Local administration of granulocyte macrophage colony-stimulating factor induces local accumulation of dendritic cells and antigen-specific CD8+ T cells and enhances dendritic cell cross-presentation

Immunotherapy has emerged as a promising treatment strategy for the control of HPV-associated malignancies. Various therapeutic HPV vaccines have elicited potent antigen-specific CD8+ T cell mediated antitumor immune responses in preclinical models and are currently being tested in several clinical trials. Recent evidence indicates the importance of local immune activation, and higher number of immune cells in the site of lesion correlates with positive prognosis. Granulocyte macrophage colony-stimulating factor (GMCSF) has been reported to posses the ability to induce migration of antigen presentation cells and CD8+ T cells. Therefore, in the current study, we employ a combination of systemic therapeutic HPV DNA vaccination with local GMCSF application in the TC-1 tumor model. We show that intramuscular vaccination with CRT/E7 DNA followed by GMCSF intravaginal administration effectively controls cervicovaginal TC-1 tumors in mice. Furthermore, we observe an increase in the accumulation of E7-specific CD8+ T cells and dendritic cells in vaginal tumors following the combination treatment. In addition, we show that GMCSF induces activation and maturation in dendritic cells and promotes antigen cross-presentation. Our results support the clinical translation of the combination treatment of systemic therapeutic vaccination followed by local GMCSF administration as an effective strategy for tumor treatment.     

Presentation of the HCV-derived lipopeptide LP20-44 by dendritic cells enhances function of in vitro-generated CD4+ T cells via up-regulation of TLR2

In vitro immunization with HCV lipopeptide aa 20-44 results in antigen-specific T cells. Here, we studied whether presentation of the lipopeptide by dendritic cells (DC) results in enhanced T cell functions. DC-immunized T cells showed significantly augmented proliferation and IFN-gamma secretion upon HCV-specific re-stimulation. This effect was related to significantly increased expression of TLR2 on DC-immunized CD4+ T cells and required TLR2 triggering during re-stimulation. In contrast, numbers of HLA-A2-pentamer-positive CD8+ T cells and granzyme B-secreting T cells remained unchanged. Thus, up-regulation of TLR2 during immunization with DC enhances functions of CD4+ T cells but not CD8+ T cells.     

PDCA循环管理法在妇幼专科医院抗菌药物管理中的应用

目的 探讨PDCA循环管理法在妇幼专科医院抗菌药物专项管理中的应用与成效。方法 运用PDCA循环法，通过多部门联合对某妇幼保健院抗菌药物临床应用的各项指标进行监管，比较应用PDCA方法前后（2014年8月—2015年7月为专项管理前，2017年8月—2018年7月为专项管理后）的效果。结果 使用抗菌药物金额比例、门急诊使用抗菌药物金额比例、住院患者使用抗菌药物金额比例和Ⅰ类切口手术预防抗菌药物使用率分别由专项管理前的16.69%、13.18%、58.63%、45.12%，下降至整治后的7.69%、6.75%、38.21%、8.13%，下降趋势均有统计学意义（均P<0.05），均符合国家要求；其他各项指标如抗菌药物使用强度、接受抗菌药物治疗患者微生物检验标本送检率等指标均达到国家要求。结论 运用PDCA循环管理法开展抗菌药物专项管理，成效明显，抗菌药物临床应用更规范、合理。     

CD4+CD25+调节性T细胞与HIV感染关系的研究进展

CD4 CD25 调节性T细胞是一类具有特殊免疫调节功能的T细胞亚群。它能够抑制自身免疫病的发生,参与肿瘤免疫的调节,在感染和移植免疫中也发挥着重要作用。T细胞的这一亚群具有免疫无能和免疫抑制特性,新近发现它亦与艾滋病的发生、发展关系密切。文中就CD4 CD25 调节性T细胞与HIV感染的研究进展进行综述。     

Peptide induces CD4(+)CD25+ and IL-10+ T cells and protection in airway allergy models

The purpose of this study was to evaluate whether a single peptide containing a major T cell epitope might induce peripheral tolerance in a complex allergen model. C57BL/6 mice were sensitized by intraperitoneal injection of house dust mite extract (HDM), and exposed to antigen via trachea instillation. Der p 1 peptide was administered by i.v. before or after sensitization. Lung lavage fluids were analyzed for cellular infiltration. Respiratory exposure of sensitized mice to antigen results in airway inflammation and eosinophilia. Intravenous administration of a single peptide protected sensitized mice from these changes. Further, the emergence of antigen-specific CD25(+)CD4+ and IL-10 secreting cell populations in DO11.10 mice was demonstrated after peptide administration. Thus, intravenous delivery of a single peptide epitope is capable of inducing peripheral tolerance and protection in a complex allergy model, possibly through regulatory T cells and bystander suppression.     

