title: Reduction of HLA class I expression by ribonucleic acid interference mitigates allogenicity of human primary and Immortalized cells.
abstract: Unmatched human leukocyte antigens (HLA) expressed by allogeneic donor cells are the major target for immunological rejection. In order to reduce the immunogenicity of allograft cells, we have developed lentiviral vectors for delivery of short hairpin ribonucleic acid (shRNA) against Class I HLA. This approach was evaluated in both an established human embryonic kidney cell line and primary human CD34+ hematopoietic stem/progenitor cells. Target cells transduced with lentiviral vectors expressing either HLA-A*0201 allele-specific or HLA-A, -B, -C consensus sequence-specific shRNA showed effective knockdown of cell surface HLA expression. Mixed lymphocyte-target cell reactions showed significantly reduced interferon-gamma production from alloreactive cytotoxic T lymphocytes and significantly reduced levels of target cell apoptosis after shRNA-mediated knockdown of HLA expression and target cell survival correlated with vector transduction efficiency. Furthermore, increasing resistance to complement-dependent cytotoxicity mediated by anti-HLA antibodies was observed to correlate with increasing levels of shRNA vector transduction in primary human CD34+ cells. Notably, non-HLA restricted killing by lymphokine-activated killer cells was not incurred after HLA knockdown. These data demonstrate the potential for genetic engineering strategies targeting incompatible HLA alleles to reduce both cellular and humoral responses and enable graft survival after transplantation of allogeneic cells and tissues.
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Lemp
authors.forename: Kazunori
authors.initials: K
authors.lastname: Haga
authors.forename: Katrin
authors.initials: K
authors.lastname: Hacke
authors.forename: Janet A
authors.initials: JA
authors.lastname: Treger
authors.forename: Brooke T
authors.initials: BT
authors.lastname: Bogan
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Faure-Kumar
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Logg
authors.forename: Carol A
authors.initials: CA
authors.lastname: Kruse
authors.forename: James C
authors.initials: JC
authors.lastname: Cicciarelli
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kasahara
mesh_terms: D000918:Antibody Specificity; D018952:Antigens, CD34 / Q000378:metabolism; D002461:Cell Line, Transformed; D005333:Fetus / Q000166:cytology / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D006084:Graft Rejection / Q000235:genetics* / Q000276:immunology*; D057809:HEK293 Cells; D015395:Histocompatibility Antigens Class I / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007519:Isoantigens / Q000276:immunology; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology; D016086:Lentivirus / Q000235:genetics; D061251:Primary Cell Culture; D034622:RNA Interference / Q000276:immunology*
chemical_list: D018952:Antigens, CD34; D015395:Histocompatibility Antigens Class I; D007519:Isoantigens