重组空肠弯曲菌PEB1蛋白的大量表达及其鉴定分析

目的 探讨基因工程菌PEB1 E.coli BL21(DE3)大量表达重组空肠弯曲菌PEB1蛋白的稳定性及其纯化方法并鉴定.方法 诱导培养工程菌E.coli BL21(DE3)表达重组空肠弯曲菌PEB1蛋白,经镍琼脂糖凝胶层析柱纯化,透析后通过SDS-PAGE分析纯化后的重组空肠弯曲菌PEB1蛋白分子量大小.结果 挑取的PEB1 E.coli BL21(DE3)单个菌落,经总共17 h的扩增,细菌总数可达1×1010以上.在咪唑浓度为100,200,300 mmol/L时,洗脱液中重组蛋白的含量较高,分子量在31KD左右.结论 基因工程菌PEB1 E.coli BL21(DE3)可大量稳定表达重组空肠弯曲菌PEB1蛋白,经鉴定与空肠弯曲菌外膜蛋白-PEB1蛋白分子量相同.     

HIV-1中国流行株gag-hIL-2重组痘苗病毒与pIRES-gag-hIL-2核酸疫苗联合免疫的实验研究

目的　将HIV 1中国流行株B亚型gag和hIL 2基因在天坛株痘苗病毒中进行共表达 ,与核酸疫苗联合免疫 ,评价免疫效果 ,为新型艾滋病疫苗开发研制奠定基础。方法　将HIV 1中国流行株gag hIL 2基因片段插入到痘苗病毒载体pJ38启动子下游 ,经同源重组和血凝素阴性空斑筛选重组痘苗病毒 ,SDS PAGE、Westernblot检测目的蛋白。以重组病毒和核酸疫苗免疫BALB c小鼠 ,进行淋巴细胞转化实验及血清抗体的细胞免疫和体液免疫指标检测。结果　获得了重组痘苗病毒vJ38gag IL 2 ,具有更好的免疫原性 ,3rVV效果好于 2rVV ,以 2rVV DNA混合免疫效果最好。结论　重组痘苗病毒vJ38gag IL 2能够表达外源蛋白并诱导机体产生细胞免疫和体液免疫     

大气颗粒物宫内暴露对出生后幼鼠免疫功能的影响

目的观察小鼠胚胎大气颗粒物(PM)亚急性宫内暴露对出生后幼鼠免疫功能的影响。方法 ICR雌、雄小鼠按2∶1合笼,将妊娠的雌鼠随机分为对照组(A)和低(B)、中(C)、大(D)、超大(E)剂量PM暴露组(n=8～11),分别向A～E组小鼠咽后壁滴注载液(PBS)或0.09、0.28、1.85、6.92μg/μl的PM标准品SRM1649a混悬液各30μl,从妊娠d0至d19,每隔3 d暴露一次,共7次。幼鼠于出生后30 d龄处死,检测脾淋巴细胞增殖活性、血浆和脾组织IFN-γ和IL-4的含量及脾GATA-3和T-bet mRNA的表达,观察脾脏和胸腺的脏体系数及病理改变。结果幼鼠脾脏系数及胸腺系数,各组间比较差异均无统计学意义(P>0.05)。脾淋巴细胞增殖活性,C(0.8852±0.0413)、D(0.2172±0.0523)、E(0.0543±0.0292)组低于A组(0.9465±0.0564),差异有统计学意义(P<0.05)。血浆和脾组织IFN-γ含量,C[(20.96±1.59)pg/ml、(149.79±7.16)pg/ml]、D[(18.27±1.57)pg/ml、(125.88±9.81)pg/ml]、E[(9.32±2.62)pg/ml、(87.15±9.76)pg/ml]组低于A[(24.69±2.23)pg/ml、(175.52±11.39)pg/ml]、B[(25.44±1.85)pg/ml、(171.93±11.03)pg/ml]组,差异均有统计学意义(P<0.05);血浆和脾组织IL-4含量,C[(20.07±1.73)pg/ml、(158.65±9.43)pg/ml]、D[(35.24±4.14)pg/ml、(189.93±14.19)pg/ml]、E[(62.59±8.60)pg/ml、(256.04±17.07)pg/ml]组高于A[(13.89±2.16)pg/ml、(108.10±5.51)pg/ml]、B[(14.48±2.08)pg/ml、(110.97±5.99)pg/ml]组,差异均有统计学意义(P<0.05)。T-bet mRNA表达C(0.35±0.10)、D(0.26±0.07)、E(0.15±0.03)组低于A(0.49±0.15)、B(0.47±0.12)组,GATA-3 mRNA表达C(0.55±0.16)、D(0.63±0.05)、E(0.81±0.18)组高于A(0.37±0.06)、B(0.40±0.13)组,差异均有统计学意义(P<0.05)。E组胸腺和脾脏组织结构损伤较重,C、D组病变程度次之,B组变化轻微。结论妊娠期宫内PM暴露可影响胎鼠免疫系统的正常发育,造成出生后幼鼠免疫功能紊乱,其作用机制可能与上调GATA-3 mRNA和下调T-bet mRNA的表达、诱导Th1向Th2偏移有关。     

