Abstract
Nosema bombycis, the first identified microsporidium, is a destructive pathogen of the silkworm Bombyx mori and causes severe worldwide economic losses in sericulture. Major microsporidian structural proteins, such as the spore wall protein (SWP), are known to be involved in host invasion. In this study, the reactivity of the monoclonal antibody 2B10 was tested against an endospore protein of N. bombycis with a molecular weight size at 50-kDa, using Western blotting. The antigen was purified after immunoprecipitation and was further identified as EOB13320 according to MALDI-TOF MS assay. We found that EOB13320 locates to the surface of the different developmental stages of the parasite, mostly the sporoblast stage and the mature spore after immunoelectron microscopy examination. EOB13320 was also widely distributed in the developing endospore, especially at the sporoblast stage. This endospore protein also accumulated in the cytoplasm of both the merogony and sporoblast stages. These results imply that EOB13320 detected by monoclonal antibody 2B10 is expressed throughout the life cycle of the parasite, notably during the stage when the endospore is formed, and that this protein is important for spore-coat formation and parasite maintenance. Our study could be instrumental in the understanding of spore wall formation and will help to gain greater insight into the biology of this parasite.
Highlights
Microsporidia are obligate intracellular spore-forming parasites that are widely acknowledged as highly reduced and derived and are most closely aligned with fungi [1,2,3,4,5]
A Monoclonal Antibody That Tracks Endospore Formation stages of N. bombycis are in direct contact with the host cell cytoplasm [10], and the internalized parasite cycle generally includes a meront cell division phase, a spore production phase, and a mature spore or infective phase [11,12]
We developed two monoclonal antibodies that recognize spore wall proteins of N. bombycis
Summary
Microsporidia are obligate intracellular spore-forming parasites that are widely acknowledged as highly reduced and derived and are most closely aligned with fungi [1,2,3,4,5]. A Monoclonal Antibody That Tracks Endospore Formation stages of N. bombycis are in direct contact with the host cell cytoplasm [10], and the internalized parasite cycle generally includes a meront cell division phase (or merogony), a spore production phase (or sporogony), and a mature spore or infective phase [11,12]. The sporonts undergo continuous transition into sporoblasts, whose most distinguishing features are the formation of the early extrusion apparatus (polar tube) and the assembly of the final spore shape. These sporoblasts develop into mature spores, which have polarized organelles and possess thick walls
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