no immune responses by the expression of the yeast ndi1 protein in rats没有免疫反应的表达式酵母ndi1蛋白质的老鼠.pdfVIP

No Immune Responses by the Expression of the Yeast Ndi1 Protein in Rats 1. 1.¤ 2 1 1 Mathieu Marella , Byoung Boo Seo , Terence R. Flotte , Akemi Matsuno-Yagi , Takao Yagi * 1 Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, United States of America, 2 Gene Therapy Center and Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America Abstract Background: The rotenone-insensitive internal NADH-quinone oxidoreductase from yeast, Ndi1, has been shown to work as a replacement molecule for complex I in the respiratory chain of mammalian mitochondria. In the so-called transkingdom gene therapy, one major concern is the fact that the yeast protein is foreign in mammals. Long term expression of Ndi1 observed in rodents with no apparent damage to the target tissue was indicative of no action by the host’s immune system. Methodology/Principal Findings: In the present study, we examined rat skeletal muscles expressing Ndi1 for possible signs of inflammatory or immune response. In parallel, we carried out delivery of the GFP gene using the same viral vector that was used for the NDI1 gene. The tissues were subjected to HE staining and immunohistochemical analyses using antibodies specific for markers, CD11b, CD3, CD4, and CD8. The data showed no detectable signs of an immune response with the tissues expressing Ndi1. In contrast, mild but distinctive positive reactions were observed in the tissues expressing GFP. This clear difference most likely comes from the difference in the location of the expressed