乙型肝炎表面抗原对BALB/c小鼠细胞免疫应答的影响

目的研究不同来源乙肝表面抗原(HBsAg)对BALB/c小鼠的某些细胞免疫应答的影响.方法采用血源、CHO和酵母HBsAg分别免疫BALB/c小鼠,于免疫后不同时间制备脾脏单个核细胞(MNC),测定MNC对刀豆蛋白(ConA)和细菌脂多糖(LPS)的增殖反应性;采用绵羊红细胞(SRBC)作为致敏原,测定不同HBsAg免疫小鼠后迟发性超敏反应(DTH)的发生程度.结果不同来源HBsAg对ConA或LPS刺激的反应指数(SI)表现出不同的时间曲线,一般于免疫20?d和25?d时显著升高,其中血源HBsAg较高,酵母较低.H.M.-HBsAg在免疫5?d时即显出较高的SI值.CHO-HBsAg对SRBC诱导的DTH反应具有显著的抑制作用(P＜0.05).结论不同来源HBsAg诱导细胞免疫反应的类型和程度存在差异,CHO细胞来源的HBsAg对TH1细胞介导的DTH反应有抑制作用.     

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。     

不同来源乙肝表面抗原的细胞免疫学对比研究

目的　研究不同来源乙肝表面抗原诱导CD8 抗原特异性的细胞毒T淋巴细胞反应(CTL)的差异及T细胞产生细胞因子的能力 ,从而对各种来源乙型肝炎表面抗原 (HBsAg)的免疫原性有更充分的评价。方法　不同来源HBsAg分别免疫BALB c小鼠 ,于免疫后不同时间制备脾脏单个核细胞 (MNC) ,再以相应的HBsAg体外刺激 ,酶联免疫方法 (ELISA)测定细胞因子分泌水平 ;以Na2 5 1CrO4标记靶细胞 ,测定CTL反应活性 ;应用流式细胞仪分析T细胞亚群。结果　HM HBsAg免疫小鼠 10d后的脾细胞产生IFN γ水平最高 ,而在免疫后 2 0d和 2 5d ,IL 2的水平显著高于其它各组 ,CD3 分子脾淋巴细胞显著高于其它各实验组 (P <0 .0 5) ;plasma HBsAg免疫小鼠的脾淋巴细胞产生IFN γ和IL 2水平较低 ,但在免疫 2 5d对P815 HBs的杀伤性最强 ;HM HBsAg和SSC HBsAg免疫后CD4 CD8- 脾淋巴细胞比例显著增高 (P <0 .0 5) ;CD4- CD8 细胞所占百分比各组间及与空白对照组间差异无显著性 (P >0 .0 5)。结论　不同来源HBsAg诱导的细胞免疫反应类型和程度不同     

CpG ODN佐剂对乙肝疫苗抗原免疫原性的影响

目的探究CpG ODN佐剂对不同抗原含量乙肝疫苗免疫效果的影响。方法在体液免疫方面不同抗原浓度的CpG ODN与Al(OH)3佐剂乙肝疫苗免疫Balb/c小鼠,于免疫后第2、4、6、8、10周收集血清,检测小鼠体内相对效力ED50和血清中的anti-HBs抗体水平;在细胞免疫方面测定诱导产生IFN-γ水平及IgG2a应答水平。结果 CpG ODN与Al(OH)3佐剂可以有效协同HBs Ag诱导机体产生的抗体滴度达49 427 mIU/ml,抗体效价随时间延长而增加。乙肝抗原减半后双佐剂乙肝疫苗诱导机体产生的特异性抗体滴度可达41 225 mIU/ml,高于无CpG ODN佐剂的铝佐剂疫苗。在诱导细胞分泌方面,对照组诱导产生IFN-γ水平及IgG2a应答水平明显低于所有双佐剂疫苗组。结论 CpG ODN对HBsAg具有良好佐剂活性,并与铝佐剂有协同作用,二者联合应用可以降低乙肝抗原用量并提高疫苗免疫原性。     

