Altered decamer and nonamer from an HLA-A0201-restricted epitope of Survivin differentially stimulate T-cell responses in different individuals

Survivin is a universal tumor antigen that is being currently targeted in vaccine approaches against cancer. Our study here examined the immunogenicity of a novel variant of an HLA-A0201-binding decamer peptide from region 95 to 104 of Survivin (ELMLGEFLKL) with a T → M modification at position 3 in the peptide. We found that this new modified 10-mer peptide had enhanced HLA-A0201 binding and induced a stronger T-cell response over its wild type counterpart peptide (ELTLGEFLKL) in select HLA-A0201⁺ normal donors. In addition, when compared to the previously characterized altered 96-104 peptide (LMLGEFLKL) from the same region of Survivin currently used in vaccine trials, we found that both peptides had similar immunogenicity, but donor T cells preferentially reacted strongly to either one or the other, but not strongly to both. These results suggest that these two closely related Survivin peptides yield distinct T-cell responses and that most individuals dominantly respond to one or the other altered peptide. We also found a novel association between positive reactivity to the new altered decamer Survivin peptide in some individuals and their expression of the HLA-C0701 allele along with HLA-A0201. Thus, vaccinating with both the 10-mer and 9-mer peptides would be required to immunize a maximum number of individuals in the HLA-A0201⁺ population and could lead to more consistent T-cell responses against this region of Survivin.     

CD8 T cell response in HLA-A*0201 transgenic mice is elicited by epitopes from SARS-CoV S protein

Cytotoxic CD8⁺ T lymphocytes (CTLs) play an important role in antiviral immunity. Several human HLA-A*0201 restricted CTL epitopes of severe acute respiratory syndrome (SARS) spike (S) protein have been identified in HLA-A*0201 transgenic (Tg) mice, but the mechanisms and properties of immune responses are still not well understood. In this study, HLA-A*0201 Tg mice were primed intramuscularly with SARS S DNA and boosted subcutaneously with HLA-A*0201 restricted peptides. The lymphocytes from draining lymph nodes, spleens and lungs were stimulated with the cognate peptides. Three different methods (ELISA, ELISPOT and FACS) were used to evaluate the immune responses during short and long periods of time after immunization. Results showed that peptide-specific CD8⁺ T cells secreted IFN-γ, TNF-α and IL-2 and expressed CD107a/b on cell surface. IFN-γ⁺CD8⁺ T cells and CD107a/b⁺CD8⁺ T cells distributed throughout the lymphoid and non-lymphoid tissues, but the frequency of peptide-specific CD8⁺ T cells was higher in lungs than in spleens and lymph nodes. The phenotype of the CD8⁺ T cells was characterized based on the expression of IFN-γ. Most of the HLA-A*0201 restricted peptide-specific CD8⁺ T cells represented a memory subset with CD45RB^high and CD62L^low. Taken together, these data demonstrate that immunization with SARS S DNA and HLA-A*0201 restricted peptides can elicit antigen-specific CD8⁺ T cell immune responses which may have a significant implication in the long-term protection. We provide novel information in cellular immune responses of SARS S antigen-specific CD8⁺ T cells, which are important in the development of vaccine against SARS-CoV infection.     

Synthetic peptides coupled to the surface of liposomes effectively induce SARS coronavirus-specific cytotoxic T lymphocytes and viral clearance in HLA-A*0201 transgenic mice

We investigated whether the surface-linked liposomal peptide was applicable to a vaccine based on cytotoxic T lymphocytes (CTLs) against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV). We first identified four HLA-A*0201-restricted CTL epitopes derived from SARS-CoV using HLA-A*0201 transgenic mice and recombinant adenovirus expressing predicted epitopes. These peptides were coupled to the surface of liposomes, and inoculated into mice. Two of the liposomal peptides were effective for peptide-specific CTL induction, and one of them was efficient for the clearance of vaccinia virus expressing epitopes of SARS-CoV, suggesting that the surface-linked liposomal peptide might offer an effective CTL-based vaccine against SARS.     

