表达水平均明显降低,miR-200b-3p、miR-30c-5p及miR-30b-5p在CIA小鼠外周血、脾脏、淋巴结CD4~+T细胞和CD8~+T细胞中的表达与正常对照相比未见明显差异,而CIA小鼠外周血、脾脏CD4~+T细胞中miR-23b-3p的表达水平显著低于正常对照小鼠,外周血CD4~+T细胞中的miR-23b-3p来源于脾脏。结论 miR-23b-3p可能参与Notch信号对CIA小鼠T细胞的免疫调控。     

外周血循环滤泡辅助性T 细胞及IL-21 在银屑病中的表达及意义

目的:探讨外周血循环滤泡辅助性T 细胞(Follicular helper T cells,Tfh)及相关细胞因子IL-21 在银屑病患者中的表达水平及其与疾病活动度的关系。方法:收集38 例银屑病患者及32 例健康对照者,流式细胞术检测外周血循环CD4+CXCR5+ Tfh 和CD4+ CXCR5+ ICOS+ Tfh 细胞比例以及Th17 细胞比例;ELISA 检测血清IL-21 浓度;分析这些指标间及与银屑病疾病活动度评分PASI 间的相关性。结果:与健康对照者相比,银屑病患者外周血循环CD4+ CXCR5+ Tfh 和CD4+ CXCR5+ ICOS+Tfh 细胞比例更高,血清IL-21 浓度和Th17 细胞比例亦显著高于对照组(P<0.05);IL-21 浓度与CD4+ CXCR5+ Tfh 和CD4+CXCR5+ ICOS+ Tfh 细胞比例均显著正相关,而与Th17 细胞比例间无显著相关性(P>0.05);且CD4+ CXCR5+ ICOS+ Tfh 细胞比例和血清IL-21 与银屑病疾病活动度PASI 评分显著正相关,而CD4+ CXCR5+ Tfh 则与之无显著相关性(P>0.05)。结论:银屑病患者外周血循环Tfh 比例、IL-21 浓度上调与银屑病患者疾病活动度密切相关,提示Tfh 可能参与银屑病的发生发展过程,这一效应可能是通过分泌高水平IL-21 实现;IL-21 浓度与Tfh 细胞比例相关,而与Th17 细胞比例无相关性提示银屑病患者IL-21可能主要由Tfh 细胞分泌。     

类风湿性关节炎患者外周血CD3~+CD8~+效应记忆T细胞和滤泡辅助性T细胞ICOS、 PD-1表达上调且与病情相关

目的研究类风湿性关节炎(RA)患者外周血T淋巴细胞程序性死亡蛋白1(PD-1)和可诱导共刺激分子(ICOS)的表达,并确定其与疾病活动度之间的关系。方法募集RA患者30例、健康对照26例。流式细胞术检测外周血CD3~+CD8~+的效应记忆T细胞(Tem)和滤泡辅助性T(Tfh)细胞比例,而后检测淋巴细胞亚群中PD-1和ICOS阳性的细胞比例;通过Spearman相关性分析评估其与28个关节疾病活动度评分(DAS28)之间的相关性。结果与健康对照组相比, RA组患者外周血Tem和Tfh细胞绝对数增高, Tem和Tfh细胞的ICOS与PD-1表达增加; RA组患者外周血CD3~+CD8~+ Tem和Tfh细胞ICOS和PD-1表达水平与DAS28之间呈正相关。结论外周血CD3~+CD8~+ Tem和Tfh细胞PD-1和ICOS可能参与RA的发生发展,并可能作为RA活动度的评价指标。     

