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Wanchai Maleewong - One of the best experts on this subject based on the ideXlab platform.
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molecular phylogenetic confirmation of Gnathostoma Spinigerum owen 1836 nematoda Gnathostomatidae in laos and thailand
Folia Parasitologica, 2016Co-Authors: Jurairat Jongthawin, Penchom Janwan, Pewpan M Intapan, Oranuch Sanpool, Tongjit Thanchomnang, Lakkhana Sadaow, Sakhone Laymanivong, Wanchai MaleewongAbstract:We report the molecular-phylogenetic identification of larvae of the nematode genus Gnathostoma Owen, 1836 collected from a snake, Ptyas koros Schlegel, in Laos and adult worms from the stomach of a dog in Thailand. DNA was extracted and amplified targeting the partial cox1 gene and the ITS-2 region of ribosomal DNA. Phylogenetic analyses indicated that all five advanced third-stage larvae and seven adult worms were Gnathostoma Spinigerum Owen, 1836. This is also the first molecular evidence of infection with G. Spinigerum in a snake from Laos.
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a dot elisa test using a Gnathostoma Spinigerum recombinant matrix metalloproteinase protein for the serodiagnosis of human gnathostomiasis
Southeast Asian Journal of Tropical Medicine and Public Health, 2014Co-Authors: S Saenseeha, Wanchai Maleewong, Porntip Laummaunwai, Chatchai Tayapiwatana, Hiroshi Yamasaki, Amnat Kitkhuandee, J Penchom, Pewpan M IntapanAbstract:Gnathostomiasis caused by Gnathostoma Spinigerum, is a hazardous food- borne helminthic zoonosis, and is endemic especially in developing countries in Asia. Definitive diagnosis, relying upon identification of worms from human tissues or body, is rarely accomplished. Consequently, sensitive supporting tools such as serological tests have been used widely. But these methods are time con- suming, need sophisticated equipment and are impractical in some settings. In the present study a dot enzyme-linked immunosorbent assay (dot-ELISA), using G. Spinigerum recombinant matrix metalloproteinase protein as the antigen, was developed and assessed using sera of gnathostomiasis and other parasitosis pa- tients as well as healthy controls. The accuracy, sensitivity, specificity, positive and negative predictive values were 97.4%, 100%, 96.1%, 92.9%, and 100%, respectively. The dot-ELISA appears to be a suitable rapid test for diagnostic purpose as well as epidemiological studies.
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detection of Gnathostoma Spinigerum antibodies in sera of non traumatic subarachnoid hemorrhage patients in thailand
Korean Journal of Parasitology, 2013Co-Authors: Amnat Kitkhuandee, Penchom Janwan, Wanchai Maleewong, Kittisak Sawanyawisuth, Waranon Munkong, Pewpan M IntapanAbstract:Gnathostoma Spinigerum can cause subarachnoid hemorrhage (SAH). The detection of specific antibodies in serum against G. Spinigerum antigen is helpful for diagnosis of neurognathostomiasis. There is limited data on the frequency of G. Spinigerum infection in non-traumatic SAH. A series of patients diagnosed as non-traumatic SAH at the Srinagarind Hospital, Khon Kaen University, Thailand between January 2011 and January 2013 were studied. CT or MR imaging of the brain was used for diagnosis of SAH. Patients were categorized as aneurysmal subarachnoid hemorrhage (A-SAH) or non-aneurysmal subarachnoid hemorrhage (NA-SAH) according to the results of cerebral angiograms. The presence of specific antibodies in serum against 21- or 24-kDa G. Spinigerum antigen was determined using the immunoblot technique. The detection rate of antibodies was compared between the 2 groups. Of the 118 non-traumatic SAH patients for whom cerebral angiogram and immunoblot data were available, 80 (67.8%) patients had A-SAH, whereas 38 (32.2%) had NA-SAH. Overall, 23.7% were positive for specific antibodies against 21- and/or 24-kDa G. Spinigerum antigen. No significant differences were found in the positive rate of specific antibodies against G. Spinigerum in both groups (P-value=0.350).
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modulation of antibody responses against Gnathostoma Spinigerum in mice immunized with crude antigen formulated in cpg oligonucleotide and montanide isa720
Korean Journal of Parasitology, 2013Co-Authors: Pewpan M Intapan, Penchom Janwan, Chakrit Hirunpetcharat, Churairat Kularbkaew, Wiboonchai Yutanawiboonchai, Wanchai MaleewongAbstract:This study aimed to investigate the antibody responses in mice immunized with Gnathostoma Spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. Spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. Spinigerum infection potently induces a Th2-type biased response.
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growth and development of Gnathostoma Spinigerum nematoda Gnathostomatidae larvae in mesocyclops aspericornis cyclopoida cyclopidae
Parasites & Vectors, 2011Co-Authors: Penchom Janwan, Pewpan M Intapan, Oranuch Sanpool, Luxkhana Sadaow, Tongjit Thanchomnang, Wanchai MaleewongAbstract:Background Gnathostoma Spinigerum larva is pathogenic, causing gnathostomiasis in humans and certain animals, and is prevalent mainly in Asia. Growth and development of Gnathostoma Spinigerum larvae in the cyclopoid copepod Mesocyclops aspericornis, the first intermediate host, were examined.
Pewpan M Intapan - One of the best experts on this subject based on the ideXlab platform.
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Molecular identification and genetic diversity of Gnathostoma Spinigerum larvae in freshwater fishes in southern Lao PDR, Cambodia, and Myanmar
Parasitology Research, 2019Co-Authors: Patcharaporn Boonroumkaew, Oranuch Sanpool, Porntip Laummaunwai, Lakkhana Sadaow, Sakhone Laymanivong, Rutchanee Rodpai, Chalermchai Somboonpatarakun, Win Pa Pa Aung, Mesa Un, Pewpan M IntapanAbstract:Gnathostomiasis, an emerging food-borne parasitic zoonosis in Asia, is mainly caused by Gnathostoma Spinigerum (Nematoda: Gnathostomatidae). Consumption of raw meat or freshwater fishes in endemic areas is the major risk factor. Throughout Southeast Asia, including Thailand, Lao PDR, Cambodia, and Myanmar, freshwater fish are often consumed raw or undercooked. The risk of this practice for gnathostomiasis infection in Lao PDR, Cambodia, and Myanmar has never been evaluated. Here, we identified larvae of Gnathostoma species contaminating freshwater fishes sold at local markets in these three countries. Public health authorities should advise people living in, or travelling to, these areas to avoid eating raw or undercooked freshwater fishes. Identification of larvae was done using molecular methods: DNA was sequenced from Gnathostoma advanced third-stage larvae recovered from snakehead fishes ( Channa striata ) and freshwater swamp eels ( Monopterus albus ). Phylogenetic analysis of a portion of the mitochondrial cytochrome c oxidase subunit I gene showed that the G. Spinigerum sequences recovered from southern Lao PDR, Cambodia, and Myanmar samples had high similarity to those of G. Spinigerum from China. Sequences of the nuclear ribosomal DNA internal transcribed spacer 2 region closely resembled sequences of G. Spinigerum from Thailand, Indonesia, the USA, and central Lao PDR. This is the first molecular evidence of G. Spinigerum from freshwater fishes in southern Lao PDR, Cambodia, and Myanmar.
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molecular phylogenetic confirmation of Gnathostoma Spinigerum owen 1836 nematoda Gnathostomatidae in laos and thailand
Folia Parasitologica, 2016Co-Authors: Jurairat Jongthawin, Penchom Janwan, Pewpan M Intapan, Oranuch Sanpool, Tongjit Thanchomnang, Lakkhana Sadaow, Sakhone Laymanivong, Wanchai MaleewongAbstract:We report the molecular-phylogenetic identification of larvae of the nematode genus Gnathostoma Owen, 1836 collected from a snake, Ptyas koros Schlegel, in Laos and adult worms from the stomach of a dog in Thailand. DNA was extracted and amplified targeting the partial cox1 gene and the ITS-2 region of ribosomal DNA. Phylogenetic analyses indicated that all five advanced third-stage larvae and seven adult worms were Gnathostoma Spinigerum Owen, 1836. This is also the first molecular evidence of infection with G. Spinigerum in a snake from Laos.
