Mohammadi Akram Miramin, Javadi Amir, Firooz Alireza, Khamesipour Ali
Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran.
Department of Social Medicines, Qazvin University of Medical Sciences, Qazvin, Iran.
Iran J Microbiol. 2019 Oct;11(5):431-439.
The surrogate marker (s) of cure and protection in intracellular pathogens is not yet well defined. The aim of this study was to compare the cytokine profile using whole blood cells (WBC) vs. peripheral blood mononuclear cells (PBMC) in healthy and cutaneous leishmaniasis (CL) volunteers.
In this study, WBC and PBMC of the volunteers with history of CL (HCL), Active lesion (ACL) and healthy volunteers were collected. The WBC and PBMC were cultured and stimulated with either PHA or soluble antigens (SLA), after 72 hours, the supernatants were collected and the levels of IFN-γ, IL-5 and IL-10 were titrated using ELISA method.
The mean ± SD of cytokines using WBC and PBMC in cutaneous leishmaniasis volunteers stimulated with phytohemagglutin (PHA) or SLA are as follow, PHA, IFN-γ=2295±995 vs. 2339±1115, IL-10=853±309 vs. 1330±966, and IL-5=299±136 vs. 352+156, SLA, IFN-γ, 931±824 vs. 825±532, IL-10, 233±78 vs. 408±381, and IL-5, 185±59 vs. 217±76, respectively. There was no significant difference between the IFN-γ, IL-5 and IL-10 levels using WBC vs. PBMC. There was a strong correlation between the cytokine profiles using WBC and PBMC in cutaneous leishmaniasis volunteers.
There was no significant difference between IFN-γ, IL-10, IL-5 levels in whole blood and PBMC of volunteers with active lesion or history of CL. Whole-blood culture which is easier, cheaper and more convenient could be used instead of PBMC to evaluate the cytokine profile in field conditions.
细胞内病原体治愈和保护的替代标志物尚未明确界定。本研究的目的是比较健康志愿者和皮肤利什曼病(CL)志愿者使用全血细胞(WBC)与外周血单个核细胞(PBMC)时的细胞因子谱。
在本研究中,收集了有CL病史(HCL)、活动性病灶(ACL)的志愿者以及健康志愿者的WBC和PBMC。将WBC和PBMC进行培养,并用PHA或可溶性抗原(SLA)刺激,72小时后收集上清液,采用ELISA法测定IFN-γ、IL-5和IL-10的水平。
在皮肤利什曼病志愿者中,用植物血凝素(PHA)或SLA刺激时,使用WBC和PBMC的细胞因子的均值±标准差如下:PHA刺激时,IFN-γ分别为2295±995和2339±1115,IL-10分别为853±309和1330±966,IL-5分别为299±136和352+156;SLA刺激时,IFN-γ分别为931±824和825±532,IL-10分别为233±78和408±381,IL-5分别为185±59和217±76。使用WBC与PBMC时,IFN-γ、IL-5和IL-10水平无显著差异。皮肤利什曼病志愿者中使用WBC和PBMC的细胞因子谱之间存在强相关性。
活动性病灶或有CL病史的志愿者的全血和PBMC中的IFN-γ、IL-10、IL-5水平无显著差异。在现场条件下,全血培养更简便、便宜且方便,可用于替代PBMC来评估细胞因子谱。