Tfh及IL-2与IL-17在老年2型糖尿病患者外周血中的变化及其临床意义

探讨老年2型糖尿病(type 2diabetes mellitus,T2DM)患者滤泡辅助性T细胞(follicular helper T cell,Tfh)、CD3~+T细胞、CD4~+T细胞、CD8~+T细胞、NK细胞的数量变化以及IL-2和IL-17等细胞因子含量差异并与患者血糖水平做相关性分析。采用流式细胞仪检测100例老年T2DM患者外周血CD3~+T细胞、CD4~+T细胞、CD8~+T细胞、NK细胞水平并与150例老年非糖尿病患者进行比较。分析淋巴细胞亚群与患者空腹血糖(fasting plasma glucose,FPG)、餐后2h血糖(2hour plasma glucose,2hPG)、糖化血红蛋白(glycosylated hemoglobin,HbA1C)、胰岛素抵抗指数(homeostasis model assessment-insulin resistance,HOMA-IR)及胰岛β细胞功能指数(homeostasis model assessment beta,HOMA-β)之间的相关性;采用ELISA方法检测外周血IL-2和IL-17水平。老年T2DM患者外周血中CD3~+T细胞、CD4~+T细胞、CD4~+T细胞/CD8~+T细胞的比值以及NK细胞均低于健康对照组,并与血糖呈负相关。老年T2DM患者外周血Tfh、CD8~+T细胞以及IL-17水平均高于健康对照组。老年T2DM的发病机制可能是随着年龄增高与长期高血糖导致免疫功能异常,外周血CD4~+T细胞/CD8~+T细胞水平下降以及Tfh和IL-17水平增高进一步加剧了细胞和体液免疫调节紊乱,致使胰岛细胞不断被破坏。     

哮喘患者急性加重期滤泡辅助型T细胞的分化水平及临床意义

目的观察滤泡辅助型T细胞(Tfh细胞)在支气管哮喘患者急性发作期和治疗后的分化水平,探讨Tfh细胞是否在哮喘中存在分化异常及其临床意义。方法采用流式细胞术检测健康志愿者、哮喘患者急性发作期及治疗后外周血单个核细胞中CD4~~+CXCR5~~+T细胞、CD4~~+ICOS~+T细胞、CD4~~+CXCR5~~+ICOS~~+T细胞的比例;反转录PCR法检测CD4~+T细胞中Bcl-6mRNA的水平;ELISA检测血浆白细胞介素21(IL-21)的水平;采用直线回归统计学分析IL-21与患者第一秒用力呼气容积/用力肺活量百分比(FEV1/FVC)及第一秒用力呼气容积/预计值百分比(FEV1/Pre)之间的相关性。结果哮喘患者CD4~~+CXCR5~~+细胞、CD4~~+CXCR5~~+ICOS~~+T细胞比例、Bcl-6 mRNA表达水平、IL-21水平均显著高于正常对照组;治疗后哮喘患者CD4~~+CXCR5~~+细胞、CD4~~+ICOS~~+细胞、CD4~~+CXCR5~~+ICOS~~+T细胞表达比例、Bcl-6 mRNA表达水平、IL-21水平均显著低于急性发作时;哮喘患者IL-21表达水平与患者FEV1/FVC、FEV1/Pre存在负相关。结论 Tfh细胞在哮喘急性期存在分化增强,治疗后Tfh细胞分化减弱,提示Tfh细胞可能参与哮喘的炎症反应,其水平可能反映哮喘患者气道受限程度。     

EP_2A体外可促进人CD34~+细胞归巢而不能促进其增殖

目的:探讨前列腺素E_2受体2激动剂(EP_2A)在体外对人CD34~+细胞的归巢与增殖作用。方法:收集健康供者经粒细胞集落刺激因子动员后的外周血,免疫磁珠法分选出人CD34~+细胞;同时收集健康供者动员前骨髓液,分离单个核细胞,并行骨髓间充质干细胞(BMMSC)培养。人CD34~+细胞和BMMSC经前列腺素E_2(阳性对照)、DMSO(阴性对照)、EP_2A和EP_2A+前列腺素E_2受体2拮抗剂(EP_2AA)处理后,对人CD34~+细胞用CCK-8法检测细胞活力,集落形成实验检测集落形成数目,流式细胞术检测G_2/M期细胞比例,Western blot检测细胞中survivin、β-catenin及CXC趋化因子受体4(CXCR4)的蛋白表达;ELISA法检测BMMSC中基质细胞衍生因子1α(SDF-1α)的含量。结果:EP_2A组与阴性对照组相比,人CD34~+细胞在细胞活力、集落形成数目、G_2/M期比例及survivin和β-catenin蛋白表达方面均无明显差别。但EP_2A组人CD34~+细胞CXCR4及BMMSC SDF-1α的表达均明显高于阴性对照组。结论:EP_2A体外可促进人CD34~+细胞归巢但不能促进其增殖。     