增强核酸疫苗免疫应答的策略及方法

核酸疫苗又称DNA疫苗,是指将编码某种目的抗原蛋白的外源基因插入真核细胞表达质粒,直接导入动物体细胞内,并通过宿主细胞的表达系统合成抗原蛋白,诱导宿主产生对该抗原蛋白的免疫应答,从而达到预防和治疗疾病的目的.核酸疫苗不仅能刺激机体产生特异性的体液免疫,还能诱导产生特异性的具有细胞毒杀伤性功能的CTL效应（细胞免疫）,并可直接清除带有目的抗原的靶细胞,因此对病毒、细胞内寄生的细菌和寄生虫所引起的传染病具有广阔的治疗前景[1-6].     

HCV核心区DNA疫苗的构建及初步鉴定

目的 构建HCV核心蛋白DNA疫苗,观察其诱导BALB/C小鼠产生体液免疫应答的效果,为研制应用于人类的HCVDNA疫苗提供实验依据.方法 将HCVC基因片段插入真核表达载体pCDNA3.1(+),构建真核表达载体pCDNA3.1HCVcore,并用酶切分析鉴定;肌注BALB/C小鼠,以ELISA法检测小鼠血清HCV抗体.结果 经酶切分析证实重组质粒pCDNA3.1HCVcore构建正确;9只小鼠经3次DNA免疫后均出现抗HCV抗体.结论 成功地构建出HCVC区DNA疫苗,并可诱导BAIB/C小鼠体液免疫应答.     

再生障碍性贫血患者血清中IL-18及IL-18BP的表达水平及其临床意义

目的 探讨白介素18(IL-18)及IL-18结合蛋白(IL-18BP)在再生障碍性贫血(AA)患者血清中的表达及其临床意义.方法 采用ELISA法检测29例AA患者和22名正常人血清IL-18和IL-18BP水平;实时定量RT-PCR方法检测IL-18及IL-18BP的mRNA表达.结果 AA患者血清IL-18水平为(365.5±160.6) pg/ml,较正常对照组[(175.9 ±92.8) pg/ml]明显升高(P＜0.01);重型AA患者的IL-18水平[(441.3±116.9) pg/ml]较慢性AA患者的IL-18水平[(326.4±167.0)pg/ml]升高更为明显(P＜0.05);AA患者血清IL-18BP[( 1788.6±523.8)pg/ml]较正常对照组[(1083.6±489.6)pg/ml]明显升高(P＜0.05),IL-18/IL-18BP比值在AA患者明显升高.RT-PCR结果证实AA患者IL-18和IL-18BP较正常对照组均明显升高,IL-18/IL-18BP比值明显升高.结论 IL-18及IL-18BP在AA患者的免疫紊乱和发生、发展中起着一定的作用,重塑IL-18/IL-18BP平衡可能是AA治疗一个新的策略和方向.     

