肠道厌氧菌失调加重流感病毒感染小鼠肺部免疫病理损伤

目的探讨肠道厌氧菌失调与流感病毒感染之间的关系,以及对小鼠肺部免疫细胞中炎性因子的影响。方法将36只BALB/c小鼠随机分为正常对照组、病毒对照组、甲硝唑组,每组12只。甲硝唑组小鼠用18 mg/m L甲硝唑灌胃8 d,造成小鼠肠道厌氧菌菌群失调;再予每只小鼠50μL/d FM1株流感病毒连续滴鼻4 d,建立厌氧菌失调小鼠感染流感病毒的动物模型。观察各组小鼠精神状态、肺指数、肺组织病理形态变化、盲肠及肠黏膜病理形态变化,ELISA测定小鼠肺组织匀浆中白细胞介素4(IL-4)、γ干扰素(IFN-γ)、IL-10和IL-17含量的变化。结果甲硝唑能使该组小鼠肺指数明显升高,其肺组织及盲肠组织病理改变较病毒对照组小鼠而言,表现出更为严重的炎症损伤。FM1甲型流感病毒感染后,甲硝唑组小鼠肺组织匀浆中IFN-γ、IL-17水平与病毒对照组相比显著降低,IL-4、IL-10含量虽有升高,但差异无统计学意义。结论肠道厌氧菌失调可能通过调节小鼠肺部炎性因子的分泌,抑制其适应性免疫应答,影响FM1流感病毒在体内复制及清除的时间,从而进一步加重流感病毒引起的免疫病理损伤。     

肺炎支原体P1C-IL-2融合基因疫苗鼻饲对小鼠免疫效果的检测

目的 研究肺炎支原体(Mp) P1C-IL-2融合基因疫苗经鼻饲免疫小鼠后的免疫应答水平和免疫保护作用,了解IL-2对P1C核酸疫苗的免疫佐剂效应.方法 将构建的P1C-IL-2核酸疫苗鼻饲免疫BALB/c鼠,ELISA检测免疫小鼠血清IgG滴度、IgG亚类和支气管肺泡灌洗液中IgA及IFN-γ、IL-4的水平;建立小鼠Mp感染模型,观察Mp攻击后小鼠肺组织炎症情况和支气管肺泡灌洗液中Mp菌落数的变化.结果 P1C-IL-2双基因疫苗组小鼠血清中的总IgG、IgG1、IgG2a亚类和支气管肺泡灌洗液中IFN-γ和IL-4水平均较PIC疫苗组小鼠显著增高(P＜0.05),但两组支气管肺泡灌洗液IgA差异无显著性(P＞0.05).用Mp滴鼻感染免疫小鼠,第1、3、6天P1C-IL-2双基因融合疫苗组小鼠肺组织炎症病理评分显著高于P1C单基因疫苗免疫组小鼠,两组小鼠支气管肺泡灌洗液中的Mp菌落数差异无显著性(P＞0.05).结论 IL-2能显著增强PIC疫苗的免疫应答水平,但在感染早期也激发了较强的肺组织炎症.     

IL-2增强肺炎支原体P1C核酸疫苗的肌注免疫效果

目的:研究IL-2对肺炎支原体(Mycoplasma pneumoniae,Mp)P1C核酸疫苗经肌注免疫BALB/c小鼠后的免疫应答水平和免疫保护作用.方法:将P1C-IL-2核酸疫苗肌注免疫BALB/c鼠,ELISA检测疫苗免疫后56天小鼠血清IgG和IgG亚类、支气管灌洗液中SIgA、IFN-γ和IL-4的水平;用2×107 Mp菌落形成单位鼻饲感染BALB/c鼠,建立感染小鼠模型,病理切片检测Mp感染后小鼠肺部炎症病理改变;将系列10倍稀释的支气管灌洗液接种于SP4固体平板,并进行菌落计数.结果:P1C-IL-2核酸疫苗免疫组小鼠血清中的总IgG、IgG1、IgG2a、IFN-γ和IL-4水平均较P1C单基因疫苗组显著增高(P＜0.05),但两组支气管灌洗液中SIgA差异无显著性(P＞0.05).Mp感染后第1、3、6天P1C-IL-2双基因疫苗组小鼠肺组织病理评分(HPS)较P1C单基因疫苗免疫组显著增高,但支气管灌洗液中的Mp菌落数明显减少;第9天后两组HPS和Mp菌落数差异无显著性.结论:IL-2能显著增强P1C疫苗肌注的免疫保护作用和免疫应答水平,但同时在Mp感染早期激发了较重的肺组织炎症.     