弗氏佐剂与氢氧化铝佐剂对诱导小鼠获得性免疫应答作用的比较

为研究、比较不同佐剂对诱导小鼠产生获得性免疫应答的不同作用,以卵清白蛋白(OVA)为抗原,分别混合完全弗氏佐剂(CFA)或Al(OH)3佐剂,对C57BL/6小鼠进行常规免疫,采用流式细胞技术对细胞内细胞因子IFN-γ和IL-4进行检测;ELISA方法对特异性抗OVA抗体滴度及抗体亚型进行了检测。结果显示在免疫后CFA组产生以IFN-γ为主的细胞因子而Al(OH)3组产生以IL-4为主的细胞因子;两组中均产生特异性抗OVA IgG抗体,但CFA组以IgG2a亚型为主,而Al(OH)3组则以IgG1亚型为主,不产生IgG2a亚型抗体。实验表明,经CFA加抗原免疫后机体产生的免疫应答以Th1型细胞免疫为主,抗体类型为IgG2a;而Al(OH)3佐剂则诱导机体产生Th2型细胞免疫应答,抗体类型为IgG1。     

分子佐剂IL-17与IL-2对铜绿假单胞菌外膜蛋白OprI免疫效果的影响

目的探讨分子佐剂IL-17、IL-2对铜绿假单胞菌(Pa)外膜蛋白OprI免疫效果的影响。方法分别以IL-17、IL-2作为分子佐剂用Pa外膜蛋白OprI于第0、2、4周免疫雌性C57BL/6小鼠,同时设立单独的佐剂与蛋白对照组。末次免疫2周后,ELISA检测各组小鼠血清IgG、IgA以及IgG亚类水平;ELISA法检测小鼠脾细胞上清中Th1/Th17型细胞因子INF-γ、IL-2、IL-17以及Th2型细胞因子IL-4、IL-6水平;MTT法检测IL-17、IL-2分子佐剂对小鼠脾细胞增殖情况的影响,并以ConA作为阳性对照;末次免疫2周后Pa鼻内感染小鼠,感染后10 d计算各组小鼠死亡率并取小鼠肺组织检测菌体载量。结果 ELISA结果显示,OprI-IL-17、OprI-IL-2组小鼠血清中IgG、Ig A以及IgG亚类水平均显著高于各对照组(P0.05),且其中各实验组IgG2a/IgG1的比值均大于1;IL-17、IL-2佐剂能够显著提高小鼠脾细胞上清中Th1/Th17型细胞因子INF-γ、IL-2、IL-17水平(P0.05),而Th2型细胞因子IL-4、IL-6水平则无显著差异(P0.05);此外,OprI-IL-17、OprI-IL-2组的淋巴细胞增殖最为显著(P0.05);相较于各对照组,OprI-IL-17、OprI-IL-2组能够显著降低小鼠肺组织菌体载量以及小鼠死亡率(P0.05),保护效果显著提高。结论 IL-17、IL-2佐剂均能有效增强Pa外膜蛋白OprI的体液、细胞免疫水平以及抗感染保护作用。     

