title: Differential expression pattern of the antiapoptotic proteins, Bcl-2 and FLIP, in experimental arthritis.
abstract: OBJECTIVE
  To examine the relationship between apoptosis and the expression of antiapoptotic proteins in the pathogenesis of experimental inflammatory arthritis.
  
  
  METHODS
  Clinical and histologic assessment of adjuvant-induced arthritis (AIA) was performed over a 42-day period. The induction of apoptosis was measured by TUNEL analysis, and the antiapoptotic proteins, Bcl-2 and FLIP, were examined by immunohistochemistry with the use of monospecific antibodies. The percentage of Bcl-2- and FLIP-positive cells was correlated with histologic markers of AIA.
  
  
  RESULTS
  Arthritis developed by day 14 following adjuvant injection. Few TUNEL-positive cells were observed between days 0 and 21, indicating that apoptosis did not occur at these time points. An increase in the number of TUNEL-positive cells was observed at day 28, particularly outside sites of cartilage or bone erosion, which dramatically declined by day 35. Immunohistochemical analyses of Bcl-2 and FLIP revealed that the synovium was positive for Bcl-2 and FLIP on day 0. On day 14, Bcl-2 was present at the sites of early erosions and correlated with the erosion and inflammation scores. FLIP was also highly expressed at sites of erosion and was localized to the pannus starting on day 21. Although TUNEL positivity peaked at day 28, a time point in which Bcl-2 and FLIP were present, the areas that displayed intense positivity for expression of Bcl-2 and FLIP were TUNEL negative. In addition, the number of neutrophils in the synovial lining and pannus significantly decreased from day 28 to day 35, suggesting that the cells undergoing apoptosis were neutrophils. Furthermore, at day 42 when TUNEL-positive cells were absent, Bcl-2 expression was diminished, while FLIP remained highly expressed in the pannus.
  
  
  CONCLUSIONS
  The overall percentage of TUNEL-positive cells in the ankle was <1% except on days 28 and 35 post-adjuvant injection, suggesting that in AIA, similar to rheumatoid arthritis, a lack of apoptosis may contribute to disease progression. Furthermore, Bcl-2 and FLIP are temporally and differentially expressed during the pathogenesis of AIA. Inhibition of these molecules may augment synovial apoptosis and ameliorate the disease.
authors.affiliation: Northwestern University Medical School and the Veterans Administration Chicago Healthcare System, Lakeside Division, IL 60611, USA.
authors.forename: H
authors.initials: H
authors.lastname: Perlman
authors.forename: H
authors.initials: H
authors.lastname: Liu
authors.forename: C
authors.initials: C
authors.lastname: Georganas
authors.forename: A E
authors.initials: AE
authors.lastname: Koch
authors.forename: E
authors.initials: E
authors.lastname: Shamiyeh
authors.forename: G K
authors.initials: GK
authors.lastname: Haines
authors.forename: R M
authors.initials: RM
authors.lastname: Pope
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D001169:Arthritis, Experimental / Q000473:pathology / Q000503:physiopathology*; D001172:Arthritis, Rheumatoid / Q000473:pathology / Q000503:physiopathology; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; D002352:Carrier Proteins / Q000235:genetics*; D004195:Disease Models, Animal; D005260:Female; D015870:Gene Expression / Q000502:physiology; D020287:In Situ Nick-End Labeling; D047908:Intracellular Signaling Peptides and Proteins; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D051381:Rats; D011917:Rats, Inbred Lew; D013583:Synovial Membrane / Q000473:pathology
chemical_list: D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D019253:Proto-Oncogene Proteins c-bcl-2