中性粒细胞CD11b、1,25-(OH)2 VitD3和γ干扰素在小儿轮状病毒性肠炎中的表达及与免疫功能的关系

目的探究中性粒细胞CD11b、1,25-(OH)_2Vit D_3、γ干扰素(INF-γ)在小儿轮状病毒性肠炎(PRE)中的表达及与免疫功能的关系。方法采用前瞻性研究方法,选取2017年9月至2019年9月琼海市人民医院收治的90例PRE患儿作为观察组,另选择同期健康体检儿童104例作为对照组。检测对比两组患儿血清CD11b、1,25-(OH)_2Vit D_3、INF-γ水平,Logistic回归分析探究各血清指标与PRE的关系。根据脱水程度分为轻度患儿28例、中度患儿42例、重度患儿20例,比较不同病情患儿血清CD11b、1,25-(OH)_2Vit D_3、INF-γ水平、细胞免疫指标(CD3~+、CD4~+、CD4~+/CD8~+)、体液免疫指标[免疫球蛋白A(Ig A)、Ig G、Ig M],采用Pearson相关性分析评价血清CD11b、1,25-(OH)_2Vit D_3、INF-γ与细胞免疫、体液免疫的关系。结果观察组患儿的血清CD11b[(86.24±7.16)%]、INF-γ[(3.62±0.72)pg/ml]高于对照组[(69.35±4.42)%、(1.15±0.20)pg/ml],1,25-(OH)_2Vit D_3[(20.75±5.03)pg/ml]低于对照组[(35.66±7.14)pg/ml],差异具有统计学意义(P0.05);Logistic回归分析显示,血清CD11b、1,25-(OH)_2Vit D_3、INF-γ与PRE显著相关,差异具有统计学意义(P0.05);重度患儿血清CD11b[(92.71±6.35)%]、INF-γ[(5.23±1.04)pg/ml]高于中度[(86.23±5.79)%、(3.72±0.65)pg/ml]、轻度患儿[(81.63±4.90)%、(2.32±0.49)pg/ml],1,25-(OH)_2Vit D_3[(18.10±2.16)pg/ml]低于中度[(20.42±2.77)pg/ml]、轻度患儿[(23.14±3.05)pg/ml],差异具有统计学意义(P0.05);重度患儿外周血CD3~+[(48.81±3.44)%]、CD4~+[(22.39±1.80)%]、CD4~+/CD8~+(0.52±0.04)、IgA[(0.92±0.21)g/L]、IgG[(6.48±1.02)g/L]、IgM[(0.48±0.10)g/L]低于中度[(51.06±4.02)%、(23.97±2.25)%、(0.58±0.05)、(1.18±0.26)g/L、(7.75±1.44)g/L、(0.61±0.13)g/L]及轻度患儿[(54.11±4.50)%、(26.08±2.73)%、(0.63±0.07)、(1.47±0.30)g/L、(9.22±1.93)g/L、(0.74±0.15)g/L],差异具有统计学意义(P0.05);血清CD11b(r_1=-0.400;r_2=-0.492;r_3=-0.431)、INF-γ(r_1=-0.406;r_2=-0.529;r_3=-0.577)与CD3~+、CD4~+、CD4~+/CD8~+呈负相关,1,25-(OH)_2Vit D_3(r_1=0.514;r_2=0.568;r_3=0.502)与CD3~+、CD4~+、CD4~+/CD8~+呈正相关(P0.05);血清CD11b(r_1=-0.519;r_2=-0.518;r_3=-0.506)、INF-γ(r_1=-0.616;r_2=-0.660;r_3=-0.692)与Ig A、Ig G、Ig M呈负相关,1,25-(OH)_2Vit D_3(r_1=0.669;r_2=0.551;r_3=0.663)与Ig A、Ig G、Ig M呈正相关(P0.05)。结论中性粒细胞CD11b、1,25-(OH)_2Vit D_3、INF-γ与PRE发生及其免疫功能显著相关,可作为预测疾病发生、评估免疫紊乱及病情程度的重要因子。     

