Kassa Desta, Ran Leonie, Geberemeskel Wudneh, Tebeje Mekashaw, Alemu Amelewerk, Selase Alemayehu, Tegbaru Belete, Franken Kees L M C, Friggen Annemieke H, van Meijgaarden Krista E, Ottenhoff Tom H M, Wolday Dawit, Messele Tsehaynesh, van Baarle Debbie
Infectious and Noninfectious Diseases Research Directorate, Ethiopian Health and Nutrition Research Institute, Addis Ababa, Ethiopia.
Clin Vaccine Immunol. 2012 Dec;19(12):1907-15. doi: 10.1128/CVI.00482-12. Epub 2012 Sep 26.
Characterizing host immune responses to molecular targets of Mycobacterium tuberculosis is essential to develop effective immunodiagnostics and better vaccines. We investigated the immune response against a large series of M. tuberculosis antigens, including 5 classical and 64 nonclassical (39 DosR regulon-encoded, 4 resuscitation-promoting factor [RPF], and 21 reactivation-associated) antigens in active-pulmonary-tuberculosis (TB) patients. Whole blood from TB patients (n = 34) was stimulated in vitro with M. tuberculosis antigens. Gamma interferon (IFN-γ) was measured after 7 days of stimulation, using an enzyme-linked immunosorbent assay (ELISA). The majority of the study participants responded to the classical M. tuberculosis antigens TB10.4 (84.8%), early secreted antigenic target-6 kDa (ESAT-6)/CFP-10 (70.6%), and purified protein derivative (PPD) (55.9%). However, only 26.5% and 24.2% responded to HSP65 and Ag85A/B, respectively. Of the 64 nonclassical antigens, 23 (33.3%) were immunogenic (IFN-γ levels, >62 pg/ml) and 8 were strong inducers of IFN-γ (IFN-γ levels, ≥100 pg/ml). The RPF antigens were the most immunogenic. In addition, we observed distinct cytokine expression profiles in response to several M. tuberculosis antigens by multiplex immunoassay. Tumor necrosis factor alpha (TNF-α), interleukin 10 (IL-10), and IL-6 were commonly detected at high levels after stimulation with 4/15 latency antigens (Rv0081, Rv2006, Rv2629, and Rv1733c) and were found especially in supernatants of the three strong IFN-γ inducers (Rv2629, Rv1009, and Rv2389c). IL-8, IL-6, and IL-17 were exclusively detected after stimulation with Rv0574c, Rv2630, Rv1998, Rv054c, and Rv2028c. In conclusion, in active-pulmonary-TB patients, we identified 23 new immunogenic M. tuberculosis antigens. The distinct expression levels of IFN-γ, TNF-α, IL-6, and IL-10 in response to specific subsets of M. tuberculosis antigens may be promising for the development of immunodiagnostics.
表征宿主对结核分枝杆菌分子靶点的免疫反应对于开发有效的免疫诊断方法和更好的疫苗至关重要。我们研究了活动性肺结核（TB）患者针对一系列结核分枝杆菌抗原的免疫反应，这些抗原包括5种经典抗原和64种非经典抗原（39种DosR调控子编码抗原、4种复苏促进因子[RPF]和21种再激活相关抗原）。用结核分枝杆菌抗原体外刺激34例TB患者的全血。刺激7天后，使用酶联免疫吸附测定（ELISA）测量γ干扰素（IFN-γ）。大多数研究参与者对经典结核分枝杆菌抗原TB10.4（84.8%）、早期分泌抗原靶标6 kDa（ESAT-6）/CFP-10（70.6%）和纯化蛋白衍生物（PPD）（55.9%）有反应。然而，分别只有26.5%和24.2%的参与者对HSP65和Ag85A/B有反应。在64种非经典抗原中，23种（33.3%）具有免疫原性（IFN-γ水平>62 pg/ml），8种是IFN-γ的强诱导剂（IFN-γ水平≥100 pg/ml）。RPF抗原的免疫原性最强。此外，我们通过多重免疫测定观察到针对几种结核分枝杆菌抗原的不同细胞因子表达谱。在用4种潜伏期抗原（Rv0081、Rv2006、Rv2629和Rv1733c）刺激后，肿瘤坏死因子α（TNF-α）、白细胞介素10（IL-10）和IL-6通常被检测到处于高水平，并且特别在三种强IFN-γ诱导剂（Rv2629、Rv1009和Rv2389c）的上清液中发现。在用Rv0574c、Rv2630、Rv1998、Rv054c和Rv2028c刺激后，仅检测到IL-8、IL-6和IL-17。总之，在活动性肺结核患者中，我们鉴定出23种新的具有免疫原性的结核分枝杆菌抗原。针对结核分枝杆菌抗原特定亚群的IFN-γ、TNF-α、IL-6和IL-10的不同表达水平可能对免疫诊断的开发具有前景。
