Sauzullo Ilaria, Mengoni Fabio, Mascia Claudia, Rossi Raffaella, Lichtner Miriam, Vullo Vincenzo, Mastroianni Claudio M
Department of Public Health and Infectious Diseases, Sapienza University, Piazzale Aldo Moro 1, 00185, Rome, Italy.
Infectious Diseases Unit, Fondazione Eleonora Lorillard Spencer Cenci, Sapienza University, Latina, Italy.
Med Microbiol Immunol. 2016 Feb;205(1):37-45. doi: 10.1007/s00430-015-0424-z. Epub 2015 Jun 25.
To ascertain whether multiparametric flow cytometry assessment of multifunctional Mycobacterium tuberculosis (Mtb)-specific CD4(+) and CD8(+) T cells can distinguish between untreated and treated patients with latent tuberculosis infection (LTBI), we enrolled 14 LTBI subjects treated with isoniazid (INH) therapy, 16 untreated LTBI patients, and 25 healthy controls. The analysis of mono-functional CD4(+) and CD8(+) T cells producing single cytokines showed significant differences only between uninfected and infected LTBI subjects (both treated and untreated). Conversely, the analysis of multifunctional CD4(+) T cells revealed a significant reduction in the frequency of two CD4(+) T cells subsets, those producing IFN-γ, IL-2, and TNF-α simultaneously (triple positive; p = 0.005) and those producing IL-2 alone (p = 0.0359), as well as a shift towards T cells producing only one cytokine in treated as compared to untreated LTBI subjects. Assigning a triple-positive CD4(+) T cells a cut-off >0.082 %, 94 % of untreated LTBI patients were scored as positive, as compared to only 28 % of treated LTBI patients and none of the healthy controls. No significant differences between untreated and treated LTBI subjects in terms of Mtb-specific CD8(+) T cell cytokine profiles (p > 0.05) were identified. The significant changes in the cytokine profiles of Mtb-specific T cells after INH therapy suggest that analysis of multifunctional T cells may be a promising means for the monitoring of LTBI treatment success.
为确定对多功能结核分枝杆菌（Mtb）特异性CD4(+)和CD8(+) T细胞进行多参数流式细胞术评估能否区分未治疗和已治疗的潜伏性结核感染（LTBI）患者，我们招募了14名接受异烟肼（INH）治疗的LTBI受试者、16名未治疗的LTBI患者和25名健康对照。对产生单一细胞因子的单功能CD4(+)和CD8(+) T细胞的分析显示，仅在未感染和感染的LTBI受试者（包括治疗和未治疗的）之间存在显著差异。相反，对多功能CD4(+) T细胞的分析显示，两个CD4(+) T细胞亚群的频率显著降低，即同时产生IFN-γ、IL-2和TNF-α的亚群（三阳性；p = 0.005）和仅产生IL-2的亚群（p = 0.0359），并且与未治疗的LTBI受试者相比，治疗后的LTBI受试者中产生单一细胞因子的T细胞出现了偏移。将三阳性CD4(+) T细胞的临界值设定为>0.082%，94%的未治疗LTBI患者被判定为阳性，而治疗后的LTBI患者中这一比例仅为28%，健康对照中则无阳性。在Mtb特异性CD8(+) T细胞细胞因子谱方面，未治疗和治疗后的LTBI受试者之间未发现显著差异（p > 0.05）。INH治疗后Mtb特异性T细胞细胞因子谱的显著变化表明，对多功能T细胞的分析可能是监测LTBI治疗成功的一种有前景的方法。