病毒性肝炎核酸疫苗研究的现状与问题

核酸疫苗又称ＤＮＡ疫苗 ,是指将含有编码某种目的抗原蛋白基因序列的真核细胞表达质粒作为疫苗 ,直接接种机体 ,在体内合成抗原蛋白 ,诱导宿主产生对该抗原蛋白的免疫应答 ,达到免疫的目的。乙肝ＤＮＡ疫苗的抗原基因片段 ,主要是编码表面抗原的Ｓ区 ,也可包括ｐｒｅＳ1 、ｐｒｅＳ2 区。此外 ,Ｇｅｉｓｓｌｅｒ等〔1〕将包含ｐｒｅＳ1 、ｐｒｅＳ2 的Ｓ区基因以及ＨＢｃＡｇ编码基因的重组质粒接种小鼠 ,诱导产生了高滴度抗 -ＨＢｓ及抗 -ＨＢｃ。93 %的小鼠产生了强烈的对病毒蛋白的ＣＤ4+ Ｔ细胞介导的体液免疫的ＣＤ8+ Ｔ细胞介导的Ｃ…     

PRMT1对哮喘小鼠的影响以及作用机制分析

目的探讨蛋白质精氨酸甲基转移酶1(PRMT1)对哮喘的影响及作用机制。方法构建小鼠哮喘模型,然后分别利用氨茶碱(AMI)和磷酸盐缓冲液(PBS)处理两组小鼠,收集并用ELISA试剂盒检测小鼠支气管肺泡灌洗液中炎症因子白细胞介素(IL)-1β、IL-4和肿瘤坏死因子α(TNF-α)的水平。运用Bradford方法检测小鼠支气管肺泡灌洗液中蛋白含量。Western blotting检测肺脏组织中PRMT1的含量以及NF-κB信号通路下游分子P65的变化。结果哮喘模型组IL-4,IL-1β和TNF-α的水平较对照组明显升高(25.48±4.96 pg/ml vs.4.01±0.56 pg/ml,23.48±6.11 pg/ml vs.3.77±0.27 pg/ml,29.45±6.98 pg/ml vs.5±1.84 pg/ml);经AMI处理的哮喘模型组IL-4,IL-1β,TNF-α的水平较PBS处理的哮喘模型组明显降低(12.14±3.87 pg/ml vs.25.48±4.96 pg/ml,11.95±2.55 pg/ml vs.23.48±6.11 pg/ml,16.81±7.49 pg/ml vs.29.45±6.98 pg/ml),差异有统计学意义(P0.05)。哮喘模型组小鼠支气管肺泡灌洗液中蛋白渗出量较对照组明显增加,差异有统计学意义(P0.05),同时对比PBS处理组和AMI处理组发现,AMI处理后小鼠支气管肺泡灌洗液中蛋白渗出含量明显减少(P0.05)。Western blotting检测了四组小鼠肺脏组织中PRMT1和P65的蛋白含量,结果显示在患有哮喘的小鼠中PRMT1和P65的表达水平较对照组显著升高,无论是对照组还是哮喘模型组,AMI处理后的PRMT1和P65的表达水平较PBS处理组降低。结论 PRMT1通过上调NF-κB信号通路促进肺脏炎性因子IL-1β、IL-1和TNF-α的产生以及蛋白渗出,诱导哮喘的发生。     

小鼠呼吸机相关肺损伤过程中免疫机制的初步研究

目的建立小鼠呼吸机相关性肺损伤模型,并初步探讨呼吸机相关性肺损伤的免疫损伤机制。方法小鼠呼吸机维持通气潮气量(Vt)12ml/kg,维持呼吸频率130次/min,持续通气时间3h,流式细胞仪检测小鼠外周血及肺组织中CD11b+细胞、粒细胞、单核细胞等的比例,流式细胞仪CBA方法检测小鼠血浆中IL-6、TNF-α、IFN-γ、巨噬细胞趋化蛋白-1(MCP-1)、IL-12、IL-10等细胞因子浓度。结果机械通气3h后小鼠心率[(521±13.40)次/min]较对照组心率[(431±20.14)次/min]明显升高(P<0.05)、R波振幅(0.44±0.0194)较对照组(0.47±0.0239)明显降低(P<0.05);机械通气组小鼠CD11b+髓系细胞比例[CD11b+CD19-/CD45+,外周血(33.2±7.75)%,肺组织(37.9±8.51)%]较对照组[外周血(22.9±5.07)%,肺组织(16.2±3.42)%]明显上升(P<0.05),升高的髓系细胞以粒细胞为主,单核细胞比例变化不明显;血浆中IL-6、TNF-α、MCP-1浓度分别为:(5.8±17.2)pg/ml、(16.8±2.16)pg/ml、(114.6±90.50)pg/ml,而对照组相应细胞因子的浓度均小于浓度标准曲线最小值(记为0pg/ml)。结论机械通气后小鼠出现明显缺氧的肺损伤表现,其血浆中炎性细胞因子IL-6、TNF-α、MCP-1浓度明显升高,肺组织的免疫损伤可能主要为髓系细胞中粒细胞介导。     

