Dengue (DENV) and chikungunya (CHIKV) viral infections trigger high patient morbidity and mortality. Mono-/co-infection of these viruses activates innate immune response, triggering Toll-like receptor (TLR) pathways. The present study investigated the differential role of TLR3, 7 and 8 single-nucleotide polymorphisms (SNPs) between mono- and co-infected Eastern Indian patients. Interaction of TLR polymorphic variants with signal peptidase complex (SPC18) was explored which might affect immune signalling against DENV/CHIKV infections. Out of 550 febrile symptomatic patients, 128 DENV-CHIKV co-infected samples were genotyped for eight SNPs of TLR3 (rs3775290-chr4:186083063), TLR7 (rs179008-chrX:12885540, rs5741880-chrX:12869297, rs179010-chrX:12884766, rs3853839-chrX:12889539) and TLR8 (rs5744080-chrX:12919685, rs3764879-chrX:12906578, rs3764880-chrX:12906707) by PCR-RFLP along with 157 healthy individuals. Statistical analysis established genotypic association of TLR SNPs with DENV-CHIKV co-infection, and difference between mono- and co-infected patients and their role in determining high viral load (HVL) during competitive viral replication among co-infected patients. In silico protein-protein docking evaluated interactive effect of TLR variants with SPC18. The findings revealed patients with CC genotypes of TLR7 and 8 SNPs were significantly susceptible towards co-infection, whereas specific genotypes of TLR7 and 8 imparted protection against co-infection. Differential analysis between mono-/co-infected patients revealed distinct genotypic distribution of TLR3, 7 and 8 SNPs. Co-infected patients with TT-rs179010 exhibited DENV-HVL, whereas CHIKV-HVL was detected among patients with other genotypes. Molecular docking of TLR7-rs179008 Q variant and TLR8-rs3764880 V variant with SPC18 generated better free binding energy. This study underlined the importance of TLR7 and 8 SNPs towards mono-/co-infection of DENV/CHIKV, with certain genotypes associated with co-infection susceptibility. Moreover, it suggested a probable role of specific genotypes of TLR7 and 8 polymorphisms imparting high dengue/chikungunya viral load among co-infected patients.
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The authors are extremely grateful to the Director, Calcutta School of Tropical Medicine, Kolkata, India, for his support, inspiration and providing of necessary facilities for this study. The authors are grateful to Indian Council of Medical Research, India, for granting fellowship to the first author [5/3/8/6/ITR-F/2018-ITR]. The authors also acknowledge Mr. Advaita Chakraborty, Lab Manager, Department of Biomedical Sciences, College of Health Sciences, Marquette University, Milwaukee, WI, for providing his email ID for using QUARK server and ClusPro 2.0 server.
All procedures performed in this study involving collection of blood from human participants as well as healthy controls were in accordance with ethical standards of Clinical Research Ethical Committee of Calcutta School of Tropical Medicine (CREC-STM/53 dated 26 September 2013).
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Sengupta, S., Mukherjee, S., Bhattacharya, N. et al. Differential genotypic signatures of Toll-like receptor polymorphisms among dengue-chikungunya mono- and co-infected Eastern Indian patients. Eur J Clin Microbiol Infect Dis (2021). https://doi.org/10.1007/s10096-020-04125-x
- Single-nucleotide polymorphism
- Signal peptide