Japanese Encephalitis Vaccine Research Articles

Efficacy and Safety of a Tetravalent Dengue Vaccine (TAK-003) in Children With Prior Japanese Encephalitis or Yellow Fever Vaccination.

We explored the impact of prior Yellow fever (YF) or Japanese encephalitis (JE) vaccination on the efficacy of Takeda's dengue vaccine candidate, TAK-003 (NCT02747927). Children 4-16 years of age were randomized 2:1 to receive TAK-003 or placebo and were under active febrile surveillance. Symptomatic dengue was confirmed by serotype-specific RT-PCR. YF and JE vaccination history was recorded. Of the 20,071 children who received TAK-003 or placebo, 21.1% had a YF and 23.9% had a JE vaccination history at randomization. Fifty-seven months after vaccination, vaccine efficacy was 55.7% (95% CI, 39.7%-67.5%) in those with YF vaccination, 77.8% (70.8%-83.1%) for JE vaccination, and 53.5% (45.4%-60.4%) for no prior YF/JE vaccination. Regional differences in serotype distribution confound these results. The apparent higher vaccine efficacy in the JE vaccination subgroup could be largely explained by serotype-specific efficacy of TAK-003. Within 28 days of any vaccination, the proportions of participants with serious adverse events in the YF/JE prior vaccination population were comparable between the TAK-003 and placebo groups. The available data do not suggest a clinically relevant impact of prior JE or YF vaccination on TAK-003 performance. Overall, TAK-003 was well-tolerated and efficacious in different epidemiological settings.

Awareness About Child Immunization among Women in Rural Area of District Jalaun

Introduction; Universal Immunization Programme (UIP) is one of the largest public health programmes targeting close of 2.67 crore newborns and 2.9 crore pregnant women annually. It is one of the most cost-effective public health interventions and largely responsible for reduction of vaccine preventable under-5 mortality rate. Under UIP, immunization is providing free of cost against 12 vaccine preventable diseases. Mission Indra dhanush (MI) was launched in December 2014 and aims at increasing the full immunization coverage to children to 90 % (1). Mother play a very important role in child immunization, so awareness and correct knowledge about immunization is very essential in women, keeping this thing in mind, the present study was undertaken. Aims and objectives;1. To assess the awareness about child immunization among women in rural area.2. To know the correct knowledge of vaccine given in children among women .Material and methods;A cross sectional study was carried out in a rural area of district Jalaun. Aata block was selected randomly for study area. RHTC Aata was selected and women attending the OPD during month of August to October were included in study. Sample size were calculated and 140 mothers were interviewed by using predesigned and pretested questionnaire after taking verbal consent and data was collected and analyzed using appropriate statistical tool. Results; mostly women had heard about immunization in children (82.1%) and most of the women got information from health workers like ASHA/ANM followed by family and friends. vaccine given to child that shows knowledge about BCG/POLIO/MMR was found good but average about DPT and HEPATITIS. Very few women had knowledge about HIB, JE and ROTA vaccine. Conclusion; need to focus on education and awareness programme on women regarding child immunization in rural areas.

A Mathematical modelling for co-infection dynamics of Japanese encephalitis-Dengue and influence of JE Vaccine on Dengue disease

A non-linear deterministic mathematical model has been described with the co-infection dynamics of Japanese encephalitis(JE) and dengue disease, incorporating the JE vaccine. A basic reproduction number is discussed to study transmission potential of the co-infection model. In co-infection model, disease-free equilibrium points of Japanese encephalitis and dengue, along with endemic, are presented in system and investigated their stability with the help of their specified method. Our analysis suggests that vaccination against Japanese encephalitis positively affect control of co-infection. By using Center Manifold Theory, the model undergoes backwards bifurcation phenomenon and this has been occurred when basic reproduction number is smaller than unity. It is shown that by taking simultaneous preventive steps, the basic reproduction number of co-infection can be reduced to less than one after eliminating both infections. Sensitivity analysis has been performed to determine which parameters significantly affect disease dynamics. The effects of these parameters on transmission of disease were investigated using a numerical simulation. According to the findings, we obtain that JE-dengue co-infection can be managed with use of JE vaccine, also minimize JE transmission rate rapidly.