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proteomic analysis identification of antigenic proteins in Gnathostoma Spinigerum larvae
Experimental Parasitology, 2015Co-Authors: Penchom Janwan, Pewpan M Intapan, Tongjit Thanchomnang, Chaisiri Wongkham, Porntip Laummaunwai, Rutchanee Rodpai, Tonkla Insawang, Oranuch SanpoolAbstract:Abstract Gnathostoma Spinigerum is the causative agent of human gnathostomiasis. The advanced third stage larva (AL3) of this nematode can migrate into the subcutaneous tissues, including vital organs, often producing severe pathological effects. This study performed immuno-proteomic analysis of antigenic spots, derived from G. Spinigerum advanced third stage larva (GSAL3) and recognized by human gnathostomiasis sera, using two-dimensional (2-DE) gel electrophoresis based-liquid chromatography/tandem mass spectrometry (LC/MS–MS), and followed by the aid of a database search. The crude GSAL3 extract was fractionated using IPG strips (pH 3-11NL) and followed by SDS-PAGE in the second dimension. Each gel was stained with colloidal Coomassie blue or was electro-transferred onto a nitrocellulose membrane and probed with gnathostomiasis human sera by immunoblotting. Individual Coomassie-stained protein spots corresponding to the antigenic spots recognized by immunoblotting were excised and processed using LC/MS–MS. Of the 93 antigenic spots excised, 87 were identified by LC/MS–MS. Twenty-seven protein types were found, the most abundant being Ascaris suum37. Six spots showed good quality spectra, but could not be identified. This appears to be the first attempt to characterize antigenic proteins from GSAL3 using a proteomic approach. Immuno-proteomics shows promise to assist the search for candidate proteins for diagnosis and vaccine/drug design and may provide better understand of the host-parasite relationship in human gnathostomiasis.
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a dot elisa test using a Gnathostoma Spinigerum recombinant matrix metalloproteinase protein for the serodiagnosis of human gnathostomiasis
Southeast Asian Journal of Tropical Medicine and Public Health, 2014Co-Authors: S Saenseeha, Wanchai Maleewong, Porntip Laummaunwai, Chatchai Tayapiwatana, Hiroshi Yamasaki, Amnat Kitkhuandee, J Penchom, Pewpan M IntapanAbstract:Gnathostomiasis caused by Gnathostoma Spinigerum, is a hazardous food- borne helminthic zoonosis, and is endemic especially in developing countries in Asia. Definitive diagnosis, relying upon identification of worms from human tissues or body, is rarely accomplished. Consequently, sensitive supporting tools such as serological tests have been used widely. But these methods are time con- suming, need sophisticated equipment and are impractical in some settings. In the present study a dot enzyme-linked immunosorbent assay (dot-ELISA), using G. Spinigerum recombinant matrix metalloproteinase protein as the antigen, was developed and assessed using sera of gnathostomiasis and other parasitosis pa- tients as well as healthy controls. The accuracy, sensitivity, specificity, positive and negative predictive values were 97.4%, 100%, 96.1%, 92.9%, and 100%, respectively. The dot-ELISA appears to be a suitable rapid test for diagnostic purpose as well as epidemiological studies.
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modulation of antibody responses against Gnathostoma Spinigerum in mice immunized with crude antigen formulated in cpg oligonucleotide and montanide isa720
Korean Journal of Parasitology, 2013Co-Authors: Pewpan M Intapan, Penchom Janwan, Chakrit Hirunpetcharat, Churairat Kularbkaew, Wiboonchai Yutanawiboonchai, Wanchai MaleewongAbstract:This study aimed to investigate the antibody responses in mice immunized with Gnathostoma Spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. Spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. Spinigerum infection potently induces a Th2-type biased response.
Yaowapa Maneerat - One of the best experts on this subject based on the ideXlab platform.
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Effects of Gnathostoma Spinigerum infective stage larva excretory-secretory products on NK cells in peripheral blood mononuclear cell culture: focused on expressions of IFN-γ and killer cell lectin-like receptors
Parasitology Research, 2020Co-Authors: Thanyathon Khetsuphan, Paron Dekumyoy, Surachet Benjathummarak, Wilanee Dechkhajorn, Urai Chaisri, Sumate Ampawong, Parnpen Viriyavejakul, Yaowapa ManeeratAbstract:Human gnathostomiasis is mainly caused by third-stage larvae of Gnathostoma Spinigerum ( G. Spinigerum L3). Excretory-secretory products (ES) released from infective helminthic larvae are associated with larval migration and host immunity modulation. Natural killer (NK) cells have important immune functions against helminth infection. Currently, the effects of ES from G. Spinigerum L3 ( G. Spinigerum ES) on NK cell activity are unclear. This study investigated whether G. Spinigerum ES affected human NK cells. Human normal peripheral blood mononuclear cell (PBMC) cultures were used to mimic immune cells within the circulation. PBMC were co-cultured with G. Spinigerum ES (0.01–0.05 μg/ml) for 5 or 7 days. Levels of IFN-γ in cultured supernatants were measured by enzyme-linked immunosorbent assay. The expressions of mRNA encoding NK cell receptors, especially the C type killer cell lectin-like family (KLR; NKG2A, NKG2C, and NKG2D) and IFN-γ in ES induced PBMC were determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). ES induced PBMC markedly decreased the levels of IFN-γ and increased the expressions of NKG2A and NKG2D on NK cells. In conclusion, low amounts of G. Spinigerum ES modulated NK cells by downregulating the transcription of IFN-γ and upregulating the expressions of KLR (NKG2A and NKG2D receptors) during the 7-day observation period. These findings indicate more in-depth studies of NK cell function are required to better understand the mechanism involved in immune evasive strategies of human gnathostomiasis.
