title: Transcript analysis of P2X receptors in PBMCs of chronic HCV patients: an insight into antiviral treatment response and HCV-induced pathogenesis.
abstract: BACKGROUND
  After invasion of hepatocytes and immune cells, hepatitis C virus has the ability to escape from the host immune system, leading to the progression of disease into chronic infection with associated liver morbidities. Adenosine 5'triphosphate (ATP) is released in most of the pathological events from the affected cells and acts as a signaling molecule by binding to P2X receptors expressed on the host's immune cells and activates the immune system for pro-inflammatory response. Therefore, the present study was designed to analyze the transcript expression of the ionotropic purinergic P2X receptors on peripheral blood mononuclear cells (PBMCs) of chronic HCV patients to have study the immune responses mediated by P2X receptors in chronic HCV infections.
  
  
  METHODS
  PBMCs were isolated from the collected blood samples. Transcript analysis of P2X receptors in PBMCs was done. The identity of amplified product was confirmed by sequencing PCR, while the quantification of the transcript expression was done by real time PCR. The relative expression of the P2X receptors was analyzed by unpaired Student's t test using GraphPad Prims 5 software.
  
  
  RESULTS
  We found that out of seven isoforms of P2X receptors, P2X1, P2X4, P2X5, and P2X7 receptors are expressed on the PBMCs. P2X1 and P2X7 are significantly upregulated in treatment-naïve chronic HCV patients by 2.2- and 2.5-fold, respectively. However, only P2X7 expression is found increased by 2.7-fold in patients achieving sustained virological response (SVR) after antiviral treatment compared to healthy controls. The expression of P2X receptors remained unaltered in chronic HCV patients not responding to the treatment.
  
  
  CONCLUSIONS
  The present study confirms the significant involvement of P2X receptors in the immune responses mediated by the PBMCs in the chronic HCV infection, which should be further investigated to devise strategies to augment the immune system against this chronic viral disease.
authors.affiliation: 1 Atta ur Rahman School of Applied Bio-Sciences, National University of Sciences & Technology, Islamabad.
authors.forename: Waseem
authors.initials: W
authors.lastname: Ashraf
authors.forename: Sobia
authors.initials: S
authors.lastname: Manzoor
authors.forename: Javed
authors.initials: J
authors.lastname: Ashraf
authors.forename: Qazi Laeeque
authors.initials: QL
authors.lastname: Ahmed
authors.forename: Madiha
authors.initials: M
authors.lastname: Khalid
authors.forename: Muqddas
authors.initials: M
authors.lastname: Tariq
authors.forename: Muhammad
authors.initials: M
authors.lastname: Imran
authors.forename: Hafsa
authors.initials: H
authors.lastname: Aziz
mesh_terms: D000328:Adult; D000998:Antiviral Agents / Q000627:therapeutic use; D016207:Cytokines / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D019698:Hepatitis C, Chronic / Q000188:drug therapy* / Q000276:immunology*; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008297:Male; D011092:Polyethylene Glycols / Q000627:therapeutic use; D020033:Protein Isoforms / Q000378:metabolism; D058469:Receptors, Purinergic P2X / Q000235:genetics* / Q000276:immunology*; D011994:Recombinant Proteins / Q000627:therapeutic use; D012254:Ribavirin / Q000627:therapeutic use
chemical_list: D000998:Antiviral Agents; D016207:Cytokines; D016898:Interferon-alpha; D020033:Protein Isoforms; D058469:Receptors, Purinergic P2X; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D012254:Ribavirin; C100416:peginterferon alfa-2a