灵芝浸液对小鼠免疫功能的调节作用

报道了灵芝对Ｂａｌｂ／ｃ小鼠产生抗绵羊红细胞（ＳＲＢＣ）抗体的能力及对迟发性变态反应（ＤＨＴ）等免疫功能的影响。实验小鼠分为高、中、低剂量组，经胃灌注不同剂量的灵芝浸液，每天灌注１次，连续灌１５ｄ，对照组自由摄水。用ＳＲＢＣ免疫小鼠，免疫后第５ｄ用溶血空斑实验（ＰＦＣ）进行抗体生成细胞检测，以反映Ｂ细胞生成ＩｇＭ抗体的能力；于免疫后第４ｄ用足底增厚法测定ＤＨＴ强度，以检测Ｔ细胞功能；用碳粒廓清实验检测巨噬细胞吞噬功能。结果：高、中剂量组溶血空斑数目明显高于对照组，ｐ＜０．０５；高、中剂量组能明显提高小鼠ＤＨＴ反应程度，ｐ＜０．０５。表明灵芝能提高小鼠Ｔ细胞和Ｂ细胞的功能。碳粒廓清实验用药组与对照组无显著差异。     

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。     

Adenovirus-mediated<Emphasis Type="Italic">CTLA4-FasL</Emphasis> gene transfer prevents autoimmune diabetes in mice induced by multiple low doses of streptozotocin

Type 1 diabetes is the result of a selective destruction of insulin-producing β cells in pancreatic islets by autoreactive T cells. Depletion of autoreactive T cell through apoptosis may be a potential strategy for the prevention of autoimmune diabetes. Simultaneous stimulation of Fas-mediated pathway and blockade of costimulation by a CTLA4-FasL fusion protein has been reported to lead to substantial inhibition of mixed lymphocyte reaction and enhanced in vitro apoptosis of peripheral lymphocytes. To test the feasibility of CTLA4-FasL-based gene therapy to prevent autoimmune diabetes, we developed recombinant adenovirus containing human CTLA4-FasL gene (AdCTLA4-FasL). A single injection of 2×10~8 plaque forming units (PFU) of AdCTLA4-FasL via tail vein dramatically reduced the incidence of autoimmune diabetes in mice induced by multiple low doses of streptozotocin. AdCTLA4-FasL administration maintained islet insulin content, significantly increased apoptosis of pancreatic lymphocytes, quantitatively     

免疫核糖核酸作用机制的研究──免疫核糖核酸传递抗绵羊红血球免疫力

本文用热酚法从 ＳＲＢＣ致敏的豚鼠的牌和淋巴结提取了免疫核糖核酸（简称ｉＲＮＡ）。该ｉＲＮＡ能够传递抗ＳＲＢＣ的体液免疫反应。核糖核酸酶能破坏其活性，但蛋白酶和脱氧核糖核酸酶对其活性没有影响。用抗 ＳＲＢＣ的 ｉＲＮＡ和抗 ＣＲＢＣ（鸡红血球）的 ｉＲＮＡ的实验表明，ｉＲＮＡ的免疫功能传递作用既有抗原特异性也有非抗原特异性。对ｉＲＮＡ作用的特异性进行了讨论。     

Carboxymethytl Pachymaram Up-Regulates Dendritic Cell＇s Function in Hepatitis B Virus Transgenic Mice

The effect of carboxymethytl pachymaram (CMP) on the function of dendritic cells(DCs) derived from spleens of hepatitis B virus transgenic mice are studied in vitro. The phenotypes of DCs are tested by flow cytometry (FCM), cytokines measured by ELISA. The expression of DCs’ phenotypes in HBV transgenic mice are low (CD80⁺CD11c⁺:59.12±11.53 vs 9.60±4.53, p<0.01; CD80⁺ MHC-II⁺: 44.86±12.31 vs 9.80±5.72, p<0.01, normal mice vs HBV transgenic mice), the ability of DCs stimulating T lymphocytes proliferation decreases (0.37±0.11 vs 0.20±0.11, p<0.05, normal mice vs HBV transgenic mice), levels of IL-12 and IFN-γ decrease whereas the level of IL-10 increases; CMP can enhance DCs’ ability of stimulating T lymphocytes proliferation, facilitate the secretion of IL-12 and IFN-γ, inhibit the secretion of IL-10, thus up regulates DCs function. The results show a good prospective use of CMP on the treatment of chronic hepatitis B. Biography: HOU Anji (1963–), male, Ph.D. candidate, Associate professor, research direction: clinical virology.     

