CD4^＋T淋巴细胞表面CD4^＋CD45RA和CD4^＋CD45RO在支气管哮喘患儿外周血中的表达

目的探讨CD4^＋CD45RA和CD4^＋CD45RO分子在支气管哮喘患儿中的表达及其意义。方法分别收集支气管哮喘发作期28例、支气管哮喘缓解期27例、健康对照儿童20例抗凝静脉血100μL，采用异硫氰酸荧光素（FITC）标记的抗CIM单抗、藻红蛋白（PE）标记的抗CD45RA单抗和PE-菁蓝色素荧光素（PE—Cy5）标记的抗CD45RO单抗，流式细胞仪检测各组儿童外周血CD4^＋T淋巴细胞表面CD45RA和CD45RO的表达。采用SPSS13．0软件进行统计学分析。结果与健康对照和支气管哮喘缓解组患儿比较，支气管哮喘发作组患儿CD4^＋CD45RA^＋T细胞明显减少（q=12．47，8，39Pa〈0．05），CD4^＋CD45RO^＋T细胞显著升高（q=9．50，8．30Pa〈0．05），CD4^＋CD45RA^＋／CD4^＋CD45RO^＋细胞比值显著降低（q=8．96，6．21P。〈0．05）；支气管哮喘缓解组患儿CD4^＋CD45RA^＋T细胞较健康对照组明显升高（q=3．08P〈0．05），CD4^＋CD45RO^＋T细胞及CD4^＋CD45RA^＋／CD4^＋CD45RO^＋细胞比值与健康对照组比较差异无显著性（q=0．45，2．02Pa〉0．05）。结论外周血CD4^＋CD45RA^＋和CD4^＋CD45RO^＋T淋巴细胞平衡失调可能参与支气管哮喘的发病。     

过敏性紫癜患儿急性期细胞免疫功能变化

目的 研究过敏性紫癜患儿急性期外周血淋巴细胞免疫的变化及其临床意义。方法 采用流式细胞术(FCM)对23例过敏性紫癜急性期患儿外周血淋巴细胞及其亚群：CD3^ 、CD4^ 、CD8^ 、CD19^ 、CD16 56^ 进行定量检测，并用^3H—TdR标记法测定非杀伤(NK)细胞活性。结果 过敏性紫癜急性期患儿外周血CD4^ ,CD4^ /CD8^ ,CD16 56^ 、NK细胞活性均显著低于健康对照组(P＜0.01)，而CD8^ 活性显著升高(P＜0．05)，CD19^ 显著升高(P＜0.01)，均有统计学意义，CD3^ 稍升高，无明显差异(P＞0．05)。结论 过敏性紫癜患儿急性期存在细胞免疫功能失调，主要表现在T细胞亚群紊乱，B细胞呈多克隆活化，NK细胞数量及活性降低。     

过敏性紫癜患儿急性期淋巴细胞凋亡与血清可溶性Fas和FasL变化的关系

目的 探讨过敏性紫癜(HSP)患儿急性期外周血淋巴细胞凋亡与血清可溶性Fasm(sFas)、可溶性FasL(sFasL)水平变化及其相互关系。方法 选择HSP患儿及健康儿童各33例。分别应用形态法、间接免疫荧光法、双抗夹心ABC-ELISA法检测外周血淋巴细胞凋亡率、CD25^ 细胞百分率及血清sFas及sFasL水平。结果 HSP患儿急性期外周血淋巴细胞培养0、48h凋亡率均显著高于正常对照组(P均＜0．01)；HSP组培养48h CD25^ 细胞百分率显著低于正常对照组(P＜0．01)，HSP患儿外周血淋巴细胞培养48h凋亡率与CD25^ 细胞百分率呈负相关(r＝-0．61 P＜0．05)；血清sFas及sFasL水平较正常对照组明显升高(P均＜0．01)；HSP患儿外周血淋巴细胞培养48h凋亡率与sFasL水平呈正相关(r＝0．69 P＜0．05)，与sFas水平呈负相关(r＝-0．58 P＜0．05)。结论 HSP患儿外周血淋巴细胞凋亡过度和血清sFas、sFasL水平升高，与患者免疫功能紊乱关系密切。血清sFasL升高是HSP患儿淋巴细胞凋亡过度的重要原因之一。     