川崎病患儿外周血 CD4^＋CD25^＋调节性 T细胞与 Th17细胞的变化

目的 探讨川崎病（ KD）患儿治疗前后外周血中CD4^＋CD25^＋调节性T细胞（ Treg）与Th17细胞比例的变化,以指导临床治疗及评估预后。方法 选择KD患儿32例（ KD组）及健康幼儿28例（对照组）;抽取对照组幼儿和KD组患儿使用免疫球蛋白＋阿司匹林治疗前及治疗后热退3天时清晨空腹静脉血,采用ELISA双抗体夹心法检测血清中白细胞介素－15（ IL-15）、白细胞介素－17（IL-17）和白细胞介素－23（IL-23）水平;采用流式细胞术检测外周血中Treg细胞和Th17细胞比例。结果 与对照组相比,KD组患儿治疗前血清中IL-15、IL-17和IL-23水平均显著增高（t值分别为－4．35、－3．98和－4．11,P＜0．05）;经治疗后,血清中IL-15、IL-17和IL-23水平均有显著降低（t值分别为3．74、2．94和3．06,P＜0．05）。 KD组患儿外周血中Treg细胞比例明显降低（t＝2．54,P＜0．05）,经治疗后可基本回升至对照组水平,与治疗前相比差异有显著性（t＝－2．13,P＜0．05）;Th17细胞比例明显升高（t＝－1．87,P＜0．05）,治疗后与治疗前相比有明显降低（t＝2．71,P＜0．05）。结论 Treg细胞与Th17细胞之间的失衡可能在川崎病的发病中起重要作用,其比例的变化对指导临床用药及评估病情和预后具有一定的意义。     

CD4+CD25+ regulatory T cell-mediated changes in the expression of endocytic receptors and endocytosis process of human dendritic cells

CD4⁺CD25⁺ regulatory T cells (Tregs) are known to inhibit immune responses to antigens. Since, the process of antigen uptake by dendritic cells (DC) is central to induction of immune responses, we analyzed the effect of Tregs on the expression of endocytic receptors on DC and its repercussion on antigen uptake. Our results demonstrate that Tregs down-regulate the expression and uptake of antigens via C-type lectin-like receptors CD206 and DC-SIGN, restrain the pinocytosis process of DC and augment the expression of FcγRIIB, an inhibitory Fcγ receptor the engagement of which by IgG-bound antigens leads to inhibition of DC activation. Our results thus provide an additional insight on the pertinence of strategies aimed at blocking Treg functions towards improved vaccination protocols.     

HBV宫内感染新生儿外周血调节性T细胞表达

目的 探讨CD4⁺CD25⁺调节性T细胞(Treg)在乙肝病毒(HBV)宫内感染新生儿外周血中的表达和意义。方法 选择乙肝表面抗原(HBsAg)阳性孕妇及其分娩新生儿79例,用酶联免疫吸附试验检测孕妇和新生儿外周血HBV标志物,实时荧光定量PCR 检测母亲和新生儿外周血 HBV DNA 含量,用流式细胞仪检测新生儿外周血CD4⁺CD25⁺ Treg和CD4⁺CD25⁺Foxp3⁺ Treg水平。结果 宫内感染HBV新生儿外周血CD4⁺CD25⁺ Treg和CD4⁺CD25⁺Foxp3⁺ Treg占CD4⁺T细胞比例分别为(10.57±2.25)%和(1.67±0.37)%,均高于未感染新生儿,差异有统计学意义(P<0.05),随产妇HBV DNA载量增加,新生儿外周血CD4⁺CD25⁺ Treg和CD4⁺CD25⁺Foxp3⁺ Treg增加,CD4⁺CD25⁺ Treg比例高于阴性组(P<0.05);CD4⁺CD25⁺ Treg和CD4⁺CD25⁺Foxp3⁺ Treg均与产妇HBV DNA载量呈正相关(r=0.430、0.409,P<0.05)。结论 Treg可能通过抑制机体细胞免疫反应影响乙肝病毒清除,新生儿HBV宫内感染可能与Treg表达上调有关。     