Inhibition of suppressor T-cell development following deoxyguanosine administration

The expression of immunodeficiency in patients with specific purine enzyme defects indicates a crucial role of the purine salvage pathway in the acquisition and expression of normal immune function. One current hypothesis links the failure of normal lymphocyte development in these diseases to the accumulation of deoxynucleotide triphosphates. In our studies of human in vitro IgM responses, we observed that antigen-induced T-suppressor cell activity was abrogated in the presence of micromolar concentrations of deoxyguanosine (dGuo). In contrast, more than 1,000-fold higher resistance to dGuo was found for both noin-proliferative T-helper cell activity and the differentiation and proliferation of the precursor B lymphocytes for direct haemolytic plaque forming cells (PFC). To determine whether these observations could have in vivo relevance, we monitored the generation of murine T-suppressor cells, capable of abrogating a primary IgM response. It was found that dGuo (but not guanosine) selectively inhibited the in vivo development of T-suppressor cells.     

灵芝对小鼠免疫功能调节作用的研究

报道了灵芝对ＢＡＬＢ／ｃ小鼠产生抗绵羊红细胞（ＳＲＢＣ）抗体的能力,对 性变态反应（ＤＨＴ）等免疫功能的影响。实验小鼠分为高、中、低剂量组、经胃灌注不同剂量的发芝浸液,每天五次连续灌１５ｄ,对照组自由摄水。用ＳＲＢＣ免疫小鼠,在免疫后第５ｄ用溶血空斑试验（ＰＦＣ）进行体生成细胞检测,以反映Ｂ细胞生成ＩｇＭ抗体的能力;于免疫后第４ｄ用足底厚法测定ＤＨＴ强度,以检测Ｔ细胞功能,用碳粒廓清检测巨噬细胞蚕     

汉滩病毒G1重组腺病毒的表达及其免疫学特性研究

将汉滩病毒(HTNV)囊膜糖蛋白G1重组腺病毒(Adeno-G1)感染VeroE6细胞,用IFA法检测其表达产物;并进一步将其免疫Balb/c小鼠。结果可检测到HTNV糖蛋白G1在VeroE6细胞中表达;用该重组腺病毒免疫小鼠,结果表明免疫小鼠体内可诱导产生抗汉滩病毒G1特异性抗体,同时微量细胞培养中和实验结果表明重组腺病毒还可刺激机体产生低水平的中和抗体,但淋巴细胞增殖反应不明显。说明Adeno-G1免疫小鼠后,主要刺激机体产生特异性的抗HTNV体液免疫应答,但刺激机体产生特异性的细胞免疫应答不明显,为HTNV基因工程疫苗的研究提供了实验基础。     

Pam3CSK4与黄连素联合用药对小鼠抗MRSA感染的作用

耐药型金黄色葡萄球菌感染的预防和治疗是临床的难题.该文考察了一种新的用药方法，将免疫激动剂Pam3CSK4和黄连的主要抗菌成分黄连素联合使用，通过激活机体先天性免疫反应和中药抗菌两个方面相结合来对抗耐甲氧西林的金黄色葡萄球菌(MRSA).探讨了Pam3CSK4和黄连素在单独或联合使用时，对小鼠受MRSA攻击的保护能力，小鼠对MRSA的清除能力，小鼠感染后分泌IgG和IgM的水平，体外二次感染后淋巴细胞的增殖能力.结果显示Pam3CSK4和黄连素联合使用可以提高小鼠在MRSA致死攻击下的的存活率，增强小鼠对MRSA的清除能力，Pam3CSK4对小鼠的特异性IgG的分泌具有抑制作用，对总IgM的分泌具有促进作用，Pam3CSK4可以促进CD4＋ 和CD8＋T细胞以及B细胞在体外的二次增殖，而黄连素可以促进对CD4＋ T细胞和B细胞的增殖.     

重组金针菇免疫调节蛋白的纯化及生物活性研究

目的研究重组的金针菇免疫调节蛋白(re FIP-fve)对小鼠脾淋巴细胞的增殖作用、诱导小鼠脾淋巴细胞对白细胞介素-2(IL-2)的促分泌作用及对血细胞的凝集作用.方法采用离子交换色谱法,利用SP Sepharose XL色谱柱纯化重组的免疫调节蛋白;将纯化后的蛋白作用于小鼠脾淋巴细胞和血细胞,用MTT法检测蛋白对脾细胞增殖和血细胞凝集的影响;用ELISA法检测蛋白对淋巴细胞分泌细胞因子的作用.结果经过SP Sepharose XL色谱柱纯化后可获得纯度为86%的重组蛋白;纯化蛋白浓度在80μg/m L时最大限度促进细胞增殖;蛋白浓度在2μg/m L时开始出现凝血;ELISA检测结果显示:重组的金针菇免疫调节蛋白能明显增强小鼠脾淋巴细胞分泌白细胞介素2(IL-2)的水平.结论经过SP Sepharose XL色谱柱纯化后可获得纯度较高的蛋白;纯化后的蛋白与小鼠脾淋巴细胞共同培养,不仅可以促进脾细胞的增殖,而且能增强小鼠脾淋巴细胞分泌白细胞介素2的水平;蛋白与血细胞共同培养能够促进血细胞的凝集.     

海参寡肽:免疫调节作用及机制研究

 为研究海参寡肽对小鼠的免疫调节作用及机制,试验选取250只SPF级雌性BALB/c小鼠,随机分为5组:空白对照组,乳清蛋白组(0.30 g/kg),0.15、0.30、0.60 g/kg海参寡肽组。连续灌胃30 d后,通过测定ConA诱导的小鼠淋巴细胞转化实验、迟发型变态反应、抗体生成细胞、血清溶血素水平、小鼠碳廓清实验、小鼠腹腔巨噬细胞吞噬鸡红细胞实验、自然杀伤(NK)细胞活性,观察海参寡肽对小鼠细胞免疫、体液免疫、单核-巨噬细胞吞噬和NK细胞活性的影响,并通过流式细胞术对脾脏T淋巴细胞亚群进行分析。结果表明:海参寡肽显著提高了小鼠细胞免疫、体液免疫、单核-巨噬细胞吞噬功能及NK细胞活性(P<0.05),且效果优于乳清蛋白。通过T淋巴细胞亚群分析表明,海参寡肽显著提高了脾脏CD3⁺百分比和CD4⁺百分比(P<0.05)。由此可知,海参寡肽可能通过增加T淋巴细胞数和Th细胞比例,介导细胞免疫、体液免疫功能、单核-巨噬细胞吞噬能力和NK细胞活性的增强作用,起到增强免疫功能的效果。     

CpG ODN增强rHBsAg免疫小鼠抗体产生及淋巴细胞增殖效应

目的：探讨合成含CpG基序的寡核苷酸（CpG ODN）对重组乙型肝炎表面抗原（rHBsAg ）增强小鼠特异性抗体产生及淋巴细胞增殖反应的效应.方法：采用BaIb/c小鼠作为免疫实验动物,经后腿胫骨前肌免疫2次,ELISA法检测血清乙型肝炎表面抗体（抗-HBs）效价;3H-TdR掺入法测定淋巴细胞增殖反应.结果：加CpG ODN组的抗-HBs效价较rHBsAg组明显增高,且维持时间长;淋巴细胞增殖反应显著增强.结论：CpG ODN对小鼠抗-HBs产生具有明显的增强作用,且可以有效刺激小鼠淋巴细胞的增殖反应.     