NKT细胞在MRSA感染性肺炎中作用的初步研究北大核心CSCD

目的:初步探讨NKT细胞在耐甲氧西林金葡菌(MRSA)感染性肺炎模型中的作用。方法:MRSA经鼻黏膜感染C57BL/6野生型(WT)小鼠和CD1d^(-/-)小鼠。观察小鼠生存率,检测肺组织中细菌载量,ELISA检测肺组织中细胞因子TNF-α、IFN-γ、IL-17、IL-6含量,流式细胞术检测肺组织中CD3^(+)CD8^(+)T细胞、NK细胞和NKT细胞百分比。结果:MRSA致死剂量感染5 d,WT小鼠生存率显著低于CD1d^(-/-)小鼠(46.2%vs 90.0%,P<0.05)。感染12~48 h,WT小鼠肺组织中细菌载量显著高于CD1d^(-/-)小鼠(P<0.01);WT小鼠肺组织中TNF-α、IFN-γ、IL-6和IL-17含量较CD1d^(-/-)小鼠明显增高(P<0.01)。MRSA感染24 h,WT小鼠肺组织中NKT细胞、NK细胞百分比显著高于CD1d^(-/-)小鼠(P<0.01),而CD3^(+)CD8^(+)T细胞百分比低于CD1d^(-/-)小鼠(P<0.01)。结论:在MRSA肺炎中,NKT细胞被激活,从而加重肺部炎症反应,降低生存率,减缓细菌清除。     

肺炎支原体感染在BALB/c小鼠肺组织不同时间炎症的变化

目的了解肺炎支原体感染的不同时期肺组织的病理变化、用PCR的方法检测肺炎支原体出现时间、阳性率的变化情况。方法建立肺炎支原体经呼吸道感染的BALB/c小鼠模型,对感染后3、7、14、21 d的小鼠的肺组织进行病理变化、肺湿重、PCR检测肺炎支原体出现时间的研究。结果肺炎支原体经呼吸道感染的BALB/c小鼠肺组织变大重量增加,肺指数同正常对照组相比,P﹤0.01,差异有统计学意义;呼吸道感染的BALB/c小鼠肺组织在3、7 d时,肺组织的病理变化最明显,14 d炎症减轻,21 d炎症基本消退;用PCR的方法检测支气管肺泡灌洗液肺炎支原体7 d时全部为阳性。结论肺炎肺炎支原体感染BALB/c小鼠模型制作成功,感染后7 d时肺部炎症最明显。     

IFN-γ在沙眼衣原体呼吸道感染中免疫防御机制的探讨

目的:检测与IFN-γ作用相关的酶吲哚胺2,3二氧化酶(IDO)、诱导性一氧化氮合成酶(iNOS)和NADPH氧化酶(ox)gp91在沙眼衣原体呼吸道感染中的表达及与机体防御的关系,探讨衣原体感染中IFN-γ免疫防御作用的机制.方法:用沙眼衣原体小鼠肺炎株(MoPn)通过鼻腔感染C57BL/6(H-2b)小鼠,用过氧化物酶连接的鼠抗衣原体脂多糖单抗染色HeLa 229细胞,检测衣原体在肺组织的生长;用RT-PCR检测衣原体感染后第7及14天小鼠肺组织IFN-γ、IDO、iNOS和gp91NADPH ox mRNA表达.结果:MoPn呼吸道感染后小鼠肺组织匀浆衣原体活性测定,于感染后第2天,HeLa 229细胞内可见有衣原体包涵体生长,IFU值增高,于感染后第7天IFU达最高水平,以后逐渐下降,至感染后21天基本恢复到基线水平.与未感染的对照组比较,Th1细胞因子IFN-γ于感染后第7天表达显著增高,感染后14天有所降低,但仍维持较高水平;同时衣原体感染可显著诱导与IFN-γ作用相关的三种酶IDO、iNOS和gp91 NADPH ox在小鼠肺组织的表达,感染后第7及14天,IDO,iNOS及gp91 NADPH ox的表达与对照组比较均有显著差异,其中IDO和gp91 NADPH ox于感染后第7天mRNA表达增高显著(P＜0.01),14天略有下降(P＜0.05).结论:衣原体呼吸道感染诱导Th1细胞因子IFN-γ mRNA高表达,参与宿主对衣原体的清除及机体免疫防御,此作用可能与其相应的酶IDO、iNOS和gp91 NADPH ox表达增高有关.     

