Love or conflict: A qualitative study of the human-long tailed macaque interface in Nakhon Sawan Province, Thailand.

A wide range of zoonotic pathogens can be transmitted during human-wildlife interactions. Few qualitative studies have been conducted on human-nonhuman primate interfaces in Thailand, notably direct and indirect contact. Since Long-tailed macaques (LTMs) are prevalent in Thailand's Banphot Phisai district, part of Nakhon Sawan province, this qualitative study was conducted in 2019 to determine in-depth contact characteristics between humans and LTMs in the communities. Key informant interviews (KIIs) and focus group discussions (FGDs) were conducted with 35 villagers who reported close contact with LTMs in this study location. The results showed that villagers had different levels of contact with LTMs, depending on their occupations, perceptions, beliefs, religions, previous experiences, and local regulations. Monks in temples and vendors selling food for LTMs were reported to have the closest contact with them. LTMs have been reported to destroy personal property, houses, buildings, and crops. However, the villagers do not hurt them due to their religious beliefs relating to a respected abbot (a man who headed an abbey of monks). Even community members have had extensive interaction with LTMs, but they lacked awareness and information regarding diseases transmitted to humans directly or indirectly by non-human primates. Therefore, individuals who have frequent and close contact with LTMs should be provided health education, and appropriate behavioral change communication interventions should be performed. Furthermore, the results could be used to develop future disease prevention strategies and public awareness campaigns in the area.

Rodent-Human Interface: Behavioral Risk Factors and Leptospirosis in a Province in the Central Region of Thailand.

This sequential explanatory mixed-method study consisted of analytical, cross-sectional, and qualitative studies. The research was conducted in the Khao Nor and Khao Kaew areas of the Banphot Pisai districts of Nakhon Sawan Province in 2019. Here, we examined the rodent contact characteristics of villagers in these areas and determined the potential characteristics/risk factors associated with rodents using a semi-structured questionnaire, key informant interview (KII), and focus group discussion (FGD). Results of the quantitative study (N1 = 372) characterized participants that contacted rodents per gender, age, occupation, knowledge, attitude, and practice (KAP), including their cultural contexts, and beliefs. Ninety participants (24.2%) reported contact with rodents, and the reasons for their direct physical rodent contact were hunting (35, 9.4%), killing (41, 11.0%), preparing rodents as food (33, 8.9%), consuming cooked meats (12, 3.2%), feeding food (4, 1.1%), cleaning feces (17, 4.6%), and cleaning carcasses (33, 8.9%). Moreover, logistic regression results showed that males encountering rodents were statistically significant (Adjusted OR = 3.137, 95% CI 1.914-5.139, p < 0.001). Low monthly household income (

Evaluation of a genus-specific rGroEL1-524 IgM-ELISA and commercial ELISA kits during the course of leptospirosis in Thailand.

In the present study, we developed a genus-specific rGroEL1-524 IgM-ELISA assay for use in screening diagnosis of suspected leptospirosis among acute undifferentiated febrile illness patients during acute fever. The diagnostic accuracies of the rGroEL1-524 IgM-ELISA, commercial Panbio IgM-ELISA, and Virion-Serion Classic IgG-ELISA were evaluated using 133 Thai leptospirosis sera and 210 controls. Sensitivities were 91.7%, 59.6%, and 17.7% for acute infection, and the specificities were 92.6%, 90.2%, and 88.3% for the non-leptospirosis control, respectively. The rGroEL1-524 IgM-ELISA had high sensitivity, at 92.3% and 91.7%, among culture-positive and MAT-negative cases at 1-3 days post-onset of symptoms (DPO1-3), respectively. Impaired specificity on scrub typhus was found, possibly from antibody cross-reaction to ortholog GroEL. Commercial Panbio IgM-ELISA had sensitivities at DPO1-3 of 30.8% and 41.7% for culture-positive and MAT-negative cases whereas Virion-Serion IgG-ELISA showed sensitivities of 5.9% and 13.3%, respectively. The rGroEL1-524 IgM-ELISA could be useful as a screening test for early diagnosis. The performance of the commercial ELISA suggests the applicability of IgM-ELISA for diagnosis, while IgG-ELISA is useful for seroprevalence surveys. However, confirmation by reference tests is recommended.

Immunodominance of LipL3293-272 peptides revealed by leptospirosis sera and therapeutic monoclonal antibodies.

