Separation of adherent and non-adherent cells
Peripheral blood mononuclear cells were harvested with the COBE Spectra Apheresis System every 2 to 4 weeks. PBMCs from 2,500 ml of blood were enriched by density gradient centrifugation with Ficoll-Paque. The PBMCs were incubated for 45 min in a 5% CO2 atmosphere at 37°C in serum-free AIM-V medium. Plastic-adherent cells were used for generation of DCs,
while non-adherent cells were used for generation of CTLs.
Generation of MUC1-CTLs
The PBMCs harvested from each patient were incubated for 45 min in a plastic plate. Non-adherent cells were cultured in AIM-V with the MUC1-expressing pancreatic cancer cell line YPK-1
inactivated with 0.2 mg/mL mitomycin C. After 3 days of culture, the cells were cultured with 10 Japan Reference Units (JRU)/mL recombinant human interleukin (rhIL)-2 in a 5% CO2 atmosphere at 37°C for 7 days. These cells were termed MUC1-CTLs. On day 10, MUC1-CTLs were washed 3 times with saline, suspended in 100 mL saline and administered intravenously as maximum available cell products.
Patients were treated from 2 to 15 times with both cell types. Total numbers of MUC1-DCs and
7889 MUC1- CTLs administered were 1.1×10 to 3.1×10 and 5.0×10 to 6.8×10 , respectively.
One of the 20 patients (patient 2), who had multiple lung metastases after curative surgery, had a complete response. After AIT, multiple lung nodules disappeared completely.
Five of twenty patients had stable disease. The duration of stable disease is range from 6 to 24 months as time to progression (TTP). Patient 1, who had unresectable pancreatic cancer with multiple liver metastases, had stable disease for at least 6 months. After AIT, there was no progression of the primary pancreatic cancer. One hepatic metastasis decreased in size, although another increased.
The survival times of the 20 pancreatic cancer patients who underwent AIT with MUC1-CTLs and MUC1-DCs ranged from 2 to 75 months, with a mean survival time of 9.8 months. One-, two-, and three-year survival rates after AIT were 20.0%, 10.0%, and 5.0%, respectively.
No grade II-IV toxicity according to CTCAE was observed in any patient after injection of MUC1-DC and MUC1-CTL. Only one patient developed transient systemic itching.
Adoptive immunotherapy with MUC1- DC and MUC1-CTL may be feasible and effective for pancreatic cancer.
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Scientific article publishing date: 1/1/2008
Immucura identifier BSC22_380EN