IFN-γ上调Th17/IL-17应答介导衣原体呼吸道感染免疫保护

目的 探讨IFN-γ在小鼠沙眼衣原体感染中对Th17/IL-17应答的调节作用.方法 利用沙眼衣原体鼠肺炎株小鼠呼吸道感染模型,用抗鼠IFN-γ单克隆抗体吸入中和肺组织IFN-γ,对照组给予同等剂量的独特型抗体IgG2a,于感染后7 d处死小鼠.免疫酶法检测小鼠肺组织衣原体生长;利用RT-PCR技术检测衣原体感染小鼠肺组织中Th17相关因子IL-17及其上游因子IL-23 mRNA的表达;细胞内细胞因子染色检测衣原体感染小鼠脾脏IL-17-CD4+T细胞的扩增.结果 与对照组相比,IFN-γ抗体中和小鼠有严重的疾病状态,包括明显的体重下降、肺组织更高的衣原体负荷和肺组织更严重的病理损伤;肺组织IL-17和IL-23 mRNA的表达水平显著降低;脾脏IL-17-CD4+T细胞百分率也显著降低.结论 小鼠衣原体感染中,IFN-γ通过上调Th17/IL-17应答起保护作用.

Abstract：

Objective To investigate the regulation of IFN-γ to Th17 response in Chlamydia muridarum (Cm) lung infection in mice. Methods A murine model of pneumonia induced by intranasal inoculation of Cm was used for this study. Anti-mouse IFN-γ McAbs were used to neutralize endogenous IFN-γfollowing Cm lung infection. Control group received the same dose of isotype antibody (IgG2a). Mice were sacrificed at day 7 postinfection. Chlamydial growth in the lung was assessed by immunoenzyme technique.IL-17 and IL-23 mRNA expression in the lung was assayed by RT-PCR and the proliferation of IL-17 + CD4 +T cells in the spleen was assayed by intracellular cytokine staining. Results IFN-γ-neutralized mice exhibited serious disease course, include greater body weight loss, higher organism growth and much more severe pathological changes in the lung compared with control mice. The mRNA expression of IL-17 and IL-23 in the lung and the proliferation of IL-17 + CD4 + T cells in the spleen significantly decreased in the IL-17- neutralized mice. Conclusion IFN-γ was protective in Cm lung infection through up-regulating the antigen specific Th17 responses.     

心房钠尿肽下调呼吸道合胞病毒毛细支气管炎小鼠肺组织T-bet水平

目的研究T-bet在呼吸道合胞病毒毛细支气管炎小鼠肺组织中的表达及心房钠尿肽刺激下T-bet的变化。方法建立毛细支气管炎小鼠模型,观察心房钠尿肽的变化;同时利用肺组织病理学、ELISA法和实时定量PCR法检测心房钠尿肽干预后气道炎症浸润、T-bet mRNA及蛋白的表达和IFN-γ等的变化。结果与正常对照组相比,呼吸道合胞病毒组小鼠肺组织中心房钠尿肽水平明显升高;而经鼻吸入心房钠尿肽可使气道炎症细胞浸润加剧,T-bet及IFN-γ表达水平均降低;而利用心房钠尿肽拮剂预处理后,心房钠尿肽的上述效应可被明显逆转。结论心房钠尿肽在毛细支气管炎肺组织中存在一定量的表达,同时可下调T-bet水平。     

