Dendritic/pancreatic carcinoma fusions for clinical use: Comparative functional analysis of healthy- versus patient-derived fusions.
Fetal calf serum (FCS)-independent pancreatic cancer cells were established in plasma protein fraction (PPF)-supplemented medium that is an agent of good manufacturing practice (GMP) grade. Dendritic cells (DCs) were activated with the Toll-like receptor agonist, penicillin-inactivated Streptococcus pyogenes (OK-432) that is also a GMP grade agent. Therefore, sufficient amounts of FCS-independent fusions were successfully generated with decreased potential hazards of FCS. The FCS-independent fusions expressed tumor-associated antigens, HLA-DR, costimulatory molecules, IL-12, and IL-10. Stimulation of T cells with fusions from healthy donors resulted in proliferation of T cells with high expression levels of perforin/granzyme B and IFN-gamma and efficient induction of antigen-specific cytotoxic T lymphocytes (CTLs). Selection and expansion of T-cell clones were confirmed by TCR Vbeta analysis. However, fusions from patients with metastatic pancreatic cancer induced increased expression levels of TGF-beta1 in CD4+ CD25high T cells and low levels of CTLs with decreased IFN-gamma production.
Division of Gastroenterology and Hepatology, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan. shigeo_koido@jikei.ac.jp
Shigeo
S
Koido
Eiichi
E
Hara
Sadamu
S
Homma
Yoshihisa
Y
Namiki
Hideo
H
Komita
Akitaka
A
Takahara
Eijiro
E
Nagasaki
Masaki
M
Ito
Yukiko
Y
Sagawa
Makoto
M
Mitsunaga
Kan
K
Uchiyama
Kenichi
K
Satoh
Seiji
S
Arihiro
Toshifumi
T
Ohkusa
Jianlin
J
Gong
Hisao
H
Tajiri
D000951:Antigens, Neoplasm / Q000276:immunology / Q000378:metabolism; D002459:Cell Fusion / Q000379:methods; D002469:Cell Separation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D006822:Hybrid Cells / Q000276:immunology*; D007167:Immunotherapy / Q000379:methods*; D008213:Lymphocyte Activation / Q000276:immunology; D010190:Pancreatic Neoplasms / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology
D000951:Antigens, Neoplasm