Prediction and identification of novel IBV S1 protein derived CTL epitopes in chicken

Infectious bronchitis virus (IBV) is a major pathogen common in the poultry industry. Broad cytotoxic T lymphocyte (CTL) response against IBV is one of the crucial factors that help to control viral replication. Spike glycoproteins on the surface of the IBV virion harbor major T cell epitopes. In this study, based on the peptide-binding motifs of chicken MHC I molecules for the BF2*4, BF2*12, BF2*15, and BF2*19 haplotypes, potential CTL epitopes were predicted using S1 proteins from different IBV strains. Twenty-one peptides were predicted to be potential CTL epitopes; they were manually synthesized and the CTL responses to them tested in vitro. Spleen lymphocytes were collected from specific-pathogen free (SPF) chicken that had been immunized with the S1 protein expression plasmid, pV-S1, and were stimulated by the synthesized peptides. IFN-γ secretion and CD8⁺ T cell proliferation in chickens were tested by ELISpot array and flow cytometry, respectively. Four epitopes (P8_SRIQTATDP, P9_SRNATGSQP, P18_GAYAVVNV, and P19_SRIQTATQP) were identified to stimulate CD8⁺ T cell proliferation and IFN-γ secretion, indicating their efficacy as CTL epitopes in chicken. Poly-CTL-epitope DNA vaccine (pV-S1T) was constructed by inserting nucleotide sequences encoding the P8, P9, P18, and P19 CTL epitopes into the pVAX1 vector. Chickens were vaccinated with either pV-S1, pV-S1T, or pVAX1 and the protection efficacy was analyzed, revealing that ninety percent of chickens immunized with pV-S1T were protected after challenge with 10⁶ ELD₅₀ of IBV, demonstrating that these novel CTL epitopes were effective against IBV challenge. This study provides a new method to screen virus CTL epitopes in chicken and to develop poly-CTL-epitope DNA vaccines.     

Characterization of immune response to novel HLA-A2-restricted epitopes from zinc transporter 8 in type 1 diabetes

ObjectiveZnT8-specific CD8+ T cells in human type 1 diabetes (T1D) have been reported recently, although the results from different laboratories are inconsistent. We aimed to characterize these ZnT8 specific CD8+ T cells and validate assays to screen peptide libraries.MethodsWe screened HLA-A2-restricted T cell candidate peptides of ZnT8 with different methods including computer algorithms, MHC-peptide binding and dissociation assays in T2 cell line, identification in HLA-A2 transgenic (Tg) mice and in vivo CTL assays. Then ELISpot assay was used to measure peptide-reactive T cell responses in 49 HLA-A2-restricted T1D patients.ResultsWe demonstrated that ZnT8_{107–116(115)}, ZnT8_110–118, and ZnT8_177–186 were novel HLA-A*0201-restricted CTL epitopes in T1D patients. ZnT8_{107–116(115)}, ZnT8_115–123, ZnT8_153–161, ZnT8_177–186 and ZnT8_291–300 represent potentially major biomarkers for T1D. T cell responses against these epitopes showed different distributions between recently diagnosed and long-standing patients. Furthermore, they displayed discriminating performance among different ethnicities. We also compared the performance of the epitope identification strategies used herein. The epitopes which exhibited strong immunogenicity in HLA-A2 Tg mice were also well recognized by T1D patients.ConclusionsThe differences in autoimmune T cell responses among T1D individuals may open new avenues toward T1D prediction and prevention. It also provides efficient strategies for immune intervention.     

我国北方地区小儿哮喘CTLA-4基因多态性的相关研究

目的:探讨细胞毒T淋巴细胞相关抗原4(CTLA-4)启动子-318C/T基因多态性与我国北方地区儿童哮喘的关系及其临床意义。方法:采用PCR-RFLP法检测90例哮喘患儿和100例健康儿童的CTLA-4-318C/T的基因多态性。结果:CTLA-4-318位点存在基因多态性;重度哮喘组基因型分布和T等位基因频率明显高于轻、中度哮喘组及正常对照组,具有显著性差异(P均<0.05)。结论:CTLA-4-318位点存在基因多态性,T等位基因可能是重度哮喘的候选基因。     

Development of a novel ZIKV vaccine comprised of immunodominant CD4+ and CD8+ T cell epitopes identified through comprehensive epitope mapping in Zika virus infected mice

