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Cellular Mechanism of Newly Synthesized Indoledione Derivative-induced Immunological Death of Tumor Cell.

Oh SJ, Ryu CK, Baek SY, Lee H - Immune Netw (2011)

Bottom Line: Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC.EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation.The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation).

View Article: PubMed Central - PubMed

Affiliation: Office of Biomedical Science, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul 135-710, Korea.

ABSTRACT

Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell.

Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC.

Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and 100 µM) with carleticulin induction. And EY-6 induced the secretion of IFN-γ but not of TNF-α from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation).

Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

No MeSH data available.


Related in: MedlinePlus

Effect of EY-6 on the DC for the OVA-specific cross presentation to CD4+(MHC II) and CD8+(MHC I) T cells. Cultured BM-DC was treated with EY-6 25 µM for overnight, then introduced with OVA before start the co-culture with MHC-restricted and OVA-specific T cell hybridomas. Details of methods are described in the "Materials and Methods" section. (A) IL-2 (pg/ml) secretion from the MHCII-restricted DOBW cells were measured by ELISA. (B) Color change induced by lacZ activity in the MHC I-restricted B3Z cells was measured. Numbers in Y-axes represent the absorption at OD580 nM. None of the responses were statistically significant.
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Figure 6: Effect of EY-6 on the DC for the OVA-specific cross presentation to CD4+(MHC II) and CD8+(MHC I) T cells. Cultured BM-DC was treated with EY-6 25 µM for overnight, then introduced with OVA before start the co-culture with MHC-restricted and OVA-specific T cell hybridomas. Details of methods are described in the "Materials and Methods" section. (A) IL-2 (pg/ml) secretion from the MHCII-restricted DOBW cells were measured by ELISA. (B) Color change induced by lacZ activity in the MHC I-restricted B3Z cells was measured. Numbers in Y-axes represent the absorption at OD580 nM. None of the responses were statistically significant.

Mentions: Cultured-DC presented OVA antigen to the OVA-specific T cell hybridomas either by MHC I-restricted (B3Z, CD8+ cell) and MHC II-restricted (DOBW, CD4+ cell) manner. MHC II-restricted OVA-specific IL-2 secretion from the DOBW cell was not significantly changed by the co-culture with EY-6 treated and OVA-introduced DC (270.4, 277.5±35.3, and 295.6±5.5 pg/ml with non-treated DC control, EY-6 25 µM and 50 µM treated-DC, respectively) (Fig. 6A). Also MHC I restricted OVA-specific B3Z cell response by EY-6 treated DC was not significant, neither (Fig. 6B). As whole, EY-6 treatment did not alter the cross-presentation ability of DCs for both MHC I and MHC II-restricted antigens.


Cellular Mechanism of Newly Synthesized Indoledione Derivative-induced Immunological Death of Tumor Cell.

Oh SJ, Ryu CK, Baek SY, Lee H - Immune Netw (2011)

Effect of EY-6 on the DC for the OVA-specific cross presentation to CD4+(MHC II) and CD8+(MHC I) T cells. Cultured BM-DC was treated with EY-6 25 µM for overnight, then introduced with OVA before start the co-culture with MHC-restricted and OVA-specific T cell hybridomas. Details of methods are described in the "Materials and Methods" section. (A) IL-2 (pg/ml) secretion from the MHCII-restricted DOBW cells were measured by ELISA. (B) Color change induced by lacZ activity in the MHC I-restricted B3Z cells was measured. Numbers in Y-axes represent the absorption at OD580 nM. None of the responses were statistically significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3275708&req=5

Figure 6: Effect of EY-6 on the DC for the OVA-specific cross presentation to CD4+(MHC II) and CD8+(MHC I) T cells. Cultured BM-DC was treated with EY-6 25 µM for overnight, then introduced with OVA before start the co-culture with MHC-restricted and OVA-specific T cell hybridomas. Details of methods are described in the "Materials and Methods" section. (A) IL-2 (pg/ml) secretion from the MHCII-restricted DOBW cells were measured by ELISA. (B) Color change induced by lacZ activity in the MHC I-restricted B3Z cells was measured. Numbers in Y-axes represent the absorption at OD580 nM. None of the responses were statistically significant.
Mentions: Cultured-DC presented OVA antigen to the OVA-specific T cell hybridomas either by MHC I-restricted (B3Z, CD8+ cell) and MHC II-restricted (DOBW, CD4+ cell) manner. MHC II-restricted OVA-specific IL-2 secretion from the DOBW cell was not significantly changed by the co-culture with EY-6 treated and OVA-introduced DC (270.4, 277.5±35.3, and 295.6±5.5 pg/ml with non-treated DC control, EY-6 25 µM and 50 µM treated-DC, respectively) (Fig. 6A). Also MHC I restricted OVA-specific B3Z cell response by EY-6 treated DC was not significant, neither (Fig. 6B). As whole, EY-6 treatment did not alter the cross-presentation ability of DCs for both MHC I and MHC II-restricted antigens.

Bottom Line: Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC.EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation.The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation).

View Article: PubMed Central - PubMed

Affiliation: Office of Biomedical Science, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul 135-710, Korea.

ABSTRACT

Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell.

Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC.

Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and 100 µM) with carleticulin induction. And EY-6 induced the secretion of IFN-γ but not of TNF-α from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation).

Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

No MeSH data available.


Related in: MedlinePlus