结核分枝杆菌海藻糖磷酸磷酸酶诱导小鼠体液和细胞免疫应答

目的 研究结核分枝杆菌(M．tuberculosis)海藻糖磷酸磷酸酶(TPP)诱导小鼠体液和细胞免疫。方法 差速离心分离结核分枝杆菌H37Rv和卡介苗(BCG)的各细胞组分,通过Western杂交检测抗原TPP在结核分枝杆菌H37Rv和BCG中的亚细胞定位情况。分别用5×10~6CFU的BCG和50μg的TPP蛋白免疫C57BL/6小鼠,检测小鼠血清中抗TPP的IgG1和IgG2a抗体效价。取免疫小鼠的脾细胞,体外抗原刺激,用酶联免疫斑点试验(ELISPOT)检测γ干扰素(IFN-γ)分泌细胞。结果 TPP亚细胞定位于结核分枝杆菌H37Rv和BCG的胞壁和细胞膜组分。TPP蛋白免疫后小鼠产生的TPP特异性IgG1和IgG2a抗体效价明显高于BCG免疫小鼠,并且IgG2a的抗体效价高于IgG1。体外抗原刺激TPP蛋白和BCG免疫小鼠的脾细胞,都能诱导较高的IFN-γ分泌。结论 结核分枝杆菌细胞壁蛋白TPP能诱导小鼠Ⅰ型辅助性T细胞介导的免疫反应,可作为抗结核疫苗的候选抗原。     

IL-17作为分子佐剂增强蛋白疫苗细胞免疫应答的研究

目的:为了进一步增强重组蛋白质疫苗的细胞免疫应答,利用重组的IL-17作为分子佐剂,与卵清蛋白(ovalbumin,OVA)一起免疫小鼠,研究IL-17作为分子佐剂对适应性免疫的影响,探索IL-17对蛋白疫苗诱导的免疫反应,特别是细胞免疫应答的影响.方法:用OVA作为特异性蛋白疫苗,与不同剂量的IL-17联合免疫C57...     

流行性乙型脑炎活疫苗细胞免疫检测方法的探索

本文建立了流行性乙型脑炎活疫苗细胞免疫检测方法。采用纯系Ｂａｌｂ／ｃ（Ｈ－２ｄ）小鼠制备效应细胞,以同一品系Ｂａｌｂ／ｃ（Ｈ－２ｄ）小鼠原代肾细胞为靶细胞。当效靶细胞比例为２０∶１,杀伤时间为４小时,ＭＴＴ反应时间为４小时,细胞毒性Ｔ淋巴细胞（ＣＴＬ）杀伤作用最强。利用该法对乙脑活苗和死苗免疫小鼠进行了ＣＴＬ活性测定,结果表明该法重复性好,活苗诱导ＣＴＬ杀伤作用强于死苗     

IL-12增强HBV融合抗原DNA疫苗在小鼠诱导的细胞免疫应答

乙型肝炎病毒(hepatitis B virus,HBV)极易形成慢性感染,主要机制在于感染者不能产生强有力的细胞免疫应答以清除病毒[1].慢性HBV感染者体内虽然存在HBV抗原特异性T淋巴细胞,但对HBV抗原的反应性较低.研究发现,增强这类T淋巴细胞的反应性,可以促进HBV的清除[2].     

分析CpG ODN佐剂对呼吸道合胞病毒重组疫苗诱导的细胞免疫应答的作用

目的:对CpG ODN佐剂对呼吸道合胞病毒重组疫苗诱导的细胞免疫应答的作用进行分析。方法:选取BALB/c小鼠作为试验对象,将小鼠随机分为6组,之后分别制备G1F/M2蛋白,用LDH释放法对CTL的活性进行检测,分析T细胞分化情况,并应用流式细胞仪分析LDH的记忆细胞及CD8+/CD4+记忆细胞。结果:将CpG2216混合G1F/M2后,通过腹腔诱导,其特异杀伤的活性显著好于单独采用G1F/M2进行腹腔注射诱导杀伤活性。并且将两者混合的杀伤活性显著强于常规佐剂Al(OH)3。G1F/M2+Al+CpG(i.p.)组的诱导杀伤活性显著优于CpG+G1F/M2(i.p.)组。结论:将CpG ODN当做呼吸道合胞病毒重组疫苗G1F/M2佐剂,能够将细胞免疫应答显著增强。     

