Willott E, Lowenberger C, Christensen B M, Kanost M R
Department of Biochemistry, Kansas State University, Manhattan 66506, USA.
Dev Comp Immunol. 1995 Nov-Dec;19(6):451-61. doi: 10.1016/0145-305x(95)00034-q.
The mechanisms by which hemocytes mediate a mosquito's defense response to parasites or pathogens are not well understood. This is due in part to difficulty in collecting intact mosquito hemocytes for experiments and to a lack of reagents, such as antibodies. Our objectives were to collect adult Aedes aegypti hemocytes under conditions suitable for immunofluorescence microscopy, and to test whether monoclonal antibodies, generated against larval Manduca sexta hemocytes, bind adult Ae. aegypti hemocytes. We present immunofluorescence micrographs of M. sexta and Ae. aegypti hemocytes stained by six monoclonal antibodies. Two antibodies, MS11 and MS32, immunolocalized to hemocyte nuclei in both species. On Western blots, these antibodies generate one signal at approximately 40 kDa and four others between 10 and 25 kDa. Immunofluorescence staining patterns of the other four antibodies were more complex. That these antibodies bind hemocytes from both species suggests significant molecular similarities exist between hemocytes from evolutionarily divergent species.
血细胞介导蚊子对寄生虫或病原体防御反应的机制尚未完全明确。部分原因在于难以收集完整的蚊子血细胞用于实验,以及缺乏诸如抗体等试剂。我们的目标是在适合免疫荧光显微镜观察的条件下收集成年埃及伊蚊的血细胞,并测试针对幼虫烟草天蛾血细胞产生的单克隆抗体是否能与成年埃及伊蚊的血细胞结合。我们展示了用六种单克隆抗体染色的烟草天蛾和埃及伊蚊血细胞的免疫荧光显微照片。两种抗体,MS11和MS32,在两个物种的血细胞细胞核中均有免疫定位。在蛋白质免疫印迹中,这些抗体在约40 kDa处产生一个信号,在10至25 kDa之间产生另外四个信号。其他四种抗体的免疫荧光染色模式更为复杂。这些抗体能与两个物种的血细胞结合,表明在进化上不同的物种的血细胞之间存在显著的分子相似性。