辛伐他汀对小鼠巨细胞病毒肺炎Toll样受体2表达的影响

目的:观察辛伐他汀对小鼠巨细胞病毒(MCMV)肺炎小鼠肺组织Toll样受体2(Toll-like receptor2,TLR-2)、干扰素γ(IFN-γ)和单核细胞趋化蛋白1(MCP-1)表达的影响,并探讨辛伐他汀干预MCMV肺炎的可能机制。方法:将40只6~8周龄BALB/c小鼠随机分成5组:正常对照(NC)组、MCMV感染组和辛伐他汀干预(SMV1、SMV2和SMV3)组。SMV1、SMV2和SMV3组分别在腹腔注射MCMV前7 d、与MCMV同时和MCMV感染后3 d给予辛伐他汀(50 mg·kg~(-1)·d~(-1),均连续给药7 d)灌胃,NC组及MCMV组分别予以同体积的生理盐水灌胃。HE染色观察小鼠肺组织的病理改变,real-time PCR检测MCMV DNA的变化,免疫组化和Western blot检测肺组织中TLR-2的表达,ELISA检测肺组织中IFN-γ和MCP-1的变化。结果:与NC组相比,MCMV组肺组织病理染色显示肺泡间质水肿,肺泡壁增宽,内可见大量炎性细胞浸润;肺组织中TLR-2的表达增加,MCMV DNA含量升高,肺组织中炎症因子IFN-γ和MCP-1明显增多(P0.05)。辛伐他汀干预后,与MCMV组相比,肺组织病理改变较前减轻,TLR-2的表达下降(P0.05),MCMV DNA含量降低(P0.05),炎性因子IFN-γ和MCP-1明显下降(P0.05),且SMV1组TLR-2的表达量及MCMV DNA含量下降较SMV2和SMV3组更明显(P0.05),而SMV2和SMV3两组相比差异无统计学显著性。结论:辛伐他汀干预通过下调TLR-2信号通路而降低TLR-2的表达,减少IFN-γ和MCP-1的分泌,并抑制MCMV DNA的复制,从而使肺组织的病理损害得以减轻;且提前给予辛伐他汀干预对预防MCMV的感染及减轻炎症反应具有重要作用。     

重组BCG联合抗痨药物对小鼠结核病治疗作用的实验研究

目的：探讨携带编码人天然颗粒溶素（Granulysin，GLS）和小鼠IL-12基因的重组BCG与抗痨药物对结核分枝杆菌感染的协同治疗效果。方法：将结核分枝杆菌H37Rv感染的BALB／c小鼠40只随机分成4组。感染4周后分别用生理盐水、重组卡介苗（BCG）、INH＋PZA和重组BCG联合INH＋PZA治疗，于第一次治疗后3个月，处死小鼠，检测器官荷菌量、脾淋巴细胞IFN-γ和TNF-α的分泌水平、血清IL-12和IFN-7．肺、脾组织中GLS表达，同时观察小鼠肺、脾组织病理改变情况。结果：重组BCG联合INH＋PZA治疗组肺、脾组织荷菌量（10gCFU／g）比重组BCG和INH＋PZA治疗组显著降低；重组BCG和INH＋PZA治疗组肺、脾组织荷菌量比生理盐水组显著降低。重组BCG组经PPD刺激后IFN-γ和TNF-α分泌水平明显高于其它组。重组BCG联合INH＋PZA治疗组和重组BCG组血清IL-12和IFN-γ水平明显高于生理盐水组。重组BCG组用免疫组化检测到小鼠肺及脾组织中GLS的表达。重组BCG和INH＋PZA组病变轻且局限，生理盐水组肺组织病理改变以渗出为主，病变广泛。重组BCG联合INH＋PZA治疗组肺组织病变最轻。结论：GLS／IL-12重组BCG对小鼠结核病有一定免疫治疗作用；重组BCG联合临床常用的抗痨药可增强抗结核病的疗效。     

Antibiotic use during pregnancy is linked to offspring gut microbial dysbiosis,barrier disruption,and altered immunity along the gut–lung axis

Antibiotic use during pregnancy is associated with increased asthma risk in children. Since approximately 25% of women use antibiotics during pregnancy, it is important to identify the pathways involved in this phenomenon. We investigate how mother-to-offspring transfer of antibiotic-induced gut microbial dysbiosis influences immune system development along the gut–lung axis. Using a mouse model of maternal antibiotic exposure during pregnancy, we immunophenotyped offspring in early life and after asthma induction. In early life, prenatal-antibiotic exposed offspring exhibited gut microbial dysbiosis, intestinal inflammation (increased fecal lipocalin-2 and IgA), and dysregulated intestinal ILC3 subtypes. Intestinal barrier dysfunction in the offspring was indicated by a FITC-dextran intestinal permeability assay and circulating lipopolysaccharide. This was accompanied by increased T-helper (Th)17 cell percentages in the offspring's blood and lungs in both early life and after allergy induction. Lung tissue additionally showed increased percentages of RORγt T-regulatory (Treg) cells at both time points. Our investigation of the gut–lung axis identifies early-life gut dysbiosis, intestinal inflammation, and barrier dysfunction as a possible developmental programming event promoting increased expression of RORγt in blood and lung CD4⁺ T cells that may contribute to increased asthma risk.     

