title: Protein kinase C overexpression does not enhance immune-stimulatory surface markers of vaccinia-infected dendritic cells and DC cell lines.
abstract: One of the shortcomings of vaccinia virus (VACV) as immunization vector is the down-regulation of HLA and costimulatory molecules in antigen presenting cells. To overcome this problem we investigated the use of protein kinase C (PKC) as immune stimulatory agent. Thus several classical and atypical PKCs were inserted into wild-type or attenuated VACV using recombination into the hemagglutinin gene and the expression driven by the VACV 7,5K-IE gene promoter. Recombinant constructs expressing PKC-alpha, -beta, -theta as well as wild-type, constitutive active or dominant negative PKC-zeta constructs were generated. Additional constructs expressing PKB/Akt1 and ICAM-1 were used for comparison. Immature and mature peripheral blood derived-dendritic cells (DC) as well as lymphoid cell lines capable of obtaining a DC-like phenotype upon mitogen stimulation were infected. Disappointingly, VACV-driven PKC overexpression did not significantly enhance expression of various activation markers or costimulatory molecules tested. Neither CD86 nor HLA-DR expression was upregulated and also no influence on the maturation of DC, as measured by DC-SIGN and CD83, was observed. However, VACV did not interfere with LPS induced up-regulation of CD83 and did not lead to substantial apoptosis of infected DC within the first 24 hours.
authors: Department of Hygiene, Microbiology & Social Medicine, Medical University Innsbruck, Innsbruck, Austria. hartwig.huemer@i-med.ac.at
authors: Hartwig P
authors: HP
authors: Huemer
authors: Markus
authors: M
authors: Geiger
authors: Wilfried
authors: W
authors: Posch
authors: Nina
authors: N
authors: Krumböck
authors: Friedrich
authors: F
authors: Fresser
authors: Doris
authors: D
authors: Wilflingseder
authors: Florian
authors: F
authors: Uberall
mesh_terms: D000818:Animals; D051940:B7-2 Antigen / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism / Q000821:virology*; D005822:Genetic Vectors; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D011493:Protein Kinase C / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011995:Recombination, Genetic; D015854:Up-Regulation; D014616:Vaccinia virus / Q000201:enzymology / Q000235:genetics
chemical_list: D051940:B7-2 Antigen; C497964:CD86 protein, human; D006684:HLA-DR Antigens; D011493:Protein Kinase C