Specific anti-tumor effect induced by attenuated <Emphasis Type="Italic">Salmonella typhimurium</Emphasis> vaccine expressing extracellular region of vascular endothelial growth factor receptor 2

The purposes of this research were to study the stable expression of exogenous gene encoding therapeutic protein in attenuated Salmonella typhimurium, observe the metabolism of oral gene vaccine carried by attenuated Salmonella typhimurium in BALB/c mouse, and investigate the feasibility of prevention and treatment of tumors by the recombinant bacteria. Recombinant plasmid pcDNA3.1＋ VEGFR2（n1-7） was transformed into competent attenuated Salmonella typhimuriurn SL3261 to develop oral DNA vaccine SL3261-pcDNA3.1＋VEGFR2（n1-7）. To observe whether the exogenous gene can be expressed in the recombinant bacteria, PCR was performed to amplify the CMV promoter of the eukaryotic expression vector as the proof of stable expression of exogenous protein; transmission elec- tron microscopy （TEM） was applied to observe the morphology of the recombinant bacteria to confirm that the exogenous gene has no impact on the growth of the bacteria, and then BALB/c mice were immunized with the gene vaccine. After inoculation of the gene vaccine, the recombinant bacteria SL3261 could be detected in the tissues such as small intestine, colon, liver and spleen. And then, mice in each group were challenged with tumor cells. The results of animal experiment showed that tumor growth of the mice in experimental group was inhibited and survival time of immunized mice was prolonged compared with control groups. A higher lymphocyte infiltration in tumors from animals treated with DNA vaccine was observed. Immunohistochemical analysis of tumor samples revealed an enhanced accumulation of CD8^＋ cytotoxic T lymphocytes, as well as an increase in CD4^＋ cells in the tumore of animals treated with the oral gene vaccine compared to tumors from control group mice. UI- trestructure of the tumor tissue showed that tumor cells in the samples of the immunized mice were well-differentiated. Our research confirmed that the exogenous gene can be stably expressed in the attenuated Salmonella typhimurium and has no impact on the growth of the r     

IL-10修饰的树突状细胞的体外生物学效应研究

探讨lL-10修饰的树突状细胞（DC）对T细胞增殖和细胞毒性T细胞（CTL）胞毒活性的影响。采用乳酸脱氢酶法和ELISA法，分别测定细胞毒活性及T细胞凋亡。结果表明，IL-10对同种细胞刺激的增殖反应和特异性CTI．细胞毒活性有明显的抑制作用，修饰的DC诱导淋巴细胞增殖反应明显降低，而未修饰的DC诱导淋巴细胞增殖反应较强；IL-10和修饰的DC可诱导淋巴细胞凋亡．可见，稳定表达IL-10的DC，对T细胞增殖和对胞毒活性有明显的抑制作用，并可诱导T细胞凋亡。     

Effects of soy isoflavone extracts on the growth of estrogen-dependent human breast cancer (MCF-7) tumors implanted in ovariectomized nude mice

In this study, we explored the effects of soy isoflavone extracts on the growth of estrogen-dependent human breast cancer (MCF-7) tumors implanted in ovariectomized athymic mice. The ovariectomized athymic mice were implanted with MCF-7 cells. They were fed with low, moderate and high doses of soy isoflavone extracts, at dietary concentrations of 6.25, 12.5 and 25 g/kg, respectively. The expression of ki-67 was detected by immunohistochemistry. The pS ₂ expression in tumors was analyzed by real-time PCR. Estrogen level in the serum was measured by chemiluminescence enzyme immunoassay. Compared with the control group, dietary soy isoflavone extracts had a significant stimulatory effect on MCF-7 tumor growth in mice (P < 0.05). The ki-67 and pS ₂ mRNA expressions in tumors were significantly increased by 6.25 and 12.5 g/kg dose of soy isoflavone extracts (P < 0.05). And, estrogen level in serum of 6.25 and 12.5 g/kg dose groups was higher than that of control group (P > 0.05). In conclusion, in the tested dietary concentration range soy isoflavone extracts had a stimulatory effect on tumor growth. 6.25 and 12.5 g/kg doses of soy isoflavone extracts can increase the cell proliferation in tumors and induce estrogen-responsive pS2 expression. Supported by National Natural Science Foundation of China (Grant No. 30572133)     