Role of novel type I interferon epsilon in viral infection and mucosal immunity

Intranasal infection with vaccinia virus co-expressing interferon epsilon (VV-HIV-IFN-ɛ) was used to evaluate the role of IFN-ɛ in mucosal immunity. VV-HIV- IFN-ɛ infection induced a rapid VV clearance in lung that correlated with (i) an elevated lung VV-specific CD8⁺CD107a⁺IFN-γ⁺ population expressing activation markers CD69/CD103, (ii) enhanced lymphocyte recruitment to lung alveoli with reduced inflammation, and (iii) an heightened functional/cytotoxic CD8⁺CD4⁺ T-cell subset (CD3^hiCCR7^hiCD62L^lo) in lung lymph nodes. These responses were different to that observed with intranasal VV-HA-IFN-α₄ or VV-HA-IFN-β infections. When IFN-ɛ was used in an intranasal/intramuscular heterologous HIV prime-boost immunization, elevated HIV-specific effector, but not memory CD8⁺T cells responses, were observed in spleen, genito-rectal nodes, and Peyer’s patch. Homing marker α4β7 and CCR9 analysis indicated that unlike other type I IFNs, IFN-ɛ could promote migration of antigen-specific CD8⁺T cells to the gut. Our results indicate that IFN-ɛ has a unique role in the mucosae and most likely can be used to control local lung and/or gut infections (i.e., microbicide) such as tuberculosis, HIV-1, or sexually transmitted diseases.     

Immunotherapeutical Potential of Mycobacterium Vaccae on M. Tuberculosis Infection in Mice

Tuberculosis remains the worldwide infectious disease. To identify the therapeutic potential of M. vaccae in treating tuberculosis, M. vaccae was injected into Mycobacterium tuberculosis （M. tuberculosis） infected mice. The optimal dose of M. vaccae （22.5 μg/mouse） treated mice showed lower pathological change index, spleen weight index, lung weight index and vital M. tuberculosis count than those of the untreated group. Treatment with M. vaccae enhanced the percentages of CD3^＋ and CD4^＋ T cells, IFN-γ^＋CD4^＋ T cells, innate immune cells including NK cells, NK1.1^＋ T cells and γδ T cells, and reduced the percentage of IL-4^＋CD4^＋ T cells. Therefore, M. vaccae could protect the mice from M. tuberculosis infection and improved mouse innate and adaptive cell-mediated immunity, suggesting that M. vaccae is a potential immunotherapeutic agent in pulmonary tuberculosis. Cellular ＆ Molecular Immunology.     

Pulmonary Aspergillus fumigatus infection in rats affects gastrointestinal homeostasis

Microbiota inhabiting mucosal tissues is involved in maintenance of their immune homeostasis. Growing body of evidence indicate that dysbiosis in gut influence immune responses at distal sites including lungs. There are also reports concerning gut involvement with pulmonary injury/inflammation in settings of respiratory viral and bacterial infections. The impact of infections with other microorganisms on gut homeostasis is not explored. In this study, the rat model of sublethal pulmonary infection with Aspergillus fumigatus was used to investigate the effect of fungal respiratory infection on gut immune-mediated homeostasis. Signs of intestinal damage, intestinal and gut-draining lymphoid tissue cytokine responses and gut bacterial microbiota diversity were examined. Intestinal injury, inflammatory cell infiltration, as well as increased levels of intestinal interferon-γ (IFN-γ) and interleukin-17 (IL-17) (as opposed to unchanged levels of anti-inflammatory cytokine IL-10) during the two-week period depict intestinal inflammation in rats with pulmonary A. fumigatus infection. It could not be ascribed to the fungus as it was not detected in the intestine of infected rats. Increased production of pro-inflammatory cytokines by major gut-draining mesenteric lymph nodes point to these lymphoid organs as places of generation of cytokine-producing cells. No changes in spleen or systemic cytokine responses was observed, showing lack of the effects of pulmonary A. fumigatus infection outside mucosal immune system. Drop of intestinal bacterial microbiota diversity (disappearance of several bacterial bands) was noted early in infection with normalization starting from day seven. From day three, appearance of new bacterial bands (unique to infected individuals, not present in controls) was seen, and some of them are pathogens. Alterations in intestinal bacterial community might have affected intestinal immune tolerance contributing to inflammation. Disruption of gut homeostasis during pulmonary infection might render gastrointestinal tract more susceptible to variety of physiological and pathological stimuli. Data which showed for the first time gut involvement with pulmonary infection with A. fumigatus provide the baseline for future studies of the impact of fungal lung infections to gut homeostasis, particularly in individuals susceptible to these infections.     

