title: Tissue MicroArray (TMA) analysis of normal and persistent Chlamydophila pneumoniae infection.
abstract: BACKGROUND
  Chlamydophila pneumoniae infection has been implicated as a potential risk factor for atherosclerosis, however the mechanism leading to persistent infection and its role in the disease process remains to be elucidated.
  
  
  METHODS
  We validated the use of tissue microarray (TMA) technology, in combination with immunohistochemistry (IHC), to test antibodies (GroEL, GroES, GspD, Ndk and Pyk) raised against differentially expressed proteins under an interferon-gamma (IFN-gamma) induced model of chlamydial persistence.
  
  
  RESULTS
  In the cell pellet array, we were able to identify differences in protein expression patterns between untreated and IFN-gamma treated samples. Typical, large chlamydial inclusions could be observed in the untreated samples with all antibodies, whereas the number of inclusions were decreased and were smaller and atypical in shape in the IFN-gamma treated samples. The staining results obtained with the TMA method were generally similar to the changes observed between normal and IFN-gamma persistence using proteomic analysis. Subsequently, it was shown in a second TMA including archival atheromatous heart tissues from 12 patients undergoing heart transplantation, that GroEL, GroES, GspD and Pyk were expressed in atheromatous heart tissue specimens as well, and were detectable morphologically within lesions by IHC.
  
  
  CONCLUSIONS
  TMA technology proved useful in documenting functional proteomics data with the morphologic distribution of GroEL, GroES, GspD, Ndk and Pyk within formalin-fixed, paraffin-embedded cell pellets and tissues from patients with severe coronary atherosclerosis. The antibodies GroEL and GroES, which were upregulated under persistence in proteomic analysis, displayed positive reaction in atheromatous heart tissue from 10 out of 12 patients. These may be useful markers for the detection of persistent infection in vitro and in vivo.
authors.affiliation: Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerland. n.borel@access.unizh.ch
authors.forename: Nicole
authors.initials: N
authors.lastname: Borel
authors.forename: Sanghamitra
authors.initials: S
authors.lastname: Mukhopadhyay
authors.forename: Carmen
authors.initials: C
authors.lastname: Kaiser
authors.forename: Erin D
authors.initials: ED
authors.lastname: Sullivan
authors.forename: Richard D
authors.initials: RD
authors.lastname: Miller
authors.forename: Peter
authors.initials: P
authors.lastname: Timms
authors.forename: James T
authors.initials: JT
authors.lastname: Summersgill
authors.forename: Julio A
authors.initials: JA
authors.lastname: Ramirez
authors.forename: Andreas
authors.initials: A
authors.lastname: Pospischil
mesh_terms: D002460:Cell Line; D023501:Chlamydophila / Q000302:isolation & purification / Q000378:metabolism*; D023521:Chlamydophila Infections / Q000175:diagnosis / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D046888:Tissue Array Analysis / Q000379:methods*