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Excretory-secretory product of third-stage Gnathostoma Spinigerum larvae induces apoptosis in human peripheral blood mononuclear cells
Parasitology Research, 2017Co-Authors: Nareerat Viseshakul, Supaporn Nuamtanong, Surachet Benjathummarak, Wilanee Dechkhajorn, Yaowapa ManeeratAbstract:Human gnathostomiasis caused by third-stage Gnathostoma Spinigerum larvae ( G. Spinigerum L3) is an important zoonotic disease in tropical areas of the world. The excretory-secretory products (ES) that are excreted by infective larva play a significant role in host immune evasion and tissue destruction. To investigate the poorly understood mechanisms of G. Spinigerum L3 pathogenesis, we focused on the potential effect of ES on inducing apoptosis in human immune cells by using human peripheral blood mononuclear cells (PBMCs) as a model. Early and late apoptosis of PBMCs were assessed following the exposure of these cells to G. Spinigerum L3 ES (0.1, 0.5, and 1.0 μg/ml) for 6–48 h. The apoptotic cells were identified by flow cytometric staining of PBMC with FITC-annexin V and propidium iodide. The expression of regulatory genes related to apoptosis mechanisms in ES-treated PBMCs was investigated using a Human Apoptosis RT^2 Profiler™ PCR Array. The results showed significant levels of early phase apoptosis at 18 h and of late phase apoptosis at 24 h. We speculate that this apoptosis in PBMCs occurs via the extrinsic pathway. Apoptosis in the ES-induced PBMCs was observed as quickly as 90 min after exposure, and the highest effect was observed at 18–24 h. Furthermore, ES can trigger apoptosis lasting for 48 h. Our findings expand the understanding of one of the mechanisms involved, immune-evasive strategy mechanism used by G. Spinigerum larvae during human gnathostomiasis.
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third stage Gnathostoma Spinigerum larva excretory secretory antigens modulate function of fc gamma receptor i mediated monocytes in peripheral blood mononuclear cell culture
Tropical Medicine and Health, 2016Co-Authors: Surachet Benjathummarak, Jitra Waikagul, Supaporn Nuamtanong, Thareerat Kalambaheti, Ratchanok Kumsiri, Nareerat Viseshakul, Yaowapa ManeeratAbstract:Background Third (infective)-stage Gnathostoma Spinigerum larvae (L3) mainly cause human gnathostomiasis. G. Spinigerum L3 migrate throughout the subcutaneous tissues, vital organs, and central nervous system and can cause various pathogenesis including sudden death. Interestingly, G. Spinigerum L3 can survive and evade host cellular immunity for months or years. The effects of G. Spinigerum excretory-secretory (ES) products involved in larval migration and immune-evasive strategies are unknown. Monocytes are innate immune cells that act as phagocytic and antigen-presenting cells and also play roles against helminthic infections via a complex interplay between other immune cells. Fc gamma receptor I (FcγRI) is a high-affinity receptor that is particularly expressed on monocytes, macrophages, and dendritic cells. The cross-linking of FcγRI and antigen-antibody complex initiates signal transduction cascades in phagocytosis, cytokine production, and antibody-dependent cell-mediated cytotoxicity (ADCC). This study investigated whether ES antigen (ESA) from G. Spinigerum L3 affects monocyte functions.
