Montgomery Benjamin C S, Mewes Jacqueline, Davidson Chelsea, Burshtyn Deborah N, Stafford James L
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada.
Dev Comp Immunol. 2009 Apr;33(4):570-82. doi: 10.1016/j.dci.2008.10.006. Epub 2008 Nov 12.
Channel catfish leukocyte immune-type receptors (IpLITRs) are immunoglobulin superfamily (IgSF) members believed to play a role in the control and coordination of cellular immune responses in teleost. Putative stimulatory and inhibitory IpLITRs are co-expressed by different types of catfish immune cells (e.g. NK cells, T cells, B cells, and macrophages) but their signaling potential has not been determined. Following cationic polymer-mediated transfections into human cell lines we examined the surface expression, tyrosine phosphorylation, and phosphatase recruitment potential of two types of putative inhibitory IpLITRs using 'chimeric' expression constructs and an epitope-tagged 'native' IpLITR. We also cloned and expressed the teleost Src homology 2 domain-containing protein tyrosine phosphatases (SHP)-1 and SHP-2 and examined their expression in adult tissues and developing zebrafish embryos. Co-immunoprecipitation experiments support the inhibitory signaling potential of distinct IpLITR-types that bound both SHP-1 and SHP-2 following the phosphorylation of tyrosine residues within their cytoplasmic tail (CYT) regions. Phosphatase recruitment by IpLITRs represents an important first step in understanding their influence on immune cell effector functions and suggests that certain inhibitory signaling pathways are conserved among vertebrates.
斑点叉尾鮰白细胞免疫型受体(IpLITRs)是免疫球蛋白超家族(IgSF)成员,被认为在硬骨鱼的细胞免疫反应的控制和协调中发挥作用。推定的刺激性和抑制性IpLITRs由不同类型的鮰免疫细胞(如自然杀伤细胞、T细胞、B细胞和巨噬细胞)共同表达,但其信号传导潜力尚未确定。在通过阳离子聚合物介导转染到人类细胞系后,我们使用“嵌合”表达构建体和表位标记的“天然”IpLITR,检测了两种推定的抑制性IpLITRs的表面表达、酪氨酸磷酸化和磷酸酶募集潜力。我们还克隆并表达了硬骨鱼含Src同源2结构域的蛋白酪氨酸磷酸酶(SHP)-1和SHP-2,并检测了它们在成年组织和发育中的斑马鱼胚胎中的表达。免疫共沉淀实验支持了不同类型IpLITRs的抑制性信号传导潜力,这些IpLITRs在其细胞质尾部(CYT)区域的酪氨酸残基磷酸化后与SHP-1和SHP-2结合。IpLITRs对磷酸酶的募集是理解它们对免疫细胞效应功能影响的重要第一步,并表明某些抑制性信号通路在脊椎动物中是保守的。