Enhanced induction of SARS-CoV nucleocapsid protein-specific immune response using DNA vaccination followed by adenovirus boosting in BALB/c mice

OBJECTIVE: To investigate immunogenicity in the induction of humoral and cellular immune responses to genetic vaccines of the recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV)-N gene expressing the same protein plasmid, pcDNA3.1-N, and replication-defective adenoviral vector, rAd-N, in a pcDNA3.1-N prime-rAd-N boost regimen and the reverse sequence in a rAd-N prime-pcDNA3.1-N boost regimen. METHOD: After the mice had been immunized intramuscularly and/or intraperitoneally with pcDNA3.1-N and rAd-N in prime-triple boost immunization, humoral and cellular immune responses were detected. RESULTS: After detection, different levels of anti-N humoral and cellular responses are shown compared to controls. The humoral immune response was induced more effectively by the DNA priming and recombinant adenovirus boosting regimen and the reverse sequence of heterogeneous combinations. There is a significant difference between heterogeneous and homologous vaccinations. However, the cytotoxic T lymphocyte (CTL) response was not significantly altered by the different prime-boost immunizations or the recombinant adenovirus of pcDNA3.1-N prime-rAd-N boost regimen alone, but lymphoproliferation and interferon-gamma (IFN-gamma) secretion were all enhanced by heterologous combination immunizations compared to homologous combinations. For the reverse sequence immunization regimen, lymphoproliferation, IFN-gamma and CTL responses were all significantly weaker compared with pcDNA3.1-N prime-rAd-N boost regimen. CONCLUSION: Taken together, of all the combinations, the prime-triple boost immunization of pcDNA3.1-N/pcDNA3.1-N/pcDNA3.1-N/rAd-N can effectively induce SARS-CoV-N-specific and strong humoral and cellular immune responses in mice. The present results suggest that DNA immunization followed by recombinant adenovirus boosting could be used as a potential SARS-CoV vaccine in the induction of an enhanced humoral and cellular immune response.     

DNA immunization of neonates induces immunity despite the presence of maternal antibody.

Neonatal animals were not considered as suitable vaccine recipients either because of immune immaturity or because passively delivered antibody interferes with immune induction. In this report, we evaluated the response of neonatal mice to immunization with naked DNA encoding a herpes simplex virus (HSV) protein, and determined if maternally derived HSV antibody interfered with immunogenicity. Our results show that neonatal mice develop effective humoral and T cell responses after immunization with either DNA or inactivated vaccines. The nature of the responses to HSV immunization, however, was more Th2-like in neonates than in adults. Whereas neonatal mice from HSV-naive mothers responded well to both DNA and inactivated vaccines, only DNA immunization induced effective immunity in neonates born to immune mothers. Our results indicate that DNA vaccines might provide a useful means of immunizing young animals that still possess high levels of potentially interfering maternal antibody.     