化疗对乳腺癌患者外周血CD4~+CD25~+CD127~-调节性T细胞的影响

目的观察化疗对乳腺癌患者外周血免疫指标的变化并探讨其影响。方法采用流式细胞术分析129例乳腺癌患者化疗前后CD4+CD25+CD127-调节性T细胞占CD4+T细胞的比率及T淋巴细胞亚群CD3、CD4、CD8;B淋巴细胞CD19;NK细胞(CD16+CD56)和CD4/CD8比值的动态变化。结果化疗后的乳腺癌患者CD4+CD25+CD127-调节性T细胞占CD4+细胞比率较化疗前降低,T淋巴细胞亚群中CD3、CD4、CD4/CD8增高,CD8、NK和CD19降低;乳腺癌患者外周血调节性T细胞的数量与临床分期和组织分化程度相关。结论乳腺癌患者化疗后调节性T细胞的数量降低,免疫调节抑制功能下降,免疫功能得到提高,但体液免疫及NK功能未见提高。     

Activation of dendritic cells and induction of CD4(+) T cell differentiation by aluminum-containing adjuvants

Aluminum-containing adjuvants are widely used in licensed human and veterinary vaccines. However, the mechanism by which these adjuvants enhance the immune response and predominantly stimulate a T(H)2 humoral immune response is not well understood. In this study, the effects of aluminum hydroxide and aluminum phosphate adjuvants on antigen presentation, expression of costimulatory molecules and cytokines by mouse dendritic cells (DCs) and the ability of DCs to induce T helper cell differentiation were investigated. Dendritic cells pulsed with ovalbumin (OVA) adsorbed to aluminum-containing adjuvants activated antigen-specific T cells more effectively than DCs pulsed with OVA alone. Aluminum hydroxide adjuvant had a significantly stronger effect than aluminum phosphate adjuvant. Both aluminum-containing adjuvants significantly increased the expression of CD86 on DCs but only aluminum hydroxide adjuvant also induced moderate expression of CD80. Aluminum-containing adjuvants stimulated the release of IL-1beta and IL-18 from DCs via caspase-1 activation. DCs incubated with LPS and OVA induced T(H)1 differentiation of na?ve CD4(+) T cells. In contrast, DCs incubated with aluminum/OVA activated CD4(+) T cells to secrete IL-4 and IL-5 as well as IFN-gamma. Addition of neutralizing anti-IL-1beta antibodies decreased IL-5 production and addition of anti-IL-18 antibodies decreased both IL-4 and IL-5 production. Inhibition of IL-1beta and IL-18 secretion by DCs via inhibition of caspase-1 also led to a marked decrease of IL-4 and IL-5 by CD4(+) T cells. These results indicate that aluminum-containing adjuvants activate DCs and influence their ability to direct T(H)1 and T(H)2 responses through the secretion of IL-1beta and IL-18.     

CD4~+ Notch1~+调节性T细胞和CD4~+ FoxP3~+调节性T细胞 在不明原因复发性流产中的表达

目的:探讨CD4+Notch1+调节性T细胞和CD4+FoxP3+调节性T细胞在不明原因复发性流产(URSA)中的表达。方法:流式细胞术分析复发性流产和正常妊娠动物模型中雌性小鼠CBA/J脾细胞CD4+CD25+Treg、CD4+FoxP3+Treg和CD4+Notch1+Treg的表达百分率变化。结果:流式细胞术分析(FACS)结果显示,相比于正常妊娠组,在复发性流产中CD4+CD25+Treg表达百分率减少、CD4+FoxP3+Treg表达百分率减少、CD4+Notch1+Treg表达百分率增加。结论:Notch1增加与FoxP3表达减少相关,CD4+Notch1+Treg的表达百分率增加可能是URSA的机制之一。     

