Identification and characterization of HLA-class-I-restricted T-cell epitopes in the putative tumor-associated antigens P21-activated serin kinase 2 (PAK2) and cyclin-dependent kinase inhibitor 1A (CDKN1A).
Multiple myeloma (MM) is one of the most common hematological malignancies. Despite a variety of therapeutical approaches including high-dose cytostatic treatment with subsequent autologous or allogeneic stem cell transplantation, as well as vaccination, cures remain rare exceptions. An important issue for future immunological treatments is the identification and characterization of appropriate tumor-associated antigens. However, the number of tumor-associated antigens in MM is limited. PBK/TOPK and activated serin kinase 2 (PAK2) are novel serin kinases that have recently been identified. PBK/TOPK is overexpressed in Burkitt lymphoma, acute lymphoblastic leukemia, and MM; PAK2 is expressed in malignant lymphatic cells. The cyclin kinase inhibitor 1A (CDKN1A) is overexpressed in MM compared to normal plasma cells. We hereby identified and characterized for the first time HLA-class-I-restricted immunogenic peptides in the amino acid sequences of PAK2 and CDKN1A. Using two independent prediction algorithms, we identified two peptides in PAK2 and three peptides in CDK1NA with high binding to HLA-A2. Using an IFN-gamma ELISPOT assay, we could demonstrate the presence and functional activity of CD8-peptide-specific T cells with all tested peptides. To show HLA-A2-restricted antigen recognition, the specific inhibition of T cell recognition was demonstrated with an anti-HLA-A2-blocking antibody. By analysis of peripheral blood of 34 healthy donors for the presence and functional activity of CD8 T cells specific for these peptides, we could demonstrate that peptide T-cell precursors specifically recognizing at least one of the tested peptides are present in 50-60% of the tested donors and that these T-cell precursors can be expanded in vitro. We conclude that PAK2- and CDKN1A-derived peptides can elicit a strong and consistent CD8 T-cell response in an in vitro model. Further investigations will examine the presence and functionality of such T cells in the tumor-bearing host.
Li
Guzi
G
Department of Hematology and Oncology, University Hospital Heidelberg, INF 410, 69120 Heidelberg, Germany.
Hundemer
Michael
M
Wolfrum
Sonja
S
Ho
Anthony D
AD
Goldschmidt
Hartmut
H
Witzens-Harig
Mathias
M
D000951:Antigens, Neoplasm / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002478:Cells, Cultured; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000276:immunology*; D019337:Hematologic Neoplasms / Q000276:immunology* / Q000628:therapy; D006801:Humans; D018448:Models, Immunological; D010950:Plasma Cells / Q000276:immunology; D017346:Protein Serine-Threonine Kinases / Q000276:immunology*; D054462:p21-Activated Kinases
D000951:Antigens, Neoplasm; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; C515907:PAK2 protein, human; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases