Overexpression of PGE2 synthase by in vivo transient expression enhances immunocompetency along with fitness cost in a lepidopteran insect

Abstract

Prostaglandins (PGs) mediate various physiological functions in insects. Especially, PGE2 is known to mediate immunity and egg-laying behavior in the beet armyworm, Spodoptera exigua. A PGE2 synthase 2 (Se-PGES2) has been identified to catalyze the final step to produce PGE2 in S. exigua. Its expression is inducible in response to immune challenge. Inhibition of the gene expression results in immunosuppression. On the other hand, any physiological alteration induced by its uncontrolled overexpression was not recognized in insects. This study used in vivo transient expression (IVTE) technique to induce overexpression and assessed subsequent physiological alteration in S. exigua. Se-PGES2 was cloned into a eukaryotic expression vector and transfected to Sf9 cells to monitor its heterologous expression. The Sf9 cells expressed the recombinant Se-PGES2 (rSe-PGES2) at an expected size (∼47 kDa), which was localized in cytoplasm. The recombinant expression vector was then used to transfect larvae of S. exigua. Hemocytes collected from the larvae treated with IVTE expressed rSe-PGES2 gene for at least 48 h. The larvae treated with IVTE exhibited an enhanced competency in cellular immune response measured by hemocyte nodule formation. In addition, IVTE treatment of Se-PGES2 induced gene expression of antimicrobial peptides without any immune challenge. The larvae treated with IVTE became significantly resistant to infection of an entomopathogenic nematode, Steinernema monticolum or to infection to its symbiotic bacterium, Xenorhabdus hominickii. However, IVTE-treated S. exigua larvae suffered from reduced pupal size and fecundity.