滤泡辅助性T细胞及亚群紊乱参与儿童过敏性紫癜发病机制初探

目的探讨外周血滤泡辅助性T细胞(T follicular helper cells,Tfh细胞)及其亚类失衡在儿童过敏性紫癜(HenochSch?nlein purpura,HSP)及紫癜性肾炎(HSP nephritis,HSPN)发生发展中的作用及可能机制。方法收集确诊的HSP患儿45例,根据肾脏受累情况分为3组:无肾脏受累组(A组)21例,孤立性血尿组(B组)12例,血尿合并蛋白尿组(C组)12例,另收集同期在我院健康体检儿童17例为对照组。采用流式细胞术检测外周血中Tfh(CD3~+CD4~+CXCR5~+)细胞及其亚类Tfh1(CCR6~-CXCR3~+Tfh),Tfh2(CCR6~-CXCR3~-Tfh),Tfh17(CCR6~+CXCR3~-Tfh)比例,同时检测ICOS及OX40在Tfh细胞及其亚类上表达情况。结果与对照组相比,A组和C组Tfh细胞及表达ICOS水平都明显增高(P0.05);C组Tfh2细胞、(Tfh2~+Tfh17)/Tfh1、Tfh细胞OX40和ICOS表达、Tfh2细胞ICOS表达均上调,而Tfh1细胞水平下调,差异有统计学意义(P0.05);A组和B组Tfh1细胞、Tfh2细胞、Tfh17细胞及这些亚类细胞上ICOS和OX40表达,均无统计学差异(P0.05)。HSP各组间比较,C组Tfh细胞比例较A、B组有增高的趋势,但差异无统计学意义,A组和B组间Tfh细胞无差异(P0.05)。进一步发现外周血Tfh细胞、Tfh2细胞、Tfh细胞上ICOS和OX40表达与血清IgA水平成正相关,而Tfh1细胞与IgA水平成负相关,Tfh17细胞与IgA无相关性。结论 Tfh亚类紊乱(Tfh2增加,Tfh1降低)参与了HSP及HPSN发生。而Tfh可能通过ICOS和OX40信号通路参与HSP高IgA血症,但其具体机制尚待进一步研究。     

C5aR在慢性移植物抗宿主病中的表达及其作用机制

目的:探讨补体5a(C5a)及其受体(C5aR)在慢性移植物抗宿主病(cGVHD)中的表达及其作用机制。方法:用流式细胞术检测20例cGVHD患者及9例健康供者外周血淋巴细胞中C5aR的表达及CD4~+CD25~+Foxp3~+调节性T细胞(Tregs)在CD4~+T细胞中的比例,并分析两者的相关性;将体外分离培养小鼠脾细胞分为对照组及重组小鼠C5a蛋白(rmC 5a)刺激组,用流式细胞术检测2组Tregs在CD4~+T细胞中的表达比例;另外,提取患者外周血单个核细胞进行体外培养,分为空白对照组及C5aR拮抗剂(C5aRA)组,用流式细胞术检测2组Tregs在CD4~+T细胞中的表达比例。结果:cGVHD患者外周血淋巴细胞表面C5aR的表达明显增多,而Tregs在CD4~+T细胞中的比例明显减少,两者呈显著负性相关(P0.05);体外培养小鼠脾细胞结果显示C5a下调Tregs在CD4~+T细胞中的比例;而体外培养患者外周血单个核细胞显示阻断C5aR可上调Tregs在CD4~+T细胞中的比例。结论:C5a/C5aR可能通过抑制Tregs的分化来介导cGVHD的发生发展。     