手足口病患儿外周血淋巴细胞亚群变化的临床研究

目的 探讨手足口病(HFMD)患儿外周血T淋巴细胞亚群、B淋巴细胞、NK细胞的变化特点.方法 将41例HFMD患儿分为一般病例组23例,重症病例组18例,选择20例健康儿童作为对照组,采用流式细胞仪检测外周血T淋巴细胞亚群、B淋巴细胞、NK细胞.结果 CD3~+与CD4~+细胞表达:HFMD重症组、一般病例组、对照组依次显著降低(P＜0.01);CD8~+细胞表达:HFMD重症组、一般病例组、对照组依次显著降低(P＜0.05);NK细胞表达:HFMD重症组与一般病例组相似(P＞0.05),皆高于对照组(P＜0.05);B淋巴细胞表达:HFMD重症组、一般病例组、对照组依次升高(P＜0.01).结论 HFMD患儿免疫系统功能紊乱,其中细胞免疫有不同程度的抑制,体液免疫反应亢进.     

肺炎患儿血CD16+CD56+、CD16+自然杀伤细胞占淋巴细胞比例的变化

目的探讨肺炎患儿CD16^＋CD56^＋、CD16^＋自然杀伤细胞（NK细胞）占淋巴细胞比例的动态变化及其意义。方法采用流式细胞术动态测定急性肺炎患儿32例治疗第2．5、10天及30例健康对照组儿童外周血NK细胞CD16^＋CD56^＋、CD16^＋占淋巴细胞比例的变化。结果肺炎患儿CD16^＋CD56^＋、CD16^＋NK细胞占淋巴细胞的比例在肺炎急性期、恢复期和康复期均显著低于健康对照组（Pa〈0．01），急性期、恢复期和康复期3组比较，杀伤活性强的NK细胞逐渐升高，各组比较有显著差异（Pa〈0．01）。结论动态监测肺炎患儿血CD16^＋CD56^＋、CD16^＋NK细胞占淋巴细胞比例对预测其病程及预后有重要意义。     

新生儿窒息外周血T细胞亚群动态观察

为探讨新生儿窒息与免疫的关系，该院于1995年5月～1996年7月，对25例窒息新生儿外周血T淋巴细胞亚群进行了动态监测，结果新生儿窒息后CD：细胞减少（P＜0．01），CD：细胞增加（P＜0．01），CD4／CD8比值降低（P＜0．0025），而且重度窒息患儿CD细胞亦明显减少（P＜0．05），提示窒息可导致新生儿外周血T细胞亚群紊乱。复苏后其T细胞亚群可恢复正常。并对其发生机理及临床意义进行了讨论。     

新生儿血清锌水平与T细胞亚群分布的探讨

探讨不同胎龄新生儿的血清锌水平与T细胞亚群分布的关系。应用原子吸收法测定血清锌；流式细胞仪测定T淋巴细胞亚群。结果显示：28－32周早产儿组（I组）血清锌水平明显低于33－36周早产儿（Ⅱ组）及足月儿组（Ⅲ组）（P＜0．05），且I组低锌发生率为35％。I组T淋巴细胞亚群CD4^ /CD8^ 比率明显低于Ⅱ、Ⅲ组（P＜0．05）；Ⅰ、Ⅱ组淋巴细胞及T淋巴细胞绝对值明显低于Ⅲ组（P＜0．05）。12例低血锌新生儿CD4^ T细胞水平明显低于正常组，CD8^＋T细胞水平明显高于正常组（P＜0．05），CD4^ /CD8^ 比率明显下降，显著低于正常组（P＜0．001）。提示血清锌水平与胎龄有关, 胎龄越小，锌水平越低，缺锌的发生率越高；而T细胞亚群分布不仅与胎龄有关，与锌水平高低也密切相关。建议加强对≤32周的早产儿进行锌水平的监测。     