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Third-stage Gnathostoma Spinigerum larva excretory secretory antigens modulate function of Fc gamma receptor I-mediated monocytes in peripheral blood mononuclear cell culture
Tropical Medicine and Health, 2016Co-Authors: Surachet Benjathummarak, Jitra Waikagul, Supaporn Nuamtanong, Thareerat Kalambaheti, Ratchanok Kumsiri, Nareerat Viseshakul, Yaowapa ManeeratAbstract:Background Third (infective)-stage Gnathostoma Spinigerum larvae (L3) mainly cause human gnathostomiasis. G. Spinigerum L3 migrate throughout the subcutaneous tissues, vital organs, and central nervous system and can cause various pathogenesis including sudden death. Interestingly, G. Spinigerum L3 can survive and evade host cellular immunity for months or years. The effects of G. Spinigerum excretory-secretory (ES) products involved in larval migration and immune-evasive strategies are unknown. Monocytes are innate immune cells that act as phagocytic and antigen-presenting cells and also play roles against helminthic infections via a complex interplay between other immune cells. Fc gamma receptor I (FcγRI) is a high-affinity receptor that is particularly expressed on monocytes, macrophages, and dendritic cells. The cross-linking of FcγRI and antigen-antibody complex initiates signal transduction cascades in phagocytosis, cytokine production, and antibody-dependent cell-mediated cytotoxicity (ADCC). This study investigated whether ES antigen (ESA) from G. Spinigerum L3 affects monocyte functions. Results Cultures of normal peripheral blood mononuclear cells (PBMC) separated from healthy buffy coats were used as a human immune cell model. ESA was prepared from G. Spinigerum L3 culture. Using Real-Time quantitative reverse transcription-polymerase chain reaction (qRT-PCR), the effect of ESA to down-regulate FcγRI mRNA expression in monocytes during 90 min of observation was not well delineated. Flow cytometry analysis revealed a significant phenotypic-decreased FcγRI expression on the monocyte surface at 12 hours (h) of cultivation with the ESA ( p = 0.033). Significantly reduced monocyte-mediated phagocytosis capacity was consistently observed after 12 h of ESA pretreatment ( p = 0.001). Conclusions Our results suggest that G. Spinigerum ESA modulates monocyte function via depletion of FcγRI expression. This study provides preliminary information for future in-depth studies to elucidate mechanisms of the immune-evasive strategy of G. Spinigerum larvae.
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roles of partially purified antigens from Gnathostoma Spinigerum larvae on antibody production by human b cell culture
Southeast Asian Journal of Tropical Medicine and Public Health, 2011Co-Authors: Kasem Somthana, Paron Dekumyoy, Jitra Waikagul, Yuki Eshita, Ratchanok Kumsisri, Thareerat Kalambaheti, Yaowapa ManeeratAbstract:A 24 kDa protein from advanced third stage Gnathostoma Spinigerum larvae (GsAL3) is used for gnathostomiasis serodiagnosis. This study investigated whether partially purified protein antigen (Ag) from GsAL3 (Gnath Ag), prepared by simple gel filtration chromatography, could be used for serodiagnosis. Using DNA microarray analysis, significant gene expression related to immunoreactivity was examined in peripheral blood mononuclear cells (PBMC) cocultured with Gnath Ag. Antigenicity was then determined by its capacity to induce antibody production among purified naive B cells stimulated with Gnath Ag and anti- CD40. Seven and 14 days post-exposure, immunoglobulin levels (Igs) in culture supernatants were determined by enzyme-linked immunosorbent assay. The Gnath Ag stimulated PBMC had a significant increase in gene expression related to an innate immune response and decreased cell mediated immunity, but the expression of gene related antibody production was not markedly increased. The Gnath Ag stimulated naive B cells or lipopolysaccharide primed B cells to produce low levels of specific antibody. Our findings support the assertion that partially purified Gnath Ag possess low antigenicity for Ig induction. Further studies are needed to improve G. Spinigerum larva Ag for serodiagnosis.
Penchom Janwan - One of the best experts on this subject based on the ideXlab platform.
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molecular phylogenetic confirmation of Gnathostoma Spinigerum owen 1836 nematoda Gnathostomatidae in laos and thailand
Folia Parasitologica, 2016Co-Authors: Jurairat Jongthawin, Penchom Janwan, Pewpan M Intapan, Oranuch Sanpool, Tongjit Thanchomnang, Lakkhana Sadaow, Sakhone Laymanivong, Wanchai MaleewongAbstract:We report the molecular-phylogenetic identification of larvae of the nematode genus Gnathostoma Owen, 1836 collected from a snake, Ptyas koros Schlegel, in Laos and adult worms from the stomach of a dog in Thailand. DNA was extracted and amplified targeting the partial cox1 gene and the ITS-2 region of ribosomal DNA. Phylogenetic analyses indicated that all five advanced third-stage larvae and seven adult worms were Gnathostoma Spinigerum Owen, 1836. This is also the first molecular evidence of infection with G. Spinigerum in a snake from Laos.
