冬虫夏草菌丝体对小鼠体液免疫的影响

本文采用血清溶血素测定新方法,对兰州三爱堂解放军第一医院人工发酵培养的冬虫夏草菌丝体进行实验分析。结果表明,对正常小鼠的体液免疫功能无影响,但对由环磷酰胺引起的体液免疫功能低下有明显的恢复,说明该菌丝体与天然虫草有一致的功能。     

藏产红景天混合提取物对BALB／c小鼠免疫功能的影响

选择淋巴器官脏器系数,迟发型变态反应,脾脏抗体生成细胞数,巨噬细胞蚕噬指数等免疫免疫学指标观察藏产多种红景天对ＢＡＬＢ／ｃ小鼠免疫功能的影响。结果表明,在５００－２０００ｍｇ／ｋｇ剂量范围内能增加ＢＡＬＢ／ｃ小鼠胸腺的相对重量,足跖厚度及脾脏抗体生成细胞的数量。     

肝炎患者红细胞免疫功能及与体液免疫功能的关系

检测１２１例各型肝炎患者红细胞免疫功能，以及大部分患者的体液免疫功能指标，并探讨两者间的相关性。结果显示：（１）各型肝炎患者及慢性乙肝病毒携带者ＲＢＣＣ３ｂＲＲ均明显低于正常人。急性肝炎急性期和中－重度慢性肝炎患者ＲＢＣ－ＩＣＲ均明显高于正常人。中－重度慢性肝炎患者ＲＢＣＣ３ｂＲＲ明显低于轻度患者，前者ＲＢＣ－ＩＣＲ明显高于后者；急性肝炎恢复期患者ＲＢＣＣ３ｂＲＲ明显高于急性期患者，前者的ＲＢＣ－ＩＣＲ却明显低于后者；（２）急性和慢性肝炎患者血清ＣＩＣ水平均明显升高，急性肝炎恢复期患者血清ＣＩＣ水平较急性期降低。慢性肝炎患者血清ＩｇＭ、ＩｇＧ、ＩｇＡ水平均有不同程度的升高。中－重度慢性肝炎患者血清Ｃ３、Ｃ４水平均明显下降；（３）急性肝炎急性期和恢复期患者ＲＢＣＣ３ｂＲＲ与血清ＣＩＣ水平均呈负相关，而他们的ＲＢＣ－ＩＣＲ与之呈正相关。中－重度慢性肝炎患者ＲＢＣＣ３ｂＲＲ与血清ＣＩＣ水平呈负相关，与血清Ｃ３、Ｃ４水平呈正相关。相反，其ＲＢＣ－ＩＣＲ与血清ＣＩＣ水平呈正相关，与Ｃ３、Ｃ４水平呈负相关。故临床上动态观察肝炎患者的红细胞免疫功能变化，有助于评估病情转归和病变程度。     

Construction of multivalent DNA vaccines forMycobacterium tuberculosis and its immunogenicity

The coding regions of Ag85B MPT-64, and ESAT-6 secreted proteins were cloned initially into the eukaryotic expression vector pJW4303, then transformed to E. coli Top 10 strain for plasmid DNA extraction and further analysis. Plasmids containing the right insertion were sequenced to confirm their identity. COS7 cells were transfected with a mixture containing serially diluted plasmid DNA encoding three secreted proteins and Lipofectin (Gibco). The supernatants and pellets prepared from various cell lines were run on SDS-PAGE gel and the expression of these proteins in COS7 cells were demonstrated by immunoblot using polyclonal or monoclonal antiserum of M.TBH37Rv. 21 days after first vaccination of C57BL-6 mice by all three recombinant eukaryotic expressing vectors, antibody titer for Ag85B reached 1∶3200. 21 days after second vaccination, the antibody titer reached 1∶102400. The highest antibody levels induced by multivalent vaccines after the second injection were equal to or even greater than the highest antibody levels of single DNA vaccine reported in literature after third injections. Antibody titer of MPT-64 was 1∶50 after the first injection and it reached 1∶200 after the second injection. No antigen-specific antibody against ESAT-6 was detected in sera harvested from immunized mice 21 days after both injections. Antigen-specific IFN-g level of Ag85B was 110 pg/mL while no antigen-specific IFN- g level of ESAT-6 and MPT-64 was detected even after third injections. To our knowledge, it is the first time that studies of polyvalent recombinant DNA vaccines against TB were carried out in C57BL-6 mice. Our results indicated that multiple DNA vaccines could be used to enhance protective responses against M.TB.     

Schistosome vaccines

Summary Schistosomiasis control currently relies primarily on chemotherapy which is both expensive and temporary. There is an urgent need for an effective vaccine. Studies in animal models and man have demonstrated the existence of protective immunity. Antibody-dependent cell-mediated cytotoxicity mechanisms involving eosinophils and macrophages have been implemented in destruction of the parasites. Antigens expressed on the surface of the schistosomulum are among the targets of protective immune responses. Vaccines comprising recombinant antigens are now being tested in vivo for their capacity to evoke protective responses. Live oral vaccines based on attenuatedSalmonella expressing schistosomular surface antigens are being developed.     

