title: Monocytes transduced with lentiviral vectors expressing hepatitis C virus non-structural proteins and differentiated into dendritic cells stimulate multi-antigenic CD8(+) T cell responses.
abstract: Halting the spread of hepatitis C virus (HCV) and also eradicating HCV in subjects with chronic infection are major goals for global health. To this end, several years of research on HCV vaccine development have led to the conclusion that multi-antigenic and multi-functional vaccine types are necessary for effectiveness against HCV infection. In this study, we evaluated lentiviral vectors (LV) expressing clusters of HCV structural (LV-HCV-S) and non-structural (LV-HCV-NS) genes for future vaccine development. Batches of high titer LV were used to transduce differentiated dendritic cells (DC) and monocytes. We report successful delivery of HCV gene clusters, particularly into monocytes, leading to >80% LV-HCV-NS and >70% LV-HCV-S and transduced cells, respectively. Intracellular expression of HCV proteins in monocyte-derived DC resulted in immunophenotypic changes, such as downregulation of CD83 and CD86. Monocytes expressing NS proteins and differentiated into DC stimulated allogeneic and autologous CD8(+) and CD4(+) T cells in vitro and resulted in antigen-specific CD8(+) T cell responses against NS3, NS4a and NS5b. Hence, lentiviral-mediated expression of the multi-antigenic HCV-NS cluster in monocytes subsequently differentiated into DC is a novel potential anti-HCV vaccine modality.
authors.affiliation: Lymphatic Cell Therapy Laboratory, Clinic of Hematology, Hannover Medical School, Hannover, Germany.
authors.forename: Adan C
authors.initials: AC
authors.lastname: Jirmo
authors.forename: Richard C
authors.initials: RC
authors.lastname: Koya
authors.forename: Bala Sai
authors.initials: BS
authors.lastname: Sundarasetty
authors.forename: Mudita
authors.initials: M
authors.lastname: Pincha
authors.forename: Guann-Yi
authors.initials: GY
authors.lastname: Yu
authors.forename: Michael
authors.initials: M
authors.lastname: Lai
authors.forename: Rakesh
authors.initials: R
authors.lastname: Bakshi
authors.forename: Verena
authors.initials: V
authors.lastname: Schlaphoff
authors.forename: Jan
authors.initials: J
authors.lastname: Grabowski
authors.forename: Georg
authors.initials: G
authors.lastname: Behrens
authors.forename: Heiner
authors.initials: H
authors.lastname: Wedemeyer
authors.forename: Renata
authors.initials: R
authors.lastname: Stripecke
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D051940:B7-2 Antigen / Q000032:analysis; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology*; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D016086:Lentivirus / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000032:analysis; D009000:Monocytes / Q000276:immunology*; D014161:Transduction, Genetic; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology; D055815:Young Adult; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D051940:B7-2 Antigen; D007136:Immunoglobulins; D008562:Membrane Glycoproteins; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins; D000097802:CD83 Antigen