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proteomic analysis identification of antigenic proteins in Gnathostoma Spinigerum larvae
Experimental Parasitology, 2015Co-Authors: Penchom Janwan, Pewpan M Intapan, Tongjit Thanchomnang, Chaisiri Wongkham, Porntip Laummaunwai, Rutchanee Rodpai, Tonkla Insawang, Oranuch SanpoolAbstract:Abstract Gnathostoma Spinigerum is the causative agent of human gnathostomiasis. The advanced third stage larva (AL3) of this nematode can migrate into the subcutaneous tissues, including vital organs, often producing severe pathological effects. This study performed immuno-proteomic analysis of antigenic spots, derived from G. Spinigerum advanced third stage larva (GSAL3) and recognized by human gnathostomiasis sera, using two-dimensional (2-DE) gel electrophoresis based-liquid chromatography/tandem mass spectrometry (LC/MS–MS), and followed by the aid of a database search. The crude GSAL3 extract was fractionated using IPG strips (pH 3-11NL) and followed by SDS-PAGE in the second dimension. Each gel was stained with colloidal Coomassie blue or was electro-transferred onto a nitrocellulose membrane and probed with gnathostomiasis human sera by immunoblotting. Individual Coomassie-stained protein spots corresponding to the antigenic spots recognized by immunoblotting were excised and processed using LC/MS–MS. Of the 93 antigenic spots excised, 87 were identified by LC/MS–MS. Twenty-seven protein types were found, the most abundant being Ascaris suum37. Six spots showed good quality spectra, but could not be identified. This appears to be the first attempt to characterize antigenic proteins from GSAL3 using a proteomic approach. Immuno-proteomics shows promise to assist the search for candidate proteins for diagnosis and vaccine/drug design and may provide better understand of the host-parasite relationship in human gnathostomiasis.
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a recombinant matrix metalloproteinase protein from Gnathostoma Spinigerum for serodiagnosis of neurognathostomiasis
Korean Journal of Parasitology, 2013Co-Authors: Penchom Janwan, Pewpan M Intapan, Chaisiri Wongkham, Porntip Laummaunwai, Viraphong Lulitanond, Chatchai Tayapiwatana, Hiroshi Yamasaki, Kittisak Sawanyawisuth, Amnat Kitkhuandee, Yukifumi NawaAbstract:Neurognathostomiasis is a severe form of human gnathostomiasis which can lead to disease and death. Diagnosis of neurognathostomiasis is made presumptively by using clinical manifestations. Immunoblotting, which recognizes antigenic components of molecular mass 21 kDa and 24 kDa in larval extracts of Gnathostoma Spinigerum (Gs 21/24), has high sensitivity and specificity for diagnosis of neurognathostomiasis. However, only very small amounts of the Gs 21/24 antigens can be prepared from parasites harvested from natural or experimental animals. To overcome this problem, we recently produced a recombinant matrix metalloproteinase (rMMP) protein from G. Spinigerum. In this study, we evaluated this rMMP alongside the Gs 21/24 antigens for serodiagnosis of human neurognathostomiasis. We studied sera from 40 patients from Srinagarind Hospital, Khon Kaen University, Thailand, with clinical criteria consistent with those of neurognathostomiasis, and sera from 30 healthy control adults from Thailand. All sera were tested for specific IgG antibodies against both G. Spinigerum crude larval extract and rMMP protein using immunoblot analysis. The sensitivity and specificity for both antigenic preparations were all 100%. These results show that G. Spinigerum rMMP protein can be used as an alternative diagnostic antigen, in place of larval extract, for serodiagnosis of neurognathostomiasis.
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detection of Gnathostoma Spinigerum antibodies in sera of non traumatic subarachnoid hemorrhage patients in thailand
Korean Journal of Parasitology, 2013Co-Authors: Amnat Kitkhuandee, Penchom Janwan, Wanchai Maleewong, Kittisak Sawanyawisuth, Waranon Munkong, Pewpan M IntapanAbstract:Gnathostoma Spinigerum can cause subarachnoid hemorrhage (SAH). The detection of specific antibodies in serum against G. Spinigerum antigen is helpful for diagnosis of neurognathostomiasis. There is limited data on the frequency of G. Spinigerum infection in non-traumatic SAH. A series of patients diagnosed as non-traumatic SAH at the Srinagarind Hospital, Khon Kaen University, Thailand between January 2011 and January 2013 were studied. CT or MR imaging of the brain was used for diagnosis of SAH. Patients were categorized as aneurysmal subarachnoid hemorrhage (A-SAH) or non-aneurysmal subarachnoid hemorrhage (NA-SAH) according to the results of cerebral angiograms. The presence of specific antibodies in serum against 21- or 24-kDa G. Spinigerum antigen was determined using the immunoblot technique. The detection rate of antibodies was compared between the 2 groups. Of the 118 non-traumatic SAH patients for whom cerebral angiogram and immunoblot data were available, 80 (67.8%) patients had A-SAH, whereas 38 (32.2%) had NA-SAH. Overall, 23.7% were positive for specific antibodies against 21- and/or 24-kDa G. Spinigerum antigen. No significant differences were found in the positive rate of specific antibodies against G. Spinigerum in both groups (P-value=0.350).
