Table 3.
Preclinical studies in cancer DNA vaccination (2015–2018)
| Cancer type | Animal | DNA vaccine | Combination therapies | Protocol | DNA vaccine delivery | Results | Year, ref. |
|---|---|---|---|---|---|---|---|
| Cervical cancer (TC-1 cells) | C57BL/6 mice | HPV plasmid encoding E6 and E7 antigens | pVAX1-ISG15 encoding an optimized mouse adjuvant ISG15 | Therapeutic vaccination Tumor: 5 × 104 TC-1 tumor cells, SC Vaccine and adjuvant: 7 D after tumor implantation, followed by 3 boosts weekly |
IM EP, in the tibialis anterior muscle | - Strong HPV E7-specific CD8+ T cell immune response - Increase in INF-γ secretion - 6/10 mice were tumor-free at D 42 |
2015 [90] |
| Cervical cancer (TC-1 cells) | C57BL/6 mice | pcDNA3.1-E7, encoding E7 antigen of HPV16 | Monophosphoryl lipid A (MPL, TLR4 agonist) and α-galactosylceramide (GalCer) | Therapeutic vaccination Tumor: 2 × 105 TC-1 tumor cells, SC Vaccine: 100 μg, 7 D after tumor implantation, followed by 2 boosts weekly Adjuvant: 25 μg of MPL and 1 μg of GalCer, SC with the vaccine |
SC | - CTL-specific cytolytic activity - Higher INF-γ, IL-4 and IL-12 production - Decrease in tumor growth if both adjuvants were administered |
2016 [136] |
| Cervical cancer (TC-1 cells) | C57BL/6 mice | HELP-E7SH, encoding E7 antigen of HPV16 and a helper epitope to stimulate CD4+ response | Abs against CD70, CTLA-4, PD-1; agonistic Ab to CD27 | Therapeutic vaccination Tumor: 1 × 105 TC-1 tumor cells, at D 0 before the vaccination, SC Vaccine: 15 μl of a 2 mg/ml DNA solution, on D 0, 3 and 6 Abs: 100 μg, at D 0, 3 and 6 (CD70 also at D 9), IP |
Tattoo (intraepidermal vaccination) | - Help epitopes increased E7-specific CD8+ response in lymph nodes and spleen - CTLA4 and PD-1 did not promote CTL priming, if not combined with CD27 or the vaccine - CD27 agonism + anti-PD1 improved mice survival, albeit CD27 + anti-CTLA4 further increased the CTL response |
2016 [35] |
| Cervical cancer (TC-1 cells) | C57BL/6 mice | pVAX1-gDE7, encoding HPV-16 E7 protein fused to HSV-1 gD protein | pcDNA3-IL2 encoding murine IL-2; anti-Gr1 Ab | Therapeutic vaccination Tumor: 7.5 × 104 TC-1 tumor cells, at D 0 before the vaccination, SC Vaccine and adjuvant: 50 μg alone or in combination with 50 μg of the adjuvant, at D 3, 2 doses, weekly Anti-Gr1: 200 μg, once/W for 6 IP injections, at D 7–10, 5 doses, weekly |
IM | - Vaccination with the 2 plasmids avoided MDSC accumulation - Combination of the vaccines and anti-Gr1 antibody increased mice survival, completely eradicating the tumor |
2016 [95] |
| Cervical cancer (TC-1 cells) | C57BL/6 mice | pcDNA3.1-E7, encoding HPV-16 E7 antigen | melatonin | Therapeutic vaccination Tumor: 2 × 105 TC-1 cells at D0, before the vaccination, SC Vaccine: 90 μg, 3 times, at 7 D interval Melatonin: 50 or 100 mg/kg |
SC | - Production of HPV16 E7-specific CTL - Increase of IFNg and TNFa in the TME - Tumor volume reduction |
2018 [34] |
| Cervical cancer (TC-1 cells) | C57BL/6 mice | dbDNA, encoding HPV16 E6 and E7 | / | Therapeutic vaccination Tumor: 5 × 104 TC-1 cells, SC, at D0 Vaccine: at D3, 25 μg/hind, boost after 7 W |
IM in the anterior tibialis and IM EP in the quadriceps | - Delay in the tumor growth - High levels of IFNg-secreting Th cells - Production of IgG1 - IL-12 production and low IL10 |
2018 [137] |
| Cervical cancer (HPV) | BALB/c mice | pNGVL4a-hCRTE6E7L2, expressing the HPV16 E6, E7 and L2 antigens | / | Prophylactic vaccination Vaccine: 3 injections, biweekly Tumor: 12 μl of HPV16 PsV, 19 D after the last vaccination |
