Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
Immunol Res. 2009;45(1):1-12. doi: 10.1007/s12026-008-8024-2.
Purified protein derivative (PPD) or tuberculin skin testing is used to identify infected individuals with Mycobacterium tuberculosis (Mtb) and to assess cell-mediated immunity to Mtb. In the present study, we compared PBMC cultures in the presence of tuberculin or Candida antigens using cytokine bead arrays and RNA microarrays. Measurements of different cytokines and chemokines in supernatants of PMBC cultures in the presence of PPD showed increased levels of interferon (IFN)-gamma in active tuberculosis infection (ATBI) and latent TB infected (LTBI) compared to controls, and increased levels of TNF-alpha in ATBI compared with LTBI. Also, we found increase of IL-6 in cultures of PPD positive and controls but not in the cultures with Candida. We also report the molecular signature of tuberculosis infection, in ATBI patients, the following genes were found to be up-regulated and absent in LTBI individuals: two kinases (JAK3 and p38MAPK), four interleukins (IL-7, IL-2, IL-6, and IFNbeta1), a chemokine (HCC-4) a chemokine receptor (CxCR5), two interleukin receptors (IL-1R2 and IL-18R1), and three additional ones (TRAF5, Smad2, CIITA, and NOS2A). By contrast, IL-17 and IGFBP3 were significantly up-regulated in LTBI. And, STAT4, GATA3, Fra-1, and ICOS were down-regulated in ATBI but absent in LTBI. Conversely, TLR-10, IL-15, DORA, and IKK-beta were down-regulated in LTBI but not in ATBI. Interestingly, the majority of the up-regulated genes found in ATBI were found in cultures stimulated with tuberculin (PPD) or Candida antigens, suggesting that these pathogens stimulate similar immunological pathways. We believe that the molecular signature distinguishing active from latent tuberculosis infection may require using cytokine bead arrays along with RNA microarrays testing cell cultures at different times following in vitro proliferation assays using several bacterial antigens and PPD.
纯化蛋白衍生物（PPD）或结核菌素皮肤试验用于识别感染结核分枝杆菌（Mtb）的个体，并评估对 Mtb 的细胞介导免疫。在本研究中，我们比较了在结核菌素或念珠菌抗原存在下的 PBMC 培养物，使用细胞因子珠阵列和 RNA 微阵列。在 PPD 存在下 PMBC 培养物上清液中不同细胞因子和趋化因子的测量显示，与对照组相比，活动性结核感染（ATBI）和潜伏性结核感染（LTBI）中干扰素（IFN）-γ水平升高，与 LTBI 相比，ATBI 中 TNF-α水平升高。此外，我们发现 PPD 阳性和对照组培养物中 IL-6 增加，但念珠菌培养物中没有增加。我们还报告了结核感染的分子特征，在 ATBI 患者中，以下基因被发现上调，而 LTBI 个体中不存在：两种激酶（JAK3 和 p38MAPK）、四种白细胞介素（IL-7、IL-2、IL-6 和 IFNbeta1）、一种趋化因子（HCC-4）、一种趋化因子受体（CxCR5）、两种白细胞介素受体（IL-1R2 和 IL-18R1）和另外三个（TRAF5、Smad2、CIITA 和 NOS2A）。相比之下，LTBI 中 IL-17 和 IGFBP3 显著上调。并且，ATBI 中 STAT4、GATA3、Fra-1 和 ICOS 下调，但 LTBI 中不存在。相反，LTBI 中 TLR-10、IL-15、DORA 和 IKK-beta 下调，但 ATBI 中不存在。有趣的是，在 ATBI 中发现的大多数上调基因在刺激结核菌素（PPD）或念珠菌抗原的培养物中发现，这表明这些病原体刺激相似的免疫途径。我们认为，区分活动性和潜伏性结核感染的分子特征可能需要使用细胞因子珠阵列和 RNA 微阵列，同时测试不同时间的细胞培养物，在体外增殖试验后使用几种细菌抗原和 PPD。
