A novel culture method of canine peripheral blood lymphocytes with concanavalin a and recombinant human interleukin-2 for adoptive immunotherapy.
This study was designed to develop a novel culture method for the efficient proliferation of canine peripheral blood lymphocytes (cPBL) for adoptive immunotherapy. When cPBL were cultured in the presence of concanavalin A (Con A), proliferation of cPBL was induced and expression of interleukin-2 receptor (IL-2R) which enables to respond to exogenously added IL-2 was upregulated. And then, when cPBL were cultured with recombinant human interleukin-2 (rhIL-2) in addition to Con A, proliferation was accelerated and increased to about 10-fold after 1 week. The phenotypic analysis showed that the main population of the cultured cPBL was consisted of CD8+ positive lymphocytes. Among them, CD4+CD8+ double positive (DP) lymphocytes had significantly increased, and the ratio of CD4+ single positive (SP) lymphocytes to CD8+ SP lymphocytes (CD4+SP/CD8+SP) was decreased as compared to before culturing. To evaluate the cytotoxic activity of cPBL cultured with Con A and rhIL-2, furthermore, cytotoxic assay was carried out against xenogeneic melanoma cell line (MeWo), which resulted in MHC-unrestricted cytokilling. These results suggest that the culture method of cPBL by the use of Con A and rhIL-2 may be useful for generating lymphokine activated killer cells, and also this may be beneficial for adoptive immunotherapy of tumor-bearing dogs.
Kato
Masahiro
M
Laboratory of Veterinary Immunology, Division of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka, Japan.
Watarai
Shinobu
S
Nishikawa
Shigeru
S
Iwasaki
Tadashi
T
Kodama
Hiroshi
H
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary*; D045744:Cell Line, Tumor; D003208:Concanavalin A / Q000276:immunology; D003601:Cytotoxicity Tests, Immunologic / Q000662:veterinary; D004285:Dogs / Q000097:blood / Q000276:immunology*; D053804:Granzymes / Q000235:genetics; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000379:methods / Q000662:veterinary*; D007376:Interleukin-2 / Q000276:immunology; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D007376:Interleukin-2; D003208:Concanavalin A; D053804:Granzymes