IM EP | - Production of Ab against E6, E7 and L2; - Protection of 3/5 of mice from the challenge, but without a significant difference compared to the control group |
2017 [138] |
| Lobular carcinoma (TUBO cells) | BALB/c mice | pAmot, coding human p80 Amot (Angiomotin), antiangiogenic | / | Therapeutic vaccination Tumor: 105 TUBO cells, SC Vaccine: 50 μg, at D 7 |
IM EP in the quadriceps muscle | - Delay in tumor progression - Heterogeneous changes in the tumor region following antiangiogenetic treatment |
2015 [139] |
| Murine breast cancer (D2F2 cells) | BALB/c mice | pVAX-E2A, encoding Her2/neu antigen | pVAX-CCL4, encoding CCL4, chemoattractant for immune effector cells | Prophylactic vaccination Vaccine: 2 × 100 μg, on D 1 and 15 Tumor: 2 × 105 Her2/neu+ cells, on D 25, SC |
IM | - With the combined therapy, 26% of mice remained tumor-free (CCL4 improved tumor protection) - CCL4 produced a Th1 anti-Her2/neu response |
2016 [140] |
| Murine breast cancer (4 T1 cells) | BALB/c mice | CpVR-FAP, encoding fibroblast associated protein (FAP) | Cyclophosphamide, chemotherapy agent | Therapeutic vaccination Tumor: 2 × 104 4 T1 cells, at D 0, SC Vaccine: 100 μg, on D 2, 9 and 16 after tumor injection Cyclophosphamide: 50 mg/kg, on D 1, 8, 15, IP |
IM in the tibialis anterior muscle | - Combination therapy increased median survival time of mice - Suppression of IL-10, VEGFα and CXCL12 mRNA expression |
2016 [115] |
| Murine breast cancer (4 T1 cells) | BALB/c mice | CpVR-FAP, encoding FAP | / | Prophylactic and therapeutic vaccination Tumor: 2 × 104 4 T1 cells, SC Vaccine: 100 μg, on D 2, 9 and 16 after tumor injection or 3 times every 2 W before tumor injection |
IM in the tibialis anterior muscle | - Specific CTL response against FAP - Increased IL-2 production - Delay of the tumor growth also in therapeutic setting - Decrease in FAP expression without impairing wound healing |
2016 [141] |
| Murine breast cancer (4 T1 cells) | BALB/c mice | pVAX1-mCr-1, encoding mouse Cripto-1 oncofetal protein | / | Prophylactic vaccination Vaccine: 40 μg Tumor: 2 × 105 4 T1 mCr-1 cells, W12 |
ID EP | - Humoral response against Cr-1 - Protective immune response against cancer stem cells - Reduced lung metastasis |
2018 [142] |
| Colon cancer (colon 26/β-gal cells) | BALB/c mice | pcDNA3/β-gal encoding β-galactosidase | pCAGGS/FasL encoding Fas ligand | Prophylactic vaccination Vaccine and adjuvant: 50 μg + 1 μM cardiotoxin to facilitate DNA uptake Tumor: 106 Colon 26/β-gal cells, 21 D after vaccine injection, SC |
/ | - The combined therapy decreased tumor growth rate - Production of Abs anti-β-gal |
2015 [143] |
| Colon cancer (CT26/HER2 cells) | BALB/c mice | pVAX1-HER2, coding HER2 antigen | Gemcitabine, chemotherapy agent; anti-Gr1 antibody; anti-PD-L1 Ab | Prophylactic and therapeutic vaccination Tumor: 3–5 × 105 CT26/HER2 cells, SC Vaccine: 50 μg Anti-PD-L1 and Gr-1 Ab: 200 μg and 250 μg, respectively, IP Gemcitabine: 75 μg/g, 2 times/W, IP |
IM EP | - In prophylactic vaccination, the combination of vaccine + anti-PD-L1 Ab failed to delay tumor growth - The addition of anti-Gr1 or gemcitabine delayed tumor growth |
2017 [144] |
| Colon cancer (CT26 cells) | Balb/c mice | CpVR-MS and CpDV-IL2-MS, encoding a fusion gene of human surviving S8 and human 33 MUC1, plus IL2) | Ad-MS (Adenovirus) | Therapeutic vaccination Tumor: 106 CT26 cells, SC Vaccine: 100 μg, twice Ad-MS: 108 pfu, D1, 15 and 29 |
