Cellular immune responses to recombinant Plasmodium vivax tryptophan-rich antigen (PvTRAg) among individuals exposed to vivax malaria

Plasmodium vivax , the most widespread species of human malaria parasite responsible for 70–80 million cases each year requires a vaccine. In recent years, many potential vaccine candidate antigens have been identified from P. vivax including PvTRAg. We describe here cellular immune response to recombinant PvTRAg expressed in Escherichia coli . The in vitro stimulation of PBMCs derived from P. vivax -exposed individuals ( n  = 16) showed strong proliferative response (SI > 2·2) to PvTRAg as compared to PBMCs from normal healthy controls ( n  = 8). Although both Th1 (IFN-γ, TNF-α and IL-12) and Th2 (IL-4 and IL-10) cytokines were secreted by the PBMCs of the P. vivax -exposed individuals in response to PvTRAg, the overall response was more inclined towards Th2. In conclusion, recombinant PvTRAg was found to elicit strong cellular immune response among the P. vivax -exposed individuals.     

Antibody and Th1/Th2-type responses in BALB/c mice inoculated with live or dead Echinococcus granulosus protoscoleces

The aims of this study were to investigate whether a Th1- or a Th2-type response is stimulated in the first stages of experimental infection with Echinococcus granulosus, and to determine whether live or dead protoscoleces equally contribute to such Th1/Th2-type polarization. Live parasites stimulated the production of IL-10, IL-4 and IL-5 as early as week 1 postinoculation. The levels of IL-10 and IL-4 decreased towards week 4 p.i. and that of IFN gamma increased. The production of specific antibodies was characterized by high levels of systemic IgG1 and local IgM and IgG3 (measured in peritoneal lavages). In contrast, dead parasites induced elevated levels of IL-4, IFN gamma, IL-10 and IL-5 on week 1 postinoculation followed by a decrease of IFN gamma and an increase of IL-4. Low levels of specific antibodies were stimulated by dead parasites both systemically and in the peritoneal cavity. These results show that E. granulosus infection induced an early Th2-type response and that live parasites stimulated stronger antibody responses than dead parasites. In addition, they strongly suggest that both phenomena were modulated by live protoscoleces.     

Influence of adjuvants on murine immune responses against the C-terminal 19 kDa fragment of Plasmodium vivax merozoite surface protein-1 (MSP-1)

The immunogenicity of a yeast-expressed 19 kDa fragment of P vivax MSP-1 in the presence of different adjuvant formulations was evaluated. ICR mice were immunized with the 19 kDa antigen, using Freund's, alum, and block copolymer P1005 in water-in-oil (W/O) or oil-in-water (O/W) emulsions with or without detoxified lipopolysaccharide (RaLPS) as adjuvants. Five weeks following immunization with the antigen, mice were boosted with asexual blood-stage antigens. Three weeks after the last immunization with the 19 kDa antigen, mice from the Freund's group and most groups that received P1005 as adjuvant had higher total IgG titres than those that received alum as adjuvant or antigen alone. Antibody responses after the antigen immunization were predominantly of the IgG1 isotype, but mice in the Freund's and P1005 (W/O or O/W emulsion with or without RaLPS) groups also had high titres of IgG2a and IgG2b. Antibody titres against merozoites increased in all groups after the parasite antigen boost. IgG2a levels in the group that received antigen in P1005 plus RaLPS in the W/O emulsion were higher than those receiving Freund's, alum or the other copolymer adjuvants. The high IgG2a titres in this group were associated with reduced IL-10 production.     

钥孔戚血蓝蛋白诱导Balb/c小鼠脾细胞Th1/Th2失衡的研究

目的 探讨钥孔戚血蓝蛋白(KLH)诱导Balb/c小鼠脾细胞Th1/Th2失衡的特点,为药物干预Th1/Th2失衡奠定基础.方法 以KLM 完全弗氏佐剂(CFA)免疫Balb/C小鼠并分离其脾细胞,ELISA法检测脾细胞上清中Th1型细胞因子IFN-γ、IL-2、IL-12p40和Th2型细胞因子IL-4、IL-5,以及血清Thl型抗体IgG2a和Th2型抗体IgG1水平.结果 ①免疫小鼠出现脾肿大、脾细胞计数升高.②免疫小鼠脾细胞IL-2于24h明显升高,48h达高峰;IL-4、IFN-γ和IL-5于24h轻度升高,96h达高峰;各时间点IL-12p40均较低.③不同剂量基础或强化免疫均可致细胞因子升高,IL4/IFN-γ升高.④免疫小鼠血清IgG1、IgG2a升高,以IgG1明显.结论 KLH CFA可诱导Balb/c小鼠脾细胞发生Th2型优势反应.     