CD+8 CD-28调节性T细胞在传染性单核细胞增多症中的作用初探

目的观察传染性单核细胞增多症（IM）患儿CD8^＋CD28^-调节性T（Tr）细胞动态改变,探讨急性EB病毒感染的免疫调控机制。方法观察25例IM患儿及相同数量同龄对照组。用流式细胞术检测外周血CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD8^＋CD28^＋表面标志;用逆转录-聚合酶链反应和实时定量聚合酶链反应检测CD8^＋CD28^-Tr细胞中IL-6、IL-10、IFN—γ及MC中ILT-3、ILT-4mRMA表达。结果IM患儿急性期CD8^＋CD28^-Tr细胞阳性表达率明显高于同龄对照组（P〈0．01）,恢复期阳性表达率较急性期有所下降,但仍高于对照组（P〈0．01）;IM患儿CD8^＋CD28^-Tr细胞和MC效应分子IL-6、IL-10、IFN-γ、ILT-3、ILT-4表达明显增高（P〈0．01）。结论IM患儿CD8^＋CD28^-Tr细胞增多可能是机体调控病毒感染的适应性免疫反应过程,其数量（及活性）降低或增高是否与EB病毒感染相关性疾病的发生发展有关尚需进一步探讨。     

系统性红斑狼疮患儿PD-1分子的表达

目的 研究PD-1分子的表达与系统性红斑狼疮（SLE）的发病机制及病情的关系。方法 应用流式细胞仪检测各期SLE患儿（活动期、缓解期）外周血CD3^＋、CD4^＋、CD8^＋T细胞上PD-1分子的表达。结果 在各期SLE患儿中外周血CD3^＋、CD4^＋、CD8^＋淋巴细胞上PD-1分子的表达均明显高于正常对照（P〈0.05）。结论 在系统性红斑狼疮患儿外周血淋巴细胞上PD-1分子表达明显高于正常,各组SLE患儿存在B淋巴细胞的异常增殖,推测 PD-1通路在SLE的免疫病理中发挥重要作用。     

癫痫患儿外周血T淋巴细胞亚群及催乳素变化

采用APAAP法和放射免疫分析法检测癫痫患儿外用血T淋巴细胞亚群及血清催乳素（PRL）水平的变化。结果癫痫患儿CD4细胞及CD4／CD8比值明显低于对照组（P＜0．01）；CD8细胞和PRL水平显著高于对照组（P＜0．01），且上述指标的改变与癫痫的发作频率有明显的关系；PRL与CD4／CD8比值之间呈显著负相关，提示癫痫患儿体内神经内分泌免疫调节网络功能的紊乱在癫痫的发病中起着重要的作用。     

Cytotoxic Function of Umbilical Cord Blood Natural Killer Cells: Relevance to Adoptive Immunotherapy

Decreased graft-versus-host disease (GVHD), ease of accessibility, and sustained engraftment encourage the use of umbilical cord blood (UCB) as an alternative source to bone marrow for immune reconstitution in children with leukemia. Natural killer (NK) cells rapidly expand after stem cell transplantation and are important for regulating GVHD and providing graft-versus-leukemia (GVL) effects. This review highlights the phenotypic and functional differences between UCB NK cells and adult peripheral blood (APB) NK cells, and discusses the possible therapeutic benefit of using UCB NK cells for adoptive immunotherapy in leukemia. Alloreactive NK cells show potent cytotoxic activities against human leukocyte antigen (HLA)-nonidentical leukemic cells and reduce leukemia relapses. The higher numbers of NK progenitors in UCB makes it a convenient source for ex vivo expansion of UCB NK cells for posttransplant treatment. UCB NK cells readily respond to interleukin-15, which may greatly enhance their antitumor effect. Activation and expansion protocols for UCB NK cells are currently being developed.     

