S-O_2-1菌苗诱生干扰素的研究 Ⅰ小鼠体内诱生

本文研究了免疫增强剂——S-O_2-1菌苗在小鼠体内诱生干扰素的能力。结果表明,给(57BL/6小鼠注射菌苗1.5×1O~9菌/只后,血清中干扰素活性明显升高,于尾静脉注射后2小时,干扰素滴度达高峰(9.06±0.34log_2U/ml)。该干扰素具有小鼠I-型干扰素相同的理化特性。实验还表明,由S-O_2-1菌体提取的核糖体和脂多糖(LPS)亦均具有诱生干扰素的活性。因此我们推测,s-O_2-1菌苗诱生干扰素的能力可能对其抗肿瘤活性的发挥起一定的怍用。     

婴儿白细胞体外诱生干扰素水平的研究

本文观察脐血、婴儿(8～10月龄)及成人静脉血白细胞经不同诱生剂作用后,产生干扰素的水平。发现人出生时诱生α干扰素的水平随诱生剂不同而有所差异,NDV诱导脐血产生的α干扰素效价与成人血相当;但SPA菌体诱导的脐血α干扰素(4.36±2.45log_2u/50μl)明显低于成人者(9.73±1.93log_2u/50μl)。PHA或ConA诱导脐血产生的γ干扰素量都明显低于成人水平。于8～10月龄时SPA菌体诱生α干扰素水平已无异于成人,而PHA或ConA作用下γ干扰素的效价仍逊于成人。这种干扰素,特别是γ干扰素,产生不足可能是新生儿易患严重胞内病原体感染的一个重要的免疫因素。     

S-O_2-1菌苗对小鼠淋巴细胞的作用

动物实验和初步临床应用表明有抑制肿瘤生长作用的S-O_2-1菌苗,在体外能直接刺激小鼠脾细胞淋巴细胞的增殖,并协同增强脾细胞的Con A增殖反应、S-O_2-1菌苗诱导和增强小鼠腹腔粘附细胞的抑制活性,后者表现在直接抑制脾细胞和肠系膜淋巴结细胞对Con A和S-O_2-1菌苗的增殖反应。腹腔转移菌苗活化的腹腔粘附细胞能抑制受体小鼠对SRBC的PFC应答。提示:S-O_2-1菌苗诱导增强巨噬细胞的抑制活性,以致间接抑制淋巴细胞的免疫应答。     

我国厌氧棒菌菌苗在小鼠体内抗流感病毒、诱生干扰素及调节免疫的作用

我国分离的厌氧棒菌制成的菌苗北京7903和上海7503两批样品,静脉注入小鼠20mg/kg一次,对6～48小时后经静脉感染流感病毒所致死亡有保护作用,可减少小鼠死亡率。最佳保护时间为注射后24小时,保护率87.6～100％。静脉注射菌苗后6小时出现血清干扰素,高峰达320～640u/ml,18小时降低,持续24小时以上,其性质可能为γ-干扰素。静脉注射菌苗6～72小时间,激活巨噬细胞活性,高峰时间在注射后24小时。腹腔巨噬细胞吞噬鸡红细胞的％和吞噬指数,比对照组高,P<0.01。静脉注射菌苗对流感病毒抗体生成有促进作用,可使血清流感病毒血凝抑制抗体增高3～4倍。腹腔及皮下注射菌苗保护率低,腹腔注射后未测出血清干扰素。 小鼠静脉注射棒菌菌苗后抗流感病毒的最佳保护时间,与血清干扰素高峰时间不一致,但与腹腔巨噬细胞吞噬功能高峰一致。给小鼠腹腔注射石英粉尘,破坏巨噬细胞吞噬功能,也降低菌苗对流感病毒感染的保护作用,但对血清干扰素滴度无显著影响,提示厌氧棒菌菌苗抗流感病毒作用,与激活巨噬细胞的关系密切。     

