Hepatitis E rORF2p Stimulated and Unstimulated Peripheral Expression Profiling in Patients with Self-Limiting Hepatitis E Infection

Abstract

To improve the current knowledge on the involvement of peripheral lymphocytes in hepatitis E virus (HEV) associated pathogenesis, we analyzed alterations in (1) immunophenotypic expressions (by flow cytometry) and (2) gene expression patterns (by TaqMan Low Density Array) of activatory, inhibitory, integrin, homing, ectonucleotidase machinery, costimulatory, inflammatory markers, and T regulatory cells (Treg) associated cytokines on HEV rORF2p stimulated and unstimulated PBMCs of 43 acute HEV patients, 30 recovered individuals, and 43 controls. The phenotypic expressions of key molecules CTLA-4, GITR, CD103, CD25, CD69, IL10 and TGF-β 1 in the acute patients and TGF-β 1 in the recovered individuals were significantly elevated on both unstimulated and stimulated PBMCs. Gene expression array data revealed upregulations of CD25, PD1, CD103, CCR4, IL10, and TGF-β 1 on both unstimulated and HEV rORF2p stimulated PBMCs of acute patients. The observed upregulations of inhibitory, integrin, activatory, and Treg-associated cytokine genes on the PBMCs of acute HEV patients complemented by their frequency data suggest them as the major players in the fine-tuning of immune response in self-limiting hepatitis E infection.

Peripheral lymphocyte subset alteration in patients with COVID-19 having differential clinical manifestations

“…Based on the reduced percentages of NK cells in COVID-19 patients and the well-known antiviral role of NK cells, it has been speculated that the restoration of NK cells and their function may be a near-future solution for COVID-19 patients 1 15 18 23 . In our study asymptomatic, mild symptomatic patients and recovered individuals demonstrated a similar low NK cell profile, consistent with other reports.…”

“…Natural Killer (NK) and NKT cells, T-cells and subsets, B and B memory cells, T regulatory cells (Tregs) enumeration : Freshly isolated peripheral blood mononuclear cells (PBMCs) (0.5 X 10 5 ) from all study participants were stained with antibodies specific for NK and NKT, T and subsets, B and B memory, T regulatory (Tregs) cells using anti-human CD3PerCP (cloneSK7), CD56APC (cloneNCAM16.2), CD4PE (cloneRPAT4), CD8APCH7 (cloneRPAT8), CD19PECy7 (cloneSJ25C1), CD27APCH7 (cloneM-T271), CD45ROAPC (cloneUCHL1) monoclonal antibodies and Treg cocktail [CD127Alexa Fluor 647 (clone HIL-7R), CD25PE-Cy7, CD4FITC (clone7RM21|2A3|SK3)] as previously described 12 13 14 15 16 . PE-Cy™7 Mouse IgG1 κ Isotype Control (BD Biosciences, San Jose, CA, USA) was used as negative control.…”

“…PBMCs isolation, surface staining and acquisition in flow cytometer : It was carried out following the methods described previously 12 13 14 15 16 . Data were analyzed using FACS Diva software (Version 8.14 BD Biosciences, San Jose, CA, USA) and results are expressed as the percentage of positive cells in the gated population ( Fig.…”

Beyond the Usual Suspects: Hepatitis E Virus and Its Implications in Hepatocellular Carcinoma

“…So far, comprehensive in vitro and in vivo studies have explored the mechanisms of inflammation and host immune response in acute hepatitis E patients [ 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 ]. For instance, immunohistochemistry staining of liver biopsy samples of patients diagnosed with HEV-related acute liver failure showed an increased infiltration of cytotoxic T-cells (CD3, CD8), natural killer cells (NK-cells; CD56) and helper T-cells (CD4) compared to liver biopsies of healthy patients [ 95 ].…”

Increased Mast Cell Activation in Mongolian Gerbils Infected by Hepatitis E Virus

“…Hepatitis E virus (HEV) is a non-enveloped, positive sense single-stranded RNA virus that belongs to the family Hepeviridae ( Smith et al, 2014 ), which is one of the main causes of acute hepatitis in humans and many other species ( Martin-Latil et al, 2016 ; Donnelly et al, 2017 ). Although, hepatitis E is largely a self-limiting disease ( Rathod and Tripathy, 2014 , 2016 ), pregnant women and fetals are exceptionally susceptible to HEV infection ( Nan et al, 2017 ; Verma et al, 2017 ). The mortality rate of pregnant women in the third trimester of pregnancy infected with HEV is as high as 25%, extremely easy to death of the mother and fetus, abortion, premature delivery or stillbirth ( Mancinelli et al, 2017 ; Naru et al, 2017 ).…”