• Korean journal of applied entomology
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• v.51 no.4
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• pp.349-356
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• 2012

Eicosanoid mediates various cellular immune responses in insects. This study aimed to discover its novel action on the modulation of hemocyte populations in response to an immune challenge. Upon bacterial challenge, the last instar larvae of the beet armyworm, Spodoptera exigua, increased their total hemocyte density in 2 h, and then decreased it to a basal hemocyte density level. This rapid increase in total hemocyte density was explained by an increase of plasmatocyte and spherulocyte densities. When larvae were treated with dexamethasone (a specific phospholipase $A_2$ ($PLA_2$) inhibitor), they did not show any increase in hemocyte density in response to bacterial challenge. However, the addition of arachidonic acid (a catalytic product of $PLA_2$) to larvae treated with dexamethasone recovered the up-regulation of hemocyte density in response to bacterial infection. Among eicosanoid, cyclooxygenase (COX), but not lipoxygenase (LOX), products seemed to mediate the increase of hemocyte density in response to bacterial infection because naproxene (a COX inhibitor) inhibited the hemocyte density increase, though esculetin (a LOX inhibitor) did not. Prostaglandin $E_2$, a COX product, significantly increased the hemocyte density even without bacterial infection. These results suggest that eicosaniod mediates a rapid increase in total hemocyte density in response to immune challenge.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2000.05a
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• pp.449-450
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• 2000

이매패의 hemocytes는 상처 치유, shell repair, 영양물질의 소화, 수송 그리고 분비에 관여한다고 알려져 있으며, 체내 방어 기능에서 필수적이다(Carmen et al., 1997). 패류의 hemocytes에 관해서는 주로 형태, 세포화학적 특징이 연구되어 왔으며(Pipe, 1990 등), 밀도차원심분리법(Bachere et al., 1988)으로 분리된 hemocytes를 면역학적 방법에 의해(Cheng et al., 1980; Yoshino and Granath, 1983) 여러 가지 subpopulatione으로 구분하지만, 현재로서는 같은 종의 패류에서도 분류 방법에 따라 다양하게 분류되고 있다. (중략)

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.spc1
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• pp.123-131
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• 2006

Immmune parameters of the Manila clam Ruditapes philippinarum collected from four tidal flats, Nudong, Gonam, Hwangdo and Bangpo on Anmyeon-do, Korea were optimized and evaluated at the single cell level using flow-cytometry Hemocytes were withdrawn from the sinus of each clam, and total hemocyte counts (THC), phagocytosis rate, hemocyte mortality (HM) and DNA damage of hemocyte were analyzed. The highest hemocyte counts was recorded from the clams collected from Gonam, followed by Hwangdo, Nudong and Bangpo (P<0.001). Phagocytosis rate and hemocyte mortality of Gonam and Nudong clams were significantly higher than those of clams from Hwangdo and Bangpo (P<0.001). DNA damage in the clams from Nudong was higher twice than that of clams from Gonam (P<0.05). We suggest that the flow-cytometry has a high potential for evaluation of immunity of marine bivalves.

• International Journal of Industrial Entomology and Biomaterials
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• v.23 no.1
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• pp.137-141
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• 2011

A previous data was provided information for tissuespecific expression genes by means of whole-genome oligonucleotide microarray in the silkworm. We analyzed the tissue-specific expression patterns in the hemocyte tissue on 5 days of 5th instar larvae during the development of $B.$ $mori$. Total 5 candidates pick out from the $Bombyx$ $mori$ Microarray Database (BmMDB; http://silkworm.swu.edu.cn/microarray). To verify the hemocyte-specific expression, we analyzed by semi-quantitative and real-time quantitative RT-PCR using the highly expressed endogenous $Actin$ RNA as an intrinsic reference. In this study, we confirmed that one gene-sw17255- out of 5 candidates expressed in the hemocyte tissue, which was consistent with the previous data. Circulating hemocytes in the body fluid of the $B.$ $mori$ are most powerful target organ for producing biomaterials. We need further studies to find hemocyte-specific promoter region from sw17255 gene. Finally, this result can be applied in creating transgenic silkworms as a biomedical insect.

• International Journal of Industrial Entomology and Biomaterials
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• v.5 no.2
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• pp.189-194
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• 2002

Silkworm diseases are major constraint in silk cocoon production. Among silkworm diseases, white muscar-dine is highly contagious and most common in winter and rainy seasons. It is suggested that hemocytes involve in defense against invasion of Beauveria bassi-ana and systemic fungicide/chemicals prevent the proliferation of fungi in the hemolymph or preventing the growth of the fungi in the body cavity through enhancing the hemocyte mediated defense response. In the present study the influence of systemic fungicide on hematological changes in silkworms infected with Beauveria bassiana was reported. It is observed that the total hemocyte counts increased in the hemolymph up to 5 days post inoculation in systemic fungicide treated batches while in the inoculated control the increase was up to 3 days indicating the positive hemocyte mediated response in silkworm treated with systemic fungicide. After 2 days in the inoculated control as the multiplication and growth of mycelia increased, defense capacity of the silkworm was decreased. The biochemical changes were also observed in the hemolymph of silkworm infected with B. bassiana. In silkworm infected with Beauveria bassi-ana, the total protein content increased whereas total carbohydrate and total lipids decreased as the infection progresses. In the case of systemic fungicide treated batches the increase in total protein content was comparatively higher and decrease in total carbohydrate and lipids were comparatively lower than the inoculated control.

• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.1
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• pp.11-14
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• 2003

Systemic fungicides viz., Bavistin and Beyleton are reported to have curative effect against the infection of Beauveria bassiana in silkworm Bombyx mori L. To understand the influence of the systemic fungicides on the disease suppression/development, hematological studies were carried out. There was an increase in the percent total hemocyte count, granulocyte and plasmatocyte in silkworm treated with the systemic fungicides. It possibly indicates the influence of systemic fungicide on the hemocyte mediated defense system leading to the higher resistance and suppression of disease development.

• Parasites, Hosts and Diseases
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• v.49 no.3
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• pp.229-234
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• 2011

In order to assess changes in the activity of immunecompetency present in Crassostrea gigas infected with Marteilioides chungmuensis (Protozoa), the total hemocyte counts (THC), hemocyte populations, hemocyte viability, and phagocytosis rate were measured in oysters using flow cytometry. THC were increased significantly in oysters infected with M. chungmuensis relative to the healthy appearing oysters (HAO) (P<0.05). Among the total hemocyte composition, granulocyte levels were significantly increased in infected oysters as compared with HAO (P<0.05). In addition, the hyalinocyte was reduced significantly (P<0.05). The hemocyte viability did not differ between infected oysters and HAO. However, the phagocytosis rate was significantly higher in infected oysters relative to HAO (P<0.05). The measurement of alterations in the activity of immunecompetency in oysters, which was conducted via flow cytometry in this study, might be a useful biomarker of the defense system for evaluating the effects of ovarian parasites of C. gigas.

• Journal of Microbiology and Biotechnology
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• v.19 no.6
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• pp.610-615
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• 2009

As a provirus, polydnavirus has a segmented DNA genome on chromosome(s) of host wasp. It contains several genes in each segment that presumably play critical roles in regulating physiological processes of target insect parasitized by the wasp. A cysteine-rich protein 1 (CRP1) is present in the polydnavirus Cotesia plutellae bracovirus (CpBV) genome, but its expression and physiological function in Plutella xylostella parasitized by the viral host C. plutellae is not known. This CpBV-CRP1 encoding 189 amino acids with a putative signal peptide (20 residues) was persistently expressed in parasitized P. xylostella with gradual decrease at the late parasitization period. Expression of CpBV-CRP1 was tissue-specific in the fat body/epidermis and hemocyte, but not in the gut. Its physiological function was analyzed by inducing transient expression of a CpBV segment containing CpBV-CRP1 and its promoter, which caused significant reduction in hemocyte -spreading and delayed larval development. When the treated larvae were co-injected with double-stranded RNA of CpBV-CRP1, the expression of CpBV-CRP1 disappeared, whereas other genes encoded in the CpBV segment was expressed. These co-injected larvae significantly recovered the hemocyte-spreading capacity and larval development rate. This study reports that CpBV-CRP1 is expressed in P. xylostella parasitized by C. plutellae and its physiological function is to alter the host immune and developmental processes.

• BMB Reports
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• v.31 no.4
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• pp.364-369
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• 1998

Insect plasma protein is abundant in the hemolymph of holometabolous insect larvae and is used as a source of amino acids and energy for construction of adult structures during metamorphosis. In order to understand the mechanism of decomposition of larval plasma proteins by hemocyte protease, we tried to purify a cysteine protease from the hemocyte lysate by using Carbobenzoxy-L-Phenylalanyl-L-Arginine-4-Methyl-Coumaryl-7-Amide (Z-Phe-Arg-MCA) as substrate and to identify plasma proteins that are selectively susceptible to the purified protease. Here, we describe the purification and characterization of a cysteine protease that specifically hydrolyzes the plasma protein of the coleopteran insect, Tenebrio molitor, larvae. The molecular mass of this enzyme was 25 kDa, as determined by SDS-PAGE under reducing conditions. The amino acids sequence of its $NH_{2}-terminus$ was determined to be Leu-Pro-Gly-Gln-Ile-Asp-Trp-Arg-Asp-Lys-Gly. This sequence contained Pro, Asp, and Arg residues, conserved in many papain superfamily enzymes. The specific cysteine protease inhibitors, such as E-64 and leupetin, inhibited its hydrolytic activity. One plasma protein with a molecular mass of 48 kDa was selectively hydrolyzed within 3 h when the purified enzyme and plasma proteins were incubated in vitro. However, the 48 kDa protein was not hydrolyzed by the purified 25 kDa protease in the presence of E-64. Western blotting analysis at various developmental stages showed that the purified enzyme was detected at larvae, pupae, and adult stages, but not the embryo stage.

• Fisheries and Aquatic Sciences
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• v.6 no.2
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• pp.74-80
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• 2003

Two cellular biomarker parameters of the farmed Pacific oyster Crassostrea gigas were studied in vivo and in vitro after exposure to concentrations of polychlorinated biphenyls in terms of neural red uptake (NRU) and lysozyme activity. The oysters exposed in vivo to the xenobiotic concentrations, 0, 30, 90, and 180 ng/g for 14 days, enhanced hemocyte NRU with occasional significant differences (P<0.05), depending on the chemical concentration and duration. An adverse tendency was manifest in the lysozyme activities both in the hemocyte and serum of the oyster treated with the chemical in a same manner, rendering these two cellular parameters as biomarker candidates against the chemical. The oysters exposed in vitro to the chemical concentrations, 0, 1, 5, 10, 100, 1,000, and 10,000 ng/g for 24 hrs at $10^{\circ}C$ showed a similar tendancy as those exposed in vivo to the chemical. Unlike in vivo response, however, the in vitro NRU was first influenced by very low concentration of the chemical. In in vitro results, marked but not significant increase of hemocyte NRU was noticed at the chemical concentration of 5 ng/g, where the value was almost as high as those exposed to higher chemical concentrations, up to 10,000 ng/g. An unusual result was observed in the in vitro lysozyme activity of hemocyte in which significant decrease was first noticed at the chemical concentration of 100 ng/g.