Zika virus (ZIKV) caused over two million human infections in more than 80 countries around 2015–2016. Current vaccines under development are mostly focused on inducing antibodies that despite capable of inhibiting the virus, may have the potential to trigger antibody dependent enhancement (ADE). T cell vaccines that do not induce antibodies targeting viral surface will unlikely cause ADE, but be capable of potentiating the effectiveness of an antibody-inducing vaccine. To develop such a protective T cell vaccine, we first examined the repertoire of antigen-specific T cells in immunocompetent mice that have been transiently infected by ZIKV. Through epitope mapping using 427 overlapping peptides spanning the entire length of ZIKV polyprotein, we discovered 27 immunodominant epitopes scattered throughout the virus on C, E, NS1-NS5 proteins. Among them, 8 were confirmed as CD4+ T cell epitopes, and 16 as CD8+ T cell epitopes, while 3 for both T cell subsets. From these 27 newly identified epitopes, the top 10 epitopes were selected to formulate three T cell vaccines comprised of either CD4+ T cell epitopes, or CD8+ T cell epitopes, or a mixture of both. Immunization with these T cell epitopes induced T cell-mediated cytotoxicity and cytokine production, and conferred varying degrees of protection against ZIKV challenge. Moreover, these new T cell vaccines also improved the protective efficacy of a neutralizing antibody-inducing recombinant E80 protein vaccine. Together, our results provided additional evidence in support of the protective role of ZIKV-specific CD4+ and CD8+ T cells, and laid foundation for future development of T cell vaccines for ZIKV.     

Conservation of HIV-1 T cell epitopes across time and clades: Validation of immunogenic HLA-A2 epitopes selected for the GAIA HIV vaccine

HIV genomic sequence variability has complicated efforts to generate an effective globally relevant vaccine. Regions of the viral genome conserved in sequence and across time may represent the “Achilles’ heel” of HIV. In this study, highly conserved T-cell epitopes were selected using immunoinformatics tools combining HLA-A2 supertype binding predictions with relative global conservation. Analysis performed in 2002 on 10,803 HIV-1 sequences, and again in 2009, on 43,822 sequences, yielded 38 HLA-A2 epitopes. These epitopes were experimentally validated for HLA binding and immunogenicity with PBMCs from HIV-infected patients in Providence, Rhode Island, and/or Bamako, Mali. Thirty-five (92%) stimulated an IFNγ response in PBMCs from at least one subject. Eleven of fourteen peptides (79%) were confirmed as HLA-A2 epitopes in both locations. Validation of these HLA-A2 epitopes conserved across time, clades, and geography supports the hypothesis that such epitopes could provide effective coverage of virus diversity and would be appropriate for inclusion in a globally relevant HIV vaccine.     

Identification and characterization of HIV-1-specific CD8+ T cell epitopes presented by HLA-A*2601

Since HLA-A*26 is one of the most common alleles in Asia, where approximately 20% of people have this allele, identification of HIV-1-specific epitopes presented by HLA-A*26 is necessary for studies on the immunopathogenesis of AIDS and vaccine development in Asia. As presented herein, we used the reverse immunogenetics approach to identify HIV-1 epitopes presented by HLA-A*2601, one of the major HLA-A*26 subtypes. We selected 24 HLA-A*2601-binding peptides out of 110 HIV-1 peptides by using a HLA-A*2601 stabilization assay. The ability of these HLA-A*2601-binding peptides to induce peptide-specific CD8(+) T cells was tested by stimulating PBMCs from HIV-1-infected individuals having HLA-A*2601 with these peptides. Four HLA-A*2601-binding peptides induced peptide-specific CD8 T cells. Analysis using HIV-1 recombinant vaccinia-infected C1R-A*2601 cells indicated that these four peptides were HIV-1 epitopes endogenously presented by HLA-A*2601. Two epitope-specific CD8(+) T cells were predominantly detected in HIV-1 infected individuals, suggesting that these epitopes may be useful for vaccine development.     

Induction of T lymphocyte responses to dengue virus by a candidate tetravalent live attenuated dengue virus vaccine

Development of a safe and immunogenic tetravalent dengue virus (DV) vaccine has been designated as a priority by the World Health Organization. We characterized the T cell response to DV induced by a candidate live attenuated tetravalent DV vaccine as part of a phase I study. Proliferation and cytotoxic T lymphocyte (CTL) responses to multiple DV serotypes were detected in six of six and four of four subjects studied, respectively. Proliferation responses were higher to DV serotypes 1 and 3 than to serotypes 2 and 4. CTL responses were higher to DV serotypes 2 and 3 than to serotype 1, and included serotype cross-reactive responses. Production of interferon-γ, but not IL-4, was observed in response to DV stimulation. This candidate vaccine is immunogenic for both CD4+ and CD8+ T lymphocytes. However, T cell responses to the four DV serotypes were not equivalent, suggesting that the vaccine could be further optimized.     

