Matsumoto Kotaro, Morisaki Takashi, Kuroki Hideo, Kubo Makoto, Onishi Hideya, Nakamura Katsuya, Nakahara Chihiro, Kuga Hirotaka, Baba Eishi, Nakamura Masafumi, Hirata Kazuho, Tanaka Masao, Katano Mitsuo
Department of Cancer Therapy and Research, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Cell Immunol. 2004 Sep-Oct;231(1-2):20-9. doi: 10.1016/j.cellimm.2004.11.002. Epub 2005 Jan 11.
We investigated the effect of exosomes secreted from human monocyte-derived dendritic cells (Mo-DCs), which are generated from PBMCs in response to treatment with GM-CSF and IL-4, on naive CD4+ T cell survival in vitro. Exosomes isolated from culture supernatants of Mo-DCs (>90% purity) were purified with anti-HLA-DP, -DQ, -DR-coated paramagnetic beads. Purified exosomes prolonged the survival of naive CD4+ T cells (>98% purity) in vitro. Treatment with neutralizing mAb against HLA-DR significantly decreased the supportive effect of purified exosomes on CD4+ T cell survival. Exosomes increased nuclear translocation of NF-(kappa)B in naive CD4+ T cells, and NF-(kappa)B activation was significantly suppressed by anti-HLA-DR mAb or NF-(kappa)B inhibitor pyrrolidine dithiocarbamate (PDTC). In addition, PDTC inhibited the effect of exosomes on naive CD4+ T cell survival. Thus, exosomes secreted by Mo-DCs appear to support naive CD4+ T cell survival via NF-(kappa)B activation induced by interaction of HLA-DR and TCRs.
我们研究了人单核细胞来源的树突状细胞（Mo-DC）分泌的外泌体对体外初始CD4 + T细胞存活的影响，Mo-DC由PBMC在GM-CSF和IL-4处理下产生。从Mo-DC培养上清液中分离的外泌体（纯度> 90％）用抗HLA-DP、-DQ、-DR包被的顺磁珠纯化。纯化的外泌体在体外延长了初始CD4 + T细胞（纯度> 98％）的存活时间。用抗HLA-DR中和单克隆抗体处理显著降低了纯化外泌体对CD4 + T细胞存活的支持作用。外泌体增加了初始CD4 + T细胞中NF-κB的核转位，抗HLA-DR单克隆抗体或NF-κB抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）显著抑制了NF-κB的激活。此外，PDTC抑制了外泌体对初始CD4 + T细胞存活的影响。因此，Mo-DC分泌的外泌体似乎通过HLA-DR与TCR相互作用诱导的NF-κB激活来支持初始CD4 + T细胞的存活。