A peptide fusion protein inhibits angiogenesis and tumor growth by blocking VEGF binding to KDR

Vascular endothelial growth factor (VEGF) binding to its tyrosine kinase receptors (KDR/FLK1, Flt-1) induces angiogenesis. In search of the peptides blocking VEGF binding to its receptor KDR/FLK1 to inhibit tumorangiogenesis and growth, we screened a phage display peptide library with KDR as target protein, and some candidate peptides were isolated. In this study, we cloned the DNA fragment coding the peptide K237 from the library, into a vector pQE42 to express fusion protein DHFR-K237 in E. coli M15. The affection of fusion protein DHFR-K237 on endothelial cell proliferation and angiogenesis was investigated. In vitro, DHFR-K237 could completely block VEGF binding to KDR and significantly inhibit the VEGF-mediated proliferation of the human vascular endothelial cells. In vivo, DHFR-K237 inhibited angiogenesis in chick embryo chorioallantoric membrane and tumor growth in nude mice. These results suggest that K237 is an effective antagonist of VEGF binding to KDR, and could be a potential agent for cancer biotherapy.     

Interleukin-2 programs mouse alpha beta T lymphocytes for apoptosis

Antigen receptor stimulation of mature alpha beta T lymphocytes can lead either to proliferation or death. Programmed cell death, termed apoptosis, leads to the clonal deletion of both thymocytes and mature T cells that establishes tolerance. How a mature T cell selects between proliferation and death is not understood. Here I show that interleukin-2 (IL-2) is a critical determinant of the choice between these two fates. Both CD4+ and CD8+ T cells previously exposed to IL-2 undergo apoptosis after antigen-receptor stimulation. Antibody blockade of IL-2 but not IL-4 reverses the marked reduction of lymph node V beta 8+ T cells caused in mice by the bacterial superantigen Staphylococcus aureus enterotoxin B. IL-2 may thus participate in a feedback regulatory mechanism by predisposing mature T lymphocytes to apoptosis.     

重组人碱性成纤维细胞生长因子诱导Balb/c3T3细胞增殖的实验研究

目的研究不同浓度重组人碱性成纤维细胞因子对Balb/c 3T3细胞增殖活性的影响.方法重组质粒pGEX-bFGF转入到DH5α大肠杆菌体内,增菌培养表达bFGF,亲和层析纯化,制备8种不同药物浓度的bFGF,分别进行诱导Balb/c 3T3细胞增殖实验研究,MTT法检测各组细胞增殖活性.结果 bFGF诱导Balb/c 3T3细胞增殖存在浓度依赖性,最适浓度为4～16μg/L.结论 bFGF具有较好的诱导Balb/c 3T3细胞增殖的能力,为后续研究工作奠定实验基础.     

BOG初次免疫联合结核分枝杆菌Ag85A DNA疫苗加强免疫效果的研究

为探讨BCG初次免疫,结核分枝杆菌Ag85A DNA疫苗加强免疫的序贯免疫策略对小鼠的免疫效果。采用BCG及结核分枝杆菌Ag85A DNA疫苗依次免疫小鼠,在末次免疫后的4、6、8周通过检测小鼠血清总IgG抗体、特异性淋巴细胞增殖．细胞因子的水平,观测BCG初次免疫,Ag85A DNA疫苗加强免疫的策略对小鼠的免疫效果。结果显示,采用BCG初次免疫,结核分枝杆菌Ag85A DNA疫苗加强免疫的策略组的小鼠与其它免疫方式组相比,IgG明显升高（P〈0．05）、特异性淋巴细胞明显增殖、IFN7、IL-2、IL-4水平明显增高（P〈0．05）。由此可知,在采用BCG初次免疫,结核分枝杆菌Ag85A DNA疫苗加强的免疫策略后,能增强对小鼠的免疫效应,尤其是Thl型细胞免疫反应增强明显,为进一步在动物体内进行保护性效应试验的研究提供了实验依据。