title: Increased Expression of TLR4 and TLR7 but Not Prolactin mRNA by Peripheral Blood Monocytes in Active Celiac Disease.
abstract: BACKGROUND
  Celiac disease (CD) is an organ-specific autoimmune disease, and both adaptive and innate immunity are involved in its development.
  
  
  OBJECTIVE
  The aim of the study was to determine whether the markers of intestinal mucosal inflammation in CD can be detected in peripheral blood monocytes (PBMs), and whether the immune properties of PBMs change as the clinical signs and symptoms of CD improve after the introduction of a gluten-free diet (GFD). The focus was on changes in mRNA expression of selected toll-like receptors (TLR2, TLR4, TLR7), stress cytokine prolactin (PRL), and proand anti-inflammatory cytokines (TNF-α, IL-6, IL-12, IL-10) in PBMs.
  
  
  METHODS
  The study involved 20 CD patients diagnosed according to the European Society for Pediatric Gastroenterology, Hepatology and Nutrition criteria and Marsh criteria: 10 recently-diagnosed cases (rCD) and 10 on a GFD for a minimum of one year. The control group comprised 10 ageand sex-matched healthy volunteers. PBMs from peripheral blood specimens were separated using immunomagnetic CD14+ beads. Total RNA was isolated using a standard commercial kit. Cytokine and TLR mRNA levels were quantified by relative qPCR with PGK1 as a reference gene.
  
  
  RESULTS
  Significantly higher expression of TLR4 and TLR7 mRNA was observed in PBMs from rCD patients compared to the healthy controls (1.63 times higher; p < 0.05). TLR7 mRNA levels in rCDs were also significantly elevated in comparison to the CD-GFD patients (2.11 times higher; p < 0.01). TNF-α mRNA expression tended to be higher in both groups of patients; by contrast, in IL-6 mRNA, a trend to a fourfold decrease was detected in PBMs from the CD-GFD subjects. IL-10, IL-12 and PRL levels did not differ among the groups.
  
  
  CONCLUSIONS
  The data suggest that the inflammatory process in rCD intestinal mucosa and submucosa reflecting enterocyte damage can be detected in PBMs in peripheral blood. Further, the cytokine and TLR expression profile in PBMs alters after one year of GFD treatment.
authors.affiliation: Department of Anthropology and Human Genetics, Faculty of Science, Charles University in Prague, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Brynychova
authors.affiliation: Second Department of Internal Medicine, Third Faculty of Medicine, Charles University in Prague, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Hoffmanova
authors.affiliation: First Medical Faculty, Charles University in Prague and General University Hospital in Prague, Czech Republic.
authors.forename: Milos
authors.initials: M
authors.lastname: Dvorak
authors.affiliation: Department of Anthropology and Human Genetics, Faculty of Science, Charles University in Prague, Czech Republic.
authors.forename: Pavlina
authors.initials: P
authors.lastname: Dankova
mesh_terms: D000328:Adult; D002446:Celiac Disease / Q000097:blood / Q000235:genetics*; D005260:Female; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D011388:Prolactin / Q000097:blood* / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger; D051197:Toll-Like Receptor 4 / Q000097:blood* / Q000235:genetics*; D051199:Toll-Like Receptor 7 / Q000097:blood* / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D015850:Interleukin-6; D012333:RNA, Messenger; C495345:TLR4 protein, human; C495348:TLR7 protein, human; D051197:Toll-Like Receptor 4; D051199:Toll-Like Receptor 7; D014409:Tumor Necrosis Factor-alpha; D011388:Prolactin
keywords: celiac disease; cytokines; innate immunity; monocytes; prolactin