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Vaccination with poly(IC:LC) and peptide-pulsed autologous dendritic cells in patients with pancreatic cancer

View Article: PubMed Central - PubMed

ABSTRACT

Background: Dendritic cells (DCs) enhance the quality of anti-tumor immune response in patients with cancer. Thus, we posit that DC-based immunotherapy, in conjunction with toll-like receptor (TLR)-3 agonist poly-ICLC, is a promising approach for harnessing immunity against metastatic or locally advanced unresectable pancreatic cancer (PC).

Methods: We generated autologous DCs from the peripheral blood of HLA-A2+ patients with PC. DCs were pulsed with three distinct A2-restricted peptides: 1) human telomerase reverse transcriptase (hTERT, TERT572Y), 2) carcinoembryonic antigen (CEA; Cap1-6D), and 3) survivin (SRV.A2). Patients received four intradermal injections of 1 × 107 peptide-pulsed DC vaccines every 2 weeks (Day 0, 14, 28, and 42). Concurrently, patients received intramuscular administration of Poly-ICLC at 30 μg/Kg on vaccination days (i.e., day 0, 14, 28, and 42), as well as on days 3, 17, 21, 31, 37, and 45. Our key objective was to assess safety and feasibility. The effect of DC vaccination on immune response was measured at each DC injection time point by enumerating the phenotype and function of patient T cells.

Results: Twelve patients underwent apheresis: nine patients with metastatic disease, and three patients with locally advanced unresectable disease. Vaccines were successfully manufactured from all individuals. We found that this treatment was well-tolerated, with the most common symptoms being fatigue and/or self-limiting flu-like symptoms. Among the eight patients who underwent imaging on day 56, four patients experienced stable disease while four patients had disease progression. The median overall survival was 7.7 months. One patient survived for 28 months post leukapheresis. MHC class I –tetramer analysis before and after vaccination revealed effective generation of antigen-specific T cells in three patients with stable disease.

Conclusion: Vaccination with peptide-pulsed DCs in combination with poly-ICLC is safe and induces a measurable tumor specific T cell population in patients with advanced PC.

Trial registration: NCT01410968; Name of registry: clinicaltrials.gov; Date of registration: 08/04/2011).

Electronic supplementary material: The online version of this article (doi:10.1186/s13045-017-0459-2) contains supplementary material, which is available to authorized users.

No MeSH data available.


Phenotypic characterization of dendritic cells. a Schematic diagram showing the different time points for vaccination and analysis. b Dendritic cells (DCs) were prepared from each patient (see Methods). Before treatment administration the DCs were characterized using the flurochrome-conjugated antibodies for cell surface expression of CD11c, CD86, HLA-DR, and CD14. The data was acquired using BD Accuri flow cytometer and analyzed using FlowJo. The numerical values adjacent to the histogram represent the mean fluorescence intensity (MFI)
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Fig1: Phenotypic characterization of dendritic cells. a Schematic diagram showing the different time points for vaccination and analysis. b Dendritic cells (DCs) were prepared from each patient (see Methods). Before treatment administration the DCs were characterized using the flurochrome-conjugated antibodies for cell surface expression of CD11c, CD86, HLA-DR, and CD14. The data was acquired using BD Accuri flow cytometer and analyzed using FlowJo. The numerical values adjacent to the histogram represent the mean fluorescence intensity (MFI)

