title: Activation of autophagic programmed cell death and innate immune gene expression reveals immuno-competence of integumental epithelium in Bombyx mori infected by a dipteran parasitoid.
abstract: In insects, the integument forms the primary barrier between the environment and internal milieu, but cellular and immune responses of the integumental epithelium to infection by micro- and macro-parasites are mostly unknown. We elucidated cellular and immune responses of the epithelium induced through infection by a dipteran endoparasitoid, Exorista bombycis in the economically important silkworm Bombyx mori. Degradative autophagic vacuoles, lamella-like bodies, a network of cytoplasmic channels with cellular cargo, and an RER network that opened to vacuoles were observed sequentially with increase in age after infection. This temporal sequence culminated in apoptosis, accompanied by the upregulation of the caspase gene and fragmentation of DNA. The infection significantly enhanced the tyrosine level and phenol oxidase activity in the integument. Proteomic analysis revealed enhanced expression of innate immunity components of toll and melanization pathways, cytokines, signaling molecules, chaperones, and proteolytic enzymes demonstrating diverse host responses. qPCR analysis revealed the upregulation of spatzle, BmToll, and NF kappa B transcription factors Dorsal and BmRel. NF kappa B inhibitor cactus showed diminished expression when Dorsal and BmRel were upregulated, revealing a negative correlation (R = (-)0.612). During melanization, prophenol oxidase 2 was expressed, a novel finding in integumental epithelium. The integument showed a low level of melanin metabolism and localized melanism in order to prevent the spreading of cytotoxic quinones. The gene-encoding proteolytic enzyme, beta-N-acetylglucosaminidase, was activated at 24 h post-infection, whereas chitinase, was activated at 96 h post-infection; however, most of the immune genes enhanced their expression in the early stages of infection. Thus the integument contributes to humoral immune responses that enhance resistance against macroparasite invasion.
authors.affiliation: Proteomics Division, Seribiotech Research Laboratory, CSB-Kodathi Campus, Bangalore, Karnataka, India. arpradeepnair@gmail.com
authors.forename: Appukuttan Nair R
authors.initials: AN
authors.lastname: Pradeep
authors.forename: Jayaram
authors.initials: J
authors.lastname: Anitha
authors.forename: Arvind K
authors.initials: AK
authors.lastname: Awasthi
authors.forename: Mohd A
authors.initials: MA
authors.lastname: Babu
authors.forename: Murthy N
authors.initials: MN
authors.lastname: Geetha
authors.forename: Hariharan K
authors.initials: HK
authors.lastname: Arun
authors.forename: Sagar
authors.initials: S
authors.lastname: Chandrashekhar
authors.forename: Guruprasad C
authors.initials: GC
authors.lastname: Rao
authors.forename: Nanjappa B
authors.initials: NB
authors.lastname: Vijayaprakash
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D012831:Bombyx / Q000235:genetics / Q000276:immunology* / Q000378:metabolism / Q000469:parasitology*; D016923:Cell Death / Q000502:physiology; D004175:Diptera / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D006790:Host-Parasite Interactions; D008297:Male; D015398:Signal Transduction