The influence of the site of parasite inoculation on the development of Th1 and Th2 type immune responses in (BALB/c × C57BL/6) F1 mice infected with Leishmania major

Although inbred strains of mice are classified as genetically resistant or susceptible to Leishmania major based upon their ability to control infection, other factors such as the strain, dose, and site of parasite inoculation can also affect the outcome of the disease. Here we used the Fl progeny of BALB/c (susceptible) and C57BL/6 (resistant) mice (designated CB6F1) to investigate whether mice or intermediate susceptibility to infection differed from the parental strains in their ability to control infections at different cutaneous sites. CB6F1 mice developed progressive disease when inoculated in the dorsal skin, but healed infections in the footpad. Consistent with these observations, mice inoculated in the footpad ultimately developed Th1 responses, known to be required for healing, while Th2 responses developed in mice inoculated in the dorsal skin. However, IL-4 and IFN-γ production during the first few weeks of infection was similar in CB6F1 mice inoculated at either site, suggesting that factors in addition to the relative levels of these cytokines produced early in infection may influence the nature of the antileishmanial immune response, and the eventual disease outcome. Infection in CB6F1 mice provides a model for the study of immunity to L. major in genetically identical animals, in which a prolonged mixed Thl/Th2 cytokine pattern initially develops, but ultimately diverges into more defined Th1 and Thl type responses.     

日本血吸虫不同免疫原对小鼠Th1/Th2免疫偏移的影响

目的　观察单、双性日本血吸虫感染小鼠后Th1/Th2细胞因子水平的动态变化 ,探讨日本血吸虫不同免疫原对Th1/Th2免疫偏移的影响及与虫卵肉芽肿形成的相关性。方法　用ELISA夹心法检测单、双性日本血吸虫感染小鼠及单性感染 8w双性再感染 0～ 12w ,小鼠脾淋巴细胞培养上清Th1细胞因子IL - 2、IFN -γ和Th2细胞因子IL - 4表达水平。结果　单性感染小鼠 6～ 12w在脾淋巴细胞培养上清中可测出一定量的IL - 2和IFN -γ ,但无明显动态变化 ,而IL - 4未能测出 ;双性感染小鼠IL - 2和IFN -γ在感染后 4～ 6w开始上升 ,8w达高峰 ,随后下降 ,IL - 4则在感染后 8w时迅速上升且随着感染时间延长而明显升高 ,单性感染 8w双性再感染 4w时 ,小鼠脾淋巴细胞培养上清IL - 2、IFN -γ和IL - 4表达水平即迅速升高且在双性再感染 8～ 12w逐渐增强。结论　日本血吸虫不同免疫原对Th1/Th2免疫偏移的影响作用不同 ,Th1优势应答可能主要由虫体抗原诱导 ,Th2优势应答可能主要由虫卵抗原 (SEA)所诱导 ,后者是诱导宿主产生免疫病理反应的主要因素     

Th1 immune response to Plasmodium falciparum recombinant thrombospondin‐related adhesive protein (TRAP) antigen is enhanced by TLR3‐specific adjuvant,poly(I:C) in BALB/c mice

