title: Predominance of PR3 specific immune response and skewed TH17 vs. T-regulatory milieu in active granulomatosis with polyangiitis.
abstract: OBJECTIVE
  We compared levels of Th1/Th2/Th17 cytokines and T-regulatory cells in active and remitting granulomatosis with polyangiitis (GPA).
  
  
  METHODS
  Twenty-one cases of GPA in active state as well as in remitting state and 20 healthy controls (HC) were enrolled in the study. Cytokines were detected in culture supernatants of PBMCs after stimulation with proteinase-3 (PR3) and phytohemagglutinin antigen (PHA). Serum IL-17 cytokine was studied by ELISA. T-regulatory cells (Tregs) were analyzed by flow cytometry. Gene expression of FOXP3 and ROR-γt was compared by Real Time PCR.
  
  
  RESULTS
  We observed significantly increased level of IL-17 in serum as well in culture supernatants of PBMCs after PR3 stimulation along with ROR-γt gene expression in active disease state of GPA as compared to HC. Importantly, remitting state showed low levels of serum IL-17 with decreased ROR-γt gene expression and increased FOXP3 expression. Using PR3 as an immunostimulant, we could demonstrate the generation of IL-17 and TNF-α secreting effector memory cells during remission. Reduced FOXP3 expression with reduced IL-10 levels in active disease indicated the reduced function of Tregs in active disease.
  
  
  CONCLUSIONS
  We observed Th17 dominant environment in peripheral blood of patients in active state of disease, with "hyporesponsiveness", in, in vitro stimulated PBMC-in their ability to secrete TNF-α and IL-6. Treg numbers were unaltered but function was compromised. Targeting PR3 specific effector memory cells, to prevent relapse, and instituting anti IL-17 therapy, or modulating Tregs could be newer forms of therapy for this serious autoimmune disease.
authors.affiliation: Department of Immunopathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: lekhapgi@yahoo.com.
authors.forename: Lekha
authors.initials: L
authors.lastname: Rani
authors.affiliation: Department of Immunopathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: rwminz.minz88@gmail.com.
authors.forename: Ranjana W
authors.initials: RW
authors.lastname: Minz
authors.affiliation: Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: amansharma74@gmail.com.
authors.forename: Aman
authors.initials: A
authors.lastname: Sharma
authors.affiliation: Department of Immunopathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: sanand312@gmail.com.
authors.forename: Shashi
authors.initials: S
authors.lastname: Anand
authors.affiliation: Department of Pulmonary Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: dheeraj1910@gmail.com.
authors.forename: Dheeraj
authors.initials: D
authors.lastname: Gupta
authors.affiliation: Department of Otolaryngology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: nkpanda59@yahoo.co.in.
authors.forename: N K
authors.initials: NK
authors.lastname: Panda
authors.affiliation: Department of Nephrology, Postgraduate Institute of Medical Education and Research, Chandigarh, India. Electronic address: vsakhuja2009@gmail.com.
authors.forename: V K
authors.initials: VK
authors.lastname: Sakhuja
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000276:immunology; D015870:Gene Expression / Q000276:immunology; D014890:Granulomatosis with Polyangiitis / Q000188:drug therapy / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D020381:Interleukin-17 / Q000097:blood / Q000276:immunology / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D053146:Myeloblastin / Q000276:immunology*; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000235:genetics / Q000276:immunology; D011446:Prospective Studies; D012074:Remission Induction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism; D058504:Th17 Cells / Q000276:immunology* / Q000378:metabolism; D055815:Young Adult
chemical_list: D016207:Cytokines; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D020381:Interleukin-17; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D016753:Interleukin-10; D053146:Myeloblastin
keywords: Cytokines; GPA; Immunopathology; T-cells; Tregs