S-O_2-1菌苗对S_(180)肿瘤细胞DNA合成的抑制作用

用小鼠 S_(180)肿瘤细胞作为靶细胞,以~3H-TdR 的掺入抑制百分率作为抗肿瘤活性的指标,在体外培养条件下,S-O_2-1 菌苗能刺激正常小鼠及带瘤小鼠的免疫活性细胞,释放对 S_(180)肿瘤细胞 DNA 合成有明显抑制作用的可溶性物质。实验提示,该类免疫因子的产生能增强免疫细胞对肿瘤细胞的细胞毒效应。同时发现 S-O_2-1菌苗本身对 S_(180)肿瘤细胞还具有直接杀伤作用。     

近红外信息辐照对大鼠细胞免疫功能的影响

本文观察近红外信息辐照对大鼠细胞免疫功能的调整作用。大鼠30只分为辐照组(每天辐照1h,治疗14～27天)与空白对照组(不辐照)。检测两组ConA诱生大鼠脾淋巴细胞产生IL-2活性,分别为335.07±122.77U/ml和211.01±118.01U/ml(P<0.01);检测脾脏自然杀伤细胞(NK)活性为70.80±29.85％和53.40±19.75％(P>0.05);ADCC效应为49.42±23.16％和27.34±10.12％(P<0.05)。结果表明,辐照有增强机体免疫功能的作用。     

慢性乙型肝炎病人单个核细胞γ干扰素的诱生和T淋巴细胞亚群的研究

本文对28例慢乙肝病人的外周血单个核细胞进行ConA诱生γ干扰素的测定,同时用T细胞单克隆抗体系统检测其T细胞亚群。结果发现,慢乙肝病人ConA诱生的γ干扰素效价为7.858±1.483log2U/ml,而正常对照组为7.839±0.621log2U/ml,两组间无显著性差异,但在慢乙肝病人组内个体间IFN-γ的效价差别显著大于对照组(P<0.05).慢乙肝病人组T细胞亚群的T_4/T_3比例与对照组无显著差异。但IFN-γ反应低下组的5人均有T_4/T_3比例异常。在28例病人中有14例T_4/T_3比例异常(高于或低于正常),且20例HBeAg阳性病人中有12例T_4/T_3比例异常;8例HBeAg阴性病人中仅2例T_4/T_3比例异常。     

胎盘转移因子对荷瘤小鼠脾脏IL－2诱生水平和外周血TNF活性的调节作用

应用ＣＴＬＬ２细胞短期培养３Ｈ－ＴｄＲ掺入法和Ｌ９２９细胞结晶紫染色法，观察ＮＩＨ小鼠接种Ｓ１８０实体瘤细胞前后，分别注射ＨＰ－ＴＦ０．５ｍｌ／鼠连续２０天，其脾细胞ＩＬ－２诱生水平和外周血ＴＮＦ活性的变化。结果显示，预防组和治疗组小鼠脾细胞ＩＬ－２诱生水平（ｃｐｍ值）分别为２１２８．７２±１６４．４２、３２９５．９６±２３０．７１，较荷瘤对照组（１２６０．４３±１４５．５７）明显提高（Ｐ＜０．００１）；而外周血ＴＮＦ活性（％）分别为４５．００±５．１８．５２．２９±５．２１，与荷瘤对照组（２５．８３±５．５２）比较有非常显著性差异（ｐ＜０．００１）。两组的抑瘤率分别为６８．８８％和６５．８２％。提示ＨＰ－ＴＦ能显著提高荷瘤小鼠脾细胞ＩＬ－２诱生水平和外周血ＴＮＦ活性，且能明显抑制Ｓ１８０实体瘤细胞在小鼠体内的生长。木研究结果有助于对ＨＰ－ＴＦ提高荷瘤机体细胞免疫功能的机理的探讨，并为临床应用ＨＰ－ＴＦ治疗肿瘤患者提供了实验依据。     