Sporozoite‐based malaria vaccines have provided a gold standard for malaria vaccine development, and thrombospondin‐related adhesive protein (TRAP) serves as the main vaccine candidate antigen on sporozoites. As recombinant malaria vaccine candidate antigens are poorly immunogenic, additional appropriate immunostimulants, such as an efficient adjuvant, are highly essential to modulate Th1‐cell predominance and also to induce a protective and long‐lived immune response. In this study, polyinosinic:polycytidylic acid [poly(I:C)], the ligand of TLR3, was considered as the potential adjuvant for vaccines targeting stronger Th1‐based immune responses. For this purpose, BALB/c mice were immunized with rPfTRAP delivered in putative poly(I:C) adjuvant, and humoural and cellular immune responses were determined in different immunized mouse groups. Delivery of rPfTRAP with poly(I:C) induced high levels and titres of persisted and also high‐avidity anti‐rPfTRAP IgG antibodies comparable to complete Freund's adjuvant (CFA)/incomplete Freund’s adjuvant (IFA) adjuvant after the second boost. In addition, rPfTRAP formulated with poly(I:C) elicited a higher ratio of IFN‐γ/IL‐5, IgG2a/IgG1, and IgG2b/IgG1 than with CFA/IFA, indicating that poly(I:C) supports the induction of a stronger Th1‐based immune response. This is a first time study which reveals the potential of rPfTRAP delivery in poly(I:C) to increase the level, avidity and durability of both anti‐PfTRAP cytophilic antibodies and Th1 cytokines.     

Efficient control of Plasmodium yoelii infection in BALB/c and C57BL/6 mice with pre-existing Strongyloides ratti infection

About 225 million malaria cases have been reported worldwide in 2009, and one-third of the world's population is infected with parasitic helminths. As helminths and Plasmodium are co-endemic, concurrent infections frequently occur. Helminths have been shown to modulate the host's immune response; therefore, pre-existing helminth infections may interfere with the efficient immune response to Plasmodium. To study the interaction between helminths and Plasmodium, we established a murine model of co-infection using the gastrointestinal nematode Strongyloides ratti and Plasmodium yoelii. We show that a pre-existing Strongyloides infection slightly enhanced peak parasitemia and weight loss in P. yoelii-infected BALB/c mice, while disease progression was not altered in co-infected C57BL/6 mice. The Plasmodium-induced IFN-γ production and final clearance of Plasmodium infection were not affected by S. ratti co-infection in both C57BL/6 and BALB/c mice. Interestingly, the T helper cell (Th) 2 response induced by S. ratti was significantly suppressed upon P. yoelii co-infection. This suppressed Th2 response, however, was still sufficient to allow expulsion of S. ratti parasitic adults. Taken together, we provide evidence that simultaneous presence of helminth and protist parasites does not interfere with efficient host defence in our co-infection model although changes in Th responses were observed.     

广州管圆线虫感染小鼠血清细胞因子及IgG亚类的动态观察

目的观察小鼠感染广州管圆线虫后机体免疫的动态变化。方法分别采集感染前、感染后第1、3、7、18d的小鼠血清,用ELISA方法检测血清中细胞因子IL-2、IL-4以及特异性免疫球蛋白G（IgG）亚类水平。结果广州管圆线虫感染的小鼠血清中IL-2的水平较感染前呈逐渐下降趋势,IL-4的水平与感染前小鼠相比先下降后呈升高趋势。抗体IgG1的水平与感染前小鼠相比明显升高,IgG2a的水平与感染前小鼠相比未见明显变化。结论小鼠Th1型免疫应答较弱,Th2型免疫应答增强。表明小鼠感染广州管圆线虫后机体细胞免疫较弱,体液免疫较强。     