CD4+CD25+T细胞与复发性流产

CD4+CD25+T细胞是一类具有免疫调节功能的细胞.正常情况下在体内发挥免疫抑制作用.该类细胞分泌白细胞介素10(IL-10)、转化生长因子β(TGF-β),表达CD25(IL-2Rα)、叉头蛋白(FOXP3蛋白)及细胞毒性T淋巴细胞抗原4(CTLA-4)分子等,通过细胞间直接接触或细胞因子间接方式广泛参与自身免疫耐受、肿瘤免疫、移植免疫.妊娠类似同种异体移植,妊娠时脱落人母体循环的胎儿抗原刺激CD4+CD25+T细胞的产生并在母胎界面形成免疫耐受微环境,防止胎儿受到母体排斥.CD4+CD25+T细胞数量或功能失调则会导致各种疾病的发生,如自身免疫病,移植物抗宿主病,流产等.在复发性流产患者体内CD4+CD25+T细胞数目及功能明显下降.实验表明增强CD4+CD25+T细胞数量或功能可以阻止移植物抗宿主病和治疗复发性流产.     

子痫前期患者CD4~+CD25~+Foxp~(3+)调节性T细胞检测及意义

目的:研究CD4+CD25+Foxp3+调节性T细胞在子痫前期患者(PE)外周血中的比例变化,探讨其在子痫前期发病中意义。方法:采用细胞内染色的流式细胞术检测22例子痫前期患者外周血中CD4+CD25high和CD4+CD25+Foxp3+的表达,并与正常妊娠和正常育龄妇女进行比较。结果:子痫前期患者外周血单个核细胞(PBMCS)CD4+T细胞中CD25high细胞明显低于正常妊娠和正常育龄妇女,CD4+CD25+Foxp3+的表达显著降低,而且其CD4+CD25+Foxp3+的表达与平均动脉压呈负相关。结论:子痫前期患者CD4+CD25+Foxp3+的表达明显减少,表明调节性T细胞的降低可能打破了母胎之间的耐受,参与了子痫前期的病理进程。     

CD4~+ CD25~+调节性T细胞在上皮性卵巢癌患者外周血中的表达及临床意义

目的:研究上皮性卵巢癌患者外周血中CD4+ CD25+调节性T细胞数量及其相关基因Foxp3 mRNA的表达特点,探讨CD4+CD25+调节性T细胞在诱导肿瘤免疫耐受中的作用。方法:采用流式细胞术检测卵巢浆液性囊腺癌和粘液性囊腺癌患者外周血中CD4+ CD25+调节性T细胞数量变化;采用半定量RT-PCR方法检测卵巢浆液性囊腺癌和粘液性囊腺癌患者外周血PBMC中Foxp3 mRNA表达水平。结果:卵巢浆液性囊腺癌和粘液性囊腺癌患者外周血中CD4+CD25+调节性T细胞数量均明显升高(P<0.05);Foxp3 mRNA在卵巢浆液性囊腺癌和粘液性囊腺癌患者外周血PBMC中表达明显增高(P<0.05)。结论:CD4+ CD25+调节性T细胞可能在卵巢癌免疫耐受中发挥重要作用。     

外周血调节性CD_4~+ CD_(25)+ T细胞比例在体外受精与胚胎移植中的意义

目的:研究外周血调节性CD4+CD25+T(CD4+CD25+Tr)细胞的比例变化,探讨CD4+CD25+Tr细胞在妊娠免疫母胎耐受中的作用。方法:应用流式细胞术检测试管婴儿成功孕妇外周血中CD4+CD25+Tr细胞的比例变化。结果:试管婴儿成功孕妇外周血中CD4+CD25+Tr细胞比例较高,高于同孕期的正常妊娠组,差异有统计学意义(P<0.05)。结论:CD4+CD25+Tr细胞在体外受精与胚胎移植的发生发展中发挥一定作用。该研究将有助于阐明妊娠的本质,为探讨妊娠相关免疫异常疾病的发生机制及提高试管婴儿的成功率提供理论依据。     

