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The enhanced green fluorescent protein (eGFP) is minimally immunogenic in C57BL/6 mice

Gene Therapy volume 8, pages 1813–1814 (2001)Cite this article

The development of immunologic responses to the products of transferred genes is a potentially limiting factor in the correction of genetic diseases by gene therapy. The nature of these responses depends on numerous factors which affect the immunogenicity of the transgene product, including differences between the transgene product and endogenous proteins, the type of cell which produces the gene product, and the MHC repertoire of the host. Proteins expressed as selectable markers may also be immunogenic, as described by Riddell et al.1 We previously reported that the enhanced green fluorescent protein (eGFP), which is used as a reporter gene detectable by fluorescent microscopy or flow cytometry, is immunogenic when expressed in a Balb/c-derived leukemia cell line transformed by the p185 bcr/abl fusion oncogene (BM185), administered intravenously.2 Cytotoxic T lymphocytes (CTL) were readily detectable which reacted against BM185 cells expressing eGFP (BM185/eGFP), but not against the parental BM185 cells.

During the review process for the above-referenced manuscript, we were asked by a reviewer to determine whether the immunologic responses seen against eGFP when expressed by the BM185 cell line in Balb/c mice are generalized to other cell lines and strains of mice. Therefore, a similar experiment was done using the EL-4 murine T cell lymphoma line in the syngeneic C57BL/6 host. Unmodified EL-4 cells grew into large tumors within 25 days when implanted subcutaneously, but no tumors formed from a clone of EL-4 cells expressing eGFP (EL-4/eGFP) at 30 days (Figure 4, in Stripecke et al2). This study suggested that eGFP was also immunogenic in C57BL/6 mice.

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Authors and Affiliations

  1. Division of Research Immunology/Bone Marrow Transplantation, John Connell Gene Therapy Program, Childrens Hospital Los Angeles, Los Angeles, CA, USA

    D Skelton, N Satake & DB Kohn

  2. Department of Pediatrics, USC Keck School of Medicine, Los Angeles, CA, USA

    DB Kohn

  3. Department of Molecular Microbiology and Immunology, USC Keck School of Medicine, Los Angeles, CA, USA

    DB Kohn

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  1. D Skelton
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  2. N Satake
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  3. DB Kohn
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Skelton, D., Satake, N. & Kohn, D. The enhanced green fluorescent protein (eGFP) is minimally immunogenic in C57BL/6 mice. Gene Ther 8, 1813–1814 (2001). https://doi.org/10.1038/sj.gt.3301586

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  • DOI: https://doi.org/10.1038/sj.gt.3301586

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