Cytotoxic function of umbilical cord blood natural killer cells: relevance to adoptive immunotherapy

Decreased graft-versus-host disease (GVHD), ease of accessibility, and sustained engraftment encourage the use of umbilical cord blood (UCB) as an alternative source to bone marrow for immune reconstitution in children with leukemia. Natural killer (NK) cells rapidly expand after stem cell transplantation and are important for regulating GVHD and providing graft-versus-leukemia (GVL) effects. This review highlights the phenotypic and functional differences between UCB NK cells and adult peripheral blood (APB) NK cells, and discusses the possible therapeutic benefit of using UCB NK cells for adoptive immunotherapy in leukemia. Alloreactive NK cells show potent cytotoxic activities against human leukocyte antigen (HLA)-nonidentical leukemic cells and reduce leukemia relapses. The higher numbers of NK progenitors in UCB makes it a convenient source for ex vivo expansion of UCB NK cells for posttransplant treatment. UCB NK cells readily respond to interleukin-15, which may greatly enhance their antitumor effect. Activation and expansion protocols for UCB NK cells are currently being developed.     

The regulatory function of umbilical cord blood CD4+ CD25+ T cells stimulated with anti-CD3/anti-CD28 and exogenous interleukin (IL)-2 or IL-15

The abundance of CD4⁺ CD25⁺ regulatory T cells in umbilical cord blood (UCB) might contribute to the decreased severity of graft-vs.-host disease (GVHD) for UCB transplantation. This study aims to characterize the phenotypes and suppressive function of UCB CD4⁺ CD25⁺ T cells under the influence of anti-CD3/anti-CD28 (CD3/CD28) and exogenous interleukin (IL)-2 or IL-15. Higher percentages of CD4⁺ CD25^high and FoxP3⁺ cells were detected in UCB compared to their adult counterparts. IL-15 was as effective as IL-2 in enhancing the proliferation of CD3/CD28 stimulated UCB CD4⁺ CD25⁺ T cells. Phenotypically, IL-2/IL-15-stimulated UCB CD4⁺ CD25⁺ T cells expressed higher level of CTLA-4, GITR, membrane bound transforming growth factor-β (mTGF-β), and especially Foxp-3 than controls. IL-2/IL-15-stimulated UCB CD4⁺ CD25⁺ T cells also produced much higher IL-10 and TGF-β than controls; while IL-2/IL-15-stimulated UCB CD4⁺ CD25⁻ T cells showed increased TGF-β, but not IL-10 production. IL-2/IL-15-cultured UCB CD4⁺ CD25⁺ T cells showed comparable suppressor activity on allogeneic adult CD4⁺ T-cell proliferation compared to controls, partly through a contact-dependent fashion. Taken together, IL-2/IL-15-stimulated UCB CD4⁺ CD25⁺ T cells show distinct regulatory T-cell phenotypic and functional features, and may be applied for the alleviation of GVHD severity following UCB transplantation.     

CD44分子在血细胞的表达及对T淋巴细胞增殖的影响

为了进一步探讨粘附分子CD44在血细胞的分布特点及与免疫功能的关系，利用荧光双染色一流式细胞仪技术检测各种血细胞的CD44分子的表达，利用^3H-TdR掺入法检测T细胞的DNA合成。结果显示：除血小板CD44分子的表达为阴性、胸腺细胞为弱阳性外，其他所有血细胞CD44分子的表达(包括红细胞、中性粒细胞、单核细胞、干细胞、T淋巴细胞、B淋巴细胞和NK细胞)均为阳性。单独抗CD3抗体可明显增强T细胞的DNA合成；但协同抗CD44抗体后，T细胞的DNA合成被明显抑制。单独抗CD44抗体对IL-2激活的T细胞DNA合成无明显影响；但协同抗CD2抗体后，可明显增强抗CD2抗体对IL-2激活的T细胞DNA合成的抑制。     