IM | - Specific immune response in splenocytes - Upregulation of CCL-19 and GM-CSF - Downregulation of PD-L1 and MMP-9 |
2018 [145] |
| Colorectal cancer (CT- 26/NIS cells) | BALB/c mice | pcDNA-hNIS, expressing human sodium/io- dide symporter (hNIS) | / | Vaccine: 100 μg, 3 times at 2 W intervals Tumor: 2 W after the final hNIS DNA injection, 5 × 105 (left) or 1 × 105 (right) CT-26/NIS cells, SC |
ID | - Increase of IgG2a/IgG1 ratio - Increase of INF-g secreting cells and IFN-g production - Th1 response - Slower tumor growth |
2018 [146] |
| Melanoma (B16F10-β-hCG cells) | C57BL/6 mice | CAVE = pSVK-VEGFR2-GFc-IL12, Semliki Forest Virus expressing VEGFR2 and IL-12 | CAVA = SFV replicon DNA vaccine targeting surviving and hCG antigens | Prophylactic vaccination Vaccine: 10 μg of CAVA/CAVE, 3 times at 10 D of interval Tumor: 7.5 × 104 B16F10-β-hCG cells, 7 D after the last immunization, SC |
IM EP | - Combination of the 2 vaccines delayed tumor growth more efficiently than the single vaccine and increased mice survival - CAVE + CAVA decreased microvessel density |
2015 [147] |
| Melanoma (B16F10 cells) | C57BL/6 mice | pSPD-gp100-CD40L, encoding gp100 inserted between mouse Surfactant Protein D (SPD) and CD40L | pIL-12, encoding IL-12p70; pcDNA3.1-GM-CSF encoding GM-CSF | Therapeutic vaccination Tumor: 5 × 104 B16F10 cells, ID Vaccine and adjuvants: 80 μg vaccine + 20 μg of each adjuvant plasmid, on D 3, 10 and 17 |
IM in hind quadriceps muscles | - Vaccine alone did not delay tumor growth, but the combination with the 2 adjuvants was very effective in increasing mice survival | 2015 [94] |
| Melanoma (B16F10 cells) | C57BL/6 mice | pVAX1-MUCI, encoding mucin I glycoprotein | pVAX1-Flt3L, encoding Fms-like tyrosinase 3-ligand | Therapeutic vaccination Tumor: 1 × 106 B16F10 cells, SC Vaccine + adjuvant: 50 μg, priming when tumors were palpable, boosts after 7 and 14 D |
IM EP | - Specific CTL and antibodies - Tumor growth suppression |
2018 [148] |
| Melanoma (B16 cells) | C57BL/6 mice | p-mBAP31 and p-LAMP/mBAP31 = p43 and p43- Lysosomal Associated Membrane Protein (LAMP) vectors + mouse B-cell receptor-associated protein (mBAP) | / | Therapeutic vaccination Tumor: 5 × 104 B16 cells, at D 0, SC Vaccine: 50 μg, at D 3, 10, 17 and 24 |
SC | - No evidence of autoimmune disorders - High INF-γ production, especially using LAMP vaccine - LAMP vaccine increased the CTL cytotoxicity - Suppression of tumor growth, especially using LAMP vaccine |
2015 [149] |
| Melanoma | Horses | Minimalistic immunogenically defined gene expression (MIDGE)-Th1 vector + eqIL12 and IL-1beta receptor antagonist protein (ILRAP)-eqIL18 | hgp100MIDGE-Th1; htyrMIDGE-Th1 | Therapeutic vaccination Tumor: horses were already affected by melanoma Vaccine and adjuvants: 500 μg ID peritumorally and 500 μg IM into the semimembranosus muscle, 3 times |
ID and IM | - Vaccine was safe and well-tolerated, except an increase in the body temperature on the day after injection and signs of acute inflammation - Tumor volume was reduced by 28.5%, but without significant differences if adjuvants were added |
2015 [150] |
| Melanoma (B16F10-OVA cells) | C57BL/6 mice | pVAX2-OVA, encoding ovalbumin; pVAX2-gp100, encoding gp100 | pVAX2-HIV-1 Gag | Prophylactic and therapeutic vaccination Vaccine: 1 μg (p-OVA) or 50 μg (p-gp100), at D 2, 9 and 16 (therapeutic) or 3 times every 2 W before the tumor challenge (prophylactic) Adjuvant: 1 μg, co-administered with the vaccine Tumor: 1 × 105 B16F10-OVA cells at D 0 (therapeutic) or 2 W after the last vaccine injection (prophylactic) |
