Molecular mechanisms To protect malaria infection by vaccine using antigen / hsp70 fusion protein

分子机制 利用抗原/hsp70融合蛋白疫苗保护疟疾感染

• 批准号: 13670248
• 负责人: HONMA Kiri
• 金额: $ 2.3万
• 依托单位: Nagasaki University Graduate School of Medical Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670248/
• 关键词: heat shock protein; cross-presentation; dendritic cells; マラリアワクチン

APCs can internalize some types of exogenous antigens for processing and presentation on MHC class I molecules, which is referred to as cross-presentation. It is known that peptides associated with hsp70 can be presented in this manner to induce specific CTLs. To determine the cross-presentation pathway of hsp70 associated peptides, we generated hsc70-OVA_<257-264> (hsc70-OVA) fusion protein, which could induce specific CTL in vivo. CD8^+ T cells (OT-1 cells) were isolated from K_b-restricted OVA_<257-264> specific TCR transgenic mice (OT-1). RMA, RMA-S (TAP-) or bone-marrow derived DC, which were generated from C57BL/6 or TAP KO mice, were used as ARC. To determine whether APC can cross-present hsc70-OVA, we measured IFN-γ production from OT-1 cells in response to hsc70-OVA-pulsed ARC, and the expression of K_b-OVA_<257-264> complex on APC using 25D1.16 mAb. Our results showed that, 1) OT-1 cells produced IFN-γ in response to hsc70-OVA-pulsed TAP-negative 8M-DC, 2) High levels of K_b-OVA_<257-264> complex were detected on hsc70-OVA-pulsed RMA. The levels of K_b-OVA_<257-264> complexes detected on hsc70-OVA-pulsed RMA-S were similar to RMA. 3) The expression level of K_b-OVA_<257-264> complexes on RMA was only partially Inhibited by brefeldin A. 4) The expression of K_b-OVA_<257-264> complexes on RMA was completely inhibited by pretreatment of pCMBS, which is the inhibitor of fluid phase endocytosis (macropinocytosis). However, IFN-γ production from OT-1 T cells were not inhibited when BM-DC were treated with pCMBS. 5) IFN-γ production from OT-1 T cells were reduced when BM-DC were pulsed with hsc70-OVA in the presence of hsc70. Our results suggest that there are two distinct pathways of endocytosis of hsc, macropinocytosis and receptor-mediated endocytosis, and that there existed TAP-independent cross-presentation pathway of hsc associated peptide.

APC可以将某些类型的外生元素内化，以表现为Olecuss I分子，以至于可以以这种方式呈现与Hsp70相关的肽以诱导特定的CTL来确定HSP70相关肽的交叉态度。 -ova_ <257-264>（HSC70-OVA）在Vivo中。从C57BL/6或Tap O Mice产生的RMA-S（Tap-）或骨row衍生的DC被用作APC跨至跨量的HSC70-ova k_b-ova_ <257-264 > APS 25D1的复合物在HSC70-OVA脉冲的8M-DC中产生IFN-1细胞，在HSC70-OVA型RMA上检测到高水平的-264>复合物K_B-OVA_ <257-264>在HSC70-OVA脉冲RMA-S上检测到的复合物与RMA上的K_b-overa_ <257-264>复合物相似。 RMA上的K_b-ova_ <257-264>由PCMB完全抑制，这是液相内吞作用的抑制剂（巨型细胞增多症）。在HSC70的情况下，PCMBS。