Mentions: This pilot study, with a planned sample size of 12 patients, was designed to evaluate the feasibility and the safety of systemic administration of polyinosinic-polycytidylic acid stabilized with polylysine and carboxymethylcellulose (poly(IC:LC), or Hiltonol; Oncovir, Washington, D.C.) concurrent with active vaccination of autologous peptide pulsed DCs in patients with advanced adenocarcinoma of the pancreas. The DC vaccine consisted of a pool of three aliquots of DCs pulsed with hTERT (YLFFYRKSV) [25–27], Cap1-6D (YLSGADLNL) [30], or survivin (LTLGEFLKL) [31, 32] peptides that were obtained from PolyPeptide Group (San Diego, CA). The CEF control peptide pool was obtained from AnaSpec, (cat# 61036, Fremont, CA). The CEF control peptides are 8–12 amino acids in length, with sequences derived from the human Cytomegalovirus, Epstein-Barr Virus and Influenza Virus. Eligible patients underwent leukapheresis on day -35 to generate immature DCs. At day -28 DCs were cryopreserved and subsequently tested according to lot release criteria. Patients were given a combination vaccine comprised of antigen-pulsed DCs (1 × 107 DC intradermally delivered on days 0, 14, 28, and 42) and TLR3 agonist Poly(IC:LC) (30 μg/kg intramuscularly administered on days 0, 3, 14, 17, 21, 28, 31, 37, 42, and 45), as outlined in Fig. 1a. All patients were premedicated with acetaminophen and diphenhydramine prior to injection. Comprehensive safety evaluations, including physical examination, vital signs, and clinical laboratory tests (hematology, blood chemistry, urine analysis) were performed at baseline, prior to each vaccination, at predetermined time points between vaccinations, and 2 weeks after the last vaccination. Adverse events were assessed for severity and relationship to treatment, and were graded according to NCI-CTCAE version 4.0. Baseline tumor assessment was performed within 28 days prior to day 0, and restaging assessments were performed within 7 days of day 56. Objective tumor response was evaluated according to RECIST criteria version 1.1 [33, 34]. Blood samples were drawn for immune monitoring before each vaccination and two weeks after the last vaccination (days 0, 14, 28, 42, and 56). Overall survival is defined as the time from leukapheresis until death. Patients were categorized by their response (complete/partial response, stable disease, or progression) at day 56. The study was approved by the Institutional Review Board at MUSC, and was performed in accordance with the Declaration of Helsinki, Good Clinical Practice (GCP) guidelines and applicable local regulatory requirements and laws. All patients provided their written informed consent.Fig. 1


Vaccination with poly(IC:LC) and peptide-pulsed autologous dendritic cells in patients with pancreatic cancer
Phenotypic characterization of dendritic cells. a Schematic diagram showing the different time points for vaccination and analysis. b Dendritic cells (DCs) were prepared from each patient (see Methods). Before treatment administration the DCs were characterized using the flurochrome-conjugated antibodies for cell surface expression of CD11c, CD86, HLA-DR, and CD14. The data was acquired using BD Accuri flow cytometer and analyzed using FlowJo. The numerical values adjacent to the histogram represent the mean fluorescence intensity (MFI)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5384142&req=5