Decreases of CD4- and CD8-positive T lymphocytes in retired chromate workers

To investigate the effects of chromates on the human immune system, we measured total T lymphocytes and their two major subpopulations (CD4 + and CD8 + T lymphocytes) in the peripheral blood of 19 retired male workers who had been exposed to chromate at a chemical plant. The results indicated that both CD4 + and CD8 + T lymphocytes were significantly decreased, resulting in decreases in total T lymphocytes and total lymphocytes.     

Hepatitis B virus (HBV)-derived DRB1*0101-restricted CD4 T-cell epitopes help in the development of HBV-specific CD8+ T cells in vivo

We previously identified two HLA-DRB1*0101-restricted epitopes in hepatitis B virus (HBV) X protein (HBx) and in HBV envelope proteins (preS2). To evaluated their help in the development of CD8+ T-cell responses, mice transgenic for human class I and class II HLA molecules were immunized with HBV-T helper constructs. The preS2 epitope favored a well-balanced response with CD4+ and CD8+ T cells producing IFN-γ, IL-2 and TNF-α. The response was focused on CD8+ T cells with the HBx epitope. Fine characterization of helper activities may meet clinical needs in terms of enhancing the potency of preventive or therapeutic polyepitope vaccines.     

Vaccine generated immunity targets an HPV16 E7 HLA-A2.1-restricted CD8(+) T cell epitope relocated to an early gene or a late gene of the cottontail rabbit papillomavirus (CRPV) genome in HLA-A2.1 transgenic rabbits

The newly established HLA-A2.1 transgenic rabbit model has proven useful for testing the immunogenicity of well known and computer-predicted A2-restricted epitopes. In the current study we compared the protective immunity induced to a preferred HPV16 E7 A2-restricted epitope that has been relocated to positions within the CRPV E7 gene and the CRPV L2 gene. Epitope expression from both the E7 protein and the L2 protein resulted in increased protection against viral DNA challenge of the HLA-A2.1 transgenic rabbits as compared to control-vaccinated rabbit groups. These data indicate that proteins expressed at both early and late time points during a natural papillomavirus infection can be targeted by epitope-specific immunity and indicate this immunity is increased to early rather than late expressed proteins of papillomaviruses. This study also highlights the broad utility of the HLAA2.1 transgenic rabbit model for testing numerous immunological factors involved in vaccine generated protective immunity.     

Silencing an immunodominant epitope of hepatitis B surface antigen reveals an alternative repertoire of CD8 T cell epitopes of this viral antigen

Immunodominance hierarchies operating in immune responses to viral antigens limit the diversity of the elicited T cell responses. The L^d/S_28–39-restricted CD8 T cell response to the hepatitis B surface antigen (HBsAg or S) prevents copriming of D^d- and K^b-restricted CD8 T cell responses. We exchanged L to V at position S₃₉ of HBsAg to construct mutant S_L39V. Comparable levels of wild-type S and mutant S_L39V were produced by transiently transfected cells, and mice immunized with the pCI/S and pCI/S_L39V DNA vaccines showed comparable serum antibody responses to HBsAg. The pCI/S but not pCI/S_L39V DNA vaccination induced L^d/S_28–39-specific CD8 T cell responses. However, the pCI/S_L39V DNA vaccine efficiently primed CD8 T cell responses to the subdominant D^d- and K^b-restricted epitopes, confirming the immunosuppressive phenotype of the L^d/S_28–39-specific CD8 T cell response. A single point mutation within the HBsAg can hence completely silence a ‘dominant’ CD8 T cell response thereby facilitating priming of a multispecific repertoire of suppressed, ‘subdominant’ epitopes. The data have practical implications for understanding HBV-specific CD8 T cell responses and for the design of novel vaccination strategies.     