BACKGROUND/PURPOSE: Leptospirosis is a neglected zoonosis, imposing significant human and veterinary public health burdens. In this study, recombinant LipL3293-147 and LipL32148-184 middle domain of LipL3293-184, and LipL32171-214, and LipL32215-272 of c-terminal LipL32171-272 truncations were defined for immunodominance of the molecule during Leptospira infections revealed by leptospirosis sera. RESULTS: IgM-dominant was directed to highly surface accessible LipL32148-184 and Lipl32171-214. IgG dominance of LipL32148-184 revealed by rabbit anti-Leptospira sera and convalescent leptospirosis paired sera were mapped to highly accessible surface of middle LipL32148-184 truncation whereas two LipL32148-184 and LipL32215-272 truncations were IgG-dominant when revealed by single leptospirosis sera. The IgM-dominant of LipL32148-214 and IgG-dominant LipL32148-184 peptides have highly conserved amino acids of 70% identity among pathogenic and intermediate Leptospira species and were mapped to the highly surface accessible area of LipL32 molecule that mediated interaction of host components. IgG dominance of two therapeutic epitopes located at LipL32243-253 and LipL32122-130 of mAbLPF1 and mAbLPF2, respectively has been shown less IgG-dominant (<30%), located outside IgG-dominant regions characterized by leptospirosis paired sera. CONCLUSION: The IgM- and IgG-dominant LipL32 could be further perspectives for immunodominant LipL32-based serodiagnosis and LipL32 epitope-based vaccine.

Hamster IFN-γ+CD4+ and IL-4+CD4+ T cell responses against leptospires are significantly higher than those of mice.

BACKGROUND: Leptospirosis is a bacterial disease caused by the Leptospira interrogans. The hamster is considered a susceptible host while the mouse is resistant. The knowledge of hamster T cell immunity is limited compared to the mouse. The reason why the hamster and the mouse give different responses to leptospires remains unclear. OBJECTIVE: To determine the differential responses of CD4+ T cells between hamsters and mice using Leptospira interrogans as an infectious model. METHODS: The CD4+ T-cell reactivity and their intracellular cytokine responses after infection with live L.interrogans serovar Autumnalis or leptospiral antigens, or injection with recombinant LipL32 protein (rLipL32) were elucidated. For secondary immune responses, mononuclear cells were re-stimulated with leptospiral crude antigens (LAg) or rLipL32. Intracellular cytokines and CD4+ T cells were determined using flow cytometry. RESULTS: There were no significant differences between the percentages of hamster and mouse CD4+ and CD25+CD4+ T cell responses to live bacteria. Mouse CD4+ (24.50±1.98%) and CD25+CD4+ T cells (3.83±0.88) responded significantly higher than those of hamster (15.07±2.82% and 2.00±0.37%) when infected and re-stimulated with LAg. The numbers of IFN-γ and IL-4 producing cells in hamsters at 1.76±0.10% and 0.82±0.25% for IFN-γ+CD4+ and IL-4+CD4+ T cells were significantly higher than those in resistant mice at 0.10±0.02% and 0.23±0.03% for IFN-γ+CD4+ and IL-4+CD4+ T cells. CONCLUSION: Hamsters responded significantly higher in secondary stimulation especially in the levels of the IFN-γ+ and IL-4+CD4+ T cells. The mechanisms of this dissimilarity remain to be elucidated.

Solenopsis geminata (tropical fire ant) anaphylaxis among Thai patients: its allergens and specific IgE-reactivity.

BACKGROUND: Specific IgE against Solenopsis invicta (imported fire ant) remains the current diagnostic tool for allergy to ants worldwide. However, S. invicta may not be the only cause of ant anaphylaxis in Thai patients. OBJECTIVE: To characterize ant species causing anaphylaxis in Thai patients and to test allergenic reactivity to whole body extracts (WBE) of S. geminata (tropical fire ants) in patients with evidence of IgE-mediated ant anaphylaxis. METHODS: Thirty-two patients with ant anaphylaxis were identified. The causative ants collected by the patients were subjected to species identification. Twelve patients with ant anaphylaxis and showed positive skin test or serum specific IgE to S. invicta and 14 control subjects were recruited. Whole body extraction from S. geminata was performed for protein characterization using SDS-PAGE and protein staining. IgE-immunoblotting and ELISA-specific IgE binding assays were performed on patients' sera and compared with controls. RESULTS: Of 32 patients with ant anaphylaxis, the most common causative ant identified was S. geminata (37.5%). Western blot analysis of crude S. geminata revealed 13 refined protein components that bound to patients' serum IgE. Three major allergens with molecular masses of 26, 55 and 75 kDa were identified. All 12 patients gave positive results for specific IgE to S. geminata with statistically significant higher absorbance units of 0.390 ± 0.044, compared to healthy control group (0.121 ± 0.010), P < 0.01. CONCLUSIONS: S. geminata is identified as the most common causative ant anaphylaxis in Thai patients. Its WBE comprises of 13 IgE-binding components and 3 major allergens (26, 55 and 75 kDa), which supported possible IgE-mediated mechanism.