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modulation of antibody responses against Gnathostoma Spinigerum in mice immunized with crude antigen formulated in cpg oligonucleotide and montanide isa720
Korean Journal of Parasitology, 2013Co-Authors: Pewpan M Intapan, Penchom Janwan, Chakrit Hirunpetcharat, Churairat Kularbkaew, Wiboonchai Yutanawiboonchai, Wanchai MaleewongAbstract:This study aimed to investigate the antibody responses in mice immunized with Gnathostoma Spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. Spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. Spinigerum infection potently induces a Th2-type biased response.
Surachet Benjathummarak - One of the best experts on this subject based on the ideXlab platform.
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Effects of Gnathostoma Spinigerum infective stage larva excretory-secretory products on NK cells in peripheral blood mononuclear cell culture: focused on expressions of IFN-γ and killer cell lectin-like receptors
Parasitology Research, 2020Co-Authors: Thanyathon Khetsuphan, Paron Dekumyoy, Surachet Benjathummarak, Wilanee Dechkhajorn, Urai Chaisri, Sumate Ampawong, Parnpen Viriyavejakul, Yaowapa ManeeratAbstract:Human gnathostomiasis is mainly caused by third-stage larvae of Gnathostoma Spinigerum ( G. Spinigerum L3). Excretory-secretory products (ES) released from infective helminthic larvae are associated with larval migration and host immunity modulation. Natural killer (NK) cells have important immune functions against helminth infection. Currently, the effects of ES from G. Spinigerum L3 ( G. Spinigerum ES) on NK cell activity are unclear. This study investigated whether G. Spinigerum ES affected human NK cells. Human normal peripheral blood mononuclear cell (PBMC) cultures were used to mimic immune cells within the circulation. PBMC were co-cultured with G. Spinigerum ES (0.01–0.05 μg/ml) for 5 or 7 days. Levels of IFN-γ in cultured supernatants were measured by enzyme-linked immunosorbent assay. The expressions of mRNA encoding NK cell receptors, especially the C type killer cell lectin-like family (KLR; NKG2A, NKG2C, and NKG2D) and IFN-γ in ES induced PBMC were determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). ES induced PBMC markedly decreased the levels of IFN-γ and increased the expressions of NKG2A and NKG2D on NK cells. In conclusion, low amounts of G. Spinigerum ES modulated NK cells by downregulating the transcription of IFN-γ and upregulating the expressions of KLR (NKG2A and NKG2D receptors) during the 7-day observation period. These findings indicate more in-depth studies of NK cell function are required to better understand the mechanism involved in immune evasive strategies of human gnathostomiasis.
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Excretory-secretory product of third-stage Gnathostoma Spinigerum larvae induces apoptosis in human peripheral blood mononuclear cells
Parasitology Research, 2017Co-Authors: Nareerat Viseshakul, Supaporn Nuamtanong, Surachet Benjathummarak, Wilanee Dechkhajorn, Yaowapa ManeeratAbstract:Human gnathostomiasis caused by third-stage Gnathostoma Spinigerum larvae ( G. Spinigerum L3) is an important zoonotic disease in tropical areas of the world. The excretory-secretory products (ES) that are excreted by infective larva play a significant role in host immune evasion and tissue destruction. To investigate the poorly understood mechanisms of G. Spinigerum L3 pathogenesis, we focused on the potential effect of ES on inducing apoptosis in human immune cells by using human peripheral blood mononuclear cells (PBMCs) as a model. Early and late apoptosis of PBMCs were assessed following the exposure of these cells to G. Spinigerum L3 ES (0.1, 0.5, and 1.0 μg/ml) for 6–48 h. The apoptotic cells were identified by flow cytometric staining of PBMC with FITC-annexin V and propidium iodide. The expression of regulatory genes related to apoptosis mechanisms in ES-treated PBMCs was investigated using a Human Apoptosis RT^2 Profiler™ PCR Array. The results showed significant levels of early phase apoptosis at 18 h and of late phase apoptosis at 24 h. We speculate that this apoptosis in PBMCs occurs via the extrinsic pathway. Apoptosis in the ES-induced PBMCs was observed as quickly as 90 min after exposure, and the highest effect was observed at 18–24 h. Furthermore, ES can trigger apoptosis lasting for 48 h. Our findings expand the understanding of one of the mechanisms involved, immune-evasive strategy mechanism used by G. Spinigerum larvae during human gnathostomiasis.