共刺激分子4-1BBL基因免疫对HBsAg核酸疫苗诱导小鼠特异性免疫应答的影响

目的 研究共刺激分子4-1BBL基因免疫对HBsAg核酸疫苗诱导小鼠特异性体液和细胞免疫应答的影响.方法 将HBV表面抗原核酸疫苗pcDS2单独或联合共刺激分子4-1BBL质粒肌肉注射免疫C57BL/6小鼠;ELISA法检测小鼠血清抗-HBs IgG及亚型IgG1和IgG2a;迟发型超敏反应(DTH)反应检测体内细胞反应;流式细胞仪检测CD4+ T淋巴细胞分泌IL-4和IFN-γ及CD8+T淋巴细胞分泌IFN-γ水平;流式细胞仪检测小鼠脾细胞HBsAg特异性体外细胞毒性T淋巴细胞杀伤作用(CTL).结果 与单纯免疫核酸疫苗pcDS2组比较,pcDS2和4-1BBL联合免疫组小鼠的抗-HBs水平显著提高,抗-HBs IgG亚类以IgG2a占优;免疫小鼠经HBsAg脚掌皮下刺激后,联合免疫组小鼠脚掌的厚度显著高于pcDS2组;联合免疫组CD4+T淋巴细胞的IL-4和IFN-γ表达水平及CD8+T淋巴细胞的IFN-γ表达水平显著升高;DNA疫苗免疫的各组小鼠,HBsAg特异性体外CIL杀伤作用高于对照组,其中联合免疫组小鼠的体外CTL杀伤作用最强.结论共刺激分子4-1BBL不仅能增强HBV DNA疫苗诱导特异性体液免疫应答,还能增强特异性型细胞免疫反应,尤其增强体内CIL的杀伤活性.     

IL-23、IL-27对RSV重组蛋白疫苗免疫原性的影响

目的:以编码IL-23和IL-27的真核表达质粒pcDNA3.1-IL-23(以下简称IL-23)和pcDNA3.1-IL-27(以下简称IL-27)为佐剂,与呼吸道合胞病毒(RSV)重组蛋白疫苗G1F/M2共免疫小鼠,观察IL-23和IL-27对疫苗的免疫原性的影响。方法:以IL-23、IL-27质粒和Al(OH)3为免疫佐剂,与G1F/M2共免疫BALB/c小鼠,末次免疫后10天杀死小鼠,用ELISA检测特异性IgG、IgG1和IgG2a水平;流式细胞术检测小鼠脾细胞CD4+、CD8+细胞的变化;用乳酸脱氢酶(LDH)释放法检测特异性小鼠脾细胞杀伤活性。结果:G1F/M2+Al(OH)3+IL-23和G1F/M2+Al(OH)3+IL-27可诱导高效价的IgG、IgG1、IgG2a抗体,且显著高于这三种佐剂单独使用诱导的抗体效价;流式细胞检测结果显示G1F/M2+IL-23和G1F/M2+Al(OH)3+IL-23刺激的CD4+T细胞和CD8+T细胞水平显著高于其他组;单独的IL-23或IL-27对G1F/M2诱导的脾细胞杀伤活性没有增强作用,而Al(OH)3+IL-23和Al(OH)3+IL-27佐剂组的杀伤率显著高于单独的IL-23、IL-27或Al(OH)3组。结论:这些结果表明:IL-23或IL-27质粒与传统铝盐佐剂Al(OH)3联合使用能显著增强RSV重组疫苗G1F/M2的免疫原性。     

重组乙肝病毒核心基因DNA与表面抗原-抗体共免疫应答研究

目的:了解小鼠对乙肝病毒核心抗原(HBcAg)基因免疫及与乙肝表面抗原-抗体复合物(HBsAg-抗HBs)联合免疫的应答。方法:用表达乙肝病毒核心抗原N端144个氨基酸的重组质粒DNA(简称pHBc144)100μg及含1μgHBsAg的重组乙肝表面抗原-鼠抗体组建的复合物(简称sIC)免疫Balb/c小鼠。用ELISA法检测血清抗体IgG和IgG1、IgG2a亚类的效价。用半定量PCR法分别检测鼠脾细胞IFN-γ及IL-4的mRNA。结果:pHBc144及sIC联合免疫鼠的抗HBs IgG、IgG1、IgG2a效价均显著高于sIC组(P＜0．05)。同时小鼠还可产生抗HBc及由HBsAg、HBcAg特异诱生的IFN-γ及IL-4。但与sIC共免疫,小鼠对HBcAg诱生的IFN-γ mRNA有所降低。结论:可用HBcAg基因与sIC共免疫,以获得针对乙肝病毒2种结构蛋白的免疫应答。     