Investigating Nonspecific Effects of the Live-Attenuated Japanese Encephalitis Vaccine on Lower Respiratory Tract Infections in Children Aged 25-35 Months: Retrospective Cohort Study.

Live attenuated vaccines may be used to prevent nontargeted diseases such as lower respiratory tract infections (LRTIs) due to their nonspecific effects (NSEs). We aimed to analyze the NSEs of the Japanese encephalitis vaccine on pediatric LRTIs in children aged 25 months to 35 months. A retrospective cohort study was conducted by using a population-based electronic health record database in Zhejiang, China. Enrolled participants were children born from January 1, 2017, to December 31, 2017, and who were inoculated with the live-attenuated Japanese encephalitis vaccine (JE-L) or inactivated Japanese encephalitis vaccine (JE-I) as the most recent vaccine at 24 months of age. The study was carried out between January 1, 2019, and December 31, 2019. All inpatient and outpatient hospital visits for LRTIs among children aged 25 months to 35 months were recorded. The Andersen-Gill model was used to assess the NSEs of JE-L against LRTIs in children and compared with those of JE-I as the most recent vaccine. A total of 810 children born in 2017 were enrolled, of whom 585 received JE-L (JE-L cohort) and 225 received JE-I (JE-I cohort) as their last vaccine. The JE-L cohort showed a reduced risk of LRTIs (adjusted hazard ratio [aHR] 0.537, 95% CI 0.416-0.693), including pneumonia (aHR 0.501, 95% CI 0.393-0.638) and acute bronchitis (aHR 0.525, 95% CI 0.396-0.698) at 25 months to 35 months of age. The NSEs provided by JE-L were especially pronounced in female children (aHR 0.305, 95% CI 0.198-0.469) and children without chronic diseases (aHR 0.553, 95% CI 0.420-0.729), without siblings (aHR 0.361, 95% CI 0.255-0.511), with more than 30 inpatient and outpatient hospital visits prior to 24 months of age (aHR 0.163, 95% CI 0.091-0.290), or with 5 to 10 inpatient and outpatient hospital visits due to infectious diseases prior to 24 months old (aHR 0.058, 95% CI 0.017-0.202). Compared with JE-I, receiving JE-L as the most recent vaccine was associated with lower risk of inpatient and outpatient hospital visits for LRTIs among children aged 25 months to 35 months. The nature of NSEs induced by JE-L should be considered for policymakers and physicians when recommending JE vaccines to those at high risk of infection from the Japanese encephalitis virus.

Hesitancy towards Japanese Encephalitis vaccine and its socio-demographic correlates among parents attending to children aged

Problem consideredJapanese encephalitis (JE) is a mosquito-borne disease with a vaccine as the main preventive strategy. Vaccine hesitancy among parents can pose a threat to the success of child vaccination programmes. ObjectivesTo determine the level of awareness regarding the transmission, treatment and sequelae of JE, assess the levels and determining factors of vaccine hesitancy and acceptance among parents accompanying their children aged <15 years to the study hospitals. MethodsThis hospital-based cross-sectional study was conducted among 204 parents of children aged <15 years from February to May 2023. Data were collected using a pretested validated proforma and analysed using Jamovi software 2.3. Chi-square test and Binary Logistic Regression analyses were used. ResultsAmong, the 204 study participants (mean age 29.6 (±3.65) years), 73% (n = 129) were aged </ = 30 years, 71.1% (n = 145) were aware of the presence of JE in the study area. Awareness about transmission and consequences was less than 30%. Overall vaccine hesitancy was 84.3% (n = 172); it was higher among those >30 years (85.3% versus 83.7% among </ = 30 years, with >2 children (90% versus 84.0% among those having </ = 2 children). With overall 72.5% vaccine acceptance, participants aged ≤30 years (OR = 2.08 (95% CI: 1.01–4.34; p = 0.047) and those aware of JE cases in the area (OR 27.76 (95% CI, 12.67–65.53), p < 0001), had higher vaccine acceptance rate. ConclusionThe participants had high level of awareness about JE cases being reported; lack of knowledge regarding the JE transmission, treatment, and consequences; high JE vaccine hesitancy and low acceptance rate.