可溶性丙型肝炎病毒E2糖蛋白抑制人活化CD4~+T细胞的促炎细胞因子分泌

目的探索可溶性丙型肝炎病毒(HCV)E2糖蛋白对人活化CD4~+ T细胞的免疫调节作用。方法真核表达质粒pcDNA3.1-HCV sE2转染HEK293T细胞,细胞培养上清经Ni-NTA树酯亲和层析纯化可溶性HCV E2蛋白。应用流式细胞术分选健康捐献者外周血CD4~+ T细胞,经1.0μg/mL小鼠抗人CD3单克隆抗体、 1.0μg/mL小鼠抗人CD28单克隆抗体刺激培养48 h。刺激后的CD4~+ T细胞分成空白对照组、(2、 5、 10)μg/mL HCV E2蛋白组,后者经HCV E2糖蛋白处理。培养36 h后,流式细胞术检测CD4~+ T细胞程序性死亡蛋白1(PD-1)的表达变化, ELISA检测细胞培养上清液肿瘤坏死因子α(TNF-α)、γ干扰素(IFN-γ)的水平。结果与空白对照组比较, HCV E2蛋白组CD4~+ T细胞PD-1表达轻微降低,但差异无统计学意义。与空白对照组比较,可溶性E2蛋白组CD4~+ T细胞分泌TNF-α水平明显降低,并呈剂量依赖性;可溶性E2蛋白组CD4~+ T细胞分泌IFN-γ水平明显降低,但高低剂量组无明显差异。结论可溶性HCV E2蛋白抑制人活化CD4~+ T细胞促炎细胞因子的分泌。     

Improved bronchodilation with levalbuterol compared with racemic albuterol in patients with asthma

Background: Racemic albuterol is an equal mixture of (R)-albuterol (levalbuterol), which is responsible for the bronchodilator effect, and (S)-albuterol, which provides no benefit and may be detrimental. Objective: We sought to compare 2 doses of a single enantiomer, levalbuterol (0.63 mg and 1.25 mg), and equivalent amounts of levalbuterol administered as racemic albuterol with placebo in patients with moderate-to-severe asthma. Methods: This was a randomized, double-blind, parallel-group trial. Three hundred sixty-two patients 12 years of age or older were treated with study drug administered by means of nebulization 3 times daily for 28 days. The primary endpoint was peak change in FEV₁ after 4 weeks. Results: The change in peak FEV₁ response to the first dose in the combined levalbuterol group was significantly greater compared with the combined racemic albuterol group (0.92 and 0.82 L, respectively; P = .03), with similar but nonsignificant results after 4 weeks (0.84 and 0.74 L, respectively). Improvement in FEV₁ was similar for levalbuterol 0.63 mg and racemic albuterol 2.5 mg and greatest for levalbuterol 1.25 mg. Racemic albuterol 1.25 mg demonstrated the weakest bronchodilator effect, particularly after chronic dosing. The greatest increase in FEV₁ was seen after levalbuterol 1.25 mg, especially in subjects with severe asthma. All active treatments were well tolerated, and β-adrenergic side effects after administration of levalbuterol 0.63 mg were reduced relative to levalbuterol 1.25 mg or racemic albuterol 2.5 mg. At week 4, the predose FEV₁ value was greatest in patients who received levalbuterol or placebo when compared with those who received racemic albuterol. The difference was more evident and was statistically significant in patients who were not receiving inhaled corticosteroids. Conclusion: Levalbuterol appears to provide a better therapeutic index than the standard dose of racemic albuterol. These results support the concept that (S)-albuterol may have detrimental effects on pulmonary function. (J Allergy Clin Immunol 1998;102:943-52.)     

Infectious diseases and immunological markers associated with patients with non-Hodgkin lymphoma treated with rituximab

Background: The use of rituximab (RTX) is increasing, even in developing countries. It has become the first-line therapy or adjuvant to chemotherapy (CHOP; cyclophosphamide, hydroxydaunorubicin, oncovin and prednisone) for various diseases, including B cell lymphoma and autoimmune diseases.