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third stage Gnathostoma Spinigerum larva excretory secretory antigens modulate function of fc gamma receptor i mediated monocytes in peripheral blood mononuclear cell culture
Tropical Medicine and Health, 2016Co-Authors: Surachet Benjathummarak, Jitra Waikagul, Supaporn Nuamtanong, Thareerat Kalambaheti, Ratchanok Kumsiri, Nareerat Viseshakul, Yaowapa ManeeratAbstract:Background Third (infective)-stage Gnathostoma Spinigerum larvae (L3) mainly cause human gnathostomiasis. G. Spinigerum L3 migrate throughout the subcutaneous tissues, vital organs, and central nervous system and can cause various pathogenesis including sudden death. Interestingly, G. Spinigerum L3 can survive and evade host cellular immunity for months or years. The effects of G. Spinigerum excretory-secretory (ES) products involved in larval migration and immune-evasive strategies are unknown. Monocytes are innate immune cells that act as phagocytic and antigen-presenting cells and also play roles against helminthic infections via a complex interplay between other immune cells. Fc gamma receptor I (FcγRI) is a high-affinity receptor that is particularly expressed on monocytes, macrophages, and dendritic cells. The cross-linking of FcγRI and antigen-antibody complex initiates signal transduction cascades in phagocytosis, cytokine production, and antibody-dependent cell-mediated cytotoxicity (ADCC). This study investigated whether ES antigen (ESA) from G. Spinigerum L3 affects monocyte functions.
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Third-stage Gnathostoma Spinigerum larva excretory secretory antigens modulate function of Fc gamma receptor I-mediated monocytes in peripheral blood mononuclear cell culture
Tropical Medicine and Health, 2016Co-Authors: Surachet Benjathummarak, Jitra Waikagul, Supaporn Nuamtanong, Thareerat Kalambaheti, Ratchanok Kumsiri, Nareerat Viseshakul, Yaowapa ManeeratAbstract:Background Third (infective)-stage Gnathostoma Spinigerum larvae (L3) mainly cause human gnathostomiasis. G. Spinigerum L3 migrate throughout the subcutaneous tissues, vital organs, and central nervous system and can cause various pathogenesis including sudden death. Interestingly, G. Spinigerum L3 can survive and evade host cellular immunity for months or years. The effects of G. Spinigerum excretory-secretory (ES) products involved in larval migration and immune-evasive strategies are unknown. Monocytes are innate immune cells that act as phagocytic and antigen-presenting cells and also play roles against helminthic infections via a complex interplay between other immune cells. Fc gamma receptor I (FcγRI) is a high-affinity receptor that is particularly expressed on monocytes, macrophages, and dendritic cells. The cross-linking of FcγRI and antigen-antibody complex initiates signal transduction cascades in phagocytosis, cytokine production, and antibody-dependent cell-mediated cytotoxicity (ADCC). This study investigated whether ES antigen (ESA) from G. Spinigerum L3 affects monocyte functions. Results Cultures of normal peripheral blood mononuclear cells (PBMC) separated from healthy buffy coats were used as a human immune cell model. ESA was prepared from G. Spinigerum L3 culture. Using Real-Time quantitative reverse transcription-polymerase chain reaction (qRT-PCR), the effect of ESA to down-regulate FcγRI mRNA expression in monocytes during 90 min of observation was not well delineated. Flow cytometry analysis revealed a significant phenotypic-decreased FcγRI expression on the monocyte surface at 12 hours (h) of cultivation with the ESA ( p = 0.033). Significantly reduced monocyte-mediated phagocytosis capacity was consistently observed after 12 h of ESA pretreatment ( p = 0.001). Conclusions Our results suggest that G. Spinigerum ESA modulates monocyte function via depletion of FcγRI expression. This study provides preliminary information for future in-depth studies to elucidate mechanisms of the immune-evasive strategy of G. Spinigerum larvae.