急性白血病患儿CD4~+CD25~+调节性T细胞和NK细胞免疫作用探讨

目的观察外周血CD4+CD25+调节性T细胞(CD4+CD25+Treg)及自然杀伤细胞(na-ture killer cell,NK)在白血病患儿及非白血病患儿的不同,了解白血病患儿的免疫状态,探讨CD4+CD25+Treg细胞及NK细胞在小儿急性白血病肿瘤免疫中的意义。方法以流式细胞术检测急性白血病初诊患儿及非白血病患儿各30例的外周血CD4+CD25+Treg细胞、NK细胞的数量及比例。结果外周血CD4+CD25+CD127-细胞占CD4+T细胞的比例白血病组为(11.45±1.41)%,显著高于对照组为(6.98±1.09)%(P<0.05)。而NK细胞数量白血病组为(5.13±2.97)%,显著低于对照组为(15.06±3.91)%(P<0.05)。结论 (1)急性白血病患儿外周血CD4+CD25+Treg细胞数量升高,NK细胞数量降低,表明急性白血病患儿NK细胞免疫功能处于抑制状态。CD4+CD25+Treg细胞可能在白血病的发生、发展中起一定作用。(2)通过检测CD4+CD25+CD127-T细胞可较好的反映CD4+CD25+Treg细胞的比例,简便可行、重复性好、检测结果准确、可靠。     

Lymphocyte subsets and cytokines in adenoviral infection in children

To determine the distribution of major blood lymphocyte subsets we evaluated blood lymphocytes by flow cytometry in adenovirus-infected infants aged 30–730 d. In addition, interleukin-1-receptor antagonist, interleukin-10 and transforming growth factor-β1 were measured in serum by enzyme-linked immunosorbent assay. According to clinical parameters, mechanical ventilation and outcome, infections were classified as moderate ( n = 15), severe ( n = 11) and fatal ( n = 12). Controls were 13 healthy children. In severe and fatal infection, T cells (CD5⁺/CD19^-), NK effectors (CD16⁺), CD4⁺ T subset and B1 subset of B lymphocytes (CD5⁺/CD19⁺) were all significantly decreased. CD8⁺ cells were decreased in severe but not fatal cases. There was no difference in serum values of interleukin-10; however, fatal cases had high interleukin 1-receptor antagonist values. Interestingly, patients with moderate infection showed significantly increased values of transforming growth factor-β1. These results demonstrate that life-threatening adenoviral infection is associated with marked abnormalities in blood lymphocyte and cytokine profile.     

Contribution of B7RP‐1/ICOS co‐stimulation to lethal acute GVHD

Fujimura J, Takeda K, Kaduka Y, Saito M, Akiba H, Yagita H, Yamashiro Y, Shimizu T, Okumura K. Contribution of B7RP‐1/ICOS co‐stimulation to lethal acute GVHD.
Pediatr Transplantation 2010: 14:540–548. © 2010 John Wiley & Sons A/S. Abstract: Co‐stimulatory molecules expressed on T cells critically regulate donor T‐cell activation and are implicated in acute GVHD after allogeneic BMT. We here investigated the role of interaction between B7‐related protein‐1 (B7RP‐1) and ICOS in murine acute GVHD model that received T cell‐depleted BM cells and splenocytes. Administration of blocking anti‐B7RP‐1 mAb significantly reduced the lethality and symptoms in acute GVHD. A significant hypo‐responsiveness of splenocytes to host alloantigen was observed in the recipient mice treated with anti‐B7RP‐1 mAb. Moreover, acute GVHD was significantly reduced in the recipients of T cells composed of ICOS‐deficient CD8 T cells and WT CD4 T cells compared with that in the recipients of T cells composed of WT CD8 T cells and ICOS‐deficient CD4 T cells. These results suggested that B7RP‐1/ICOS co‐stimulatory signal plays a role in the activation of alloantigen‐reactive donor T cells, particularly in CD8 T cells, in murine acute GVHD model, and that the blockade of B7RP‐1/ICOS interaction may be useful for selectively manipulating allo‐reactive T cells in the recipients with acute GVHD.     