在人扁桃体细胞中诱生γ干扰素

本文初步研究了用人扁桃体细胞诱生γ干扰素及其影响因素。试验结果显示:细胞浓度在5×10~7/ml时产生的γ干扰素效价最高;PHA、PWM、ConA等T细胞促分裂剂诱生γ干扰素的适宜浓度范围分别为50～400μg/ml,2.5～40μg/ml,5～80μg/ml;诱生时间均以72小时为宜;2-ME对人γ干扰素的产生没有影响。本文还将人扁桃体细胞和外周血白细胞对4种γ干扰素诱生利(PHA、PWM、ConA、SEA)和3种α干扰素诱生剂(NDV、SPA菌体、CP)的反应进行了比较。     

质粒DNA促进HBsAg-抗HBs复合型疫苗诱生的免疫应答的研究

目的　研究HBsAg 抗HBs 质粒DNA复合型疫苗的免疫原性及其诱生细胞免疫应答的类型。方法　分别以HBsAg、HBsAg 抗 HBs(IC)、pI/AmpHBs、IC pI/AmpHBs及IC pI/Amp免疫小鼠 ,检测抗 HBs的效价 ,分析抗 HBsIgG亚类 (ELISA) ;取免疫小鼠脾细胞 ,体外抗原刺激 ,用竞争性RT PCR方法检测IFN γ及IL 4mRNA转录水平。结果　IC pI/AmpHBs诱生的抗 HBs效价明显高于IC或pI/AmpHBs单独免疫组 ,其IgG2a/IgG1比值高于IC免疫组 ,而低于pI/AmpHBs免疫组。IC pI/AmpHBs免疫小鼠脾细胞在HBsAg刺激下 ,IFN γmRNA转录水平明显高于其他免疫组 ,其IFN γmRNA的T/C(目的片段Target/竞争片段Competitor)比值为IC免疫小鼠脾细胞的 10倍 ;IC pI/AmpHBs免疫小鼠脾细胞IL 4mRNA转录水平亦高于其他免疫组 ,其IL 4mRNA的T/C比值为IC免疫小鼠脾细胞的 2倍。结论　HBsAg 抗HBs 质粒DNA复合型疫苗在增强体液免疫应答的同时可诱导脾细胞IFN γ的表达水平增高。     

Production of lymphokines and interferon by immune cells involved in recovery of mice from herpes simplex virus type 2 hepatitis

Adoptive transfer of spleen cells from mice 4 days after infection with herpes simplex virus type 2 (HSV-2) reduced the virus titer in the liver of recipient mice infected 24 h before transfer. Macrophage chemotactic factor (CF) and macrophage migration inhibition factor (MIF) were produced by day 3 of infection in spleen cell cultures stimulated with HSV-2, but not with control antigen, i.e. 1 day before the cells are active in adoptive transfer. Interferon was produced in cultures established throughout the infection but not in normal spleen cells. From days 1 to 5 of infection interferon was produced irrespective of in vitro restimulation, although the highest amounts were always produced after stimulation with the specific antigen. Spleen cells from mice infected for 6 days produced interferon only when stimulated with HSV-2. The cells from 6-day-immune mice active in adoptive transfer and CF and MIF production were found to be Thy 1+, Ig- and Lyt2-. Both Thy 1+ and plastic adherent cells were necessary for interferon production, whereas Ig+ and Lyt2+ cells did not produce interferon. The interferon was acid stable and neutralized by antiserum against alpha/beta-interferon and thus has the characteristics of alpha-interferon. The data indicate that a delayed type hypersensitivity reaction with lymphokine-induced macrophage recruitment into infectious foci may be a central feature of the recovery process in HSV-2-induced hepatitis. A possible role of interferon produced by the accumulated cells needs further investigation.     

