Characterization of host immunity to cytomegalovirus pp150 (UL32).
The basic phosphoprotein 150 (pp150), the product of UL32 (unique long domain 32) gene of human cytomegalovirus (CMV), is an abundant component of the viral tegument and a target of human leukocyte antigen (HLA)-restricted cytotoxic T cells (CTLs) after infection. Identification of minimal cytotoxic epitopes (MCEs) from this CMV protein is of importance for peptide-based vaccines and immunotherapeutic approaches. Several pp150-specific CTL clones were derived from peripheral blood mononuclear cells of healthy CMV-positive donors with autologous fibroblasts infected either with CMV AD169 or with a recombinant vaccinia virus expressing full-length pp150 protein. HLA A*0301- and HLA A*6801-restricted CD8+ pp150 T-cell clones derived from different donors were found to efficiently kill autologous CMV-infected fibroblasts. Fine mapping of each MCE first used a T-cell epitope prediction algorithm. Overlapping peptides within the recognized regions were screened. The analysis identified pp150(792-802) and pp150(945-955) as MCEs for the HLA A*6801 and the HLA A*0301 pp150 clones, respectively. In vitro stimulation by recombinant modified vaccinia Ankara virus expressing full-length pp150 elicited high frequencies of CMV-CTL and interferon gamma production specific for the MCE identified in all subjects. The consistent presence of pp150 T cells in CMV-exposed individuals supports a role for this antigen in shaping the antiviral CTL response and indicates that pp150 could be a pivotal constituent of prophylactic and therapeutic CMV vaccines.
La Rosa
Corinna
C
Laboratory of Vaccine Research, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.
Wang
Zhongde
Z
Lacey
Simon F
SF
Markel
Susan F
SF
Sharma
Madeva C
MC
Martinez
Joybelle
J
Lalimarmo
Maria M
MM
Diamond
Don J
DJ
D002999:Clone Cells; D003587:Cytomegalovirus / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D018604:Epitope Mapping; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D010750:Phosphoproteins / Q000235:genetics* / Q000276:immunology*; D016693:Receptors, Antigen, T-Cell, alpha-beta; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000821:virology*; D019444:Vaccines, DNA / Q000276:immunology*; D014616:Vaccinia virus; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000276:immunology*
D006680:HLA Antigens; D010750:Phosphoproteins; D016693:Receptors, Antigen, T-Cell, alpha-beta; D019444:Vaccines, DNA; D014763:Viral Matrix Proteins; C056367:pp150 protein, Cytomegalovirus