内容紹介
Comparison of the Chemoimmunotherapeutic Effect of Doxorubicin and Bafilomycin-A1 in Mouse Neuroblastoma Cells
Summary
The aim of this study was to compare the ability of the drugs doxorubicin and Bafilomycin-A1(Baf-A1)to promote an immune reaction following the induction of cell death in a mouse neuroblastoma model. Neuro-2a cells were cultured in medium containing doxorubicin or Baf-A1. Bone marrow-derived dendritic cells(BM-DCs)were co-cultured with neuro-2A cells that were grown in doxorubicin- or Baf-A1-containing media, and phagocytosis of neuro-2a cells by the BN-DCs was evaluated. Additionally, dead neuro-2a cells were co-cultured with CD8α+ lymphocytes and BM-DCs, and the proliferation of CD8α+ cells was evaluated. Interferon-γ(IFN-γ)production was used as an index of the immune response. Dead neuro-2a cells treated with doxorubicin were phagocytosed effectively compared to the cells treated with Baf-A1. However, phagocytosis of cells treated with Baf-A1 was promoted after stimulation with CpG oligodeoxynucleotide(CpG-ODN). When CD8α+ cells were co-cultured with BM-DCs and doxorubicin-treated neuro-2a cells, CD8α+ lymphocyte proliferation was observed. There was no statistical difference in IFN-γ secretion between the doxorubicin-treated and Baf-A1-treated cells. However, after stimulation by CpG-ODN, IFN-γ production was more effectively observed in the Baf-A1-treated cells. Induction of cell death by doxorubicin or Baf-A1 could possibly enhance antitumor immunity in patients receiving chemotherapy for neuroblastoma. Selection of anti-tumor agents and stimulation of BM-DCs with a toll-like receptor(TLR)agonist is considered important in promoting antitumor activity after chemotherapy.
要旨
マウス神経芽腫モデルを用いてdoxorubicinとBafilomycin-A1(Baf-A1)が腫瘍細胞に細胞死を誘導する際の免疫学的効果を比較検討した。まず,マウス由来神経芽細胞腫細胞株neuro-2a培養系にdoxorubicin,Baf-A1を添加して細胞死を誘導した。neuro-2a死細胞を骨髄由来樹状細胞(BM-DC)と混合培養し,BM-DCの死細胞貪食効果を比較した。次に,死細胞をマウスCD8α+リンパ球およびBM-DCと混合培養し,CD8α+リンパ球増殖反応をIFN-γ産生を指標に評価した。さらに,培養系にTLR agonistのCpG-ODNを付加しadjuvant効果を検討した。結果は,doxorubicinで細胞死を誘導された腫瘍細胞はBaf-A1で処理した死細胞と比較してより効率的にBM-DCに貪食され,IFN-γ産生を介したCD8α+リンパ球増殖を誘導した。しかし,CpG-ODNのadjuvant効果はBaf-A1を用いた場合より効果的だった。神経芽腫に対する化学療法による免疫学的効果を念頭に入れたプロトコール作成には,抗腫瘍薬の選択とadjuvantとしてのTLR agonistとの適切な組み合わせの選択が重要である。
目次
Summary
The aim of this study was to compare the ability of the drugs doxorubicin and Bafilomycin-A1(Baf-A1)to promote an immune reaction following the induction of cell death in a mouse neuroblastoma model. Neuro-2a cells were cultured in medium containing doxorubicin or Baf-A1. Bone marrow-derived dendritic cells(BM-DCs)were co-cultured with neuro-2A cells that were grown in doxorubicin- or Baf-A1-containing media, and phagocytosis of neuro-2a cells by the BN-DCs was evaluated. Additionally, dead neuro-2a cells were co-cultured with CD8α+ lymphocytes and BM-DCs, and the proliferation of CD8α+ cells was evaluated. Interferon-γ(IFN-γ)production was used as an index of the immune response. Dead neuro-2a cells treated with doxorubicin were phagocytosed effectively compared to the cells treated with Baf-A1. However, phagocytosis of cells treated with Baf-A1 was promoted after stimulation with CpG oligodeoxynucleotide(CpG-ODN). When CD8α+ cells were co-cultured with BM-DCs and doxorubicin-treated neuro-2a cells, CD8α+ lymphocyte proliferation was observed. There was no statistical difference in IFN-γ secretion between the doxorubicin-treated and Baf-A1-treated cells. However, after stimulation by CpG-ODN, IFN-γ production was more effectively observed in the Baf-A1-treated cells. Induction of cell death by doxorubicin or Baf-A1 could possibly enhance antitumor immunity in patients receiving chemotherapy for neuroblastoma. Selection of anti-tumor agents and stimulation of BM-DCs with a toll-like receptor(TLR)agonist is considered important in promoting antitumor activity after chemotherapy.
要旨
マウス神経芽腫モデルを用いてdoxorubicinとBafilomycin-A1(Baf-A1)が腫瘍細胞に細胞死を誘導する際の免疫学的効果を比較検討した。まず,マウス由来神経芽細胞腫細胞株neuro-2a培養系にdoxorubicin,Baf-A1を添加して細胞死を誘導した。neuro-2a死細胞を骨髄由来樹状細胞(BM-DC)と混合培養し,BM-DCの死細胞貪食効果を比較した。次に,死細胞をマウスCD8α+リンパ球およびBM-DCと混合培養し,CD8α+リンパ球増殖反応をIFN-γ産生を指標に評価した。さらに,培養系にTLR agonistのCpG-ODNを付加しadjuvant効果を検討した。結果は,doxorubicinで細胞死を誘導された腫瘍細胞はBaf-A1で処理した死細胞と比較してより効率的にBM-DCに貪食され,IFN-γ産生を介したCD8α+リンパ球増殖を誘導した。しかし,CpG-ODNのadjuvant効果はBaf-A1を用いた場合より効果的だった。神経芽腫に対する化学療法による免疫学的効果を念頭に入れたプロトコール作成には,抗腫瘍薬の選択とadjuvantとしてのTLR agonistとの適切な組み合わせの選択が重要である。