Disruption of endocytic trafficking protein Rab7 impairs invasiveness of cholangiocarcinoma cells.

BACKGROUND: Alterations and mutations of endo-lysosomal trafficking proteins have been associated with cancer progression. Identification and characterization of endo-lysosomal trafficking proteins in invasive cholangiocarcinoma (CCA) cells may benefit prognosis and drug design for CCA. OBJECTIVE: To identify and characterize endo-lysosomal trafficking proteins in invasive CCA. METHODS: A lysosomal-enriched fraction was isolated from a TNF-α induced invasive CCA cell line (KKU-100) and uninduced control cells and protein identification was performed with nano-LC MS/MS. Novel lysosomal proteins that were upregulated in invasive CCA cells were validated by real-time RT-PCR. We selected Rab7 for further studies of protein level using western blotting and subcellular localization using immunofluorescence. The role of Rab7 in CCA invasion was determined by siRNA gene knockdown and matrigel transwell assay. RESULTS: Rab7 mRNA and protein were upregulated in invasive CCA cells compared with non-treated controls. Immunofluorescence studies demonstrated that Rab7 was expressed predominantly in invasive CCA cells and was localized in the cytoplasm and lysosomes. Suppression of Rab7 translation significantly inhibited TNF-α-induced cell invasion compared to non-treated control (p= 0.044). CONCLUSIONS: Overexpression of Rab7 in CCA cells was associated with cell invasion, supporting Rab7 as a novel candidate for the development of diagnostic and therapeutic strategies for CCA.

Molecular characterization of serine protease inhibitor isoform 3, SmSPI, from Schistosoma mansoni.

Serine protease inhibitors, known as serpins, are pleiotropic regulators of endogenous and exogenous proteases, and molecule transporters. They have been documented in animals, plants, fungi, bacteria, and viruses; here, we characterize a serpin from the trematode platyhelminth Schistosoma mansoni. At least eight serpins have been found in the genome of S. mansoni, but only two have characterized molecular properties and functions. Here, the function of S. mansoni serpin isoform 3 (SmSPI) was analyzed, using both computational and molecular biological approaches. Phylogenetic analysis showed that SmSPI was closely related to Schistosoma haematobium serpin and Schistosoma japonicum serpin B10. Structure determined in silico confirmed that SmSPI belonged to the serpin superfamily, containing nine α-helices, three ß-sheets, and a reactive central loop. SmSPI was highly expressed in schistosomules, predominantly in the head gland, and in adult male and female with intensive accumulation on the spines, which suggests that it may have a role in facilitating intradermal and intravenous survival. Recombinant SmSPI was overexpressed in Escherichia coli; the recombinant protein was of the same size (46 kDa) as the native protein. Immunological analysis suggested that mice infected with S. mansoni responded to rSmSPI at 8 weeks postinfection (wpi) but not earlier. The inhibitory activity of rSmSPI was specific to chymotrypsin but not trypsin, neutrophil elastase, and porcine pancreatic elastase. Elucidating the biological and physiological functions of SmSPI as well as other serpins will lead to further understanding of host-parasite interaction machinery that may provide novel strategies to prevent and control schistosomiasis in the future.

Regulation of airway eosinophil and neutrophil infiltration by alpha-galactosylceramide in a mouse model for respiratory syncytial virus (RSV) vaccine-augmented disease.

Respiratory syncytial virus (RSV) is a leading cause of respiratory disease among infants, the elderly and immunocompromised adults. In this study, we assessed the effects of alpha-galactosylceramide, a known immunoregulatory lipid, on liposomal RSV vaccine-induced responses in BALB/c mice subsequently challenged with RSV. Liposomes containing a recombinant fragment of the RSV G protein were prepared with and without alpha-galactosylceramide and used to immunize mice by the intranasal route. The inclusion of alpha-galactosylceramide in the liposomal formulation caused a dramatic reduction in bronchoalveolar lavage neutrophils, but also an increase in eosinophils, following subsequent RSV challenge. The reduction in neutrophils was specific to mice receiving alpha-galactosylceramide-containing liposomes and was not reproduced in mice administered liposomes containing another alpha-galactosyl lipid, alpha-galactosylphosphatidylglyceroylalkylamine. Lung IL-13 mRNA levels were particularly elevated in mice administered alpha-galactosylceramide-containing liposomes followed by RSV challenge. This study demonstrates a striking ability of alpha-galactosylceramide to modulate the cellular airway infiltrate in mice immunized with liposomal RSV vaccine followed by RSV challenge.