TY - JOUR
T1 - Clinical-grade myeloma Ag pre-loaded DC vaccines retain potency after cryopreservation
AU - Szmania, S.
AU - Yi, Q.
AU - Cottler-Fox, M.
AU - Rosen, N. A.
AU - Freeman, J.
AU - Kordsmeier, B. J.
AU - Moreno, A.
AU - Shi, J.
AU - Barlogie, B.
AU - Tricot, G.
AU - van Rhee, Frits
N1 - Funding Information:
The authors thank the Immunex-Amgen for providing CD40L, GM-CSF and additional funding for these studies. This work was made possible by grant # 66543-01 Leukemia Society of America and grant # CA55819-08 (PO1) from the NIH .
PY - 2005
Y1 - 2005
N2 - Background: The use of myeloma Ag-loaded mature DC vaccines, cryopreserved in single-use aliquots, is an attractive immunotherapeutic strategy. In this study we investigated the retention of phenotype, viability and potency of DC vaccines after freezing and thawing. Methods: Plastic-adherent monocytes, derived from a steady-state leukapheresis, were cultured in serum free media containing GM-CSF and IL-4. DC were loaded on day 6 with myeloma lysate (ML) or idiotype (Id) Ag and keyhole limpet hemocyanin (KLH), induced to mature on day 7 with CD40-ligand and cryopreserved on day 9. Seventeen clinical-scale cultures were evaluated for DC yield, recovery and immunophenotype after potency was validated with allogeneic mixed lymphocyte culture and Ag presentation assays. Results: We produced 88 individual vaccines from 17 clinical-scale cultures. Median DC yield at harvest was 131 × 106 (range 37-375 × 106) and median recovery of viable DC after thawing was 69% (range 11-100%). We confirmed viability (7AAD-), phenotype (CD14-, CD83+/CD40+, CD83+/CD80+, CD83+/CD86+, CD83+/CD54+, HLA-DR++) and the ability of the DC to present Ag and stimulate allogeneic T cells post-thawing. Discussion: We have validated a serum free culture system for the production of DC. Cryopreservation did not interfere with DC activity, allowed time for rigorous quality control (QC) and flexible scheduling of intranodal vaccination, and reduced the time to prepare multiple vaccines.
AB - Background: The use of myeloma Ag-loaded mature DC vaccines, cryopreserved in single-use aliquots, is an attractive immunotherapeutic strategy. In this study we investigated the retention of phenotype, viability and potency of DC vaccines after freezing and thawing. Methods: Plastic-adherent monocytes, derived from a steady-state leukapheresis, were cultured in serum free media containing GM-CSF and IL-4. DC were loaded on day 6 with myeloma lysate (ML) or idiotype (Id) Ag and keyhole limpet hemocyanin (KLH), induced to mature on day 7 with CD40-ligand and cryopreserved on day 9. Seventeen clinical-scale cultures were evaluated for DC yield, recovery and immunophenotype after potency was validated with allogeneic mixed lymphocyte culture and Ag presentation assays. Results: We produced 88 individual vaccines from 17 clinical-scale cultures. Median DC yield at harvest was 131 × 106 (range 37-375 × 106) and median recovery of viable DC after thawing was 69% (range 11-100%). We confirmed viability (7AAD-), phenotype (CD14-, CD83+/CD40+, CD83+/CD80+, CD83+/CD86+, CD83+/CD54+, HLA-DR++) and the ability of the DC to present Ag and stimulate allogeneic T cells post-thawing. Discussion: We have validated a serum free culture system for the production of DC. Cryopreservation did not interfere with DC activity, allowed time for rigorous quality control (QC) and flexible scheduling of intranodal vaccination, and reduced the time to prepare multiple vaccines.
KW - Cellular immunotherapy
KW - Clinical trial
KW - DC
KW - Multiple myeloma
KW - Tumor lysate
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U2 - 10.1080/14653240510027235
DO - 10.1080/14653240510027235
M3 - Article
C2 - 16162460
AN - SCOPUS:27544502459
SN - 1465-3249
VL - 7
SP - 374
EP - 384
JO - Cytotherapy
JF - Cytotherapy
IS - 4
ER -