Effects of allergic airway disease on mouse adenovirus type 1 respiratory infection

Virus infection may contribute to asthma pathogenesis. In turn, a Th2-polarized pulmonary environment may increase host susceptibility to infection. We used a cockroach antigen (CRA) model of allergic airway disease to test the hypothesis that Th2 cytokine overproduction increases susceptibility to mouse adenovirus type 1 (MAV-1). CRA sensitization led to upregulated lung expression of IL-4 and IL-13, lung cellular inflammation, and exaggerated airway mucus production. Following intranasal MAV-1 infection, lung cellular inflammation was more pronounced in CRA-sensitized mice than in unsensitized mice at 7 days post-infection but not at a later time point. CRA sensitization did not significantly suppress lung IFN-γ expression, and lung IFN-γ expression was upregulated in both CRA-sensitized mice and unsensitized mice over the course of MAV-1 infection. Despite CRA-induced differences in pulmonary inflammation, MAV-1 viral loads in lung and spleen and MAV-1 gene expression in the lung did not differ between CRA-sensitized and unsensitized mice. Our data therefore suggest that MAV-1 pathogenesis is not affected directly or indirectly by the Th2 polarization associated with allergic airway disease.     

壳聚糖递送的结核基因疫苗诱导的肺部黏膜免疫及在抗结核免疫保护中的意义

诱导肺局部黏膜免疫对于早期控制结核分枝杆菌(Mycobacterium tuberfulosis)十分关键.在我们过去构建的结核基因疫苗pHSP65pep有效诱导脾脏IFN-γ+Th1细胞应答的基础上,为更好诱导呼吸道和肺局部黏膜免疫,采用天然生物多糖壳聚糖(chitosan)作为黏膜递送介质递送结核基因疫苗.制备壳聚...     

慢性尘螨暴露诱导小鼠气道Th1炎症相关的气道重塑

目的:观察慢性尘螨变应原暴露对小鼠气道变应性炎症及重构的影响。方法:采用α平滑肌肌动蛋白启动子驱动的Cre重组酶(α-SMA-Cre)与R26R双转基因报告小鼠,尘螨连续滴鼻60 d后测定气道阻力;肺泡灌洗并分类计数;分离肺组织并提取肺组织蛋白;制备病理组织切片;分离培养脾脏淋巴细胞。结果:慢性尘螨暴露组小鼠气道阻力明显升高(P0.01),肺泡灌洗液(BALF)中细胞总数和淋巴细胞比例显著高于正常对照组(P0.01),肺组织HE染色示:慢性尘螨暴露组小鼠气道上皮细胞炎症水肿,小血管周围淋巴细胞浸润,未见嗜酸粒细胞。X-gal染色显示:尘螨暴露组气道平滑肌细胞及上皮下肌纤维母细胞明显增生,相应Western blotting结果显示α平滑肌肌动蛋白(α-SMA)表达明显增多。尘螨暴露组肺匀浆上清γ干扰素(IFN-γ)较正常对照组升高(P0.01),白细胞介素-4(IL-4)水平两者间没有显著差异(P0.05)。流式细胞检测显示尘螨暴露组分泌IFN-γ的CD4+Th1细胞较正常对照组增多。结论:慢性尘螨气道暴露诱导了Th1炎症,与气道重构和气道高反应性密切相关。     