连续性肾替代治疗对脓毒症患者CD8+T淋巴细胞功能的影响

目的了解脓毒症患者经连续性肾脏替代治疗（CRRT）后外周血CD8+T淋巴细胞数量变化以及对免疫功能的影响。方法收集2015年10月-2016年8月入住某院急诊科的脓毒症住院患者的一般资料,并采集经单次CRRT前后脓毒症患者外周血,分别检测总CD8+T细胞的数量、分泌干扰素 γ（IFN γ）的CD8+T细胞数、CD8+T细胞产生抑制性分子水平、共刺激性分子水平以及分泌IFN γ、肿瘤坏死因子 α（TNF α）水平。结果共有37例脓毒症住院患者,所有患者均为革兰阴性（G-）菌感染,感染的细菌种类为肺炎克雷伯菌（22株）、鲍曼不动杆菌（11株）、阴沟肠杆菌（3株）。脓毒症患者经CRRT后体温、心率、白细胞计数、尿素氮、血肌酐水平较治疗前下降（均P＜0.05）。经CRRT后脓毒症患者总CD8+T细胞数量较治疗前无明显变化（P＞0.05）,但分泌IFN γ的CD8+T细胞的数量较治疗前升高（P＜0.05）。同时,经CRRT后CD8+T细胞产生抑制性分子细胞毒T淋巴细胞相关抗原4（CTLA 4）、程序性死亡受体1（PD 1）、含T细胞免疫球蛋白及黏蛋白结构域的分子3（TIM 3）的水平均较治疗前降低（均P＜0.05）,而产生共刺激分子CD28、分泌IFN γ的水平较治疗前升高（均P＜0.05）。结论CRRT不但有效改善脓毒症患者的生命体征和肾功能,还可增强CD8+T细胞免疫功能。     

Identification of B- and T-cell epitopes from glycoprotein B of herpes simplex virus 2 and evaluation of their immunogenicity and protection efficacy

Herpes simplex virus (HSV) infection is a major health concern worldwide. Evidence obtained from animals and humans indicates that B- and T-cell responses contribute to protective immunity against herpes virus infection. Glycoprotein B is a transmembrane envelope component of HSV-1 and HSV-2, which plays an important role in virion morphogenesis and penetration into host cells, and can induce neutralizing antibodies and protective T-cell response when it is used to immunize humans and animals. However, little is known about gB epitopes that are involved in B- and T-cell activities in vitro and in vivo. Thus, the HSV-2 gB sequence was screened using B- and T-cell epitope prediction systems, and the B-cell regions and the HLA-A*0201-restricted epitopes were identified. These B-cell epitopes elicited high IgG antibody titers in Balb/C mice, with a predominantly IgG1 subclass distribution, which indicated a Th2 bias. Specific IgGs induced by these two epitopes were evaluated as the neutralizing antibodies for virus neutralization. The predicted T-cell epitopes stabilized the HLA-A*0201 molecules on T(2) cells, and stimulate interferon-γ-secreting and cytotoxic CD8(+) T cells. Immunization with the predicted peptides reduced virus shedding and protected against lethal viral challenge in mice. The functional epitopes described herein, both B- and T-cell epitopes, are potentially implicated in vaccine development.     

湖南省苗族健康人群外周血CD3+、CD4+、CD8+T淋巴细胞正常参考值研究

目的了解并建立湖南省苗族健康人群外周血CD3＋、CD4＋、CD8＋T淋巴细胞正常参考值。方法应用FACSCalibur流式细胞仪检测10-19岁、20-29岁、30-39岁、40-49岁、50-59岁5个年龄组共360名健康苗族人的CD3＋、CD4＋、CD8＋T淋巴细胞数值。结果10-59岁组苗族人群外周血CD3＋、CD4＋、CD8＋T淋巴细胞绝对数和CD4＋／CD8＋比值分别为（1426．71±462．92）个／μL、（787．54±258．36）个／μL、（589．86±242．91）个／μL和1．46±0．50;10～19岁年龄组的CD3＋、CD8＋T淋巴细胞计数均明显高于30岁以上年龄组（F分别为10．80、8．51,均P〈0．05）,CD4＋T淋巴细胞计数明显高于50-59岁组（F=7．03,P（0．05）;不同性别间上述指标无差异（t分别为0．98、0．80、0．88、0．16,均P〉0．05）。结论建立不同民族的CD3＋、CD4＋、CD8＋T淋巴细胞正常参考值有助于临床诊断。     

An intact signal peptide on dengue virus E protein enhances immunogenicity for CD8 T cells and antibody when expressed from modified vaccinia Ankara

Dengue is a global public health concern and this is aggravated by a lack of vaccines or antiviral therapies. Despite the well-known role of CD8⁺ T cells in the immunopathogenesis of Dengue virus (DENV), only recent studies have highlighted the importance of this arm of the immune response in protection against the disease. Thus, the majority of DENV vaccine candidates are designed to achieve protective titers of neutralizing antibodies, with less regard for cellular responses. Here, we used a mouse model to investigate CD8⁺ T cell and humoral responses to a set of potential DENV vaccines based on recombinant modified vaccinia virus Ankara (rMVA). To enable this study, we identified two CD8⁺ T cell epitopes in the DENV-3 E protein in C57BL/6 mice. Using these we found that all the rMVA vaccines elicited DENV-specific CD8⁺ T cells that were cytotoxic in vivo and polyfunctional in vitro. Moreover, vaccines expressing the E protein with an intact signal peptide sequence elicited more DENV-specific CD8⁺ T cells than those expressing E proteins in the cytoplasm. Significantly, it was these same ER-targeted E protein vaccines that elicited antibody responses. Our results support the further development of rMVA vaccines expressing DENV E proteins and add to the tools available for dengue vaccine development.     

