• Parasites, Hosts and Diseases
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• v.61 no.4
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• pp.388-396
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• 2023

Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca²⁺ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.

• Parasites, Hosts and Diseases
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• v.62 no.1
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• pp.151-156
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• 2024

Bovine borreliosis, caused by Borrelia theileri which is transmitted via hard tick bites, is associated with mild clinical symptoms, such as fever, lethargy, hemoglobinuria, anorexia, and anemia. Borrelia theileri infects various animals, such as cattle, deer, horses, goats, sheep, and wild ruminants, in Africa, Australia, and South America. Notably, no case of B. theileri infection has been reported in Korean cattle to date. In this study, 101 blood samples were collected from a Korean indigenous cattle breed, among which 1.98% tested positive for B. theileri via nested PCR. The obtained sequences exhibited high homology with B. theileri strains identified in other regions. Phylogenetic analysis of 16S rRNA confirmed the B. theileri group affiliation; however, flagellin B sequences exhibited divergence, potentially due to regional evolutionary differences. This study provides the first molecular confirmation of B. theileri infection in Korean livestock. Further isolation and nucleotide sequence analyses are necessary to better understand the presence of B. theileri strains in cows in Korea.

• Parasites, Hosts and Diseases
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• v.62 no.1
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• pp.75-84
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• 2024

Ancylostoma ceylanicum is a zoonotic soil-derived nematode that parasitizes the intestines of humans and animals (dogs and cats), leading to malnutrition and iron-deficiency anemia. Helminth parasites secrete calreticulin (CRT), which regulates or blocks the host's immune response. However, no data on A. ceylanicum calreticulin (Ace-CRT) are available. We investigated the biological function of recombinant Ace-CRT (rAce-CRT). rAce-CRT showed reliable antigenicity and stimulated the proliferation of mouse splenocytes and canine peripheral blood mononuclear cells. Quantitative reverse-transcription PCR assays revealed that rAce-CRT primarily promoted the expression of T helper 2 cytokines, particularly IL-13, in canine peripheral blood lymphocytes. rAce-CRT inhibited complement-mediated sheep erythrocyte hemolysis in vitro. Our findings indicate that Ace-CRT plays an immunomodulatory role and may be a promising candidate molecule for a hookworm vaccine.

• Clinical and Experimental Vaccine Research
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• v.13 no.2
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• pp.146-154
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• 2024

Purpose: Infection by the intracellular apicomplexan parasite Toxoplasma gondii has serious clinical consequences in humans and veterinarians around the world. Although about a third of the world's population is infected with T. gondii, there is still no effective vaccine against this disease. The aim of this study was to develop and evaluate a multimeric vaccine against T. gondii using the proteins calcium-dependent protein kinase (CDPK)1, CDPK2, CDPK3, and CDPK5. Materials and Methods: Top-ranked major histocompatibility complex (MHC)-I and MHC-II binding as well as shared, immunodominant linear B-cell epitopes were predicted and linked using appropriate linkers. Moreover, the 50S ribosomal protein L7/L12 (adjuvant) was mixed with the construct's N-terminal to increase the immunogenicity. Then, the vaccine's physicochemical characteristics, antigenicity, allergenicity, secondary and tertiary structure were predicted. Results: The finally-engineered chimeric vaccine had a length of 680 amino acids with a molecular weight of 74.66 kDa. Analyses of immunogenicity, allergenicity, and multiple physiochemical parameters indicated that the constructed vaccine candidate was soluble, non-allergenic, and immunogenic, making it compatible with humans and hence, a potentially viable and safe vaccine candidate against T. gondii parasite. Conclusion: In silico, the vaccine construct was able to trigger primary immune responses. However, further laboratory studies are needed to confirm its effectiveness and safety.