HIV感染者NKT样细胞基线功能对疾病进程的影响

目的本实验通过检测HIV感染者NKT样细胞基线功能的变化,研究NKT样细胞对HIV感染疾病进程的影响。方法应用流式细胞术直接对HIV感染者以及健康对照外周血NKT样细胞IFN-γ分泌和CD107a表达进行研究。结果 NKT样细胞分泌IFN-γ百分比高者HIV疾病进展慢(P<0.008,P<0.001),与CD4+T细胞计数呈显著正相关(r=0.402,P=0.027),而与病毒载量呈显著负相关(r=-0.472,P=0.037)。结论 NKT样细胞功能较强,具有免疫保护作用,是延缓HIV病程的重要因素之一,可作为监测HIV疾病进展的指标。     

Tyk2信号传导在表达OVA的重组卡介菌(rBCG-OVA)所致慢性胞内细菌感染小鼠模型的CD8+Tc1反应中的作用

目的 探讨酪氨酸激酶2(tyrosine kinase 2,Tyk2)在IL-12和IFN-γ激发的信号传导中所起的作用.方法 为阐明Tyk2在CD8+Tc1反应中的作用,我们跟踪了Tyk2基因敲除(Tyk2-/-)小鼠和Tyk2野生型(Tyk2+/+)小鼠感染表达卵清蛋白的重组卡介菌(rBCG-OVA)后功能性CD8+T细胞的增殖分化过程.结果 与Tyk2+/+小鼠相比,在rBCG-OVA感染后,Tyk2-/-小鼠的OVA257-264抗原特异的CD8+T细胞能够随着感染的发生和发展开始增生和收缩,但是CD8+T细胞总数明显减少,OVA257-264/Kb-四聚物阳性的CD8+T细胞和IFN-γ产生阳性的CD8+Tc1细胞数量明显不足,动力曲线与增加的体内细菌的增长相对应.结论 Tyk2的信号缺失使rBCG-OVA致慢性细胞内病原菌感染导致的功能型CD8+Tc1反应减弱.

Abstract：

Objective To elucidate potential roles of tyrosine kinase 2 (Tyk2) in the generation and maintenance of Ag-specific CD8+ T cells. Methods We followed the fate of OVA-specific CD8 + T cells in Tyk2-deficient ( Tyk2 -/- ) mice after infection with recombinant OVA-expressing BCG ( rBCGOVA ). Because the immunostimulatory BCG-derived peptides recognized by CD8 + T cells have not been defined, and the OVA is definite peptide for specific CD8 + T cells that has been accepted widely, therefore we examine the kinetics of the OVA-Ag-specific CD8 + T cell response after rBCG-OVA infection in mice.Tyk2-/- and wild type(Tyk2+/+ ) mice were inoculated with rBCG-OVA by intra-trachea( i. t. ), after the examination of bacterial growth in the lung and spleen, the population of CD8 + T cells were detected by FACS analysis, the epitope-specific CD8 + T cells were followed with tetrameric H-2Kb molecule folding with OVA257 264 peptide, and the kinetics of Ag-specific CD8 + Tc1 cells were detected by intracellular IFN-γ production in response to OVA257-264 peptide by cytokine FACS analysis. Results After rBCG-OVA challenge,the bacteria number in spleen and lung of Tyk2 -/- mice were significantly larger than those in Tyk2 +/+ mice on days 14, 21 and 49. Almost as same as that in Tyk2+/+ mice, the size of epitope-specific CD8+ T cella with OVA257-264/Kb-tetramer-positive and the CD8 +Tc1 (T eytotoxic 1 )cells positive for intracellular IFN-γ could proliferate to its peak on day 21, then contract and maintain to the memory phase in spleen and lung of Tyk2-/- mice, but the population of CD8+ T cells in spleen and lung of Tyk2 -/- mice were significantly smaller than those in Tyk2+/+ mice on days 21 and 49, the number of epitope-specific CD8+ T cells in spleen and lung of Tyk2 -/- mice were significantly decreased and the frequency of CD8 + Tc1 cells in spleen and lung of Tyk2 -/- mice significantly reduced on day 21,49 and 70 after rBCG-OVA infection. So correspond with the larger number of bacteria in Tyk2-/- mice than those in Tyk2 +/+ mice, the expansion of OVA257-264-specific CD8 + T cells and CD8+ Tc1 response were attenuated in Tyk2 -/- mice following rBCG-OVA infection. Conclusion These results suggest that the lack of Tyk2 signaling impairs the proliferation and difference of effector CD8 + T cell to rBCG-OVA infection and at least, is partly responsible for the susceptible to the rBCG-OVA infection.