HPV-16 E6 and E7 protein T cell epitopes prediction analysis based on distributions of HLA-A loci across populations: An in silico approach

Human papillomavirus type 16 (HPV-16) is the most prevalent virus in human cervical cancers, as it is present in more than half of all cases. Many studies have found continued expression of E6 and E7 proteins in the majority of cervical cancer cases, but not in normal tissues. These results indicated that the E6 and E7 proteins could be ideal candidate therapeutic vaccines against HPV-16 infection and cervical cancer. Using the Immune Epitope Database Analysis Resource, cytotoxic T lymphocyte (CTL) epitopes of the HPV-16 E6 and E7 proteins were predicted according to worldwide frequency distributions of HLA-A alleles (HLA-A*01:01, -A*02:01, -A*02:06, -A*03:01, -A*11:01, -A*24:02, -A*26:01, -A*31:01 and -A*33:03). Our results predicted a total of 81 epitopes of HPV-16 E6 (n = 59) and E7 (n = 22). Epitope cluster analysis showed that among the 20 clusters of HPV-16 E6, cluster 3 contained the most epitopes (10 epitopes), which was represented by HLA-A*31:01 and -A*33:03. Of the 10 clusters of HPV-16 E7, cluster 3 contained the most epitopes (5 epitopes), which was represented by HLA-A*01:01 and -A*26:01. Our results indicated that the combination of epitopes FAFRDLCIVYR_52-62 of E6 (HLA-A*02:06, HLA-A*31:01, and HLA-A*33:03), PYAVCDKCLKF_66-76 of E6 (HLA-A*11:01 and HLA-A*24:02), HGDTPTLHEY_2-11 of E7 (HLA-A*01:01 and HLA-A*26:01), and YMLDLQPETT_11-20 of E7 (HLA-A*02:01) could vaccinate >50% of all individuals worldwide. Our results propose CTL epitopes or combinations of them predicted in current study for candidate therapeutic vaccines to effectively control HPV-16 infection and development of cervical cancer.     

结核分枝杆菌抗原Rv0585c人T细胞抗原表位鉴定及其免疫原性评价

目的 研究结核分枝杆菌抗原Rv0585c的抗原表位及其免疫原性,为结核病特异免疫诊断技术和疫苗的研发提供基础。方法 利用生物信息学TE-predict和IEDB人T细胞抗原表位预测软件进行结核分枝杆菌抗原Rv0585c的人T细胞抗原表位预测,根据预测结果,合成表位多肽,用ELISpot试验检测预测表位在临床结核病患者中的免疫反应性。分别采用高、低剂量（100 μg/只和50 μg/只）的Rv0585c抗原表位多肽、同时以高、低剂量（50 μg/只和20 μg/只）的Ag85B蛋白抗原为对照,对随机分组的BALB/c小鼠进行免疫。利用ELISA方法检测IFN-γ、IL-2、IL-4、IL-10的水平。结果 经生物信息学技术预测到Rv0585c 66个人T细胞抗原表位,选取合成了表位分布集中的抗原表位多肽9条。人群ELISpot试验筛选出3条阳性人T细胞表位多肽：P10110、P10112、P10117,用于肺结核检测的灵敏度分别为14.00%、12.00%和6.00%,特异度分别为100.00%、100.00%和97.96%;联合用于肺结核检测的灵敏度和特异度分别为22.00%和97.96%。动物免疫试验结果显示,P10110多肽高、低剂量刺激小鼠产生较高水平的IFN-γ、IL-2、IL-4和IL-10,P10112多肽高、低剂量刺激小鼠产生较高水平的IFN-γ、IL-2和IL-10,均高于阴性对照组,差异有统计学意义（P<0.001）。结论 Rv0585c蛋白及其T细胞表位具有较好的免疫原性及免疫反应性,能刺激机体产生较强烈的细胞免疫应答,具有潜在的结核病细胞免疫诊断和新型结核疫苗的应用价值。