HIV-1感染的T细胞免疫应答与病毒免疫逃逸

HIV-1感染可以同时活化T细胞免疫与体液免疫.所活化的免疫反应尽管对病毒有一定的抑制作用,但不能清除病毒,因而,HIV-1感染均形成慢性持续性感染,最终大部分个体以免疫系统功能耗竭、严重的免疫缺陷为结局,感染者可因并发感染或肿瘤而死亡.大量的研究表明HIV-1特异性T细胞在与HIV作斗争中起极其重要的作用.本文主要就HIV-1感染过程中T细胞的免疫作用、病毒免疫逃逸、以及不同感染时期免疫反应的特征等最新进展加以综述.     

口蹄疫细胞免疫研究进展

口蹄疫是世界性重大动物疫病之一,接种疫苗是预防该病的重要策略之一.随着对口蹄疫疫苗及其免疫特性的深入研究和探讨,许多学者对口蹄疫细胞免疫机制的研究更进了一步,就参与口蹄疫细胞免疫的各类细胞及细胞免疫与新疫苗设计的研究进展作一综述.     

疫苗诱导性抗体应答的评价:新技术的影响

<正>针对免疫接种则产生的宿主应答在历史上已经通过抗体滴度的变化而进行了评价,这种滴度的变化是通过免疫接种后与免疫接种前的滴度进行比对而发现的,抗原特异性抗体滴度呈4倍或较大提高已经被理解成在针对疫苗接种的应答中抗体产生有所增高。新的技术,如象粒珠试验能够使研究者和临床医生检测出精确的抗体水平(以每毫升的浓度进行报告),其水平范围在以往采用的如象ELISA这样的标准测定所得数值的上下范围之内,为测定     

Effect of Triphala on oxidative stress and on cell-mediated immune response against noise stress in rats

Stress is one of the basic factors in the etiology of number of diseases. The present study was aimed to investigate the effect of Triphala (Terminalia chebula, Terminalia belerica and Emblica officinalis) on noise-stress induced alterations in the antioxidant status and on the cell-mediated immune response in Wistar strain male albino rats. Noise-stress employed in this study was 100 dB for 4 h/d/15 days and Triphala was used at a dose of 1 g/kg/b.w/48 days. Eight different groups of rats namely, non-immunized: control, Triphala, noise-stress, Triphala with noise-stress, and corresponding immunized groups were used. Sheep red blood cells (5×10⁹ cells/ml) were used to immunize the animals. Biochemical indicators of oxidative stress namely lipid peroxidation, antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), ascorbic acid in plasma and tissues (thymus and spleen) and SOD, GPx and corticosterone level in plasma were estimated. Cell-mediated immune response namely foot pad thickness (FPT) and leukocyte migration inhibition (LMI) test were performed only in immunized groups. Results showed that noise-stress significantly increased the lipid peroxidation and corticosterone level with concomitant depletion of antioxidants in plasma and tissues of both non-immunized and immunized rats. Noise-stress significantly suppressed the cell-mediated immune response by decreased FPT with an enhanced LMI test. The supplementation with Triphala prevents the noise-stress induced changes in the antioxidant as well as cell-mediated immune response in rats. This study concludes that Triphala restores the noise-stress induced changes may be due to its antioxidant properties.     

Dirofilaria immitis: cell-mediated and humoral immune responses in experimentally-infected dogs.

Four dogs were experimentally infected with 30 Dirofilaria immitis infective larvae, four dogs received two such infections and four dogs served as uninfected controls. A partially-purified D. immitis antigen was used in an indirect hemagglutination assay to determine anti-D. immitis antibody titers. Anti-D. immitis antibody was first detected in infected dogs 4 weeks after infection. Titers were highest 2 weeks after the appearance of microfilariae and diminished to low levels thereafter in the single infection group. Antibody levels in the double infection group decreased similarly but were demonstrable throughout the study. Antibody titers were significantly higher in the infected dogs, but there were no differences in titers between single and double infection groups.The responses of peripheral lymphocytes to phytohemagglutinin P and pokeweed mitogen were significantly depressed in infected dogs. Peripheral blood lymphocyte transformation could not be induced with D. immitis antigens. Differences between groups in T-cell function were not demonstrated by total hemagglutinating antibody or 2-mercaptoethanol labile hemagglutinating antibody following immunization with sheep erythrocytes.     

