IL-12p70–producing patient DC vaccine elicits Tc1-polarized immunity
Beatriz M. Carreno, … , Kathryn M. Trinkaus, Gerald P. Linette
Beatriz M. Carreno, … , Kathryn M. Trinkaus, Gerald P. Linette
Published July 11, 2013
Citation Information: J Clin Invest. 2013;123(8):3383-3394. https://doi.org/10.1172/JCI68395.
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Clinical Research and Public Health Oncology

IL-12p70–producing patient DC vaccine elicits Tc1-polarized immunity

Abstract

Background. Systemic administration of IL-12p70 has demonstrated clinical activity in cancer patients, but dose-limiting toxicities have hindered its incorporation in vaccine formulations. Here, we report on the immunological and clinical outcomes upon vaccination with CD40L/IFN-γ–matured, IL-12p70–producing DCs.

Methods. 7 HLA-A*0201+ newly diagnosed stage IV melanoma patients were immunized against the gp100 melanoma antigen using autologous peptide-pulsed, CD40L/IFN-γ–matured DCs. PBMCs were taken weekly for immune monitoring by tetramer analysis and functional assays. CT imaging was performed at baseline, week 9, and week 18 for clinical assessment using RECIST.

Results. 6 of 7 treated patients developed sustained T cell immunity to all 3 melanoma gp100 antigen–derived peptides. 3 of the 6 immunological responders developed confirmed clinical responses (1 complete remission >4 years, 2 partial response). Importantly, DC vaccine–derived IL-12p70 levels positively correlated with time to progression (P = 0.019, log-rank), as did T-cytotoxic 1 (Tc1) immunity, as assessed by IFN-γ/IL-13 and IFN-γ/IL-5 ratios (P = 0.035 and P = 0.030, respectively, log-rank). In contrast, a pathway-specific defect in IL-12p35 transcription was identified upon CD40L/IFN-γ activation in clinical nonresponder patient DCs, and gp100-specific T cells from these patients displayed a Tc2 phenotype. Incorporation of TLR3 and TLR8 agonists into the CD40L/IFN-γ activation protocol corrected the IL-12p70 production defect in DCs derived from clinical nonresponder patients.

Conclusion. These findings underscore the essential role of IL-12p70 in the development of therapeutic type 1 antigen–specific CD8+ T cell immunity in humans with cancer.

Trial registration. Clinicaltrials.gov NCT00683670.

Funding. Barnes-Jewish Hospital Foundation, Siteman Cancer Frontier Fund, Washington University/JNJ Translational Medicine Award, and NCI (P30 CA91842).

Authors

Beatriz M. Carreno, Michelle Becker-Hapak, Alexander Huang, Megan Chan, Amer Alyasiry, Wen-Rong Lie, Rebecca L. Aft, Lynn A. Cornelius, Kathryn M. Trinkaus, Gerald P. Linette

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Figure 6

IL-12p70 production and IL-12p35 transcription by mDCs derived from patients with melanoma.

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IL-12p70 production and IL-12p35 transcription by mDCs derived from pati...
(A) DCs from age- and gender-matched healthy donors (H) and melanoma patients (M) were activated with CD40L/IFN-γ for 24 hours, and supernatants were harvested and assayed for IL-12p70 production by ELISA. Horizontal lines and error bars denote median and interquartile range. P value (Wilcoxon matched-pairs test) is indicated. (B) Patient DCs were activated with CD40L/IFN-γ for 24 hours, supernatants were collected, and IL-12p40 and IL-12p70 production was measured by ELISA. Results are shown for 10 melanoma patients. Horizontal lines and error bars denote median and interquartile range. (C) To examine IL-12p35 gene activation, DCs were activated with CD40L/IFN-γ for 6 hours, cells were harvested and washed, and total RNA was prepared. Total RNA was also prepared from iDCs. Using primers specific for IL-12p35 and CD11c (DC lineage marker), qRT-PCR was performed and analyzed using the relative standard method. Results were normalized to CD11c, and values are expressed as fold IL-12p35 induction in mDCs relative to iDCs. Results are representative of 2 experiments.