Zheng L, Dengler T J, Kluger M S, Madge L A, Schechner J S, Maher S E, Pober J S, Bothwell A L
Section of Immunobiology, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, CT 06520, USA.
J Immunol. 2000 May 1;164(9):4665-71. doi: 10.4049/jimmunol.164.9.4665.
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.
在急性同种异体移植排斥反应中，移植内皮细胞是宿主细胞毒性T淋巴细胞（CTL）介导损伤的主要靶标。作为减少CTL介导的内皮损伤的基因治疗体外试验，我们用抗凋亡基因Bcl-2的一种抗半胱天冬酶突变形式（D34A）稳定转导早期传代的人脐静脉内皮细胞（HUVEC）。将Bcl-2转导细胞与平行转导增强型绿色荧光蛋白（EGFP）基因的HUVEC进行比较。两种转导的HUVEC在完全培养基中具有相同的生长速率，并且都表现出生长接触抑制。然而，与转导EGFP的HUVEC相比，转导Bcl-2的细胞对血清和生长因子撤除的凋亡作用具有抗性，并且对星形孢菌素或神经酰胺诱导的凋亡也具有抗性，无论有无肿瘤坏死因子（TNF）。转导的Bcl-2并未降低TNF介导核因子κB（NF-κB）的激活或I类主要组织相容性复合体（MHC）分子的组成性表达。与表达EGFP的HUVEC相比，表达D34A Bcl-2的HUVEC对I类限制性同种异体反应性或PHA重定向CTL的裂解具有明显更高的抗性。我们得出结论，用D34A Bcl-2转导移植内皮细胞是减少同种异体移植排斥反应的一种可能方法。
