title: Mesenchymal stem cells engineered to inhibit complement-mediated damage.
abstract: Mesenchymal stem cells (MSC) preferentially migrate to damaged tissues and, due to their immunomodulatory and trophic properties, contribute to tissue repair. Although MSC express molecules, such as membrane cofactor protein (CD46), complement decay-accelerating factor (CD55), and protectin (CD59), which confer protection from complement-mediated lysis, MSC are recruited and activated by anaphylatoxins after transplantation, potentially causing MSC death and limiting therapeutic benefit. We have previously demonstrated that transduction of MSC with a retrovirus encoding HCMV-US proteins resulted in higher levels of MSC engraftment due to decreased HLA-I expression. Here, we investigate whether engineering MSC to express US2 (MSC-US2), US3 (MSC-US3), US6 (MSC-US6), or US11 (MSC-US11) HCMV proteins can alter complement recognition, thereby better protecting MSC from complement attack and lysis. HCMV-US proteins increased MSC CD59 expression at different levels as determined by flow cytometric evaluation of the median fluorescence intensity ratio (MFI). A significant increase in CD59 expression was seen in MSC-US2, MSC-US3, and MSC-US6, but not in MSC-US11. Only MSC-US2 displayed increased expression of CD46, while US2 and US3 proteins were both able to augment the percentage of MSC expressing this molecule. Regardless of the HCMV protein expressed, none changed CD55 MFI; however, expression of US6, US11, and US2 each increased the percentage of MSC that were positive for this molecule. Because US2 protein was the most efficient in up-regulating all three complement regulatory proteins, we used a functional complement-mediated cytotoxicity assay to investigate whether MSC-US2 were protected from complement-mediated lysis. We demonstrated that over-expression of the US2 protein reduced complement lysis by 59.10±12.89% when compared to untransduced MSC. This is the first report, to our knowledge, describing a role of HCMV-US proteins in complement evasion, and our data shows that over-expression of US2 protein on MSC could serve as a strategy to protect these cells from complement lysis.
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina, United States of America.
authors.forename: Melisa A
authors.initials: MA
authors.lastname: Soland
authors.forename: Mariana
authors.initials: M
authors.lastname: Bego
authors.forename: Evan
authors.initials: E
authors.lastname: Colletti
authors.forename: Esmail D
authors.initials: ED
authors.lastname: Zanjani
authors.forename: Stephen
authors.initials: S
authors.lastname: St Jeor
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Porada
authors.forename: Graça
authors.initials: G
authors.lastname: Almeida-Porada
mesh_terms: D018958:CD55 Antigens / Q000032:analysis / Q000276:immunology; D018957:CD59 Antigens / Q000032:analysis / Q000276:immunology; D060846:Cell Engineering / Q000379:methods; D002478:Cells, Cultured; D003165:Complement System Proteins / Q000276:immunology*; D003587:Cytomegalovirus / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D051916:Membrane Cofactor Protein / Q000032:analysis / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics*; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D018958:CD55 Antigens; D018957:CD59 Antigens; D006023:Glycoproteins; D017874:Immediate-Early Proteins; D051916:Membrane Cofactor Protein; D008565:Membrane Proteins; D016601:RNA-Binding Proteins; C074546:US11 protein, herpesvirus; C118670:US2 protein, Varicellovirus; C095111:US3 protein, cytomegalovirus; C432877:US6 protein, Human cytomegalovirus; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D003165:Complement System Proteins
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