Open Access, Peer-reviewed
pISSN 2093-4378
eISSN 2093-4386
    J Korean Soc Spine Surg. 2002 Jun;9(2):84-90. Korean.
    Published online Jun 30, 2002.
    https://doi.org/10.4184/jkss.2002.9.2.84
    Copyright © 2002 Korean Society of Spine Surgery
    Original Article

    Host Immune Responses to Intradiscal Gene Transfer

    Seong-Hwan Moon, M.D., Hyang Kim, B.S., Eun-Hae Kwon, M.S., Eu-Hyun Park, M.D., Hwan-Mo Lee, M.D., Soo-Bong Hahn, M.D., Nam-Hyun Kim, M.D., Lars G. Gilbertson, Ph.D.,* and James D. Kang, M.D.*
      • Department of Orthopedic Surgery, Yonsei University College of Medicine, Seoul, Korea.
      • *Department of Orthopedic Surgery, University of Pittsburgh, Pittsburgh, PA, USA.
    • Address reprint requests to Seong-Hwan Moon, M.D. Department of Orthopaedic Surgery, Yonsei University College of Medicine, #134 Shinchon-dong, Soedaemun-gu, Seoul 120-752, Korea. Tel: 82-2-361-5649, Fax: 82-2-363-1139, Email: shmoon@yumc.yonsei.ac.kr

    Abstract

    Purpose

    To elucidate host immune responses to intradiscal gene transfer.

    Materials and Methods

    Twenty rabbits were utilized. Ad/luciferase (adenovirus construct) were injected into nucleus pulposus of lumbar vertebrae. Group 1 received intradiscal injection of Ad/luciferase only, Group 2 received subcutaneous and intradiscal injection simultaneously, Group 3 received subcutaneous injection then intradiscal injection with 2 weeks interval. Blood samples were obtained serially after injection. Animals were sacrificed at 7 weeks. Antibody to adenovirus in peripheral blood was measured with ELISA and transgene expression was measured with standard luciferase kits.

    Results

    All rabbits in the Group 2 and 3 exhibited increased production of neutralizing antibody. There were clearly two subgroups in Group 1, three rabbits exhibited production of antibody but remaining three rabbits showed little or no production of antibody. All rabbits showed robust increase in transgene expression regardless of titer of neutralizing antibody.

    Conclusion

    The intervertebral disc is favorable site for adenovirus-mediated gene transfer escaping from systemic immunity.

    Keywords
    Intradiscal Gene Therapy; Immune Responses

    Figures

    Fig. 1
    Qualitative analysis of intradiscal transgene expression of A) 3 weeks, B) 6 weeks after direct application of Ad/lacZ into lumbar intervertebral discs of adult New Zealand white rabbits. There were strong transgene expresson indicated by the arrow (dark staining, originally blue color) in the nucleus pulposus without evidence of infiltration of inflamatory cell and neovascularization (X-Gal staining with counter staining with hematoxylin-eosin, Original magnification ×100).

    Fig. 2
    Sequential production of specific antibody for adenoviral proteins in peripheral blood as function of time. Transverse bar chart on the right denote intradiscal transgene expression in each individual rabbit. A: Three rabbits injected with Ad/luciferase produced little or no neutralizing antibody while remaining three rabbits showed production of antibody (likely due to leakage of adenovirus from injected disc). B: Six rabbits in Group 2 after simultaneous injection of Ad/luciferase into subcutaneous tissue and intervertebral disc produced neutralizing antibody in significant amount. C: Six rabbits in Group 3 were immunized two weeks prior to intradiscal gene transduction with injection of Ad/luciferase into subcutaneous tissues. All rabbits showed significant amount of transgene (luciferase) expression.

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    MeSH Terms
    Adenoviridae
    Animals
    Antibodies, Neutralizing
    Enzyme-Linked Immunosorbent Assay
    Injections, Subcutaneous
    Intervertebral Disc
    Luciferases
    Lumbar Vertebrae
    Rabbits
    Transgenes
    United Nations
    Metrics
    Share
    Figures
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