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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

Activation status of bone-marrow-derived murine dendritic cells after transfection with lipoplexes LF/RNA and 2D3/RNA.A. Dead cells were excluded from DC population according to size and structure of cells. B. Surface expression of CD80, CD83, CD86 and MHC II by DCs after transfection with lipoplexes LF/RNA and 2D3/RNA. DCs without any stimulation (w/s) and treated with naked RNA isolated from B16 cells served as negative controls. DCs treated with LPS served as a positive control. DCs were stained with anti-CD80 (phycoerythrin), anti-CD83 (phycoerythrin), anti-CD86 (fluorescein isothiocyanote) and anti-MHC II (phycoerythrin), and were analyzed by flow cytometry. Ten thousand cells were counted for each sample. Data are presented as percent of fluorescent cells and mean fluorescence intensity of whole population of DCs normalized to w/s group (in arbitrary units, a.u.).
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pone.0136911.g001: Activation status of bone-marrow-derived murine dendritic cells after transfection with lipoplexes LF/RNA and 2D3/RNA.A. Dead cells were excluded from DC population according to size and structure of cells. B. Surface expression of CD80, CD83, CD86 and MHC II by DCs after transfection with lipoplexes LF/RNA and 2D3/RNA. DCs without any stimulation (w/s) and treated with naked RNA isolated from B16 cells served as negative controls. DCs treated with LPS served as a positive control. DCs were stained with anti-CD80 (phycoerythrin), anti-CD83 (phycoerythrin), anti-CD86 (fluorescein isothiocyanote) and anti-MHC II (phycoerythrin), and were analyzed by flow cytometry. Ten thousand cells were counted for each sample. Data are presented as percent of fluorescent cells and mean fluorescence intensity of whole population of DCs normalized to w/s group (in arbitrary units, a.u.).

Mentions: DC phenotype after loading was studied using mAb against maturation markers CD80, CD86, CD83 and MHC II (Fig 1). Non-loaded DCs (Fig 1B, w/s panel) expressed low amounts of maturation markers and their phenotype was close to immature or slightly mature (probably due to the influence of FBS in the culture medium). In the case of DCs treated with LPS an increase of expression level of three maturation markers CD80, CD83 and MHC II was observed (Fig 1B, compare w/s and LPS panels) both in terms of fluorescent cell number and mean fluorescence intensity (MFI). It is seen that DCs stimulated by lipoplexes of LF or 2D3-DOPE with RNA-B16 expressed some maturation markers and most probably corresponded to matured DCs. Stimulation of DCs with LF/RNA caused six-fold increase of CD80+ cells together with two-fold increase of MFI, two-fold increase of CD86+ cells and slight increase of MHC II+ cells as well as MFI in comparison with non-stimulated DCs (Fig 1B, compare w/s and LF/RNA panels). Stimulation of DCs with 2D3/RNA resulted in five-fold increase of CD80+ cells together with 1.6-fold increase of MFI and three-fold increase of CD83+ cells accompanied by slight increase of MFI in comparison with non-stimulated DCs (Fig 1B, compare w/s and 2D3/RNA panels).


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)

Activation status of bone-marrow-derived murine dendritic cells after transfection with lipoplexes LF/RNA and 2D3/RNA.A. Dead cells were excluded from DC population according to size and structure of cells. B. Surface expression of CD80, CD83, CD86 and MHC II by DCs after transfection with lipoplexes LF/RNA and 2D3/RNA. DCs without any stimulation (w/s) and treated with naked RNA isolated from B16 cells served as negative controls. DCs treated with LPS served as a positive control. DCs were stained with anti-CD80 (phycoerythrin), anti-CD83 (phycoerythrin), anti-CD86 (fluorescein isothiocyanote) and anti-MHC II (phycoerythrin), and were analyzed by flow cytometry. Ten thousand cells were counted for each sample. Data are presented as percent of fluorescent cells and mean fluorescence intensity of whole population of DCs normalized to w/s group (in arbitrary units, a.u.).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0136911.g001: Activation status of bone-marrow-derived murine dendritic cells after transfection with lipoplexes LF/RNA and 2D3/RNA.A. Dead cells were excluded from DC population according to size and structure of cells. B. Surface expression of CD80, CD83, CD86 and MHC II by DCs after transfection with lipoplexes LF/RNA and 2D3/RNA. DCs without any stimulation (w/s) and treated with naked RNA isolated from B16 cells served as negative controls. DCs treated with LPS served as a positive control. DCs were stained with anti-CD80 (phycoerythrin), anti-CD83 (phycoerythrin), anti-CD86 (fluorescein isothiocyanote) and anti-MHC II (phycoerythrin), and were analyzed by flow cytometry. Ten thousand cells were counted for each sample. Data are presented as percent of fluorescent cells and mean fluorescence intensity of whole population of DCs normalized to w/s group (in arbitrary units, a.u.).
Mentions: DC phenotype after loading was studied using mAb against maturation markers CD80, CD86, CD83 and MHC II (Fig 1). Non-loaded DCs (Fig 1B, w/s panel) expressed low amounts of maturation markers and their phenotype was close to immature or slightly mature (probably due to the influence of FBS in the culture medium). In the case of DCs treated with LPS an increase of expression level of three maturation markers CD80, CD83 and MHC II was observed (Fig 1B, compare w/s and LPS panels) both in terms of fluorescent cell number and mean fluorescence intensity (MFI). It is seen that DCs stimulated by lipoplexes of LF or 2D3-DOPE with RNA-B16 expressed some maturation markers and most probably corresponded to matured DCs. Stimulation of DCs with LF/RNA caused six-fold increase of CD80+ cells together with two-fold increase of MFI, two-fold increase of CD86+ cells and slight increase of MHC II+ cells as well as MFI in comparison with non-stimulated DCs (Fig 1B, compare w/s and LF/RNA panels). Stimulation of DCs with 2D3/RNA resulted in five-fold increase of CD80+ cells together with 1.6-fold increase of MFI and three-fold increase of CD83+ cells accompanied by slight increase of MFI in comparison with non-stimulated DCs (Fig 1B, compare w/s and 2D3/RNA panels).

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