Fig1: Phenotypic characterization of dendritic cells. a Schematic diagram showing the different time points for vaccination and analysis. b Dendritic cells (DCs) were prepared from each patient (see Methods). Before treatment administration the DCs were characterized using the flurochrome-conjugated antibodies for cell surface expression of CD11c, CD86, HLA-DR, and CD14. The data was acquired using BD Accuri flow cytometer and analyzed using FlowJo. The numerical values adjacent to the histogram represent the mean fluorescence intensity (MFI)
Mentions: This pilot study, with a planned sample size of 12 patients, was designed to evaluate the feasibility and the safety of systemic administration of polyinosinic-polycytidylic acid stabilized with polylysine and carboxymethylcellulose (poly(IC:LC), or Hiltonol; Oncovir, Washington, D.C.) concurrent with active vaccination of autologous peptide pulsed DCs in patients with advanced adenocarcinoma of the pancreas. The DC vaccine consisted of a pool of three aliquots of DCs pulsed with hTERT (YLFFYRKSV) [25–27], Cap1-6D (YLSGADLNL) [30], or survivin (LTLGEFLKL) [31, 32] peptides that were obtained from PolyPeptide Group (San Diego, CA). The CEF control peptide pool was obtained from AnaSpec, (cat# 61036, Fremont, CA). The CEF control peptides are 8–12 amino acids in length, with sequences derived from the human Cytomegalovirus, Epstein-Barr Virus and Influenza Virus. Eligible patients underwent leukapheresis on day -35 to generate immature DCs. At day -28 DCs were cryopreserved and subsequently tested according to lot release criteria. Patients were given a combination vaccine comprised of antigen-pulsed DCs (1 × 107 DC intradermally delivered on days 0, 14, 28, and 42) and TLR3 agonist Poly(IC:LC) (30 μg/kg intramuscularly administered on days 0, 3, 14, 17, 21, 28, 31, 37, 42, and 45), as outlined in Fig. 1a. All patients were premedicated with acetaminophen and diphenhydramine prior to injection. Comprehensive safety evaluations, including physical examination, vital signs, and clinical laboratory tests (hematology, blood chemistry, urine analysis) were performed at baseline, prior to each vaccination, at predetermined time points between vaccinations, and 2 weeks after the last vaccination. Adverse events were assessed for severity and relationship to treatment, and were graded according to NCI-CTCAE version 4.0. Baseline tumor assessment was performed within 28 days prior to day 0, and restaging assessments were performed within 7 days of day 56. Objective tumor response was evaluated according to RECIST criteria version 1.1 [33, 34]. Blood samples were drawn for immune monitoring before each vaccination and two weeks after the last vaccination (days 0, 14, 28, 42, and 56). Overall survival is defined as the time from leukapheresis until death. Patients were categorized by their response (complete/partial response, stable disease, or progression) at day 56. The study was approved by the Institutional Review Board at MUSC, and was performed in accordance with the Declaration of Helsinki, Good Clinical Practice (GCP) guidelines and applicable local regulatory requirements and laws. All patients provided their written informed consent.Fig. 1

View Article: PubMed Central - PubMed

ABSTRACT

Background: Dendritic cells (DCs) enhance the quality of anti-tumor immune response in patients with cancer. Thus, we posit that DC-based immunotherapy, in conjunction with toll-like receptor (TLR)-3 agonist poly-ICLC, is a promising approach for harnessing immunity against metastatic or locally advanced unresectable pancreatic cancer (PC).

Methods: We generated autologous DCs from the peripheral blood of HLA-A2+ patients with PC. DCs were pulsed with three distinct A2-restricted peptides: 1) human telomerase reverse transcriptase (hTERT, TERT572Y), 2) carcinoembryonic antigen (CEA; Cap1-6D), and 3) survivin (SRV.A2). Patients received four intradermal injections of 1 × 107 peptide-pulsed DC vaccines every 2 weeks (Day 0, 14, 28, and 42). Concurrently, patients received intramuscular administration of Poly-ICLC at 30 μg/Kg on vaccination days (i.e., day 0, 14, 28, and 42), as well as on days 3, 17, 21, 31, 37, and 45. Our key objective was to assess safety and feasibility. The effect of DC vaccination on immune response was measured at each DC injection time point by enumerating the phenotype and function of patient T cells.

Results: Twelve patients underwent apheresis: nine patients with metastatic disease, and three patients with locally advanced unresectable disease. Vaccines were successfully manufactured from all individuals. We found that this treatment was well-tolerated, with the most common symptoms being fatigue and/or self-limiting flu-like symptoms. Among the eight patients who underwent imaging on day 56, four patients experienced stable disease while four patients had disease progression. The median overall survival was 7.7 months. One patient survived for 28 months post leukapheresis. MHC class I –tetramer analysis before and after vaccination revealed effective generation of antigen-specific T cells in three patients with stable disease.

Conclusion: Vaccination with peptide-pulsed DCs in combination with poly-ICLC is safe and induces a measurable tumor specific T cell population in patients with advanced PC.

Trial registration: NCT01410968; Name of registry: clinicaltrials.gov; Date of registration: 08/04/2011).

Electronic supplementary material: The online version of this article (doi:10.1186/s13045-017-0459-2) contains supplementary material, which is available to authorized users.

No MeSH data available.