稽留流产患者外周血CD~4+ CD25~+ Foxp3~+调节性T细胞的表达及意义

目的:探讨CD4+ CD25+ Foxp3+调节性T细胞在稽留流产患者外周血中的比例变化。方法:选取16例稽留流产患者,采用细胞内染色的流式细胞术及定量PCR方法分别在蛋白质和mRNA水平检测其外周血中CD4+ CD25+ Foxp3+的表达,并与正常妊娠和正常育龄妇女进行比较。结果:稽留流产患者外周血单个核细胞(PBMCS)CD4+ T细胞中CD25high细胞明显低于正常妊娠和正常育龄妇女,CD4+ CD25+ Foxp3+的表达显著降低。结论:稽留流产患者CD4+ CD25+ Foxp3+的表达明显减少,提示稽留流产的发生可能与患者外周血中CD4+ CD25+ Foxp3+下降有关;而在妊娠早期CD4+ CD25+ Foxp3+的表达明显升高,表明其可能是调控母-胎间免疫耐受形成的重要因素,参与了正常妊娠的维持。     

非酒精性脂肪性肝病患者外周血CD4~+CD25~+Foxp3~+调节性T细胞的变化及意义

目的观察非酒精性脂肪性肝病患者外周血单个核细胞(PBMC)CD4+CD25+Foxp3+调节性T细胞的变化及与胰岛素抵抗的关系,初步探讨其意义。方法 46例非酒精性脂肪性肝病患者根据体质指数分为肥胖组(21例)及非肥胖组(25例),42名健康医务工作者作为健康对照。检测空腹胰岛素及空腹血糖水平,计算胰岛素抵抗指数;采用流式细胞术检测患者外周血PBMC CD4+CD25+Foxp3+调节性T细胞数目,并计算CD4+CD25+Foxp3+调节性T细胞占CD4+T淋巴细胞的百分比;分析CD4+CD25+Foxp3+调节性T细胞与胰岛素抵抗间的关系。结果与健康对照组相比,非酒精性脂肪性肝病患者外周血PBMC CD4+CD25+Foxp3+调节性T细胞数及其占CD4+T淋巴细胞的百分比均显著下降(P<0.01,P<0.05),但在肥胖组与非肥胖组间差异均无统计学意义(P>0.05);肥胖者胰岛素抵抗指数较非肥胖者显著升高(P<0.01);肥胖组内PBMC CD4+CD25+Foxp3+调节性T细胞数值及其占CD4+T淋巴细胞的百分比均与胰岛素抵抗指数呈明显负相关(r=-0.989,P<0.01)。结论非酒精性脂肪性肝病患者PBMC CD4+CD25+Foxp3+调节性T细胞数目和比例减少,可能存在自身免疫调节功能的紊乱;PBMC CD4+CD25+Foxp3+调节性T细胞可能通过调节胰岛素抵抗参与非酒精性脂肪性肝病的形成和发展。     

CD4+T辅助细胞与常见自身免疫性疾病

CD4辅助性T细胞(T help cell/ Th)作为T细胞中的一个重要群体因分泌的细胞因子不同而被分为多个亚群,各个Th细胞亚群在自身免疫疾病发病中的作用在近十年中得到了广泛的研究及肯定.本文就Th细胞各亚群与自身免疫性疾病关系做一概述.     

CD4+T细胞、CD8+T细胞、CD57+NK细胞和Vimentin在外阴硬化性苔藓中表达及临床意义

目的:探讨外阴硬化性苔藓中CD4+、CD8+T细胞及CD57+ NK细胞和Vimentin的表达.方法:采用免疫组化S-P法分别检测CD4+T细胞、CD8+T细胞、CD57+NK细胞和VIM阳性细胞在外阴正常皮肤(15例)、外阴硬化性苔藓早期(15例)及进展期(15例)病变中的浸润及表达.结果:CD4+T细胞在外阴硬化性苔藓早期中浸润明显高于正常皮肤,CD8+T细胞在外阴硬化性苔藓早期浅层及进展期深层病变组织中浸润均高于正常皮肤,CD57+ NK细胞在外阴硬化性苔藓早期中浸润明显高于正常皮肤,Vimentin标记的成纤维细胞在外阴硬化性苔藓早期浅层及进展期深层病变中表达明显高于正常皮肤,差异均有统计学意义(P＜0.05).结论:外阴硬化性苔藓中淋巴细胞的浸润与成纤维细胞增殖有一定相关性.