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1.
Parasites Hosts Dis ; 61(1): 60-71, 2023 Feb.
Article in English | MEDLINE | ID: mdl-37170465

ABSTRACT

Cockroaches can cause allergic sensitization in humans via contact with their feces or frass. Antibiotics can affect concentration of major allergen and total bacteria production in German cockroaches (Blattella germanica). This study examined the ability of antibiotic-treated German cockroaches to induce allergic airway inflammation and the effect of antibiotics on their lipopolysaccharide and Bla g1, 2, and 5 expression levels. Specifically, we measured the ability of German cockroach extract (with or without prior antibiotic exposure) to induce allergic inflammation in human bronchial epithelial cells and a mouse model of asthma. Bacterial 16S rRNA and lipopolysaccharide levels were lower in ampicillin-treated cockroaches than in the control group. The Bla g1, Bla g2, and Bla g5 expression in ampicillin-treated cockroaches decreased at both the protein and RNA levels. In human bronchial epithelial cell lines BEAS-2B exposed to the ampicillin-treated extract, expression levels of interleukin-6 and interleukin-8 were lower than that in the control group. The total cell count and eosinophil count in bronchoalveolar lavage fluid was also lower in mice exposed to the ampicillin-treated extract than in those exposed to normal cockroach extract. Mouse lung histopathology showed reduced immune cell infiltration and mucus production in the ampicillin group. Our results showed that ampicillin treatment reduced the symbiont bacterial population and major allergen levels in German cockroaches, leading to reduced airway inflammation in mice. These results can facilitate the preparation of protein extracts for immunotherapy or diagnostics applications.


Subject(s)
Asthma , Blattellidae , Humans , Mice , Animals , Lipopolysaccharides , RNA, Ribosomal, 16S , Asthma/chemically induced , Lung , Inflammation/drug therapy , Allergens , Ampicillin/pharmacology , Anti-Bacterial Agents/pharmacology
2.
Parasitology ; 145(12): 1531-1539, 2018 10.
Article in English | MEDLINE | ID: mdl-29580315

ABSTRACT

Due to the lack of an effective prophylactic intervention and diagnosis, human liver fluke Clonorchis sinensis continues to afflict a large human population, causing a chronic inflammatory bile duct disease. With an aim to identify target antigens for sensitive serodiagnosis, adenylate kinase 3 of C. sinensis (CsAK3) was successfully expressed in soluble form in Escherichia coli by fusion to an RNA-interacting domain derived from human Lys-tRNA synthetase and purified by Ni2+-affinity chromatography. Anti-CsAK3 serum was raised by immunization of mice, and Western blotting confirmed that CsAK3 was expressed in adult-stage C. sinensis. Histochemical analysis showed that CsAK3 was localized to the subtegumental tissue of C. sinensis and was excreted into the bile duct of the host. When tested against sera from various parasite-infected patients by enzyme-linked immunosorbent assay, the recombinant CsAK3 elicited a specific response to C. sinensis-infected sera. The results suggest that CsAK3, either alone or in combination with other antigens, could be used for improving the clinical diagnosis of clonorchiasis.


Subject(s)
Adenylate Kinase/immunology , Antigens, Helminth/immunology , Clonorchiasis/diagnosis , Clonorchis sinensis/immunology , Adenylate Kinase/genetics , Animals , Antigens, Helminth/genetics , Blotting, Western , Clonorchiasis/parasitology , Clonorchis sinensis/enzymology , Clonorchis sinensis/genetics , Enzyme-Linked Immunosorbent Assay , Female , Humans , Mice , Mice, Inbred BALB C , Phylogeny , Recombinant Proteins , Serologic Tests
3.
Korean J Parasitol ; 49(2): 177-80, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21738275

ABSTRACT

Entamoeba histolytica is an enteric tissue-invading protozoan parasite that can cause amebic colitis and liver abscess in humans. E. histolytica has the capability to kill colon epithelial cells in vitro; however, information regarding the role of calpain in colon cell death induced by ameba is limited. In this study, we investigated whether calpains are involved in the E. histolytica-induced cell death of HT-29 colonic epithelial cells. When HT-29 cells were co-incubated with E. histolytica, the propidium iodide stained dead cells markedly increased compared to that in HT-29 cells incubated with medium alone. This pro-death effect induced by ameba was effectively blocked by pretreatment of HT-29 cells with the calpain inhibitor, calpeptin. Moreover, knockdown of m- and µ-calpain by siRNA significantly reduced E. histolytica-induced HT-29 cell death. These results suggest that m- and µ-calpain may be involved in colon epithelial cell death induced by E. histolytica.


Subject(s)
Calpain/metabolism , Cell Death , Entamoeba histolytica/pathogenicity , Epithelial Cells/parasitology , Calpain/antagonists & inhibitors , Calpain/genetics , Cell Line , Cell Survival/drug effects , Dipeptides/metabolism , Gene Knockdown Techniques , Humans
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