双歧杆菌对食物过敏小鼠肠道屏障功能及Th1/Th2细胞因子的影响

目的:探讨双歧杆菌在改善食物过敏动物肠道屏障功能、调整肠道菌群结构以及对免疫功能调节方面的作用及其机制.方法:无受试蛋白喂养BALB/c小鼠40只,随机分为4组: 分别于0、3、9 d腹腔注射生理盐水,金黄色葡萄球菌肠毒素B(SEB),卵清蛋白(OVA),SEB+OVA; 并于第7、14天给予OVA灌胃.在SEB+OVA致敏组的基础上设立自然恢复组、双歧杆菌作用组、思密达作用组、双歧杆菌+思密达共同作用组,第15天开始分别经灌胃给予不同的药物,连续7 d,每日1次.培养法分析粪便菌群,检测血清二胺氧化酶(DAO)含量,ELISA法测定血清IgE、IL-4、INF-γ含量.对肠系膜淋巴结(MLN)及肝、肾、肺组织进行培养以探讨有无细菌移位(BT)发生.同时,采用流式细胞术分析其脾细胞悬液中CD4+CD25+调节性T细胞的数量变化.结果:与SEB+OVA实验组相比,双歧杆菌作用组小鼠血清IgE、DAO含量(A 值)、血清IL-4(51.314±3.785 ng/L vs 69.980±9.103ng/L,P<0.05)含量显著降低; 血清INF-γ水平显著升高(194.281±12.144 ng/L vs 133.875±33.822 n g/L,P<0.05); 脾细胞悬液中CD4+CD25+T细胞数量显著升高(5.778%±0.773% vs 4.216%±0.439%,P<0.05); 肠道固有菌群中益生菌乳酸杆菌的含量(6.670±0.443 vs 5.654±0.289,P<0.05)、双歧杆菌的含量(8.611±0.295 vs 7.491±0.339,P<0.05)显著升高,条件致病菌大肠杆菌的含量(5.364±0.537 vs 6.718±0.267,P<0.05)、类杆菌的含量(7.427±0.544 vs 8.606±0.317,P<0.05)显著降低; MLN及外周器官细菌移位率显著降低(12.5% vs 37.5%,P<0.05).结论:双歧杆菌可以有效调节机体免疫功能、调整肠道菌群失调及保护肠道黏膜屏障功能.     

香菇多糖对约氏疟原虫感染BALB/c小鼠Th1型应答的调节作用

目的探讨香菇多糖(Lentinan,Lent)对致死型约氏疟原虫(Plasmodium yoelii 17XL,Py17XL)感染BALB/c小鼠Th1型细胞免疫应答的调节效应。方法对Py17XL感染的BALB/c小鼠进行不同时间点的Lent预处理,动态观察用药后各组感染小鼠原虫血症水平和生存率;于感染后第0d、1d、3d和5d分别提取小鼠脾细胞,ELISA法检测脾细胞培养上清中IL-12、IFN-γ的分泌水平,Griess反应检测脾细胞培养上清中一氧化氮(NO)含量。结果与药物未处理组相比,感染前15d 1mg/kg Lent用药组显著降低感染小鼠的原虫血症水平,提高生存率;明显增强Th1型免疫应答中关键细胞因子IL-12、IFN-γ的分泌水平,并提高NO含量。结论Lent预处理能够有效激发Py17XL感染的BALB/c小鼠Th1型细胞免疫应答的建立,提示调控免疫应答对于致死型约氏疟原虫感染早期免疫防御的重要性。     

T‐cell clones in human trichinellosis: Evidence for a mixed Th1/Th2 response

In humans, studies on the cellular immune response against Trichinella are scarce. Aim of this study was to characterize the cytokine profile of T cells specific for Trichinella britovi in trichinellosis patients. Peripheral blood mononuclear cells (PBMC) were obtained from five patients involved in a trichinellosis outbreak caused by T. britovi, which occurred in 2013 in Tuscany (Italy). All the patients resulted positive for Trichinella‐specific IgG, IgE and presented eosinophilia. T cells were investigated for their proliferation to excretory/secretory antigens from Trichinella spiralis muscle larvae (TsES) and for their cytokine profile. A total of 284 CD4+ and 42 CD8+ T‐cell clones were obtained from the TsES‐specific T‐cell lines from PBMC. All T‐cell clones proliferated in response to mitogen. Of the 284 CD4+ T‐cell clones generated from TsES‐specific T‐cell lines, 135 (47%) proliferated significantly to TsES; 26% CD8+ T‐cell clones showed proliferation to TsES. In the series of the 135 TsES‐specific CD4+ clones, 51% expressed a Th2 profile, 30% a Th0 and 19% Th1. In the series of the 11 TsES‐specific CD8+ T‐cell clones, 18% were Tc2, 45% Tc0 and 36% Tc1. In human trichinellosis, the cellular immune response is, during the chronic phase, mixed Th1/Th2.     