尿酸钠对BALB/c小鼠抗体应答及树突状细胞和迟发型超敏反应的影响

目的:探讨尿酸钠作为佐剂对BALB/c小鼠体液和细胞免疫应答的影响。方法:利用尿酸钠悬浮液为佐剂、天花粉蛋白(TCS)为免疫原对BALB/c小鼠进行免疫,以酶联免疫测定法检测特异性抗体IgG的效价。体外诱导小鼠树突状细胞(DC),流式细胞术分析DC表型,评价尿酸钠体外对DC成熟的效应。以二硝基氟苯建立迟发型超敏反应(DTH)模型,分析尿酸钠在体内对细胞免疫应答的影响。结果:传统弗氏佐剂可极大地增强小鼠对TCS的抗体应答,尿酸钠佐剂对抗体应答不但没有促进,与单独使用免疫原相比,抗体应答反而明显降低。流式细胞术分析显示,尿酸钠对DC表达CD11c和CD83没有影响,但可明显提高MHCII的表达水平。DTH模型中,尿酸钠增强致敏原引发的耳廓肿胀程度,并促进DTH小鼠淋巴细胞的体外增殖能力。结论:尿酸钠悬浮液作为佐剂,对细胞免疫有显著增强作用,而对体液免疫应答却有一定的抑制作用,提示该佐剂在疫苗研究中有潜在应用前景。     

Pathogenetic aspects of severe course of herpetic infection

We studied the state of antiviral defense in patients with severe course of herpetic infection of anogenital and labial localization and the frequency of its combination with other herpes virus infections. It was found that severe course of herpetic infection caused by herpes simplex virus occurs against the background of combined secondary immunodeficiency and its complication. We first demonstrated that severe course of the disease is associated with mixed viral infection. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Supplement 2, pp. 76–81, April, 2007     

The flip side of immune surveillance: immune dependency

Summary: The growths of many and perhaps all tumors may be stimulated rather than inhibited by a quantitatively low level of immunity. The reason tumors have antigens may be that tumors do not develop in vivo in the absence of at least a minimal immune reaction; in this sense, cancer may be considered an autoimmune disease. This review, based largely on the work of our own laboratory, outlines the data showing that the titration of anti-tumor immunity exhibits the phenomenon of hormesis, i.e. the dose–response curve is non-linear such that low levels of immunity are generally stimulatory but larger quantities of the same immune reactants may inhibit tumor growth. Evidence is also reviewed that suggests that the immune response may vary qualitatively and quantitatively during progression, such that there seems to be, during oncogenesis, a very low level of immune reaction that aids initial tumor growth, followed by a larger reaction that may cause remission of early neoplasms, followed, if the neoplasm survives, by a relative immunologic tolerance to the tumor that may be dependent, at least in part, on suppressor cells. This knowledge may help to explain some clinical observations concerning the relationships among tumor types and the organ distribution of metastases.     

The liver works as a school to educate regulatory immune cells

Because of its unique blood supply, the liver maintains a special local immune tolerogenic microenvironment. Moreover, the liver can impart this immune tolerogenic effect on other organs, thus inducing systemic immune tolerance. The network of hepatic regulatory cells is an important mechanism underlying liver tolerance. Many types of liver-resident antigen-presenting cells (APCs) have immune regulatory function, and more importantly, they can also induce the differentiation of circulating immune cells into regulatory cells to further extend systemic tolerance. Thus, the liver can be seen as a type of ‘school'', where liver APCs function as ‘teachers'' and circulating immune cells function as ‘students.''     

Asymmetry of delayed type hypersensitivity reaction in mice

The intensity of delayed type hypersensitivity reactions in the left and right paws was studied in mice divided into left- and right-pawed by the motor asymmetry of the brain. The reaction was more pronounced in the left paw in all animals irrespective of motor asymmetry. Motor asymmetry of the brain hemispheres little contributed to the manifestation of differences in the delayed type hypersensitivity reactions in the left and right paws. The authors concluded that asymmetry of cellular immunity is determined by functional asymmetry of cells in regional lymph nodes.     

