Multi-Dimensionality Immunophenotyping Analyses of MAIT Cells Expressing Th1/Th17 Cytokines and Cytotoxic Markers in Latent Tuberculosis Diabetes Comorbidity.

Gokul Raj Kathamuthu,Kadar Moideen,Subash Babu,Nathella Pavan Kumar,Paul Kumaran,Chandrakumar Dolla

doi:10.3390/pathogens11010087

Abstract

Mucosal-associated invariant T (MAIT) cells are innate like, and play a major role in restricting disease caused by Mycobacterium tuberculosis (Mtb) disease before the activation of antigen-specific T cells. Additionally, the potential link and synergistic function between diabetes mellitus (DM) and tuberculosis (TB) has been recognized for a long time. However, the role of MAIT cells in latent TB (LTB) DM or pre-DM (PDM) and non-DM (NDM) comorbidities is not known. Hence, we examined the frequencies (represented as geometric means, GM) of unstimulated (UNS), mycobacterial (purified protein derivative (PPD) and whole-cell lysate (WCL)), and positive control (phorbol myristate acetate (P)/ionomycin (I)) antigen stimulated MAIT cells expressing Th1 (IFNγ, TNFα, and IL-2), Th17 (IL-17A, IL-17F, and IL-22), and cytotoxic (perforin (PFN), granzyme (GZE B), and granulysin (GNLSN)) markers in LTB comorbidities by uniform manifold approximation (UMAP) and flow cytometry. We also performed a correlation analysis of Th1/Th17 cytokines and cytotoxic markers with HbA1c, TST, and BMI, and diverse hematological and biochemical parameters. The UMAP analysis demonstrated that the percentage of MAIT cells was higher; T helper (Th)1 cytokine and cytotoxic (PFN) markers expressions were different in LTB-DM and PDM individuals in comparison to the LTB-NDM group on UMAP. Similarly, no significant difference was observed in the geometric means (GM) of MAIT cells expressing Th1, Th17, and cytotoxic markers between the study population under UNS conditions. In mycobacterial antigen stimulation, the GM of Th1 (IFNγ (PPD and WCL), TNFα (PPD and WCL), and IL-2 (PPD)), and Th17 (IL-17A, IL-17F, and IL-22 (PPD and/or WCL)) cytokines were significantly elevated and cytotoxic markers (PFN, GZE B, and GNLSN (PPD and WCL)) were significantly reduced in the LTB-DM and/or PDM group compared to the LTB-NDM group. Some of the Th1/Th17 cytokines and cytotoxic markers were significantly correlated with the parameters analyzed. Overall, we found that different Th1 cytokines and cytotoxic marker population clusters and increased Th1 and Th17 (IL-17A, IL-22) cytokines and diminished cytotoxic markers expressing MAIT cells are associated with LTB-PDM and DM comorbidities.

Highlights

Latent tuberculosis (LTB) does not have any of the clinical symptoms of active tuberculosis (ATB): it is defined by the presence of an immunological reaction to Mycobacterium tuberculosis (Mtb) antigens as determined by a tuberculin skin test (TST) or an interferongamma release assay (IGRA) [1,2,3,4]
The multi-dimensionality representative plots of gated Mucosal-associated invariant T (MAIT) cells expressing Th1, Th17, and cytotoxic markers for LTB (DM, PDM, and NDM) comorbidities are depicted by the unsupervised clustering of flow data using two-dimensional uniform manifold approximation (UMAP) (UMAP_1 vs. UMAP_2)
We provide the UMAP statistical values of MAIT cells expressing Th1, Th17 cytokines, and cytotoxic markers in Supplementary Materials Table S1

Summary

Introduction

Latent tuberculosis (LTB) does not have any of the clinical symptoms of active tuberculosis (ATB): it is defined by the presence of an immunological reaction to Mycobacterium tuberculosis (Mtb) antigens as determined by a tuberculin skin test (TST) or an interferongamma release assay (IGRA) [1,2,3,4]. Only a small fraction (5–15%) of LTB-infected individuals have a lifetime risk of reactivation of ATB, with the majority of progression occurring within the first 2–5 years following initial exposure [5,6]. LTB serves as a key reservoir for the development of ATB [7]. Is increasing, and by the year 2030, around 470 million individuals will develop PDM; depending on the demographic and geographic situation, 5–10% of them will develop overt diabetes each year [10,11,12]. PDM was correlated with increased risk of LTB infection [7] and is known to be connected with pulmonary TB (PTB) patients with respiratory ailments [8]

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