Pitabut Nada, Dhepakson Panadda, Sakurada Shinsaku, Keicho Naoto, Khusmith Srisin
Faculty of Medicine, King Mongkut's Institute of Technology Ladkrabang, Bangkok 10520, Thailand.
Medical Biotechnology Center, Medical Life Sciences Institute, Department of Medical Sciences, Ministry of Public Health, Nonthaburi 11000, Thailand.
Pathogens. 2020 Aug 14;9(8):655. doi: 10.3390/pathogens9080655.
Granule-associated killing molecules released from cytotoxic T lymphocytes participate as a crucial step in immunity against tuberculosis (TB), but the role of coordinated production remains controversial. Coordinated release of effector molecules in vitro after stimulating peripheral blood mononuclear cells (PBMCs) of active TB or HIV/TB coinfection patients with PPD, purified protein derivative of tuberculin and avirulent , H37Ra, an attenuated strain were investigated in association with clinical outcomes. Perforin, granzyme-B, granulysin and IFN-γ were measured using ELISA. Before anti-TB treatment, PBMCs of TB stimulated with PPD or H37Ra released higher perforin, granzyme-B, and granulysin levels than in HIV/TB and released significantly higher IFN-γ ( = 0.045, = 0.022). Granulysin positively correlated with perforin in TB ( = 0.042, r = 0.385), HIV/TB coinfection ( = 0.003, r = 0.941) after PPD stimulation, and after H37Ra stimulation in TB ( = 0.005, r = 0.549), but negatively correlated with granzyme B in TB ( = 0.042, r = -0.386), HIV/TB coinfection ( = 0.042, r = 0.754) were noted. After anti-TB treatment, increased levels of perforin, granulysin and IFN-γ in TB or HIV/TB upon PPD or H37Ra stimulation, and decreased granzyme-B levels after PPD ( = 0.003) or H37Ra ( = 0.028) stimulation in TB were observed. These results suggest that granulysin may act synergistic with perforin and IFN-γ in TB, indicating its crucial function in host immunity to tuberculosis. Future studies with larger numbers of patients ought to be conducted in the future.
细胞毒性T淋巴细胞释放的颗粒相关杀伤分子参与抗结核免疫的关键步骤，但协同产生的作用仍存在争议。研究了用结核菌素纯蛋白衍生物（PPD）刺激活动性结核病或HIV/TB合并感染患者的外周血单个核细胞（PBMC）后，效应分子在体外的协同释放情况，PPD是结核菌素的纯化蛋白衍生物，无毒力的H37Ra是一种减毒株，同时研究了其与临床结果的关系。采用酶联免疫吸附测定法（ELISA）检测穿孔素、颗粒酶B、颗粒溶素和干扰素-γ。在抗结核治疗前，用PPD或H37Ra刺激的结核病患者的PBMC释放的穿孔素、颗粒酶B和颗粒溶素水平高于HIV/TB患者，且释放的干扰素-γ水平显著更高（P = 0.045，P = 0.022）。在PPD刺激后，颗粒溶素与结核病患者的穿孔素呈正相关（P = 0.042，r = 0.385），与HIV/TB合并感染患者呈正相关（P = 0.003，r = 0.94），在H37Ra刺激后，与结核病患者的穿孔素也呈正相关（P = 0.005，r = 0.549），但与结核病患者的颗粒酶B呈负相关（P = 0.042，r = -0.386），与HIV/TB合并感染患者的颗粒酶B呈负相关（P = 0.042，r = 0.754）。抗结核治疗后，在PPD或H37Ra刺激下，结核病或HIV/TB患者的穿孔素、颗粒溶素和干扰素-γ水平升高，在PPD（P = 0.003）或H37Ra（P = 0.028）刺激后，结核病患者的颗粒酶B水平降低。这些结果表明，颗粒溶素可能在结核病中与穿孔素和干扰素-γ协同发挥作用，表明其在宿主抗结核免疫中的关键作用。未来应该对更多患者进行研究。