The role of regulatory T cells during Plasmodium chabaudi chabaudi AS infection in BALB/c mice

An inappropriate immune response to parasite infection is one of the primary drivers of malaria pathogenesis. Regulatory T cells (Tregs), an important subset of CD4⁺ T cells, can maintain self‐tolerance and prevent autoimmune diseases. However, there is little consensus about their role in malaria pathogenesis. In this study, we transiently depleted Tregs (CD25⁺ T cells) using an anti‐CD25 mAb (7D4 clone) at different time points following Plasmodium chabaudi chabaudi AS infection in BALB/c mice and investigated the effect of depletion of Tregs in this model. In control mice, Tregs proliferated significantly and their suppressive function was enhanced after infection. IL‐10 was increased drastically during infection. Depletion of Tregs at various time points can lead to divergent outcomes. When Tregs were depleted prior to or during the early phase of infection, most mice survived and had a robust Th1 immune response. In contrast, when Tregs were depleted close to peak parasitemia, all mice died as a result of inflammation. Taken together, these data suggest that in P. c. chabaudi AS‐infected BALB/c mice, Tregs inhibit the Th1 response and macrophage activation, leading to increased parasite load; however, they also control inflammation‐mediated pathology by secreting high levels of IL‐10.     

Th1/Th2及Tc1/Tc2在胃癌患者外周血中的漂移及意义

目的: 研究胃癌患者外周血Th1/Th2及Tc1/Tc2漂移状况,并分析其免疫预存状态.方法: 2008-10-21/2008-12-05中国人民解放军总医院普通外科新入院的术前胃癌患者外周血样本25例,同期健康献血员正常对照组外周血样本25例.四色流式细胞仪检测胃癌患者及正常对照人群外周血激活的淋巴细胞内细胞因子(IFN-γ、IL-4),并分析Th1/Th2及Tc1/Tc2漂移状态.结果: 胃癌患者外周血Th1/PBL、Tc1/PBL、Th1/Th2、Tc1/Tc2比值均较正常对照组明显减少(6.242%±4.078% vs 3.047%±1.710%,14.171%±8.984% vs 6.393%±5.235%,3.127%±3.633% vs 1.172%±0.300%,17.200%±25.930% vs 3.252%±8.732%,均P＜0.01).结论: 胃癌患者Th1及Tc1细胞比例减少,并出现Th1/Th2及Tc1/Tc2漂移现象,提示胃癌患者预存免疫状态低下,机体抗肿瘤免疫功能抑制.     

狼疮肾炎患者血清白细胞介素-18的升高与Th1/Th2型细胞的失衡

目的测定狼疮肾炎（LN）患者血清白细胞介素-18（IL-18）的水平以及外周血单个核细胞（PBMC）内干扰素（IFN）-γ／IL-4的表达情况，探讨LN患者体内Th1/Th2型细胞的平衡情况以及IL-18与Th1/Th2型免疫反应的相关性。方法酶联免疫吸附试验（ELISA）方法检测血清IL-18的表达：佛波酯（PMA）和离子霉素联合刺激培养，流式细胞术从单个核细胞水平检测细胞内IFN-γ/IL-4的表达情况。结果LN患者血清IL-18的水平显著高于对照组（P=0．000）；Th1型（IFN—γ^＋）细胞百分率明显高于对照组（P=0．009），Th1/Th2型细胞的比率（IFN-γ／IL-4^＋）明显增高（P=0．005）；Th1/Th2型细胞比率与尿蛋白定量（24h）呈明显正相关；Th1型细胞百分率与血清IL-18的水平呈明显的正相关。结论LN患者血清IL-18水平升高，并与Th1/Th2型免疫反应失衡有关，IL-18可以促进LN患者体内Th1型免疫反应增强。从而加重蛋白尿的发展。削弱Th1型免疫反应重建Th1/Th2型免疫反应的平衡，理论上可能会阻止狼疮肾炎的发病。     

