Growth of Yersinia enterocolitica and interaction with mammalian cells in three-dimensional (3D) extracellular matrix

小肠结肠炎耶尔森菌的致病性是由毒力质粒pYV和几个染色体决定因子控制的，这些决定因子为该病原体提供了抵抗吞噬和在宿主组织细胞外繁殖的武器。为了研究在类动物条件下感染过程的动力学，我们建立了3d -胶原凝胶（3DCoG）感染模型。这项建议包括两个主要目标：(i)在3D-CoG中对耶尔森菌的生活方式进行分子表征，包括细菌间的相互作用（如簇和胶囊的形成和扩散）以及与ECM的相互作用；（ii）分析耶尔森菌与吞噬细胞的相互作用，重点关注细胞迁移、粘附、吞噬、氧化破裂、蛋白酶的释放以及3型分泌系统（T3SS）介导的耶尔森菌抗宿主效应蛋白（Yops）易位导致的细胞死亡。这些目标将通过使用选定的耶尔森菌突变体和不同的t3ss注射报告蛋白以及对耶尔森菌接触宿主细胞的功能分析来实现。

The pathogenicity of Yersinia enterocolitica is controlled by the virulence plasmid pYV and several chromosomally located determinants which provide this pathogen with the armament to resist phagocytosis and to multiply extracellularly in host tissue. To study the dynamics of the infection process under in vivo-like conditions, we established the 3D-Collagen Gel (3DCoG) infection model. This proposal comprises two major aims: (i) molecular characterization of the life style of yersiniae in 3D-CoG including interbacterial interaction (e.g. cluster and capsule formation and dispersal) and interaction with ECM and (ii) analysis of the interaction of yersiniae with phagocytic cells by focusing on cell migration, adherence, phagocytosis, oxidative burst, release of proteases and cell death due to type 3-secretion system (T3SS) mediated translocation of Yersinia anti-host effector proteins (Yops). These goals will be accomplished by using selected yersinae mutants and diverse T3SS-injected reporter proteins and by functional analysis of Yersinia-contacted host cells.