title: Expression of fibronectin splicing variants in organ transplantation: a differential pattern between rat cardiac allografts and isografts.
abstract: Allograft rejection is associated with infiltration of inflammatory cells and deposition of extracellular matrix proteins. The extent to which diversity in the extracellular matrix regulates inflammatory cell function in transplants remains unclear. One group of extracellular matrix proteins, termed fibronectins (FNs), exhibits inherent diversity as a consequence of alternative splicing in three segments: EIIIA, EIIIB, or V. Although the EIIIA segment has documented functions in mesenchymal cell differentiation, neither this segment nor the EIIIB segment have been tested for effects specific to leukocyte functions. By contrast, the V region can include the CS-1 segment to which leukocytes may adhere through alpha 4 beta 1 integrins. In this study, we demonstrate that EIIIA+, EIIIB+, and V+ FN variants are synthesized, primarily by macrophages in distinct temporal and spatial patterns in two rat cardiac transplant models: either with antigenic challenge, allografts, or without challenge, isografts. The ratio of EIIIA inclusion into FN increases by day 1 in allografts and isografts and remains high until allografts are rejected (approximately 7 days) but falls to normal levels in tolerated isografts (day 6). EIIIB+ FN ratios in allografts peak later than do EIIIA+ FNs (day 4). EIIIB+ FN ratios remain relatively low in isografts. Interestingly, EIIIA+ and EIIIB+ FNs are deposited prominently in the myocardium of rejecting allografts in close association with infiltrating leukocytes, and FN expression and deposition are prominent at sites of infarction. By contrast, these FNs are largely restricted to the epicardium and to a lesser degree in the immediately adjacent myocardium in isografts. CS-1+ FNs increase in allografts and isografts at 3 hours after transplantation but are particularly prominent in allografts 1 to 3 days before rejection. Our data suggest that FN splicing variants have a differential role in the effector functions of leukocytes in allografts and isografts and provide a foundation for testing their function on leukocytes and a rationale for FN-based therapeutics to modulate allograft rejection in transplant recipients.
authors.affiliation: Department of Pathology, Beth Israel Hospital, Boston, Massachusetts, USA.
authors.forename: A J
authors.initials: AJ
authors.lastname: Coito
authors.forename: L F
authors.initials: LF
authors.lastname: Brown
authors.forename: J H
authors.initials: JH
authors.lastname: Peters
authors.forename: J W
authors.initials: JW
authors.lastname: Kupiec-Weglinski
authors.forename: L
authors.initials: L
authors.lastname: van de Water
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D015152:Blotting, Northern; D002465:Cell Movement; D005353:Fibronectins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019085:Fluorescent Antibody Technique, Direct; D016027:Heart Transplantation / Q000276:immunology / Q000473:pathology*; D017403:In Situ Hybridization; D008297:Male; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011914:Rats, Inbred BN; D011917:Rats, Inbred Lew; D012260:Ribonucleases; D013194:Staining and Labeling; D014184:Transplantation, Homologous; D014185:Transplantation, Isogeneic
chemical_list: D005353:Fibronectins; D015347:RNA Probes; D012333:RNA, Messenger; D012260:Ribonucleases