Simser Jason A, Macaluso Kevin R, Mulenga Albert, Azad Abdu F
Department of Microbiology and Immunology, University of Maryland School of Medicine, 655 West Baltimore Street, Baltimore, MD 21201, USA.
Insect Biochem Mol Biol. 2004 Dec;34(12):1235-46. doi: 10.1016/j.ibmb.2004.07.003.
Immune-responsive lysozyme encoding cDNAs were identified from two medically important tick species by an expressed sequence tag approach of D. variabilis hemocytes (Dv Lys) and a D. andersoni embryonic derived cell line, DAE100. Comparative sequence analyses indicated the Dermacentor molecules to be products of orthologous genes and to be most similar to arthropod c-type lysozymes. Northern blotting analyses demonstrated that Dv Lys expression levels were most abundant in tick hemocytes and to a much lesser degree in the midgut while barely detectable in ovary, salivary gland, and Malpighian tubule tissues. Involvement of the Dermacentor c-type lysozymes in innate immunity was demonstrated by Escherichia coli challenges of D. variabilis ticks by injection resulting in a temporal profile of significantly elevated transcript abundances above those of naive controls that was similarly observed of the D. andersoni cells co-cultured with E. coli. In contrast to that reported of the digestive gut lysozyme of the soft tick Ornithodoros moubata, Dv Lys levels were not statistically differentially regulated by blood meal digestion. Additionally, given the differences in tissue distribution, sequence characteristics and phylogenetic placements between the Dermacentor and Ornithodoros lysozymes demonstrates that ticks possess differently adapted c-type lysozymes that are spatially and temporally differentially expressed.
通过美洲钝缘蜱血细胞(Dv Lys)的表达序列标签方法以及安德逊革蜱胚胎衍生细胞系DAE100,从两种具有医学重要性的蜱虫物种中鉴定出了免疫反应性溶菌酶编码cDNA。比较序列分析表明,革蜱属分子是直系同源基因的产物,并且与节肢动物c型溶菌酶最为相似。Northern印迹分析表明,Dv Lys表达水平在蜱血细胞中最为丰富,在中肠中的表达水平要低得多,而在卵巢、唾液腺和马氏管组织中几乎检测不到。通过向美洲钝缘蜱注射大肠杆菌进行挑战,证明了革蜱属c型溶菌酶参与天然免疫,导致转录本丰度暂时显著高于未感染对照组,这与与大肠杆菌共培养的安德逊革蜱细胞中观察到的情况相似。与软蜱莫氏钝缘蜱消化肠道溶菌酶的报道相反,Dv Lys水平在血餐消化过程中没有统计学上的差异调节。此外,鉴于革蜱属和钝缘蜱属溶菌酶在组织分布、序列特征和系统发育位置上的差异,表明蜱虫拥有适应性不同的c型溶菌酶,它们在空间和时间上差异表达。