The Role of the Immune Microenvironment in Bone Regeneration

Bone is an active tissue, being constantly renewed in healthy individuals with participation of the immune system to a large extent. Any imbalance between the processes of bone formation and bone resorption is linked to various inflammatory bone diseases. The immune system plays an important role in tissue formation and bone resorption. Recently, many studies have demonstrated complex interactions between the immune and skeletal systems. Both of immune cells and cytokines contribute to the regulation of bone homeostasis, and bone cells, including osteoblasts, osteoclasts, osteocytes, also influence the cellular functions of immune cells. These crosstalk mechanisms between the bone and immune system finally emerged, forming a new field of research called osteoimmunology. Therefore, the immune microenvironment is crucial in determining the speed and outcome of bone healing, repair, and regeneration. In this review, we summarise the role of the immune microenvironment in bone regeneration from the aspects of immune cells and immune cytokines. The elucidation of immune mechanisms involved in the process of bone regeneration would provide new therapeutic targets for improving the curative effects of bone injury treatment.     

白色念珠菌感染免疫应答的研究进展

白色念珠菌的致病是其与机体免疫系统相互作用的结果.研究发现,白色念珠菌感染人体时,刺激机体产生固有免疫及特异性细胞免疫和体液免疫应答.其中特异性细胞免疫占主导地位.了解认识白色念珠菌感染的免疫应答对诊断、预防及治疗白色念珠菌感染具有重要意义.     

Hepatitis C: the complications of immune dysfunction

Hepatitis C virus (HCV) infection has been linked to numerous diseases of immune dysfunction, including, but not limited to, essential mixed cryoglobulinemia and non-Hodgkin’s lymphoma. Clinical studies support these associations and treatment of the underlying HCV infection has been variably successful. Recent studies, focusing on the role of HCV gene products, have discovered evidence of dysregulated responses in multiple aspects of host immunity that may be contributing to the genesis of these diseases. Novel treatments that target these areas of dysregulation offer hope for improved therapy for the diseases associated with immunodysregulation by HCV.     

Phases of development of the humoral immune response

Research Institute of Clinical and Experimental Medicine, Siberian Branch, Academy of Medical Sciences of the USSR. All-Union Research Center for Maternal and Child Health Care, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. D. Ado.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 110, No. 12, pp. 635–637, December, 1990.     

Monkeypox virus and insights into its immunomodulatory proteins

Summary: Monkeypox is a disease that is endemic in Central and Western Africa. However, in 2003, there was an outbreak in the United States, representing the first documented monkeypox cases in the Western hemisphere. Although monkeypox virus is less fatal and not as transmissible as variola virus, the causative agent of smallpox, there is concern that monkeypox virus could become a more efficient human pathogen. The reason for this may lie in the virus' genetic makeup, ecological changes, changes in host behavior, and the fact that with the eradication of variola virus, routine smallpox vaccination is no longer carried out. In this review, we focus on the viral proteins that are predicted to modulate the host immune response and compare the genome of monkeypox virus with the genomes of variola virus and the vaccinia virus, the orthopoxvirus that represented the smallpox vaccine. There are differences found in several of these immune-modulating genes including genes that express proteins that affect cytokines such as interleukin-1, tumor necrosis factor, and interferon. There are also differences in genes that code for virulence factors and host range proteins. Genetic differences likely also explain the differences in virulence between two strains of monkeypox virus found in two different regions of Africa. In the current setting of limited smallpox vaccination and little orthopoxvirus immunity in parts of the world, monkeypox could become a more efficient human pathogen under the right circumstances.     

肿瘤微环境中的免疫效应抑制

肿瘤微环境内的免疫调节在肿瘤发生发展中具有重要功能。其中多个因素共同作用，形成了肿瘤局部免疫的特有模式。肿瘤微环境中局部效应细胞功能障碍，抑制性免疫调节细胞Treg，细胞因子的免疫负调节作用，抑制性配体受体反应以及肿瘤微环境中T细胞代谢活性的负调节等参与了肿瘤微环境中的免疫效应抑制。