NO介导Th1/Th2免疫偏移与日本血吸虫感染小鼠肝肉芽肿病变的作用

目的　观察日本血吸虫感染小鼠肝肉芽肿病变与一氧化氮 (NO)、Th1/Th2细胞因子水平的动态变化 ,探讨NO介导Th1/Th2免疫偏移在血吸虫卵肉芽肿病变中的作用。方法　采用石蜡切片 ,HE染色后观察感染小鼠肉芽肿病变。用硝酸还原酶法和ELISA夹心法分别检测日本血吸虫感染小鼠 0～ 12周血清及脾CD+ 4 T淋巴细胞培养上清NO、IFNγ和IL - 4表达水平 ,并进行相关分析。结果　感染小鼠 0～ 12周血清和脾CD+ 4 T淋巴细胞培养上清NO表达水平与小鼠肝肉芽肿病变动态相一致 ,并呈显著正相关 ,与IFNγ表达水平呈显著正相关 ,而与IL - 4在小鼠感染慢性期 (12周 )呈显著负相关。结论　NO在日本血吸虫感染小鼠肝肉芽肿病变过程中可能是一种与Th1/Th2细胞因子具有同样重要作用的调节因子 ,并可能是通过调节Th1/Th2细胞因子而发挥作用的 ,通过调控NO合成介导Th1/Th2免疫偏移可能是控制血吸虫卵肉芽肿病变的新途径     

小儿肾母细胞瘤Th1／Th2类细胞因子的漂移及临床意义

采用逆转录聚合酶链反应（RT－PCR）技术,检测正常儿及肾母细胞瘤患儿肿瘤组织,手术前后外周血Th1/Th2类细胞因子的基因表达,观察肾母细胞瘤患儿手术前后Th1/Th2类细胞因子的表达和漂移情况,结果显示肾母细胞瘤患儿外周血,肿瘤组织均出现明显的Th2类细胞因子偏移趋势,手术和化疗可促进Th1/Th2平衡向Th1漂移,肾母细胞瘤患者体内Th2类细胞因子的强势表达,可能与肾母细胞瘤的发生发展有关,Th1/Th2平衡漂移是观察机体抗肿瘤免疫动态变化的良好指标。     

HLA-DRB1＊15及相应位点下Th1／Th2细胞相关因子对乙肝疫苗免疫效果的影响

目的探讨广西人群HLA—DRB1＊15基因及相应位点下Th1／Th2细胞相关因子IFN-γIL-4对乙肝疫苗免疫应答水平的影响。方法选取完成重组乙型肝炎疫苗标准全程接种的广西籍汉族健康大学生中抗-HBsS／N值〈10mIU／ml的无、弱应答者80名（A组）作为研究对象,随机选取抗-HBsS／N值〉10mIU／ml的中、强应答者62名（B组）作为对照。应用PCR—SSP进行HLA-DRB1＊15等位基因的检测,采用酶联免疫吸附试验（ELISA）检测血清中Th1／Th2细胞相关因子干扰素-γ（IFN-γ）、白细胞介素-4（IL-4）。结果①A组HLA—DRB1＊15基因表达频率为11．25％,显著低于B组的36．67％（P=0．001）;②A组中IFN-γ的平均表达水平为（6．28±6．84）pg／ml,显著低于B组（16．28±10．05）pg/ml（P=0．000）;③A组的IL-4平均表达水平为（2．36±1．91）pg／ml。显著低于B组（8．50±6．68）pg／ml（P=0．000）;④HLA—DRB1＊15阳性组IFN-γ平均表达水平为（13．82±9．89）pg／ml,显著高于HLA—DRB1＊15阴性组（9．76±9．54）pg／ml（P=0．029）;⑤HLA-DRB1＊15阳性组IL-4平均表达水平为（5．81±4．27）pg／ml,阴性组IL-4平均表达水平为（4．82±5．84）pg／ml,两组间差异无统计学意义（P=0．207）。结论①HLA—DRB1＊15基因可能是促进广西人群乙肝疫苗免疫应答的相关基因;②Th1／Th2细胞相关因子IFN-1、IL-4的表达水平可能影响机体乙肝疫苗的免疫效果;③HLA—DRBB1＊15基因可能是通过影响Th1细胞相关因子IFN-γ的表达水平,而不是通过影响Th2细胞相关因子IL-4的表达水平来影响乙肝疫苗的免疫应答。     