猪肺炎支原体感染诱导的固有免疫应答研究进展

猪肺炎支原体是引起猪支原体肺炎的病原。由于缺乏成熟的猪肺炎支原体感染动物模型,使得猪肺炎支原体相关的抗感染免疫研究进展较为缓慢。本文从猪肺炎支原体感染后的炎症反应、固有免疫系统对猪肺炎支原体的识别、固有免疫细胞的作用、补体系统、抗菌肽、自噬以及细胞凋亡7个方面进行综述,旨在阐明固有免疫系统各组分在猪肺炎支原体感染中发挥的作用的研究进展,并对今后猪肺炎支原体感染的固有免疫应答研究的重点方向进行展望。     

The study of cell-mediated immune response in recurrent vulvovaginal candidiasis

The aim of this work was to examine in vitro the ability of cells from patients with recurrent vulvovaginal candidiasis (RVVC) to cell-mediated immune response. Peripheral blood mononuclear cells (PBMC) and whole blood cells (WBC) of 37 RVVC patients in acute infection and 14 in remission were examined for the ability to proliferation and cytokines production (IFN, TNF, IL-6). As a control, a group of 25 healthy women were examined. The cells were stimulated with Candida antigen (HKCA), LPS and PHA. To indicate the level of cytokines, the following cell-lines were used: A549 for IFN, WEHI 164 for TNF and 7TD1 for IL-6. The proliferation/death of cells was determined by colorimetric test using MTT. Distinct suppression of cell-mediated immune response (CMI) was shown in all patients comparing to the control. Greatest suppression was found in the acute phase of the disease. The ability of cells to proliferate and produce IFN increases only in remission. The data seem to suggest that in this phase of disease, the ability of cell-mediated immune response is restored. It was also indicated that IFN may take part in protection against Candida infection.     

Selection for brain size impairs innate,but not adaptive immune responses

Both the brain and the immune system are energetically demanding organs, and when natural selection favours increased investment into one, then the size or performance of the other should be reduced. While comparative analyses have attempted to test this potential evolutionary trade-off, the results remain inconclusive. To test this hypothesis, we compared the tissue graft rejection (an assay for measuring innate and acquired immune responses) in guppies (Poecilia reticulata) artificially selected for large and small relative brain size. Individual scales were transplanted between pairs of fish, creating reciprocal allografts, and the rejection reaction was scored over 8 days (before acquired immunity develops). Acquired immune responses were tested two weeks later, when the same pairs of fish received a second set of allografts and were scored again. Compared with large-brained animals, small-brained animals of both sexes mounted a significantly stronger rejection response to the first allograft. The rejection response to the second set of allografts did not differ between large- and small-brained fish. Our results show that selection for large brain size reduced innate immune responses to an allograft, which supports the hypothesis that there is a selective trade-off between investing into brain size and innate immunity.     

Interleukin 2 in cell-mediated immune responses

The lymphokine Interleukin 2 (IL2) restores T cell responses in a number of in vitro systems where immunogenicity has been compromised. UV irradiation of the stimulating allogeneic cells in a mixed leukocyte culture eliminates the production of cytotoxic T lymphocytes and greatly reduces the DNA synthesis response. IL2 restores both parameters. UV-irradiated stimulators are also unable to induce the normal production of IL2 which is observed in a mixed leukocyte culture. The cytotoxic activity of allogeneically stimulated thymocytes is almost completely lost within 24 hours after removal of IL2 at 5 days, indicating that the lymphokine is continuously required to maintain CTL. Thymocytes in 4-day cultures do not adsorb IL2 unless they are simultaneously activated with a mitogen. Finally, IL2 does not adequately restore a secondary response to the purified protein derivative of tuberculin (PPD) in adherent-cell-depleted cultures, indicating that macrophages, in addition to being required for IL2 production, have other functions. These probably include the presentation of soluble antigens to responding cells.     

