Identification and immuno-electron microscopy localization of p40, a protein component of immunosuppressive virus-like particles from Leptopilina heterotoma, a virulent parasitoid wasp of Drosophila.
Lamellocytes are specialized larval blood cells of Drosophila that carry out encapsulation of metazoan pathogens such as parasitoid wasps. Large virus-like particles (VLPs) from two closely related virulent parasitoid wasp species, Leptopilina heterotoma and Leptopilina victoriae, suppress the host encapsulation response by promoting lysis of lamellocytes. The molecular basis of VLP-lamellocyte interaction and lamellocyte lysis is not understood. Here, it was shown that mature VLPs are composed of at least four major proteins. Polyclonal antisera against the most abundant L. heterotoma VLP protein, p40, cross-reacted with the most abundant L. victoriae VLP protein, p47.5. Immuno-electron microscopy (EM) of the long gland-reservoir complex revealed that p40 was expressed early in VLP biogenesis and was detected along with VLP precursors within the long gland cells and lumen. In the reservoir, VLPs had an angular core, resembled mature particles and p40 was detected outside the VLP cores. Immuno-EM staining of mature VLPs from both species localized the p40 and p47.5 proteins largely to the periphery of the VLPs and along the VLP spike-like projections. p40 staining was observed in VLP-treated host haemocytes. In vitro, anti-p40 antibody almost completely blocked the ability of L. heterotoma VLPs to promote lamellocyte lysis. Anti-p40 antibody blocked lysis by L. victoriae VLPs by >50%. It is proposed that the VLP surface proteins p40 and p47.5 share antigenic determinants and significantly contribute to the strong virulence of their Hymenopteran hosts.
Chiu
Hsiling
H
Department of Biology and the Graduate Center, City College of The City University of New York, 138th Street and Convent Avenue, New York, NY 10031, USA.
Morales
Jorge
J
Department of Biology and the Graduate Center, City College of The City University of New York, 138th Street and Convent Avenue, New York, NY 10031, USA.
Govind
Shubha
S
Department of Biology and the Graduate Center, City College of The City University of New York, 138th Street and Convent Avenue, New York, NY 10031, USA.
D000818:Animals; D000906:Antibodies / Q000032:analysis; D004330:Drosophila / Q000276:immunology / Q000469:parasitology*; D006927:Hymenoptera / Q000502:physiology*; D019476:Insect Proteins / Q000302:isolation & purification* / Q000648:ultrastructure; D007814:Larva / Q000276:immunology / Q000469:parasitology; D016253:Microscopy, Immunoelectron; D014863:Wasps / Q000737:chemistry* / Q000276:immunology / Q000472:pathogenicity / Q000648:ultrastructure
D000906:Antibodies; D019476:Insect Proteins