Tumor-Intrinsic or Drug-Induced Immunogenicity Dictates the Therapeutic Success of the PD1/PDL Axis Blockade
Abstract
:1. Introduction
2. Materials and Methods
2.1. Mice
2.2. Cell lines and Reagents
2.3. Primary Cells
2.4. In Vitro Treatments
2.5. Therapeutic Protocol
2.6. IFNγ ELISpot
2.7. Flow Cytometry
2.8. Intracellular Staining
2.9. mRNA Extraction and Real-Time PCR
2.10. Morphometric Analysis of MCA205 Tumor Cells
2.11. Statistical Analysis
3. Results
3.1. PD1-PDL Axis Blockade Induces Systemic Polyclonal Immune Activation but no Tumor Specific Immune Responses or CD8+ T Cells and Mice Survival
3.2. Modulation of PD1/PDL Molecules in Tumor and Spleen of Mice Treated with CTX
3.3. Therapeutic Synergism between CTX and PDL1/2 Blockade and Immune Correlates of Therapeutic Efficacy
3.4. Immunogenic Chemotherapy Augments the Therapeutic Efficacy of PDL Blockade in Poorly Immunogenic Tumors
3.5. Chemotherapy-Induced Vaccination only Partially Explains the Therapeutic Synergism between Immunogenic Chemotherapy and PDL Blockade
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene | Forward and Reverse Primers (5′–3′) | NCBI Accession Number | Amplicon Size (Base Pairs) |
---|---|---|---|
IFN-γ | TCAAGTGGCATAGATGTGGAAGAA TGGCTCTGCAGGATTTTCATG | NM_008337.4 | 92 bp |
IL-2 | CCTGAGCAGGATGGAGAATTACA TCCAGAACATGCCGCAGAG | NM_008366.3 | 141 bp |
IL-4 | ACAGGAGAAGGGACGCCAT GAAGCCCTACAGACGAGCTCA | NM_021283.2 | 95 bp |
IL-10 | GGTTGCCAAGCCTTATCGGA ACCTGCTCCACTGCCTTGCT | NM_010548.2 | 191 bp |
IL-12 | GGAAGCACGGCAGCAGAATA AACTTGAGGGAGAAGTAGGAATGG | NM_001303244.1 | 180 bp |
IL-13 | AGACCAGACTCCCCTGTGCA TGGGTCCTGTAGATGGCATTG | NM_008355.3 | 123 bp |
IL-17 | GCTCCAGAAGGCCCTCAGA AGCTTTCCCTCCGCATTGA | NM_010552.3 | 142 bp |
IL-33 | GGGCTCACTGCAGGA GGACCAGGGCTTCGC | NM_ 001164724.2 | 147 bp |
TGF-β1 | TGACGTCACTGGAGTTGTACGG GGTTCATGTCATGGATGGTGC | NM_ 011577.2 | 170 bp |
Prf1 | GTGTCGCATGTACAG TGTGGTAAGCATGCT | NM_ 011073.3 | 116 bp |
Gzmb | GATCGGGAGTGTGAGTCCTAC GAAAGCACGTGGAGGTGAAC | NM_013542.2 | 183 bp |
TLR-3 | TTGCGTTGCGAAGTGAAGAA CAGTTGGGCGTTGTTCAAGA | NM_126166.5 | 149 bp |
HPRT | CTGGTGAAAAGGACCTCTCG TGAAGTACTCATTATAGTCAAGGGCA | NM_013556.2 | 109 bp |
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Rossi, A.; Lucarini, V.; Macchia, I.; Sestili, P.; Buccione, C.; Donati, S.; Ciccolella, M.; Sistigu, A.; D’Urso, M.T.; Pacca, A.M.; et al. Tumor-Intrinsic or Drug-Induced Immunogenicity Dictates the Therapeutic Success of the PD1/PDL Axis Blockade. Cells 2020, 9, 940. https://doi.org/10.3390/cells9040940
Rossi A, Lucarini V, Macchia I, Sestili P, Buccione C, Donati S, Ciccolella M, Sistigu A, D’Urso MT, Pacca AM, et al. Tumor-Intrinsic or Drug-Induced Immunogenicity Dictates the Therapeutic Success of the PD1/PDL Axis Blockade. Cells. 2020; 9(4):940. https://doi.org/10.3390/cells9040940
Chicago/Turabian StyleRossi, Alessandra, Valeria Lucarini, Iole Macchia, Paola Sestili, Carla Buccione, Simona Donati, Maria Ciccolella, Antonella Sistigu, Maria Teresa D’Urso, Anna Maria Pacca, and et al. 2020. "Tumor-Intrinsic or Drug-Induced Immunogenicity Dictates the Therapeutic Success of the PD1/PDL Axis Blockade" Cells 9, no. 4: 940. https://doi.org/10.3390/cells9040940