Cytoprotection of Human Umbilical Vein Endothelial Cells Against Apoptosis and CTL-Mediated Lysis Provided by Caspase-Resistant Bcl-2 Without Alterations in Growth or Activation Responses

Graft endothelial cells are primary targets of host CTL-mediated injury in acute allograft rejection. As an in vitro trial of gene therapy to reduce CTL-mediated endothelial injury, we stably transduced early passage HUVEC with a caspase-resistant mutant form (D34A) of the anti-apoptotic gene Bcl-2....

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Bibliographic Details
Published in:The Journal of immunology (1950) 2000-05, Vol.164 (9), p.4665-4671
Main Authors: Zheng, Lian, Dengler, Thomas J, Kluger, Martin S, Madge, Lisa A, Schechner, Jeffrey S, Maher, Stephen E, Pober, Jordan S, Bothwell, Alfred L. M
Format: Article
Language:English
Subjects:
Apoptosis - drug effects
Apoptosis - immunology
Caspases - physiology
Cell Division - immunology
Cell Line, Transformed
Cells, Cultured
Culture Media, Conditioned
Cytotoxicity, Immunologic
Endothelial Growth Factors - pharmacology
Endothelium, Vascular - cytology
Endothelium, Vascular - immunology
Genetic Vectors - immunology
Green Fluorescent Proteins
Humans
Luminescent Proteins - genetics
Proto-Oncogene Proteins c-bcl-2 - biosynthesis
Proto-Oncogene Proteins c-bcl-2 - genetics
Proto-Oncogene Proteins c-bcl-2 - physiology
Retroviridae - genetics
T-Lymphocytes, Cytotoxic - immunology
Transduction, Genetic - immunology
Transfection
umbilical vein
Umbilical Veins
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Summary:Graft endothelial cells are primary targets of host CTL-mediated injury in acute allograft rejection. As an in vitro trial of gene therapy to reduce CTL-mediated endothelial injury, we stably transduced early passage HUVEC with a caspase-resistant mutant form (D34A) of the anti-apoptotic gene Bcl-2. Bcl-2 transductants were compared with HUVEC transduced in parallel with an enhanced green fluorescent protein (EGFP) gene. Both transduced HUVEC have equivalent growth rates in complete medium and both show contact inhibition of growth. However, compared with EGFP-transduced HUVEC, the Bcl-2-transduced cells are resistant to the apoptotic effects of serum and growth factor withdrawal and are also resistant to the induction of apoptosis by staurosporine or by ceramide, with or without TNF. Transduced Bcl-2 did not reduce TNF-mediated NF-kappaB activation or constitutive expression of class I MHC molecules. HUVEC expressing D34A Bcl-2 were significantly more resistant to lysis by either class I-restricted alloreactive or PHA-redirected CTL than were HUVEC expressing EGFP. We conclude that transduction of graft endothelial cells with D34A Bcl-2 is a possible approach for reducing allograft rejection.
ISSN:0022-1767
1550-6606
DOI:10.4049/jimmunol.164.9.4665