title: Autologous lymphoblastoid cell lines stably transfected with Plasmodium falciparum circumsporozoite protein as targets in cytotoxic T-lymphocyte assays.
abstract: To produce cell lines that can be used as a continuous source of antigen presenting cells for stimulating T-cell lines and clones and as targets in cytotoxic T-lymphocyte (CTL) assays, we used a retroviral vector with a simian virus (SV40) early promotor to transfer a Plasmodium falciparum circumporozoite (PfCSP) gene into human EBV transformed B-lymphoblastoid cell lines (B-LCL). We herein report successful, stable transfection and cell surface expression of this gene, as confirmed by PCR, Western blot analysis and immunoelectron microscopy. One of three successfully transfected autologous cell lines expressed PfCSP on the cell surface and was lysed by CD8+ T-cell dependent CTL from a donor volunteer who had been immunized with irradiated P. falciparum sporozoites. Such cell lines should provide excellent tools for characterizing human CD8+ T-cell responses against Plasmodium sp. proteins.
authors.affiliation: Malaria Program, Naval Medical Research Institute, Bethesda, MD, USA.
authors.forename: A
authors.initials: A
authors.lastname: Kumar
authors.forename: Y H
authors.initials: YH
authors.lastname: Kang
authors.forename: K
authors.initials: K
authors.lastname: Gowda
authors.forename: C A
authors.initials: CA
authors.lastname: Mullen
authors.forename: D M
authors.initials: DM
authors.lastname: Gordon
authors.forename: S
authors.initials: S
authors.lastname: Kumar
authors.forename: S L
authors.initials: SL
authors.lastname: Hoffman
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics; D001402:B-Lymphocytes / Q000378:metabolism / Q000648:ultrastructure; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000235:genetics / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic; D005822:Genetic Vectors; D006801:Humans; D008213:Lymphocyte Activation / Q000235:genetics* / Q000502:physiology; D010963:Plasmodium falciparum / Q000737:chemistry; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection / Q000235:genetics / Q000276:immunology*
chemical_list: D000953:Antigens, Protozoan; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan