Limits...
Prophylactic Dendritic Cell-Based Vaccines Efficiently Inhibit Metastases in Murine Metastatic Melanoma.

Markov OV, Mironova NL, Sennikov SV, Vlassov VV, Zenkova MA - PLoS ONE (2015)

Bottom Line: Lipofectamine 2000 and liposomes consisting of helper lipid DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and cationic lipid 2D3 (1,26-Bis(1,2-de-O-tetradecyl-rac-glycerol)-7,11,16,20-tetraazahexacosan tetrahydrocloride) were used for RNA transfection.Antimetastatic effect of single prophylactic DC vaccination in metastatic melanoma model was accompanied by the reductions in the levels of Th2-specific cytokines however the change of the levels of Th1/Th2/Th17 master regulators was not found.In the case of therapeutic DC vaccine the polarization of Th1-response was found nevertheless the antimetastatic effect was less effective in comparison with prophylactic DC vaccine.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Sciences, Novosibirsk, Russia.

ABSTRACT
Recent data on the application of dendritic cells (DCs) as anti-tumor vaccines has shown their great potential in therapy and prophylaxis of cancer. Here we report on a comparison of two treatment schemes with DCs that display the models of prophylactic and therapeutic vaccination using three different experimental tumor models: namely, Krebs-2 adenocarcinoma (primary tumor), melanoma (B16, metastatic tumor without a primary node) and Lewis lung carcinoma (LLC, metastatic tumor with a primary node). Dendritic cells generated from bone marrow-derived DC precursors and loaded with lysate of tumor cells or transfected with the complexes of total tumor RNA with cationic liposomes were used for vaccination. Lipofectamine 2000 and liposomes consisting of helper lipid DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and cationic lipid 2D3 (1,26-Bis(1,2-de-O-tetradecyl-rac-glycerol)-7,11,16,20-tetraazahexacosan tetrahydrocloride) were used for RNA transfection. It was shown that DCs loaded with tumor lysate were ineffective in contrast to tumor-derived RNA. Therapeutic vaccination with DCs loaded by lipoplexes RNA/Lipofectamine 2000 was the most efficient for treatment of non-metastatic Krebs-2, where a 1.9-fold tumor growth retardation was observed. Single prophylactic vaccination with DCs loaded by lipoplexes RNA/2D3 was the most efficient to treat highly aggressive metastatic tumors LLC and B16, where 4.7- and 10-fold suppression of the number of lung metastases was observed, respectively. Antimetastatic effect of single prophylactic DC vaccination in metastatic melanoma model was accompanied by the reductions in the levels of Th2-specific cytokines however the change of the levels of Th1/Th2/Th17 master regulators was not found. Failure of double prophylactic vaccination is explained by Th17-response polarization associated with autoimmune and pro-inflammatory reactions. In the case of therapeutic DC vaccine the polarization of Th1-response was found nevertheless the antimetastatic effect was less effective in comparison with prophylactic DC vaccine.

No MeSH data available.


Related in: MedlinePlus

The expression level of Tbet, GATA3, RORg, and Foxp3 in spleen cells of mice after DC vaccination (qPCR data).A. Prophylactic treatment scheme: healthy mice receiving DC vaccines 1-1/2D3/RNA and 1-2/2D3/RNA (1 and 2 immunization, respectively). B. Mice with metastatic melanoma receiving saline buffer. C. Therapeutic treatment scheme: mice with metastatic melanoma receiving DC vaccines 2-1/2D3/RNA and 2-2/2D3/RNA (1 and 2 immunization, respectively). Crossed square on Y axis displays the level of gene expression in healthy intact mice (baseline). Type of DC vaccine is indicated as S-I/T/A where S—scheme of the treatment 1 or 2, I—immunization number, T—transfectant, A—antigen source. Expression of the genes in group without treatment was measured on days 9 and 16 of tumor development. Expression of the genes in prophylactic and therapeutic groups was measured on day 5 after each immunization. Data represent the mean ± SD of three experiments performed in triplicate. Data were statistically analysed using one-way ANOVA with post hoc Fisher test.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4556596&req=5

pone.0136911.g006: The expression level of Tbet, GATA3, RORg, and Foxp3 in spleen cells of mice after DC vaccination (qPCR data).A. Prophylactic treatment scheme: healthy mice receiving DC vaccines 1-1/2D3/RNA and 1-2/2D3/RNA (1 and 2 immunization, respectively). B. Mice with metastatic melanoma receiving saline buffer. C. Therapeutic treatment scheme: mice with metastatic melanoma receiving DC vaccines 2-1/2D3/RNA and 2-2/2D3/RNA (1 and 2 immunization, respectively). Crossed square on Y axis displays the level of gene expression in healthy intact mice (baseline). Type of DC vaccine is indicated as S-I/T/A where S—scheme of the treatment 1 or 2, I—immunization number, T—transfectant, A—antigen source. Expression of the genes in group without treatment was measured on days 9 and 16 of tumor development. Expression of the genes in prophylactic and therapeutic groups was measured on day 5 after each immunization. Data represent the mean ± SD of three experiments performed in triplicate. Data were statistically analysed using one-way ANOVA with post hoc Fisher test.

Mentions: Since analysis the level of Th1- and Th2-specific cytokines in blood plasma of mice with metastatic melanoma after DC vaccination did not let to determine exactly the type of Th response we studied the expression of master transcription factors Tbet (Th1), GATA3 (Th2), RORg (Th17) and Foxp3 (Treg) by real time RT-PCR in spleen cells of mice received DC vaccines (Fig 6). It was shown that both prophylactic vaccines have no effect on the expression level of GATA3 (Fig 6A): the levels of this gene did not differ from baseline (healthy animals, point on Y axis, Fig 6A–6C) and w/t group (non-treated mice with metastatic melanoma). With the increase of immunization number we observed disappearance of Tbet (p = 0.0018) and 2-fold increase of RORg (p = 0.04). Also three-fold decrease of the expression level of Foxp3 in comparison with baseline was observed but it was statistically insignificant (Fig 6A).


Prophylactic Dendritic Cell-Based Vaccines Efficiently Inhibit Metastases in Murine Metastatic Melanoma.

Markov OV, Mironova NL, Sennikov SV, Vlassov VV, Zenkova MA - PLoS ONE (2015)

The expression level of Tbet, GATA3, RORg, and Foxp3 in spleen cells of mice after DC vaccination (qPCR data).A. Prophylactic treatment scheme: healthy mice receiving DC vaccines 1-1/2D3/RNA and 1-2/2D3/RNA (1 and 2 immunization, respectively). B. Mice with metastatic melanoma receiving saline buffer. C. Therapeutic treatment scheme: mice with metastatic melanoma receiving DC vaccines 2-1/2D3/RNA and 2-2/2D3/RNA (1 and 2 immunization, respectively). Crossed square on Y axis displays the level of gene expression in healthy intact mice (baseline). Type of DC vaccine is indicated as S-I/T/A where S—scheme of the treatment 1 or 2, I—immunization number, T—transfectant, A—antigen source. Expression of the genes in group without treatment was measured on days 9 and 16 of tumor development. Expression of the genes in prophylactic and therapeutic groups was measured on day 5 after each immunization. Data represent the mean ± SD of three experiments performed in triplicate. Data were statistically analysed using one-way ANOVA with post hoc Fisher test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556596&req=5

pone.0136911.g006: The expression level of Tbet, GATA3, RORg, and Foxp3 in spleen cells of mice after DC vaccination (qPCR data).A. Prophylactic treatment scheme: healthy mice receiving DC vaccines 1-1/2D3/RNA and 1-2/2D3/RNA (1 and 2 immunization, respectively). B. Mice with metastatic melanoma receiving saline buffer. C. Therapeutic treatment scheme: mice with metastatic melanoma receiving DC vaccines 2-1/2D3/RNA and 2-2/2D3/RNA (1 and 2 immunization, respectively). Crossed square on Y axis displays the level of gene expression in healthy intact mice (baseline). Type of DC vaccine is indicated as S-I/T/A where S—scheme of the treatment 1 or 2, I—immunization number, T—transfectant, A—antigen source. Expression of the genes in group without treatment was measured on days 9 and 16 of tumor development. Expression of the genes in prophylactic and therapeutic groups was measured on day 5 after each immunization. Data represent the mean ± SD of three experiments performed in triplicate. Data were statistically analysed using one-way ANOVA with post hoc Fisher test.
Mentions: Since analysis the level of Th1- and Th2-specific cytokines in blood plasma of mice with metastatic melanoma after DC vaccination did not let to determine exactly the type of Th response we studied the expression of master transcription factors Tbet (Th1), GATA3 (Th2), RORg (Th17) and Foxp3 (Treg) by real time RT-PCR in spleen cells of mice received DC vaccines (Fig 6). It was shown that both prophylactic vaccines have no effect on the expression level of GATA3 (Fig 6A): the levels of this gene did not differ from baseline (healthy animals, point on Y axis, Fig 6A–6C) and w/t group (non-treated mice with metastatic melanoma). With the increase of immunization number we observed disappearance of Tbet (p = 0.0018) and 2-fold increase of RORg (p = 0.04). Also three-fold decrease of the expression level of Foxp3 in comparison with baseline was observed but it was statistically insignificant (Fig 6A).

Bottom Line: Lipofectamine 2000 and liposomes consisting of helper lipid DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and cationic lipid 2D3 (1,26-Bis(1,2-de-O-tetradecyl-rac-glycerol)-7,11,16,20-tetraazahexacosan tetrahydrocloride) were used for RNA transfection.Antimetastatic effect of single prophylactic DC vaccination in metastatic melanoma model was accompanied by the reductions in the levels of Th2-specific cytokines however the change of the levels of Th1/Th2/Th17 master regulators was not found.In the case of therapeutic DC vaccine the polarization of Th1-response was found nevertheless the antimetastatic effect was less effective in comparison with prophylactic DC vaccine.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Sciences, Novosibirsk, Russia.

ABSTRACT
Recent data on the application of dendritic cells (DCs) as anti-tumor vaccines has shown their great potential in therapy and prophylaxis of cancer. Here we report on a comparison of two treatment schemes with DCs that display the models of prophylactic and therapeutic vaccination using three different experimental tumor models: namely, Krebs-2 adenocarcinoma (primary tumor), melanoma (B16, metastatic tumor without a primary node) and Lewis lung carcinoma (LLC, metastatic tumor with a primary node). Dendritic cells generated from bone marrow-derived DC precursors and loaded with lysate of tumor cells or transfected with the complexes of total tumor RNA with cationic liposomes were used for vaccination. Lipofectamine 2000 and liposomes consisting of helper lipid DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine) and cationic lipid 2D3 (1,26-Bis(1,2-de-O-tetradecyl-rac-glycerol)-7,11,16,20-tetraazahexacosan tetrahydrocloride) were used for RNA transfection. It was shown that DCs loaded with tumor lysate were ineffective in contrast to tumor-derived RNA. Therapeutic vaccination with DCs loaded by lipoplexes RNA/Lipofectamine 2000 was the most efficient for treatment of non-metastatic Krebs-2, where a 1.9-fold tumor growth retardation was observed. Single prophylactic vaccination with DCs loaded by lipoplexes RNA/2D3 was the most efficient to treat highly aggressive metastatic tumors LLC and B16, where 4.7- and 10-fold suppression of the number of lung metastases was observed, respectively. Antimetastatic effect of single prophylactic DC vaccination in metastatic melanoma model was accompanied by the reductions in the levels of Th2-specific cytokines however the change of the levels of Th1/Th2/Th17 master regulators was not found. Failure of double prophylactic vaccination is explained by Th17-response polarization associated with autoimmune and pro-inflammatory reactions. In the case of therapeutic DC vaccine the polarization of Th1-response was found nevertheless the antimetastatic effect was less effective in comparison with prophylactic DC vaccine.

No MeSH data available.


Related in: MedlinePlus