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The Location in Cartilage of Infectious Retrovirus in Cats Infected with Feline Leukemia Virus
Steven P. Arnoczky, DVM1; Cheryl Swenson, DVM, PhD1; Monika Egerbacher, DVM, PhD1; Keri Gardner, MS1; Oscar Caballero, MS1; Meghan Burns, DVM1
1 Laboratory for Comparative Orthopaedic Research (S.P.A., M.E., K.G., O.C., and M.B.) and the Department of Pathobiology and Diagnostic Investigation (C.S.), College of Veterinary Medicine, G-387, Michigan State University, East Lansing, MI 48824. E-mail address for S.P. Arnoczky: arnoczky@cvm.msu.edu
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Disclosure: The authors did not receive any outside funding or grants in support of their research for or preparation of this work. Neither they nor a member of their immediate families received payments or other benefits or a commitment or agreement to provide such benefits from a commercial entity. No commercial entity paid or directed, or agreed to pay or direct, any benefits to any research fund, foundation, division, center, clinical practice, or other charitable or nonprofit organization with which the authors, or a member of their immediate families, are affiliated or associated.
Investigation performed at the Laboratory for Comparative Orthopaedic Research and the Department of Pathobiology and Diagnostic Investigation, Diagnostic Center for Population and Animal Health, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan

The Journal of Bone and Joint Surgery, Incorporated
J Bone Joint Surg Am, 2007 Sep 01;89(9):2030-2036. doi: 10.2106/JBJS.G.00054
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Abstract

Background: Previous studies have suggested that articular cartilage allografts were not likely to transmit infectious retrovirus since viral DNA could not be isolated from chondrocytes of infected individuals. However, the ability of the extracellular matrix of articular cartilage to harbor and transmit a retrovirus has not been examined. We hypothesized that articular cartilage fragments, but not isolated chondrocytes, from cats systemically infected with feline leukemia virus (FeLV) are capable of transmitting infectious retrovirus.

Methods: Fresh cartilage segments and chondrocytes isolated from cats systemically infected with feline leukemia virus were used in this study. Feline embryonic fibroblast cells were cocultured with segments of cartilage, isolated chondrocytes, or fragments of cortical bone from each infected cat. The FeLV p27 antigen was measured in the coculture media by enzyme-linked immunosorbent assay. In addition, FeLV proviral nucleic acids were quantified by real-time quantitative polymerase chain reaction with use of DNA extracted from feline embryonic fibroblast cell cocultures as well as isolated chondrocytes. Immunohistochemistry was used to assess for FeLV p27 antigen in both intact cartilage fragments and isolated chondrocytes.

Results: Feline embryonic fibroblast cells cocultured with cartilage fragments from each of the five FeLV-infected cats all demonstrated high levels of proviral DNA, indicating transmission of infective virus. In addition, media from all cocultures of feline embryonic fibroblast cells and chondral fragments became positive for p27 antigen, indicating active viral replication. In contrast, cocultures of feline embryonic fibroblast cells and isolated chondrocytes from all FeLV-infected cats were negative for proviral DNA and p27 antigen. Likewise, no proviral nucleic acids could be detected in isolated chondrocytes from any infected cats. Cocultures of feline embryonic fibroblast cells with cortical bone fragments were positive for proviral DNA and p27 antigen. Immunohistochemical staining of cartilage fragments from FeLV-infected cats demonstrated the presence of p27 antigen throughout the extracellular matrix, but the p27 antigen was not detected in isolated chondrocytes.

Conclusions: Articular cartilage fragments can readily transmit infectious retrovirus, but isolated chondrocytes were likely not the source of the infectious virus because they did not harbor proviral DNA or p27 antigen.

Clinical Relevance: Because donor-cell viability improves the long-term function of chondral allografts (precluding the use of secondary sterilization procedures employed for tendon and bone allografts), these results underscore the importance of rigorous donor screening when the use of articular cartilage allografts is being considered.

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    Accreditation Statement
    These activities have been planned and implemented in accordance with the Essential Areas and policies of the Accreditation Council for Continuing Medical Education (ACCME) through the joint sponsorship of the American Academy of Orthopaedic Surgeons and The Journal of Bone and Joint Surgery, Inc. The American Academy of Orthopaedic Surgeons is accredited by the ACCME to provide continuing medical education for physicians.
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