猪带绦虫抗原刺激囊虫病患者PBMC Th1/Th2细胞因子的表达及其免疫调节

目的检测猪带绦虫不同虫期抗原刺激囊虫病患者外周血单个核细胞(PBMC)Th1/Th2细胞因子的表达水平,并分析其在囊虫病免疫调控中的作用。方法用流式细胞仪检测囊虫病患者新鲜血T淋巴细胞亚群,并以猪带绦虫六钩蚴抗原或囊尾蚴抗原刺激囊虫病患者PBMC,分别在刺激的第0d、5d、10d和15d时检测Th1/Th2细胞因子(IFN-γ及IL-4),对分泌IFN-γ或IL-4的不同细胞群体进行数据分析。结果囊虫病患者新鲜血T淋巴细胞亚群CD3+与CD4+细胞较正常人升高,CD4+/CD8+较正常人升高,分泌IFN-γ及IL-4的CD3+细胞百分率较正常人升高;猪囊尾蚴抗原刺激囊虫病患者PB-MC第0d、5d、10d和15d时,分泌IFN-γ的CD3+细胞百分率分别为25.42%±5.53%,30.46%±4.94%,36.52%±4.73%,38.69%±5.58%;分泌IL-4的CD3+细胞百分率分别为2.52%±0.52%,3.00%±0.57%,3.81%±0.70%,5.03%±0.73%。六钩蚴抗原刺激囊虫病患者PBMC分泌IFN-γ及IL-4的CD3+细胞百分率亦呈现相同的变化趋势。结论囊虫病患者与正常人相比PBMC中存在T淋巴细胞的极化水平异常,CD3+与CD4+细胞百分率均显著升高,T细胞亚群比例失调,免疫功能紊乱;随着囊尾蚴抗原刺激外周血PBMC时间延长,分泌IFN-γ和IL-4的CD3+细胞百分率逐渐升高。     

耻垢分枝杆菌疫苗对小鼠细胞因子产生及Th1/Th2应答的影响

目的研究以耻垢分枝杆菌(M.S)制备的M.S疫苗对小鼠免疫应答的影响,验证M.S的免疫原性、探讨M.S疫苗的免疫调节作用。方法将BALB/c小鼠随机分成生理盐水对照组和M.S疫苗低、中、高剂量组,每组6只,分别注射生理盐水和不同剂量的M.S疫苗。取小鼠脾细胞或腹腔巨噬细胞体外培养,用酶联免疫吸附测定(ELISA)法检测培养上清中白细胞介素(IL)-2、IL-4、IL-12和γ干扰素(IFN-γ)的含量,分析M.S疫苗对小鼠Th1/Th2类应答的影响。结果(1)生理盐水对照组和M.S疫苗低、中、高剂量组小鼠产生的IL-12分别为(32.6±22.7)、(58.9±18.6)、(77.3±38.0)、(114.7±9.9)pg/m l,M.S疫苗中、高剂量组与生理盐水对照组比较差异有统计学意义(P<0.05)。(2)生理盐水对照组和M.S疫苗低、中、高剂量组小鼠产生的IL-2分别为(5.0±2.6)、(13.4±9.3)、(15.3±9.7)、(22.6±7.5)pg/m l,M.S疫苗高剂量组与生理盐水对照组比较差异有统计学意义(P<0.01)。(3)体外以ConA刺激时,生理盐水对照组和M.S疫苗中剂量组小鼠产生的IFN-γ分别为(662±279)和(807±163)pg/m l,IL-4分别为(407±127)和(101±26)pg/m l,但M.S疫苗组与生理盐水对照组比较差异均无统计学意义(P>0.05);体外以M.S-纯化蛋白衍生物(PPD)刺激时,生理盐水对照组和M.S疫苗中剂量组小鼠产生的IFN-γ分别为(14.0±6.31)和(55.3±32.4)pg/m l,两组比较差异有统计学意义(P<0.05),各组产生的IL-4均在检测限以下。结论M.S具有良好的免疫原性,以此制备的M.S疫苗具有促进Th1类应答、抑制Th2类应答的免疫调节作用。