衣原體抗原組合物及其用圖
【技術(shù)領(lǐng)域】
[0001] 本發(fā)明涉及細(xì)菌感染。更具體地，本發(fā)明部分地提供了用于抗衣原體（Chlamydia) 感染的融合蛋白。
【背景技術(shù)】
[0002] 沙眼衣原體（Chlamydia trachomatis)是在全世界每年引起超過九千二百萬性 傳播感染和八千五百萬眼部感染的細(xì)胞內(nèi)病原體（Starnbach，M.N.，and N.R. Roan. 2008. Conquering sexually transmitted diseases. Nat Rev Immunol8:313-317. ) 〇 性傳播的 沙眼衣原體是婦女中長(zhǎng)期疾病后遺癥（例如不孕和異位妊娠）的主要原因（Brunham，R. C, D. J. Zhang, X. Yang, and G. M. McClarty. 2000. The potential for vaccine development against chlamydial infection and disease. J Infect Dis 181Suppl 3:S538-543 ； Igietseme, J. U. , C. M. Black, and H. D. Caldwell. 2002. Chlamydia vaccines: strategies and status. BioDrugs 16:19-35)。婦女中的沙眼衣原體感染經(jīng)常被忽視，直到嚴(yán)重的生殖 損害（不孕、盆腔炎疾病、異位妊娠）已經(jīng)發(fā)生。此外，感染沙眼衣原體的婦女在暴露后感 染HIV的風(fēng)險(xiǎn)增加。
[0003] 由于患病率以及再感染率持續(xù)升高，為預(yù)防和控制沙眼衣原體的性傳播感染 的"尋找和治療"方案看來是失敗了（Brunham，R. C，B. Pourbohloul，S. Mak，R. White，and M. L. Rekart. 2005.The unexpected impact of a Chlamydia trachomatis infection control program on susceptibility to reinfection. J Infect Dis 192:1836-1844)， 可能是由于早期的治療干擾了保護(hù)性免疫反應(yīng)的發(fā)展（Su, H.，R. Morrison, R. Messer, W. Whitmire, S. Hughes, and H. D. Caldwell. 1999. The effect of doxycycline treatment on the development of protective immunity in a murine model of chlamydial genital infection. J Infect Dis 180:1252-1258)。
[0004] 先前嘗試使用無活力的（dead)原生小體（elementary bodies，EB)在人和鼠 科模型中進(jìn)行接種以抵抗沙眼衣原體和小鼠生物型沙眼衣原體（C.muridarum)感染，原 生小體是感染的細(xì)胞破裂時(shí)釋放的非復(fù)制的傳染性顆粒，提供有限的保護(hù)（Grayston，J. T. , and S. P. Wang. 1978. The potential for vaccine against infection of the genital tract with Chlamydia trachomatis. Sex Transm Dis 5:73-77 ；Grayston, J. T. , S. P. Wang, L. J. Yehj and C. C. Kuo. 1985. Importance of reinfection in the pathogenesis of trachoma. Rev Infect Dis 7 : 717-725 ；Lu, H. , Z. Xing, and R. C. Brunham. 2002. GM-CSF transgene-based adjuvant allows the establishment of protective mucosal immunity following vaccination with inactivated Chlamydia trachomatis. J Immunol 169:6324-6331 ；Schachter, J. , and H. D. Caldwell. 1980. Chlamydiae. Annu Rev Microbiol34:285-309)。但是，免疫活的小鼠生物型沙眼衣原體EB的小鼠已經(jīng)顯示產(chǎn)生 更好的保護(hù)（Lu, H.，Z. Xing，and R.C. Brunham. 2002. GM-CSF transgene-based adjuvant allows the establishment of protective mucosal immunity following vaccination with inactivated Chlamydia trachomatis. J Immunol 169:6324-6331 ；Su, H. , R. Messer,W. Whitmire, E. Fischer, J. C. Portis, and H.D. Caldwell. 1998.Vaccination against chlamydial genital tract infection after immunization with dendritic cells pulsed ex vivo with nonviable Chlamydiae. J Exp Med 188:809-818).