表达结核分枝杆菌休眠相关抗原Rv2031c-Rv2626c融合蛋白

利用大肠埃希菌-分枝杆菌穿梭表达载体pDE22,构建能够表达结核分枝杆菌（Mycobacterium tuberculosis, MTB）休眠相关抗原Rv2031c-Rv2626c融合蛋白的重组耻垢分枝杆菌（Mycobacterium smegmatis, M.s）。采用聚合酶链反应（PCR）方法,从MTB H37Rv基因组DNA中扩增MTB休眠相关抗原Rv2031c和Rv2626c基因片段并克隆入pMD18-T载体,序列测定正确后分别亚克隆入大肠埃希菌-分枝杆菌穿梭表达载体pDE22,将重组载体pDE22-Rv2031c-Rv2626c电穿孔导入M.s,42℃热诱导表达目的蛋白,并进行SDS-PAGE和Western-blot分析。利用PCR方法扩增获得MTB休眠相关抗原Rv2031c和Rv2626c基因片段,大小分别为435bp和480bp,基因测序与Genebank公布的基因序列完全一致。SDS-PAGE分析结果表明重组载体pDE22-Rv2031c-Rv2626c电穿导入M.s后能够特异性表达目的蛋白,大小约为35kDa,与理论值相符。Western-blot鉴定结果表明抗Rv2031c和抗Rv2626c的单克隆抗体均能够与目的蛋白发生特异性的反应,大小相一致。成功构建表达MTB休眠相关抗原Rv2031c-Rv2626c融合蛋白的重组M.s,并命名为Rv2031c-Rv2626c-rM.s,为其用于结核新型疫苗的研究奠定了基础。     

吉林人参低聚肽的免疫调节作用

 为研究吉林人参低聚肽(GOP)对小鼠的免疫调节作用, 选取280 只SPF 级雌性BALB/c 小鼠, 随机分为7 组:空白对照组、乳清蛋白组(150 mg/kg)及5 个GOP 组(37.5、75、150、300、600 mg/kg)。连续灌胃30 d 后, 进行免疫7 项实验测定, 观察GOP 对小鼠免疫器官相对重量、细胞免疫功能、体液免疫功能、单核-巨噬细胞功能和NK 细胞活性的影响。结果表明:GOP 显著提高了ConA 诱导的小鼠脾淋巴细胞增殖能力、迟发型变态反应能力、抗体生成细胞数、小鼠碳廓清指数、巨噬细胞吞噬率和吞噬指数、NK 细胞活性(P<0.05), 且效果优于乳清蛋白。由此可知, GOP 可以通过增强细胞免疫功能、体液免疫功能、单核-巨噬细胞吞噬能力和NK 细胞活性, 起到增强免疫力的作用。     

1-(2-(4-取代)苯氧乙基)-1H-1,2,3-三氮唑衍生物的合成及其抗菌活性研究

 设计并合成了8个未曾见文献报道的1-(2-(4-取代)苯氧乙基)-1H-1,2,3-三氮唑衍生物,并通过NMR、IR、MS确认了化合物的结构。以克拉霉素和左氧氟沙星为参照物,对8个化合物进行了金黄色葡萄球菌、大肠杆菌和绿脓杆菌的体外抑制活性测试,测试结果表明其中一种化合物对金黄色葡萄球菌表现出一定的抗菌活性(MIC₅₀＝64 μg/mL)。     

硒对免疫功能的影响

目的：了解硒对免疫功能的影响的研究进展，为今后的研究提供依据。方法：对国内外文献进行收集、整理和分析。结果：硒是一种重要的人和动物体内所必需微量元素，缺硒会损害免疫系统的功能。补充适量的硒可增强非特异免疫及细胞免疫、体液免疫功能，但补硒要适量，超过安全摄入量会引起硒中毒。结论：硒在免疫反应中发生作用的机制尚未明确，仍有待于进一步研究。     

黄粉幼虫防卫机制的研究

对黄粉（Ｔｅｎｅｂｒｉｏｍｏｌｉｔｅｒ）末龄幼虫血淋巴的防卫机制做了报道，结果表明：（１）对末龄幼虫分别进行针刺体壁、注射大肠杆菌（Ｅｓｃｈｅｒｉｃｈｉａｃｏｌｉ）、苏云金杆菌（Ｂａｃｉｌｕｓｔｈｕｒｉｎｇｉｅｎｓｉｓ）及无菌生理盐水，结果均能引起血细胞总数（ＴＨＣ）规律性的变动，其变动曲线是一致的，即处理后１５ｍｉｎＴＨＣ急剧下降，在随后的１０ｈ内ＴＨＣ出现３个高峰期，但不同的处理ＴＨＣ的变动幅度不同；（２）对幼虫分别注射大肠杆菌及苏云金杆菌后，虫体内细菌的残留量逐渐减少，ＴＨＣ高峰期出现之后，伴随一个杀菌的高峰期，其相对杀死率的高峰期出现在注射后的１～２及４～６ｈ；（３）测定了黄粉幼虫体液防卫的诱导期是在处理后的６～８ｈ，所以黄粉幼虫早期的防卫作用是由血细胞进行，６～８ｈ之后，体液的防卫作用开始参与。