• Parasites, Hosts and Diseases
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• v.62 no.2
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• pp.238-242
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• 2024

Strongyloidiasis is a chronic infection caused by the intestinal nematode parasite Strongyloides stercoralis and is characterized by a diverse spectrum of nonspecific clinical manifestations. This report describe a case of disseminated strongyloidiasis with urination difficulty, generalized weakness, and chronic alcoholism diagnosed through the presence of worms in the urinary sediment. A 53-year-old man was hospitalized for severe abdominal distension and urinary difficulties that started 7-10 days prior. The patient also presented with generalized weakness that had persisted for 3 years, passed loose stools without diarrhea, and complained of dyspnea. In the emergency room, approximately 7 L of urine was collected, in which several free-living female adult and rhabditiform larvae of S. stercoralis, identified through their morphological characteristics and size measurements, were detected via microscopic examination. Rhabditiform larvae of S. stercoralis were also found in the patient's stool. During hospitalization, the patient received treatment for strongyloidiasis, chronic alcoholism, peripheral neurosis, neurogenic bladder, and megaloblastic anemia, and was subsequently discharged with improved generalized conditions. Overall, this report presents a rare case of disseminated strongyloidiasis in which worms were detected in the urinary sediment of a patient with urination difficulties and generalized weakness combined with chronic alcoholism, neurogenic bladder, and megaloblastic anemia.

• Parasites, Hosts and Diseases
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• v.29 no.3
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• pp.235-244
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• 1991

An epidemiological study and mass treatments of Enterobius vermicularis infection among children near Wonju area of Kangwon province were carried out. The children were divided into 4 groups according to their residing localities; children in the mountainous area, rural area, urban area and in orphanage. They were examined by adhesive cellotape anal swab technique, and egg positive rates were obtained. The rates of egg reduction and re-infection rates after repeated mass treatments were also observed. The results obtained were as follows: 1. The overall egg Positive rate of E. vermicularis in the first screening was 19.9% (251 out of 1, 262 examinees; 19.7% in males and 20.1% in females). The positive rates were 13.0% in the mountainous area, 11 9% in the rural area, 15.1% in the urban (medium-sized) area and 61.9% in orphanages. 2. The highest positive rates were observed in the kindergarten children, and 1st and 2nd grade children of primary schools (26.2~32.2%), and the lowest rate (13.6%) in 6-year grade children of primary schools. 3. Cumulative detection rates from 3 repeated anal swabs at 4~5 days interval were higher (70.8%) than those from single anal swabs (50.0~59.2%). 4. Out of the examinees who showed the highest cumulative positive rate (70.8%), about 39.2% were consecutively positive in 3 anal swabs. Among different groups of children, the higher the total egg detection rates (87.5%), the higher the consecutive positive rates (71.9%) . 5. A total of 2, 609 (male : female=1 : 12.4) worms were collected from 17 egg-positive cases treated with anthelinintics. The mean number of worms per child was 153 (range: 4-824) . 6. The egg-positive cases in several studied groups (180 children) were treated with anthelmintics 6 times at 3-week intervals. In this case, the overall positive rate was decreased from 54.8% to 2.2% at 15 weeks after the treatments, but no complete negative conversion was experienced. However, in a group of children (154 children) including egg Positive and negative cases who were both treated with anthelmintics at 3-week interval, a complete egg-negative conversion was observed in the 9th week after treatments. 7. The egg-detection rate in the brothers or sisters of egg Positive children was 70.0% (28 out of 40 examined), and the egg-positive rate according to the family unit was 69.7%. In summarizing the above results, it is concluded that Enterobius vermicularis infection is still highly prevalent among children in Korea, and that repeated mass treatments of more than 3 times will be effective for control of this infection.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.57-61
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• 2006

Suppressor of cytokine signaling (SOCS) is known to be as a negative feedback regulator in Janus kinase signal transducer and activator of transcription signaling. Highly conserved SOCS box domain was cloned from a Korean malaria vector, Anopheles sinensis. Sequence analysis indicates that it has identity to Anopheles gambiae (96%), Aedes aegypti (94%), Drosophila melanogaster (78%), Mus musculus (72%) and Homo sapiens (72%), respectively. Tissue specificity RT-PCR demonstrated that the expression level of AsSOCS transcript was high at abdomen, midgut, and ovary, whereas developmental expression patterns showed that the level of AsSOCS was high at egg, early pupae, and adult female. On the other hand, RT-PCR analysis after bacterial challenge showed that SOCS mRNA was strongly induced in larvae. In addition, it was also induced by various immune elicitors such as lipoteicoic acid, CpG-DNA, and laminarin. It seems that AsSOCS, repressor of JAK-STAT pathway, is highly conserved in mosquito, and may play an important role in mosquito innate immune response.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.2
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• pp.63-67
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• 2006