Update on antiviral DNA vaccine research (1998-2000)

DNA vaccines can induce protective cellular and humoral immune responses and have therefore been used during the last decade to develop vaccines against a variety of different pathogens. Because current antiviral vaccines predominantly generate humoral immunity, DNA immunization may be especially useful to provide long-term protection against viral diseases that also require cellular immunity (e.g. HIV). A significant number of articles published in the field of DNA vaccines are dealing with viral diseases, reflecting the need for better and alternative vaccination strategies against viruses. The success of DNA immunization depends on a variety of parameters (e.g. type of antigen, method of application and usage of adjuvants). Therefore, different strategies have been explored to modulate the induced immune response with respect to the requirements necessary to protect against a specific pathogen (e.g. induction of mucosal or cell-mediated immunity). The following article provides an update on different aspects of antiviral DNA vaccine research that have previously been reviewed by others.     

2004-2007年浙江省麻疹疫苗质量及免疫成功率监测结果分析

目的分析2004-2007年浙江省麻疹高发原因。方法对2004-2007年浙江省麻疹疫苗质量及免疫成功率监测结果进行分析。麻疹疫苗质量监测采用定点和线性两种方式,免疫成功率监测通过对比8月龄儿童接种前后麻疹抗体水平的变化进行。结果从浙江省疾病预防控制中心冷库采集的31个批次的麻疹疫苗均合格,效价为（3.16±0.25）logTCID50/0.1ml;从浙江省市、县、乡各级监测点所采集的47份麻疹疫苗均合格,效价为（3.04±0.32）logTCID50/0.1ml,且各级监测点麻疹疫苗效价差异无统计学意义。8月龄接种1剂次麻疹疫苗可使95%的儿童获得保护性抗体,免疫前抗体水平越高免疫后阳转率越低。结论2004-2007年浙江省应用的麻疹疫苗处于保护效价水平,且初免效果良好,尚不能认为是近年麻疹高发的主要原因。需继续开展含麻疹成分疫苗的质量和免疫成功率监测。     

DNA vaccination for HIV-1 and SIV

Control of the worldwide AIDS epidemic will only be achieved with a safe and effective prophylactic HIV-1 vaccine. DNA vaccination has recently emerged as a promising vaccine modality that can elicit both humoral and cellular immune responses. HIV-1- and SIV-specific immune responses have been elicited by DNA vaccines in both mice and nonhuman primates. However, these immune responses have not been capable of protecting nonhuman primates against pathogenic AIDS virus challenges. A number of approaches are therefore being investigated to augment DNA vaccine-elicited immune responses.     

丙型肝炎病毒包膜区基因DNA免疫的研究

目的:构建丙型肝炎病毒包膜区基因(E1、E2)真核表达载体,命名为PCI-neo-E1和E2。方法:将真核重组体转染NIH-3T3细胞中,利用RT-PCR方法鉴定相应的基因片段,显示获得了相应的稳定转染的细胞克隆。提取转染细胞蛋白,进行WesternBlot分析。结果:HCVE1、E2基因在真核细胞中获得有效表达。将所构建的PCI-neo-E1、E2免疫Balb/C小鼠,获得特异的抗体反应。结论:通过HCV包膜区DNA免疫的研究表明能激发特异的免疫反应,为进一步进行HCV疫苗研究奠定基础。     

Immunogenicity and safety profiles of genetic vaccines against human Her-2/neu in cynomolgus monkeys

Her-2/neu is a well-characterized tumor-associated antigen, the overexpression of which in human carcinomas correlates with a poor prognosis. Here, we evaluated Her-2/neu-specific humoral and cellular immune responses in immunized monkeys after immunization with nonreplicating adenovirus (AdHM) expressing the extracellular and transmembrane domain of human Her-2/neu (HM) and/or naked DNA vaccine (pHM-hGM-CSF) expressing human granulocyte-macrophage colony-stimulating factor together with HM. Priming of monkeys with AdHM generated Her-2/neu-specific long-lasting antibody production. Furthermore, these Her-2/neu-specific antibodies produced by AdHM immunization, some of which shared epitope specificity with Herceptin, were able to induce antibody-dependent cellular cytotoxicity against Her-2-expressing target cells. Cellular immune responses were elicited in all monkeys immunized with Her-2/neu-expressing vaccine; interferon-gamma was secreted when these splenocytes were restimulated with Her-2/neu-expressing autologous cells, and immunization with AdHM induced Her-2/neu-specific lymphoproliferative responses. Further, immunization with pHM-hGM-CSF before AdHM immunization noticeably enhanced cytotoxic T-lymphocyte activity. In addition, we observed no abnormalities that would indicate that the genetic vaccines had toxic effects in the immunized monkeys. Thus, we can conclude that our genetic vaccines efficiently elicited Her-2/neu-specific humoral and cellular immune responses without causing severe adverse effects in nonhuman primates and that as such they warrant further clinical investigation.     