IM EP | - Delay of tumor growth and increase in mice survival - Codelivery of the adjuvant-encoding plasmid polarized the immune response towards a Th1-like phenotype |
2016 [40] |
| Mastocytoma (P815 cells) | DBA/2 mice | Differently optimized pVAX2-P1A vaccines, encoding P815A | / | Prophylactic and therapeutic vaccination Vaccine: 50 μg, at D 2, 9 and 16 (therapeutic) or 3 times every 2 W before the tumor challenge (prophylactic) Tumor: 1 × 106 P815 cells at D 0 (therapeutic) or 2 W after the last vaccine injection (prophylactic) |
IM EP | - Delay of tumor growth and increase in mice survival Activation of innate immunity related to the different CpG motif amount inside the P1A gene |
2017 [37] |
| Mastocytoma (P815 cells) | DBA/2 mice | Optimized pVAX2-P1A vaccine, encoding P815A | Anti-CTLA4, anti-PD1 | Therapeutic vaccination Tumor: 1 × 106 P815 cells at D 0 Vaccine: 50 μg, at D 2, 9 and 16 Anti-CTLA4, anti-PD1: 100 μg at D 3, 6 and 9 |
IM EP | - Survival reached 90% - Increase of specific T cell infiltration in the TME - Increase of IL-12 production - Decrease of metastasis formation |
2018 [109] |
| Malignant tumor | HLA-A2.1/Kb transgenic mice | p-GST-YL66, against multiepitope YL66 (from COX2 and MAGE4), linked with membrane permeable Tat-PTD and the universal Th epitope | / | Not available | / | - CTL-mediated tumor cell lysis in vitro and in vivo | 2017 [87] |
| Colorectal cancer (MC32 cells) | C57BL/6 mice | Pc-DNA3-CEA, carcinoembryonic antigen (CEA) | Ab 4-1BB | Prophylactic and therapeutic vaccination Tumor: 1–5 × 105 MC32 cells, SC Vaccine: 50 μg, at 1 W of interval Ab anti-4-1BB: 50 μg, systemically, after vaccine injection |
IM EP | - Antigen-specific CTL activity and tumor-protective immune response in prophylactic model - Ab 4-1BB increased CTL lytic activity - MC32 cells resisted to CEA DNA vaccination by loss of antigen presentation to CEA-specific CTL in therapeutic model |
2015 [151] |
| Melanoma (B16), carcinoma (3LL) | C57BL/6 mice | SCT-KDR2, encoding the mouse β2microglobulin + KDR2 (VEGFR2 antigen peptide) + MHC class I H-2Db, subcloned into pdDNA3.1 | / | Prophylactic vaccination Vaccine: 50 μg, 3 times, at 1 W of interval Tumor: 105 B16 cells or 2 × 105 3LL cells, 10 D after the last vaccination, SC |
ID | - CTL response to VEGFR2 - Inhibition of tumor-induced angiogenesis - Inhibition of tumor metastasis |
2015 [89] |
| Sarcoma | HHDII-DR1 mice | SSX2-optimized vaccine, encoding modified cancer testis antigen | Ab anti-PD-1/L1 | Prophylactic and therapeutic vaccination Vaccine: 100 μg, 6 times, every 2 W (prophylactic) or weekly the day after the tumor injection (therapeutic) Tumor: 2 × 104 SSX2-expressing sarcoma cells An anti-PD1/L1: 100 μg, IP on the day following each vaccination |
ID | - Optimized vaccine elicited inferior antitumor effect relative to the native vaccine - Increase of PD-L1 expression on tumor cells - CTL from immunized mice expressed more PD-1, increasing the antitumor efficacy of the combination with ICB |
2015 [152] |
| Kidney cancer (RenCa cells) | BALB/c mice | pVAX1-G250-F2A-CTLA4, containing the co-expression gene G250-CTLA4, linked by Furin-2A (F2A) | / | Therapeutic vaccination Tumor: 105 RenCa cells, SC Vaccine: 50 μg, at D 7, 17 and 27 |
EP | - Humoral and cellular-specific immune response against CTLA4 and G250 - Increase in INFγ and IL-4 (Th1/2 response) - Tumor growth rate decreased |
2017 [153] |
Keywords search “cancer plasmid DNA vaccine”, from 2015 to 2018. D = day, W = week, M = month