Proteomic profiling of peripheral blood mononuclear cells isolated from patients with tuberculosis and diabetes copathogenesis -A pilot study

Abstract

18Background: Diabetes is an important risk factor for developing tuberculosis. This association 19 leads to exacerbation of tuberculosis symptoms and delayed treatment of both the diseases. 20 Molecular mechanism and biomarkers/drug targets related to copathogenesis of tuberculosis and 2 21 diabetes, however, still remains to be poorly understood. In this study, proteomics based 2D-22 MALDI/MS approach was employed to identify host signature proteins which are altered during 23 copathogenesis of tuberculosis and diabetes. 24 Methods: Comparative proteome of human peripheral blood mononuclear cells (PBMCs) from 25 healthy controls, tuberculosis and diabetes patients in comparison to comorbid diabetes and 26 tuberculosis patients was analyzed. Gel based proteomics approach followed by in gel trypsin 27 digestion and peptide identification by mass spectrometry was used for signature protein 28 identification. 29 Results: Total of 18 protein spots with differential expression in TBDM patients in comparison 30 to other groups were identified. These include Vimentin, tubulin beta chain protein, superoxide 31 dismutase, Actin related protein 2/3 complex subunit 2, PDZ LIM domain protein, Rho-GDP 32 dissociation inhibitor, Ras related protein Rab, dCTPpyrophosphatase 1, Transcription initiation 33 factor TFIID subunit 12, coffilin 1, three isoforms of Peptidylprolylcis-trans isomerase A, three 34 isoforms of Protein S100A9, Protein S100A8 and SH3 domain containing protein. These 35 proteins belonged to four functional categories i.e. structural, cell cycle/growth regulation, 36 signaling and intermediary metabolism.40 42 10million fresh TB cases has been reported worldwide (1). Despite extensive research on the 3 43 biology of M. tuberculosis, exact mechanism of infection and immune evasion still remains 44 elusive. Copathogenesis with HIV and diabetes further complicates the tuberculosis control 45 measures. Although HIV infection is the topmost risk factor for development of active 46 tuberculosis but population attributable risk of diabetes is more than that of HIV infection (2, 3) 47 as diabetes is known to triple the risk of developing active tuberculosis (4). As diabetes is 48 associated with various immunological dysfunctions, diabetic patients fall prey to other co-49 infections like tuberculosis, melioidosis and other conventional hyperglycemia related 50 complications like various cardiovascular disorders, retinopathy, nephropathy and many more. 51 Diabetes mellitus affected 425 million individuals worldwide in 2017 and is predicted to reach 52 629 million by 2045, the time at which 80% of diabetics will be residents of economically 53 challenged countries where active tuberculosis (TB) prevails (5, 6). Countries with highest 54 burden of diabetes are also in the list of WHO's tuberculosis high burden countries and India and 55China for example, have first two positions for the copathogenesis of TB and diabetes (7, 8). The 56 epidemiological data for association between TB and DM is increasing significantly...

Human peripheral blood mononuclear cells: A review of recent proteomic applications

“…For example, in the 2 studies investigating the PBMCs exposure to low dose radiation [32, 33], there were seven proteins or almost half of the proposed candidates in each of the studies that overlapped between the studies exhibiting great reproducibility. Similarly, two studies investigating sepsis [18–35] and those investigating diabetes and tuberculosis‐diabetes co‐pathogenesis [11–20], also identified mutual candidates. The most promising PBMCs biomarker based on specificity and validation approaches performed, are marked with red circles in Figure 4.…”

“…It was found that the purified Rv0147 protein stimulated dendritic cells of MDR‐TB patients to increase production of INF‐γ while the production of IL‐10 was not detected supporting its potential as protein vaccine candidate against tuberculosis. In another work which also studied tuberculosis effect on PBMCs [11], the molecular mechanism and biomarkers associated with co‐pathogenesis of tuberculosis and diabetes were studied. Comparative proteomics of healthy controls, tuberculosis patients, diabetes patients and tuberculosis and diabetes co‐pathogenesis (TBDM) patients was done.…”

“…Commonly, centrifugation with 1.077 g mL –1 gradient medium (Ficoll™, Lymphoprep™) is used to isolate PBMCs from the other blood elements. After separation, PBMCs are dispensed in the buffy coat situated between plasma constituting the upper fraction, and the red blood cells and polymorphonuclear leucocytes representing the lower fraction [8–19]. When aiming to isolate PBMCs subpopulations/subsets such as CD4+ T cells, magnetic‐activated cell sorting (MACS) can be further helpful [20].…”

“…After recognition of antigen/body intruder, PBMCs turn from naïve/resting to state, undergoing differentiation and development of effector functions [8–10]. Changes associated with inflammation [11], immune‐mediated inflammation—autoimmune disorders [12, 13], genetic pathology towards neurodegeneration [14], mental disorder alternations [15, 16], cancer progression [10] or virus‐hosting—SARS‐CoV‐2 pathogenesis [17], are to name a few.…”