高复制HBV转基因小鼠模型对抗乙型肝炎病毒药物的效应研究

 目的：研究高复制HBV转基因小鼠模型对抗乙肝病毒药物的效应评价。 方法： 选用抗乙肝药物拉咪呋啶、大剂量重组乙肝蛋白疫苗、α-1b型干扰素和RNA干扰在转基因小鼠进行药效及作用机制评价。 结果：拉咪呋啶、重组乙肝疫苗、α-1b型干扰素均可使HBV转基因小鼠血清中HBV DNA滴度显著降低。其中后两者还可提高机体脾细胞IL-2和IFN-γ的水平及使分泌IFN-γ脾细胞Elispot斑点数明显增加。将RNA干扰表达载体pU6-siHBV质粒尾静脉注入小鼠体内。注射后5 d血清HBsAg下降56.7%,抑制作用持续14 d。肝脏免疫组化显示HBcAg阳性细胞明显减少，但血清HBV DNA定量无明显降低。 结论： 本近交系高复制HBV转基因小鼠模型对抗乙肝药物药效学评估是可靠、可行的。     

Phorbol ester replacement of helper cell and interleukin 2 requirements in gamma interferon production.

Immune interferon (IFN-gamma) production by mitogen-stimulated C57BL/6 mouse spleen cells required both Lyt 1+,2- and Lyt 1-,2+ lymphocytes. The Lyt 1-,2+ cells were the IFN-gamma producers, whereas the Lyt 1+,2- cells were the helpers. Interleukin 2 mediated the Lyt 1+,2- cell help. Phorbol myristic acetate (10 ng/ml) completely replaced Lyt 1+,2- helper cell or interleukin 2 requirements in IFN-gamma production. The phorbol ester exerted its helper effects by direct interaction with IFN-gamma-producing Lyt 1-,2+ cells and not through stimulation of production of interleukin 2 by Lyt 1+, 2- cells. Suppressor cell effects in IFN-gamma production were also completely abrogated by phorbol myristic acetate. The data suggest a mechanism for phorbol myristic acetate regulation of IFN-gamma production at the cellular level.     

幽门螺杆菌Lpp20核酸疫苗的构建及其免疫活性的初步研究

目的 构建幽门螺杆菌脂蛋白Lpp20基因的真核表达载体pcDNA3.1(+)/Lpp20,并在HeLa细胞中进行表达.通过肌肉注射免疫C57BL/6小鼠,观察其诱导小鼠产生的体液免疫和细胞免疫应答水平.方法 用PCR法扩增Lpp20全基因,再将Lpp20基因克隆至pcDNA3.1(+)真核细胞表达载体构建pcDNA3.1(+)/Lpp20重组体,观察其在HeLa细胞中的表达.将核酸疫苗PcDNA3.1(+)/Lpp20、对照空质粒pcDNA3.1(+)及PBS分组通过肌肉注射免疫6周龄C57BL/6小鼠.隔2周免疫一次,共免疫4次.间接ELISA法测定小鼠血清中抗Lpp20 IgG抗体水平,双抗体夹心ELISA法检测脾淋巴细胞培养上清中IFN-γ水平,MTT比色法检测脾淋巴细胞增殖反应.通过PCR法检测小鼠肌细胞中Lpp20基因的存在.结果 小鼠接种pcDNA3.1(+)/Lpp20核酸疫苗后能产生特异性IgG抗体,6周后ELISA测定血清抗体A450值为0.74,效价为1:1024.核酸疫苗免疫组小鼠脾淋巴细胞经特异性抗原刺激后,培养上清中IFN-γ含量明显升高[(410.36±56.23)ps/ml],与空质粒组[(25.26±10.85)pg/ml]之间差异有统计学意义(P<0.01).脾淋巴细胞增殖反应测定,核酸疫苗组小鼠脾淋巴细胞经特异性抗原刺激后,刺激指数(2.37±0.22)明显高于空质粒组(1.53±0.47)和PBS组(1.20±0.13),P<0.01.PCR检测Lpp20基因可在小鼠肌细胞中存在.结论 成功构建了pcDNA3.1(+)/Lpp20核酸疫苗,且其在小鼠体内可诱导较强的特异性体液免疫和细胞免疫应答.为进一步研究该疫苗的免疫保护作用提供实验依据.     

