Localization and characterization of porcine patellar tendon xenograft antigens in a rabbit model of medial collateral ligament replacement.
BACKGROUND
Ligament injuries of the knee are common and, if severe, can predispose to joint pain, instability, reinjury, and, ultimately, osteoarthritis. Xenograft replacement of ligaments could have potential; however, a limited understanding of the immunology of ligament xenograft rejection has inhibited their use. The purpose of this study was to characterize the antigenic elements of a fresh porcine tendon xenograft in a rabbit model and to provide a better understanding of what would need to be done to either block or extract these antigenic elements.


METHODS
Three experimental situations were evaluated in a pig to rabbit ligament transplantation model: subcutaneous implantation of fresh porcine patellar tendon (PPT), implantation of fresh PPT into a medial collateral ligament midsubstance gap, and replacement of the entire medial collateral ligament complex with either fresh or guanidinium hydrochloride-extracted PPT. Preimmune and immune sera were collected from rabbits and used to localize antigenic targets in PPT, meniscus, and cartilage with indirect immunofluorescence techniques. The reactivities of the same rabbit sera towards tissue extracts of PPT, meniscus, and cartilage by Western immunoblot analyses were used to characterize the antigenic components.


RESULTS
Indirect immunofluorescence with preimmune rabbit sera on PPT showed staining of tendon fibroblasts. Immune sera from rabbits transplanted with xenografts stained regions of the extracellular matrix of PPT. Fresh PPT induced antibodies that consistently recognized six extracellular matrix components with molecular masses of >200 kDa, 180 kDa, 135 kDa, 108 kDa, 63 kDa, and 59 kDa.


CONCLUSIONS
Our results suggest that naturally occurring rabbit anti-pig antibodies of the IgG isotype recognize immunogenic components on tendon fibroblasts, whereas induced rabbit anti-pig antibodies recognize a specific subset of six extracellular matrix components of PPT. PPT xenografts appeared to induce a similar humoral immune response irrespective of graft location. Finally, our results indicate that selective extraction of PPT xenograft components before implantation altered the induced rabbit anti-pig antibody response; however, such extraction did not change the ultimate fate of the transplant tissue.
Veillette
C J
CJ
McCaig Centre for Joint Injury and Arthritis Research, Faculty of Medicine, The University of Calgary, Alberta, Canada.
Cunningham
K D
KD
Hart
D A
DA
Fritzler
M J
MJ
Frank
C B
CB
D000818:Animals; D015478:Antigens, Heterophile / Q000032:analysis*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000276:immunology / Q000637:transplantation; D016326:Extracellular Matrix Proteins / Q000032:analysis*; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000276:immunology; D019084:Fluorescent Antibody Technique, Indirect; D008022:Ligaments / Q000637:transplantation*; D011817:Rabbits; D013552:Swine; D013710:Tendons / Q000637:transplantation*; D014182:Transplantation, Autologous / Q000276:immunology; D014183:Transplantation, Heterologous / Q000276:immunology* / Q000379:methods
D015478:Antigens, Heterophile; D016326:Extracellular Matrix Proteins