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目的评价免疫调节T细胞和细胞因子在再生障碍性贫血(再障)细胞免疫功能紊乱中的作用。方法2004-02—2005-06中山大学第二附属医院采用流式细胞术检测27例特发性再障患儿骨髓及外周血淋巴细胞亚群和CD 4CD2 5T细胞水平,ELISA检测骨髓转化生长因子(TGFβ-1)和F lt-3L水平,并与正常儿童对照。结果与对照组比较,初诊再障患儿外周血和骨髓CD 8T细胞均显著增高(P<0.05),重型再障(SAA)组伴外周血CD3-CD 56NK细胞及骨髓B细胞显著下降(P<0.05)。初诊SAA组骨髓CD 4CD 25T细胞[(7.5±3.4)%]显著高于对照组[(4.3±0.9)%,P<0.05],初诊SAA组及轻型再障(MAA)组骨髓CD 4CD2 5/CD 4比值分别为(28.9±11.1)%和(28.2±9.4)%,均显著高于对照组[(17.4±0.9)%,P均<0.05],骨髓TGFβ-1分别为(2.2±1.7)μg/L和(2.0±0.6)μg/L,均较对照组[(4.4±0.9)μg/L]显著降低(分别为P<0.01、P<0.05),而F lt-3L水平分别为(1031.1±321.8)ng/L和(694.7±424.7)ng/L,均较对照组[(63.0±37.5)ng/L]显著增高(P均<0.01)。缓解期SAA儿童除外周血CD8 T细胞仍较对照组显著增高外,其余上述指标均接近正常水平。相关分析显示,骨髓CD4 CD 25T细胞与CD 3CD 4T细胞呈显著正相关(r分别为0.495、0.540,P<0.01);F lt-3L与骨髓CD 3、CD 4、CD 8T细胞及CD 4CD 25T细胞均呈显著正相关(r分别为0.732、0.542、0.688、0.405,P分别<0.01、0.01、0.01、0.05),而TGFβ-1与骨髓CD 8T细胞和F lt-3L水平呈显著负相关(r分别为-0.431、-0.482,P分别<0.05、<0.01)。结论儿童再障发病与CD 4CD 25T细胞数量缺乏无关,骨髓TGFβ-1水平显著降低和F lt-3L水平显著增高可能在再障儿童T淋巴细胞数量增多和功能紊乱中起重要作用。     

胆道闭锁术后胆管炎与肝胆道组织中CD4/CD8+T细胞的相关性研究

目的 了解胆道闭锁肝脏组织和残存胆道组织中T细胞的主要类型及其与Kasai术后胆管炎的关系.方法 应用免疫组织化学的方法对22例胆道闭锁患儿(早期胆管炎和非胆管炎各11例进行配对)肝组织标本和肝门纤维块标本与13例其他肝胆道疾病患儿肝组织标本进行对比研究.同时结合胆管炎的发生与否分析免疫组化结果与Kasai术后胆管炎相关性.结果 胆道闭锁患儿肝脏组织和肝门纤维块内有大量CD4+T细胞和CD8+T细胞浸润(与对照组相比均P=0.000).胆道闭锁组与对照组中CD57+的NK细胞均为阴性.颗粒酶B在胆道闭锁组与对照胆道闭锁组中均为阴性.早期胆管炎组CD8+T细胞数量明显低于非胆管炎组(P=0.002).CD8+T评分与早期胆管炎的发生呈负相关,相关系数R=-0.674(P=0.001).结论 在Kasai手术之前患儿的肝脏组织和残存的胆道组织中有大量CD4+T和CD8+T细胞浸润,而颗粒酶B的杀伤途径不参与胆道闭锁的免疫损伤.CD8+T细胞浸润程度与胆管炎的发生有明显负相关,其对胆道闭锁肝内的损伤性炎症反应可能具有保护性免疫调节作用.