Protection of mice against infection with mouse hepatitis virus type 3 by injection of silica

Injection of silica did not brake the resistance against MHV3 conferred to C57BL/6 mice by injection of C. parvum. However, silica itself had a marked protective effect against MHV3 infection that was maximal when injecting 1 mg 2 hrs before virus infection. The protective effect of silica was observed in a number of inbred mouse strains that differ in their relative resistance to MHV3 infection. No viral titers were observed in the spleen and liver of mice which had received MHV3 plus silica, whereas high titers were observed in the virus-infected controls. Injection of silica caused a marked decrease in the number of esterase-positive macrophages in the peritoneal wash-out population, that may be compatible with the possibility that the cause of the protection is the depletion of target cells for the viral infection. This latter effect, however, was short-lived and 24-48 hrs after injection of silica, high numbers of esterase-positive cells were again observed. This may explain why only little protection was observed when silica was administered 2 days before virus infection.     

Study on immunofunction and immunoregulation post newcastle disease vaccination of chickens infected with chicken anemia virus

Study on immunofunction and immunoregulation post newcastle disease vaccination of chickens infected with chicken anemia virus     

Interleukin-2-induced activation of natural killer activity in spleen cells from old and young mice.

Generation of natural killer (NK) activity in response to a partially purified preparation of rat interleukin-2 (IL-2) was compared in spleen cells derived from young (8-10 weeks old) and old (greater than 2 years old) female C57BL/6 mice. Significant NK activation was observed in both young and old mouse spleen cells incubated with 100 U IL-2/ml for 1-4 days, but the levels of cytotoxic activity generated in old mouse spleen cells was always lower than those of similarly treated young mouse spleen cells. Differences in IL-2-induced NK activation in old and young mouse spleen cells was obtained irrespective of the concentration of IL-2 used (25-400 U/ml). Quantitative comparisons indicated that old spleen cells activated by 3 day incubation with IL-2 acquired about two-fold higher NK activity than fresh young mouse spleen cells but still had only one-fourth of the levels of NK activity attained by IL-2-activated young mouse spleen cells. Cytotoxic activity of IL-2-activated young or old mouse spleen cells were totally abrogated by anti-asialo GM-1 antiserum + C but not by anti-Ly-2 + C treatment, indicating that the activated cytotoxic cells fell in the NK cell category. An analysis of NK precursor (NK-p) frequency by limiting dilution assay indicated that the NK-p frequency was about 4-fold higher in young as compared to old mouse spleen cells. The level of cytotoxic activity attained per NK-p cell was not significantly different for NK-p cells of old or young mice.     

High rate of insufficient antibody titers after single-day immunization with human diploid-cell-strain vaccine against rabies

Summary We examined the possibility of simplifying the currently recommended immunization schedule against rabies. Four groups of 11 healthy volunteers were each immunized with either one, two, four, or eight doses of 0.1 ml human diploid cell vaccine (HDCV) intradermally on 1 single day. Antibody titers in serum were determined using a rapid fluorescent focus inhibition test. Geometric mean antibody titers 10, 30 and 90 days after immunization were all >0.5 IU/ml, which is considered protective. A dose-proportional increase in the geometric mean antibody titer was observed for all four groups. However, at each dose level, at least 1 of the 11 volunteers on day 30 and 2 of the 11 volunteers on day 90 had insufficient antibody titers <0.5 IU/ml. Single-day immunization against rabies with HDCV vaccine cannot be recommended because of the unacceptably high failure rate.Abbreviations DEV Duck-embryo-vaccine - GMT geometric mean titer - HDCV human diploid cell vaccine - IU international units     

Development of a live culture of cold-adapted reassortant influenza vaccines

Optimal conditions for the cultivation of the MDCK cell lines in the laboratory spinner or by using the Eagle-MEM with or without fetal serum were worked out. The cold-adapted reassortant vaccine strains of virus influenza A/H1N1, A/H3N2 and B are well replicated in the MDCK cells both in a monolayer and in the spinner by using the serum-free medium. A maximum virus titer depends on a multiplicity of infection used in a fetal medium and on the addition of trypsin. Under the optimal conditions, the titer of the studied cold-adapted reassortants, while using a serum-free medium, reaches as much as 10(9.0)-10(9.5) EID50/ml.