Department of Laboratory Medicine, Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Laboratory Medicine, Hubei University of Chinese Medicine, Wuhan, China.
J Clin Lab Anal. 2020 Nov;34(11):e23477. doi: 10.1002/jcla.23477. Epub 2020 Jul 16.
Mycobacterium tuberculosis (Mtb)-specific perforin were significantly increased in patients with tuberculosis. This study aims to evaluate the diagnosis value of Mtb-specific perforin in pediatric patients with tuberculosis.
Diagnostic performance of perforin levels induced by 6-kDa early secreted antigen target (ESAT6) or culture filtered protein 10 (CFP10) were evaluated in eighty-six samples from children participants by receiver operating characteristic curve analysis. Flow cytometry was used to detect the expression of perforin and INF-γ of CD4 , CD8 T cells in response to CFP10 stimulation.
After ex vivo stimulation, levels of ESAT6/CFP10-specific perforin in LTBI patients were significantly higher than active TB (ATB) patients, non-tuberculosis infection (non-TB), and health control (HC) individuals. The diagnostic efficacy of CFP10-specific perforin for TB diagnosis was significantly higher than ESAT6-specific perforin and T-SPOT assay, and when 0.74 ng/mL was taken as the cutoff value, the sensitivity, specificity, and accuracy were 97.83%, 87.5%, and 93.02%. CFP10-specific perforin in both CD4 and CD8 T cells were significantly higher in ATB patients compared to HCs and further increased in LTBI patients. However, INF-γ was mainly secreted by CD4 T cells and showed no significant difference between LTBI and ATB patients. In addition, CFP10-specific perforin can effectively distinguish between ATB and LTBI with the cutoff value of 1.80 ng/mL. Sensitivity and specificity were 88.46% and 85.62%, respectively.
CFP10-specific perforin may be used as a novel cellular immunity-based diagnostic marker of pediatric patients with tuberculosis, and with the potential for discriminating ATB from LTBI.
结核分枝杆菌(Mtb)特异性穿孔素在结核病患者中显著增加。本研究旨在评估 Mtb 特异性穿孔素在儿童结核病患者中的诊断价值。
通过受试者工作特征曲线分析,评估 86 例患儿样本中 6kDa 早期分泌抗原靶标(ESAT6)或培养过滤蛋白 10(CFP10)诱导的穿孔素水平的诊断性能。流式细胞术用于检测 CF10 刺激后 CD4、CD8 T 细胞中穿孔素和 INF-γ的表达。
在体外刺激后,LTBI 患者的 ESAT6/CFP10 特异性穿孔素水平明显高于 ATB 患者、非结核感染(non-TB)和健康对照(HC)个体。CFP10 特异性穿孔素对结核病的诊断效能明显高于 ESAT6 特异性穿孔素和 T-SPOT 检测,当以 0.74ng/mL 为截值时,其灵敏度、特异性和准确性分别为 97.83%、87.5%和 93.02%。与 HC 相比,ATB 患者的 CD4 和 CD8 T 细胞中的 CFP10 特异性穿孔素均显著升高,而 LTBI 患者的穿孔素进一步升高。然而,INF-γ主要由 CD4 T 细胞分泌,LTBI 和 ATB 患者之间无显著差异。此外,CFP10 特异性穿孔素可有效区分 ATB 和 LTBI,截值为 1.80ng/mL,灵敏度和特异性分别为 88.46%和 85.62%。
CFP10 特异性穿孔素可作为儿童结核病患者新型细胞免疫诊断标志物,具有区分 ATB 和 LTBI 的潜力。