Aim: We describe the infectious diseases and immunological markers associated with RTX treatment of patients with non-Hodgkin lymphoma (NHL).

Methods: Serum immunoglobulins were determined before and after intravenous immunoglobulin (IVIg) administration. Pneumo-23IgG-specific anti-pneumococcal antibodies were evaluated before and after vaccination. Immunophenotyping and lymphocyte proliferation were determined in the course of the treatment.

Results: Seven patients were followed and median age was 56.0?±?5.0?years (range, 41.9–71.6?years). At baseline, the mean level of IgG was 333.7?±?40.8?and IgM 40.9?±?11.3?mg/dL, respectively; immunoglobulin A and E (IgA and IgE) were under the limit of detection. Two patients had reduced or absent B cells and T cell subsets were at normal levels in five patients. All patients failed to mount an efficient post-vaccination immune response against hepatitis B virus, tetanus, diphtheria and against the 23-valent pneumococcal polysaccharide vaccine. During RTX/CHOP treatment, human-IgG-immunoglobulin (IVIg) therapy was introduced in six patients after recurrent infections, including community-acquired pneumonia (85.7%), chronic sinusitis (85.7%) and gastroenteritis (42.9%).

Conclusion: Poor response against pneumococcal vaccines increases the susceptibility of respiratory diseases in these patients. In patients with NHL treated with RTX, the benefits achieved with IVIg replacement for the control of recurrent infectious diseases is of paramount importance. Clinicians dealing with monoclonal antibodies against cancer therapy, especially RTX, should be aware of the increasing risks for symptomatic induced hypogammaglobulinemia and respiratory infections.     

Satellite RNA associated with bamboo mosaic potexvirus shares similarity with satellites associated with sobemoviruses

Summary A putative nonstructural protein encoded by a satellite RNA associated with bamboo mosaic potexvirus shares 46% identity with the capsid protein of satellite virus of panicum mosaic sobemovirus. The sequence similarity among satellite plant viruses which have no apparent relationship implies a common origin.     

拉米夫定与泛昔洛韦联合治疗乙型肝炎病毒慢性感染的临床研究

目的 观察拉米夫定与泛昔洛韦联合治疗乙型肝炎病毒(HBV)慢性感染的临床疗效。方法 慢性乙型肝炎患者90例。设联合治疗组28例，单用拉米夫定组30例，单用泛昔洛韦组32例。联合治疗组给予口服拉米夫定0．1g／d(PO)，泛昔洛韦1．5g／d(PO)，24周。拉米夫定、泛昔洛韦单用组剂量及疗程分别同联合治疗组。结果 3组均无明显副反应，丙氨酸转氨酶(ALT)复常率无差异。3组HBV DNA阴转率分别为89．3％、66．7％、40．6％，差异有显著性。乙型肝炎表面抗原(HBeAg)阴转率分别为28．6％、23．3％、21．9％，差异无显著性。结论 拉米夫定与泛昔洛韦联合用药安全、耐受性好，临床显示联合治疗对HBV DNA的抑制作用显著优于单用药。     

Gynecomastia with marked cellular atypia associated with chemotherapy

Gynecomastia is a common benign male breast disease, which may exhibit mild cellular atypia in cytology specimens. However, marked cytologic atypia can be seen in gynecomastia superimposed by chemotherapy. The case described in this report demonstrated severe cytologic atypia of gynecomastia mimicking carcinoma in a patient treated with chemotherapy for acute leukemia. A distinct cytologic feature helpful in avoiding the diagnostic error is described, namely, atypical cells admixed with bland ductal cells and appearing at a different plane. The importance of applying strict diagnostic criteria in breast cytology and clinical correlation is also emphasized.     

Intervention with epinephrine in hypotension associated with mastocytosis

The occurrence of the episodes of vasodilatory hypotension can be a life-threatening manifestation of systemic mastocytosis. This article describes the reversal by epinephrine of episodes of severe hypotension in two hospitalized patients with mastocytosis. Recognition of the efficacy of epinephrine in hypotension associated with mastocytosis can be important when other methods fail to restore hemodynamic stability.