Inhibition of host cell encapsulation through inhibiting immune gene expression by the parasitic wasp venom calreticulin.
Parasitoid wasps inject venom into the host to protect their offspring against host immune responses. In our previous study, we identified a calreticulin (CRT) in Pteromalus puparum venom. In this study, we expressed the wild-type and the coiled-coil domain deletion mutant P. puparum calreticulins (PpCRTs) in Escherichia coli and prepared polyclonal antibody in rabbit against PpCRT. Western blot analysis showed that PpCRT protein was not only present in the venom but also in all the tissues tested. Real time PCR results indicated that PpCRT mRNA was highly expressed in the venom gland. The transcript level of PpCRT in the venom gland was peaked at 2 days post-eclosion, while the PpCRT protein in the venom was maintained at a constant level. Both recombinant wild-type and mutant PpCRT proteins could bind to the surface of P. puparum eggs. Recombinant PpCRT inhibited hemocyte spreading and cellular encapsulation of the host Pieris rapae in vitro, and the coiled-coil domain is important for the inhibitory function of PpCRT. Immunocytochemistry results showed that PpCRT entered P. rapae hemocytes, and the coiled-coil domain played a role in this process. After injection of recombinant PpCRT into P. rapae pupae, real time PCR results showed that PpCRT inhibited transcript levels of host encapsulation-related genes, including calreticulin and scavenger receptor genes. In conclusion, our results suggest that P. puparum venom protects its offspring against host cellular immune responses via its functional component PpCRT to inhibit the expression of host cellular response-related genes.
State Key Laboratory of Rice Biology & Key Laboratory of Agricultural Entomology of Ministry of Agriculture, Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China; Laboratory of Sericulture, College of Life Science, Anhui Agricultural University, Hefei 230036, China.
Lei
L
Wang
Qi
Q
Fang
Cen
C
Qian
Fei
F
Wang
Xiao-Qiang
XQ
Yu
Gongyin
G
Ye
D000818:Animals; D002080:Butterflies / Q000276:immunology / Q000469:parasitology; D037282:Calreticulin / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation / Q000235:genetics* / Q000276:immunology; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology; D006434:Hemocytes / Q000276:immunology / Q000378:metabolism; D006790:Host-Parasite Interactions / Q000276:immunology; D010063:Ovum / Q000276:immunology / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D014862:Wasp Venoms / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014863:Wasps / Q000235:genetics / Q000378:metabolism*
D037282:Calreticulin; D011994:Recombinant Proteins; D014862:Wasp Venoms
Calreticulin; Encapsulation; Parasitoid; Venom