Immune response to green fluorescent protein: Implications for gene therapy

R. Stripecke, M. Del Carmen Villacres, D. C. Skelton, Noriko Satake, S. Halene, D. B. Kohn

Research output: Contribution to journalArticlepeer-review

283 Scopus citations


Green fluorescent protein (GFP) is a widely used intracellular reporter molecule to assess gene transfer and expression. A potential use for GFP is as a co-expressed marker, to select and enrich gene-modified cells by flow cytometry. Processed peptides derived from GFP and presented by the major histocompatibility complex on the cell surface could potentially induce T cell immune responses against GFP+ cells. Thus, clinical application of GFP is premature, since in vivo studies on its immunogenicity are lacking. Therefore, we investigated immune responses against EGFP (enhanced-GFP) in two transplantable murine models: the BALB/c (H-2(d)) BM185 pre-B leukemia and the C57BL/6 (H-2b) EL-4 T cell lymphoma. B185 and EL-4 cell lines modified to express high levels of EGFP showed drastic reduction of disease development when transplanted into immunocompetent mice. BM185/EGFP did lead to rapid development of disease in immunodeficient Nu/Nu mice. Mice surviving BM185/EGFP leukemia challenge developed high cytotoxic T lymphocyte (CTL) responses against EGFP-expressing cells. Furthermore, immune stimulation against BM185/EGFP cells could also be induced by immunization with EGFP+ transduced dendritic cells. The effects of the co-expression of EGFP and immunomodulators (CD80 plus GM-CSF) were also investigated as an irradiated leukemia vaccine. EGFP co-expression by the vaccine did not interfere with the development of CTLs against the parental leukemia or with the anti-leukemia response in vivo. These results indicate that the immune response against EGFP may interfere with its applicability in gene insertion/replacement strategies but could potentially be employed for leukemia cell vaccines.

Original languageEnglish (US)
Pages (from-to)1305-1312
Number of pages8
JournalGene Therapy
Issue number7
StatePublished - Jul 1999
Externally publishedYes


  • Cell vaccine
  • Dendritic cell
  • Gene therapy
  • Green fluorescent protein
  • Immune response
  • Leukemia

ASJC Scopus subject areas

  • Genetics


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