Use of a lentiviral vector encoding a HCMV-chimeric IE1-pp65 protein for epitope identification in HLA-Transgenic mice and for ex vivo stimulation and expansion of CD8(+) cytotoxic T cells from human peripheral blood cells.
H2-deleted, HLA-A2, or HLA-B7 transgenic mice were used to identify new human cytomegalovirus (HCMV)-derived major histocompatibility complex class I-restricted epitopes. Three different approaches for mice immunization were compared for their ability to induce a cytotoxic CD8(+) T cell (CTL) response: (1). inoculation of infectious HCMV, (2). injection of immunogenic synthetic peptides, and (3). infection with a newly designed HIV-derived central DNA flap positive lentiviral vector encoding the chimeric IE1-pp65 protein (Trip-IE1-pp65). Targets pulsed with either known immunogenic peptides or computer predicted ones were used to characterize CTL. Most of the mice immunized with the pp65 (495-NLVPMVATV-503) immunodominant peptide responded after one injection whereas only two of six mice responded to two successive inoculations with HCMV. Infection of mice with Trip-IE1-pp65 induced activation and expansion of CTL directed against peptides from both pp65 and IE1 and allowed identification of new epitopes. We further demonstrated the high capacity of monocyte-macrophage cells transduced with Trip-IE1-pp65 to activate and expand CTL directed against pp65 from peripheral blood mononuclear cells of HCMV-seropositive donors. Altogether these results suggest that Trip-IE1-pp65 is a powerful construct both to characterize new epitopes in combination with human leukocyte antigen-transgenic mice immunization and to provide an alternative to the use of known infectious and noninfectious approaches to expand effector T cells for adoptive immunotherapy.
Rohrlich
Pierre S
PS
Antiviral Cellular Immunity Unit, Paris, France.
Cardinaud
Sylvain
S
Lulè
Jacqueline
J
Montero-Julian
Felix A
FA
Prodhomme
Virginie
V
Firat
Hüseyin
H
Davignon
Jean-Luc
JL
Perret
Emmanuelle
E
Monseaux
Sylvain
S
Necker
Antje
A
Michelson
Susan
S
Lemonnier
François A
FA
Charneau
Pierre
P
Davrinche
Christian
C
D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D000950:Antigens, Ly / Q000235:genetics; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002460:Cell Line; D018920:Coculture Techniques; D003587:Cytomegalovirus / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000276:immunology; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000737:chemistry / Q000187:drug effects / Q000276:immunology; D009154:Mutation; D010446:Peptide Fragments / Q000276:immunology / Q000494:pharmacology; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*; D001613:beta 2-Microglobulin / Q000235:genetics
D000950:Antigens, Ly; D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I; C088645:IE1 protein, cytomegalovirus; D017874:Immediate-Early Proteins; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; D001613:beta 2-Microglobulin; C065418:cytomegalovirus matrix protein 65kDa; D007371:Interferon-gamma