Hemolin is a member of the immunoglobulin (Ig) superfamily and contains four Ig domainsthat are similar to neural cell adhesion molecules. It has been regarded as a recognition molecule at immune challenge in insects. This study showed that hemolin of Plutella xylostella was expressed during pupal and adult stages but absent in all larval instars without any immune challenge. It is, however, strongly induced by the injection of Escherichia coli or its lipopolysaccharide in hemocytes, fat body and gut. A double-stranded RNA (dsRNA) interference experiment revealedits role in activation of prophenoloxidase (PPO) in the hemolymph during bacterial infection. Also its involvement in cellular defense was investigated in its mediation of the adherence of hemocytes to rat blood erythrocytes which was knocked down by its dsRNA. Finally, its physiological significance in pupal stage was confirmed by using dsRNA, which significantly prevented adult development. Therefore, it is concluded that hemolin plays roles in both immune and adult development in P. xylostella.

A cysteine- rich protein encoded by Cotesia plutellae bacovirus (CpBV) was identified in the parasitized Plutella xylostella. The gene, called CpBV-CRP, encodes 189 amino acids with a signal peptide of 20 residues at N-terminus determined by bioinformatic analysis, suggesting a secretory protein. High CpBV-CRP expression in the parasitized P. xylostella was observed at early days after parasitization and decreased with the course of parasitization. Expression of CpBV-CRP was tissue-specific in the fat body/epidermis, but not in hemocyte and gut. Its physiological function was analyzed by transient expression of a CpBV segment containing CpBV-CRP. The treated larvae underwent an immunosuppression in terms of hemocyte-spreading behavior. When the treated larvae were also co-injected with dsRNA against CpBV-CRP, the suppressed hemocyte behavior was significantly recovered. This study reports a cysteine-rich protein encoded in CpBV genome and its physiological function to be an immunosuppressant.

DNA in the eukaryotic nucleus is packaged into highly organized chromatin. The basic structural unit of chromatin is the nucleosome, which consists of approximately 146 base pairs of DNA wrapped around a histone octamer core containing two molecules each of core histones H2A, H2B, H3, and H4. Histone covalent modification at the protruding N-terminal region from the nucleosomal core can change the chromatin conformation in order to regulate gene expression. A viral H4 was found in the genome of Cotesia plutellae bracovirus (CpBV). The obligate host of the virus is an endoparasitoid wasp, C. plutellae, which parasitizes the diamondback moth, Plutella xylostella, and interrupts host development and immune reactions. CpBV-H4 has been regarded as an immunosuppressive gene. Its extended N-terminal region contains nine lysine residues which are the target for modification. Previous report showed that CpBV-H4 inhibited hemocyte-spreading after transient expression. Here, transient expression of truncated CpBV-H4 (without N-terminal region) did not show high inhibitory effects on hemocyte-spreading. Moreover, the truncated CpBV-H4 induced acetylation of nucleus histone H4. Host H4 was found to be decreased in transcription after parasitization compared to nonparasitized larvae. Atransient expression of CpBV-H4 significantly inhibited host H4 transcription, suggesting a role of CpBV-H4 in controlling gene expression. Point mutagenesis study showed that two lysines (K6 and K16) of CpBV-H4 were found to have high inhibitory effects on hemocyte spreading. These results indicate the importance of CpBV-H4 and its N-terminal region to control gene expression and suppress host immunity.

Monoamines have been known to mediate cellular immune responses such as hemocyte nodulation and phagocytosis in some insects. Eicosanoids also mediate the cellular immune reactions responsible for clearing bacterial infection from hemolymph circulation in various insects. In this study, we investigated a functional relationship of both mediators in expressing hemocyte nodulation and phagocytosis of fifth instar larvae of the beet armyworm, Spodoptera exigua. Addition of octopamine or serotonin along with bacteria enhanced hemocyte nodulation and phagocytosis of S. exigua larvae. Phentolamine (octopamine inhibitor) and ketanserin (serotonin inhibitor) suppressed hemocyte nodulation and phagocytosis of S. exigua in response to a bacterial challenge. Each inhibitory effect of both monoamine inhibitors was recovered by addition of either octopamine or serotonin, suggesting their independent signaling pathways. Both inhibitor effects were also recovered by addition of arachidonic acid (a precursor of eicosanoid biosynthesis). Furthermore, the stimulatory effects of monoamines on the cellular immune responses were prevented by addition of dexamethasone (a specific phospholipase A2). However, the inhibitory effect of dexamethasone was not recovered by excess amounts of monoamines. These results suggest that monoamines mediate hemocyte nodulation and phagocytosis through a downstream eicosanoid signal pathway in S. exigua.