[0005] 對(duì)與死的有機(jī)體相比，由活的小鼠生物型沙眼衣原體提供的免疫的有效誘導(dǎo)的機(jī) 制的研宄暗示，暴露于活或死的小鼠生物型沙眼衣原體的樹突細(xì)胞（DC)發(fā)展為不同的表 型。具體地，暴露于活的小鼠生物型沙眼衣原體的DC變成熟，并刺激抗原特異的CD4T細(xì)胞， 而暴露于死的小鼠生物型沙眼衣原體的DC被抑制了獲得成熟的表型。用死的EB和CpG寡 脫氧核苷酸共刺激DC已經(jīng)顯示部分地克服了死的EB對(duì)DC成熟的抑制（Rey-Ladino, J.，K. M. Koochesfahani, M. L. Zaharik, C. Shenj and R. C. Brunham. 2005. A live and inactivated Chlamydia trachomatis mouse pneumonitis strain induces the maturation of dendritic cells that are phenotypically and immunologicalIy distinct. Infect Immun 73:1568-1577)。使用GeneChip微陣列對(duì)在暴露于活的和死的小鼠生物型沙眼衣原 體后骨髓衍生的DC的轉(zhuǎn)錄響應(yīng)的研宄揭示了暴露于活的或死的有機(jī)體的DC中的CXC趨 化因子譜的顯著差異（Zaharik，M. L，T. Nayar，R. White，C. Ma，B. A. Vallance，N. Straka，X. Jiang, J. Rey-Ladinoj C. Shenj and R. C. Brunham. 2007. Genetic profiling of dendritic cells exposed to live-or ultraviolet-irradiated Chlamydia muridarum reveals marked differences in CXC chemokine profiles. Immunology 120:160-172) 〇 總之，數(shù) 據(jù)顯示暴露于活的EB的DC在表型及功能上與通過暴露于死的EB產(chǎn)生的DC不同。
[0006] 認(rèn)為對(duì)小鼠生物型沙眼衣原體感染的免疫主要地由細(xì)胞調(diào)節(jié)，因此依賴于通 過抗原呈遞細(xì)胞上的MHV分子呈遞到⑶4T細(xì)胞的衣原體衍生的肽（Brunham，R. C，and J. Rey-Ladino.2005. Immunology of Chlamydia:implications for a Chlamydia trachomatis vaccine. Nat Rev Immunol 5:149-161 ；Steinman, R. M. , and M. Pope. 2002. Exploiting dendritic cells to improve vaccine efficacy.J Clin Invest 109:1519-1526 ;Su，H.，and H.D. Caldwell. 1995. CD4+T cells play a significant role in adoptive immunity to Chlamydia trachomatis infection of the mouse genital tract. Infect Immun 63: 3302-3308 ；Morrison, S. G. , H. Suj H. D. Caldwell, and R.P. Morrison. 2000.1 mmunity to murine chlamydial genital tract reinfection involves B cells and CD4 (+)T cells but not CD8 (+)T cells. Infect Immun 68:6979-6987 ;Morrison，R. P.，and H. D. Caldwell. 2002. Immunity to murine chlamydial genital infection. Infect Immun70:2741-2751 ；Igietseme,J.U. , K. H. Ramsey, D. M. Magee, D. M. Williams, T. J. Kincy, and R. G. Rank. 1993. Resolution of murine chlamydial genital infection by the adoptive transfer of a biovar-specific, Thl lymphocyte clone. Reg Immunol 5:317-324)〇
[0007] 用于識(shí)別小鼠生物型沙眼衣原體T細(xì)胞抗原的免疫蛋白組方法（Hunt,D. F. , R. A. Henderson, J. Shabanowitz, K. Sakaguchi, H. Michel, N. Sevilir, A. L. Cox, E. Appella,and V. H. Engelhard. 1992.Characterization of peptides bound to the class I MHC molecule HLA-A2. Iby mass spectrometry. Science255:1261-1263 ；de Jong,A. 1998. Contribution of mass spectrometry to contemporary immunology. Mass Spectrom Rev 17: 311-335 ；01sen, J. V. , L. M. de Godoy, G. Li, B. Macek, P. Mortensen, R. Pesch, A. Makarov, 0. Lange, S. Horning, and M. Mann. 2005. Parts per million mass accuracy on an Orbitrap mass spectrometer via lock mass injection into a C-trap.Mol Cell Proteomics4:2010-2021)基于結(jié)合在用活的 EB 脈沖（pulsed)DC 后 在DC表面上呈遞的MHC II類分子的病原體衍生的肽的分離和測(cè)序，其導(dǎo)致鑒定了許多 衍生自8種新型表位的小鼠生物型沙眼衣原體（1^1'111^1^四11，1