黄酮对免疫功能影响的体内研究

目的 通过动物实验探讨黄芪桂枝五物汤中总黄酮对机体免疫功能的影响,从而为临床合理应用及其相关产品开发提供理论基础.方法 取40只小鼠,经适应性喂养后,随机分为正常对照组,黄酮1组(低剂量),黄酮2组(中等剂量)和黄酮3组(高剂量),每组10只.总黄酮组小鼠每天灌服0.2 ml浓度为1 g/L,2 g/L,4 g/L的总黄酮溶液,正常对照组小鼠每天灌服0.2 ml生理盐水,10天后取样本测定小鼠各项免疫指标.①免疫器官:对脾脏和胸腺称重计算脾脏指数和胸腺指数;②固有免疫:检测小鼠巨噬细胞吞噬功能;③体液免疫:血凝法测定小鼠血清中溶血素含量;④细胞免疫:小鼠后足迟发型超敏反应强度.结果 总黄酮各组小鼠脾脏指数和胸腺指数均较对照组小鼠有明显提高(P＜0.01);总黄酮各组小鼠巨噬细胞吞噬率和吞噬指数明显高于对照组(P＜0.01);总黄酮各组小鼠左右后足质量差与对照组小鼠相比,有明显提高(P＜0.01);但总黄酮各组小鼠抗体积数与对照组相比差异无统计学意义(P＞0.05).结论 黄芪桂枝五物汤中总黄酮可以显著提高机体的固有免疫功能和细胞免疫功能,但对体液免疫功能的影响不明显.     

8 Br-cAMP enhances both humoral and cell-mediated immune responses induced by an HIV-1 DNA vaccine

From a series of preclinical studies and animal experiments, we have been able to demonstrate that DNA vaccines are a promising tool in strategies for protecting hosts from a variety of infectious diseases. Since the promoter activity of the human cytomegalovirus immediate-early promoter/ enhancer (CMV promoter) is known to be responsive to an elevation in the level of intracellular cAMP, we hypothesized that use of cAMP analogue (8-Bromo adenosine 3'5'-cyclic monophosphate, 8 Br-cAMP) would increase the level of transgene expression supported by the CMV, and enhance the ability of DNA vaccines to evoke an immune response against the transgene product in vivo. To evaluate this hypothesis, immune responses against HIV-1 envelope protein, gp160, an immunogenic HIV-1 component expressed under the control of the CMV promoter, were evaluated in BALB/c mice with or without stimulation by 8 Br-cAMP. DNA vaccine with 8 Br-cAMP was intramuscularly (i.m.) or intranasally (i.n.) administered to BALB/c mice twice on days 0 and 14. Regardless of which route was used, the combination increased the serum IgG antibody (Ab) titer, HIV-1-specific cytotoxic T lymphocyte (CTL) activity and the delayed-type hypersensitivity (DTH) response, compared with the effect of using the vaccine alone. When administered via the i.n. route, the combination also remarkably increased the titer of secretory IgA (sIgA). Moreover, it induced increased production of interferon-gamma with reduction in IL-4 synthesis, and decreased the ratio of serum IgG1/IgG2a. However, these enhancements were not observed when 8 Br-cAMP was coadministered with peptide vaccine or protein antigen. These data suggest that 8 Br-cAMP is able to enhance both humoral and cellular immune responses induced by the DNA vaccine. The induction of T helper type 1 (Th1) immunity against HIV-1 was also enhanced by coadministration of 8 Br-cAMP. A CAT assay study demonstrated that the adjuvant effect of 8 Br-cAMP may be due to the activation of the CMV promoter in the DNA vaccine. The virus challenge experiment in a mouse influenza model also proved our hypothesis.