The diamondback moth, Plutella xylostella, is reluctant to a baculovirus, Autographa california nucleopolyhedrosis virus (AcNPV) at its oral administration. However, parasitization by an endoparasitoid wasp, Cotesia plutellae, enhances the viral susceptibility. This study analyzed an antiviral activity of P. xylostella in response to the viral infection and determined the parasitic factor inhibiting the antiviral mechanism. For the analysis of antiviral activity of P. xylostella, a recombinant AcNPV expressing enhanced green fluorescence (AcNPV-EGFP) was orally adminstered to lavae of P. xylostella. After 24 h, EGFP expression was observed in the midgut tissue at a confocal-FITC mode. At the same time, a characteristic midgut melanotic response (MMR) was observed in some midgut regions under a phase contrast microscope. Thereafter, the EGFP signal was attenuated, while MMR spread on most midgut region. When the MMR was scored from 0 to 5 by the intensity of melanized cell density, it increased in time- and dose-dependent manners at the viral administration per os. These results suggest that the MMR is an antiviral activity of P. xylostella. This antiviral activity was significantly attenuated by C. plutellae parasitism. The parasitized P. xylostella showed significant decrease in the MMR score compared to nonparasitized larvae when they were orally administered with the same dose of AcNPV. To determine the parasitic factor(s) inhibiting the antiviral activity from the symbiotic polydnavirus of C. plutellae (C. plutellae bracovirus: CpBV), CpBV-IkB, which is a viral homolog of NFkB inhibitor and has been considered as an antiviral factor as in other polydnaviruses, was tested. A recombinant AcNPV expressing CpBV-IkB (AcNPV-IkB) was constructed and administered to P. xylostella larvae. As expected, AcNPV-IkB significantly decreased the antiviral activity measured by the MMR score compared to AcNPV-EGFP treatment. This study suggests that CpBV-IkB plays an antiviral parasitic role in the molecular interactions between P. xylostella and C. plutellae.

A comprehensive quality survey for heavy metals, organophosphorous and organochlorine residual pesticides, and coplanar PCBs in samples such as water, sediments and soils as well as spiders has been implemented. The samples were undertaken at nine stream sites and their vicinity in August 2006, representing different surrounding environments. The levels of PCBs were expressed as concentrations and WHO-TEFs. Among 12 coplanar PCBs as target compounds in this study, 2,3',4,4',5-PentaCB (IUPAC # 118) was the congener with the highest concentration. The total concentrations and TEF values of coplanar PCBs in Siheung stream sediment (heavy industrial complex site located in Ansan city) were 3915.50 pg/g and 0.8366 pg-TEQ/g on a dry weight basis, respectively. Such levels were around 40 times higher compared to sediment from Gapyung stream (green site located in the upper of Myunggi mountain). It is probably due to the direct input of PCBs from PCBs treatment materials. Organophosphorous (EPN, dementon-s-methyl, diazinon, parathion, and phenthoate) and organochlorine (alpha-BHC, aldrin, 4,4'-DDT, 4,4'-DDE, endosulfan alpha and etc.) pesticides were not detected above 5 ppb of detection limits. The concentrations of Cu and Cd in water and sediment samples from Siheung stream were 44.11 and 0.17 ug/㎖ and 713.42 ㎍/g and 3.73 ㎍/g, respectively, which contained 20～40 times higher concentrations than those from Gapyung stream. In addition to the water and sediment samples, the levels of heavy metals in spider from designated sampling sites were also determined. Heavy metals in spider collected near Siheung stream was appeared to be equivalent and/or a little higher levels with respect to other spiders. Furthermore, the ratio of relative heavy metals (Cu, Cd, and Pb) in spider from each stream site showed a correlation as similar as that of heavy metals in soil samples.