Two arms are better than one: parasite variation leads to combined inducible and constitutive innate immune responses

Parasites represent a major threat to all organisms which has led to the evolution of an array of complex and effective defence mechanisms. Common to both vertebrates and invertebrates are innate immune mechanisms that can be either constitutively expressed or induced on exposure to infection. In nature, we find that a combination of both induced and constitutive responses are employed by vertebrates, invertebrates and, to an extent, plants when they are exposed to a parasite. Here we use a simple within-host model motivated by the insect immune system, consisting of both constitutive and induced responses, to address the question of why both types of response are maintained so ubiquitously. Generally, induced responses are thought to be advantageous because they are only used when required but are too costly to maintain constantly, while constitutive responses are advantageous because they are always ready to act. However, using a simple cost function but with no a priori assumptions about relative costs, we show that variability in parasite growth rates selects for a strategy that combines both constitutive and induced defences. Differential costs are therefore not necessary to explain the adoption of both forms of defence. Clearly, hosts are likely to be challenged by variable parasites in nature and this is sufficient to explain why it is optimal to deploy both arms of the innate immune system.     

Cell-mediated immune responses (CMIR) in human rhinosporidiosis

Cell mediated immune responses (CMIR) to Rhinosporidium seeberi in human patients with rhinosporidiosis have been studied. With immuno-histochemistry, the cell infiltration patterns in rhinosporidial tissues from 7 patients were similar. The mixed cell infiltrate consisted of many plasma cells, fewer CD68+ macrophages,a population of CD3+ T lymphocytes, and CD56/57+ NK lymphocytes which were positive for CD3 as well. CD4+ T helper cells were scarce. CD8+suppressor/cytotoxic-cytolytic cells were numerous. Most of the CD8+ cells were TIA-l+ and therefore of the cytotoxic subtype. CD8+ T cells were not sub-typed according to their cytokine profile; 1L2, IFN-γ (Tcl); IL4, ILS (Tc2).In lympho-proliferative response (LPR) assays in vitro, lymphocytes from rhinosporidial patients showed stimulatory responses to Con A but lymphocytes from some patients showed significantly diminished responses to rhinosporidial extracts as compared with unstimulated cells or cells stimulated by Con A, indicating suppressor immune responses in rhinosporidiosis. The overall stimulatory responses with Con A suggested that the rhinosporidial lymphocytes were not non-specifically anergic although comparisons of depressed LPR of rhinosporidial lymphocytes from individual patients, to rhinosporidial antigen with those to Con A, did not reveal a clear indication as to whether the depression was antigen specific or non-specific. The intensity of depression of the LPR in rhinosporidial patients bore no relation to the site, duration, or the number of lesions or whether the disease was localized or disseminated. Rhinosporidial extracts showed stimulatory activity on normal control lymphocytes, perhaps indicating mitogenic activity. These results indicate that CMIR develops in human rhinosporidiosis, while suppressed responses are also induced. This revised version was published online in June 2006 with corrections to the Cover Date.     

Cellular immune responses to autologous chronic myelogenous leukaemia cells in vitro

 Using a modification of the autologous mixed lymphocyte/tumour cell culture (MLTC), it is demonstrated here that lymphocytes from chronic-phase myelogenous leukaemia (CML) patients (n = 58), but not from their HLA-identical siblings, proliferated upon coculture with autologous tumour cells. However, in most cases, the level of proliferation measured was low (stimulation index <3, n = 37). This was most likely related to the amount of interleukin-10 (IL-10) released into the culture medium by the CML cells, because addition of neutralizing anti-IL-10 serum to MLTC markedly enhanced proliferative responses. In addition, supplementation of media with IL-1α further enhanced proliferative responses and a combination of anti-IL-10 serum and IL-1α was more effective than either agent alone. Only HLA-DR-matched CML cells, but not HLA-DR-mismatched CML cells or matched or mismatched PBMC restimulated proliferation of IL-2-dependent T cell lines derived from MLTC supplemented with IL-1α and anti-IL-10 serum. The responding cells under these conditions were predominantly CD4⁺ and secreted IL-2, and interferon γ; some secreted IL-4, but none secreted IL-10. These data therefore suggest the existence of an HLA-DR-restricted DTH/Th1-type of tumour-specific immunity in CML patients, which may be down-regulated in vitro by excessive secretion of IL-10 together with depressed secretion of IL-1. Received: 9 November 1995 / Accepted: 8 February 1996