Suppressor of cytokine signaling (SOCS) is known to playa key role as a negative feedback regulator in JAK/STAT signaling cascade in innate immunity. Our laboratory has recently been interested in elucidating the interactions between Spodoptera exigua (Se) and SeNPV. This context leads us to clone and characterize SeSOCS that may have important functions in response to SeNPV infection. Using the RT-PCR and TA cloning approach, we found a partial fragment (416 bp) of SeSOCS. Blast search and multiple alignment data showed that it has a homology to various insects such as Anopheles gambiae (78%), Aedes aegypti (75%), Drosophila melanogastar (77%), Mus musculus (69%), and Homo sapiens (69%). Temporal induction patterns of SeSOCS were analysed after being immune-challenged with either NPV or laminarin. It showed that the level of SeSOCS mRNA was strongly induced in a biphasic manner in response to SeNPV and laminarin, respectively. It seems that SOCS, a negative regulator of JAK/STAT signaling system is also present in S. exigua and may playa role in innate immunity albeit its precise role should be further elucidated at the molecular and cellular level in the early phase of SeNPV infection in larvae.

• IMMUNE NETWORK
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• v.2 no.2
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• pp.91-95
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• 2002

Background: In order to characterize human antibodies with specificity for mite allergens at the molecular level, a scFv phage display library was constructed using peripheral blood mononuclear lymphocytes from an asthma patient allergic to mite as Ig gene sources. Methods: Immunoglobulin $V_H$ and V gene fragments were obtained by polymerase chain reaction, and randomly combined in pCANTAB-5E vector. The resulting human scFv phage display library had $3{\times}10^4$ independent clones, and biopanning was performed with house dust mite extracts. Results: Four scFv clones specific for house dust mite extract were isolated. Immunoblot assay showed that our clones reacted to 25 kDa and 50~60 kDa proteins with unknown identity in mite extracts. Sequence analysis indicated that two clones (b7 and c15) are identical, and all clones belong to human $V_H3$ subgroup. On the other hand, light chain usage was different in that two clones (a2 and b7 / c15) belonging to V ${\kappa}4$ subgroup, but a4 used V ${\kappa}1$ light chain gene. Conclusion: Our approach should facilitate provision of useful information on the antibody responses against allergens at the molecular level in humans.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.1
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• pp.87-90
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• 2012

Cholangiocarcinoma (CCA) is the most common cancer in northeastern Thailand. At present, effective diagnosis of CCA either in humans or animals is not available. Monitoring the development and progression of CCA in animal models is essential for research and development of new promising chemotherapeutics. Ultrasonography has been widely used for screening of bile duct obstruction in CCA patients. In this study, we preliminarily investigated the applicability of ultrasonography to monitor the development and progression of CCA in Syrian golden hamsters (n=8) induced by Opisthorchis viverrini (OV)/dimethylnitrosamine (DMN) administration. Ultrasonography and histopathological examination of hamsters was performed at week 0, 20, 24 and 28 of OV infection or at the start of water/Tween-80 administration to controls. The ultrasonographic images of liver parenchyma and gallbladders of OV/DMN-induced CCA hamsters showed sediments in gallbladder, thickening of gallbladder wall, and hypoechogenicity of liver parenchyma cells. The ultrasonographic images of liver tissues were found to correlate well with histopathological examination. Although ultrasonography does not directly detect the occurrence of CCA, it reflects the thickening of bile ducts and abnormality of liver tissues. It may be applied as a reliable tool for monitoring the development and progression of CCA in animal models in research and development of new promising chemotherapeutics for CCA.