Mass immunisation to eradicate Japanese encephalitis: Real-world evidence from Guizhou Province in 2005–2021

ObjectivesTo explore epidemiological changes of Japanese encephalitis (JE) in a long-time span and evaluate the impact of mass immunisation. MethodData on JE cases from hospitals and the county Centers for Disease Control and Prevention in Guizhou Province was collected between 2005 and 2021. Epidemiological changes were analyzed according to a series of policy implementations and the coronavirus disease 2019 (COVID-19) pandemic. ResultsA total of 5138 JE cases and 152 deaths were reported in Guizhou Province during 2005–2021. The average incidence and case fatality rates were 0.83/100,000 and 2.96%, respectively. The JE prevalence showed a declining trend over the years with the reduced incidence gap between age groups and narrowing of the high-epidemic regions. During the COVID-19 pandemic, the JE activity reached its nadir in 2020. The inclusion in the Expanded Program on Immunization of the JE vaccine and catch-up immunisations showed a significant impact on the JE declining incidence rate. ConclusionsThe implementation of JE immunisation programs has played a crucial role in controlling its spread. Continued efforts should be made to maintain high coverage of the JE vaccine and strengthen disease surveillance systems, ensuring JE effective control and eventual elimination.

A novel alternative for pyrogen detection based on a transgenic cell line

Pyrogen, often as a contaminant, is a key indicator affecting the safety of almost all parenteral drugs (including biologicals, chemicals, traditional Chinese medicines and medical devices). It has become a goal to completely replace the in vivo rabbit pyrogen test by using the in vitro pyrogen test based on the promoted ‘reduction, replacement and refinement’ principle, which has been highly considered by regulatory agencies from different countries. We used NF-κB, a central signalling molecule mediating inflammatory responses, as a pyrogenic marker and the monocyte line THP-1 transfected with a luciferase reporter gene regulated by NF-κB as an in vitro model to detect pyrogens by measuring the intensity of a fluorescence signal. Here, we show that this test can quantitatively and sensitively detect endotoxin (lipopolysaccharide from different strains) and nonendotoxin (lipoteichoic acid, zymosan, peptidoglycan, lectin and glucan), has good stability in terms of NF-κB activity and cell phenotypes at 39 cell passages and can be applied to detect pyrogens in biologicals (group A & C meningococcal polysaccharide vaccine; basiliximab; rabies vaccine (Vero cells) for human use, freeze-dried; Japanese encephalitis vaccine (Vero cells), inactivated; insulin aspart injection; human albumin; recombinant human erythropoietin injection (CHO Cell)). The within-laboratory reproducibility of the test in three independent laboratories was 85%, 80% and 80% and the interlaboratory reproducibility among laboratories was 83.3%, 95.6% and 86.7%. The sensitivity (true positive rate) and specificity (true negative rate) of the test were 89.9% and 90.9%, respectively. In summary, the test provides a novel alternative for pyrogen detection.

COVID-19 and Japanese Encephalitis Vaccination: Occurrence of Disease in Area with and without Mandatory Vaccination Policies

COVID-19 and Japanese Encephalitis Vaccination: Occurrence of Disease in Area with and without Mandatory Vaccination Policies

Japanese Encephalitis: Risk of Emergence in the United States and the Resulting Impact.

Japanese encephalitis virus is a mosquito-borne member of the Flaviviridae family. JEV is the leading cause of viral encephalitis in Asia and is characterized by encephalitis, high lethality, and neurological sequelae in survivors. The virus also causes severe disease in swine, which are an amplifying host in the transmission cycle, and in horses. US agricultural authorities have recently recognized the threat to the swine industry and initiated preparedness activities. Other mosquito-borne viruses exotic to the Western Hemisphere have been introduced and established in recent years, including West Nile, Zika, and chikungunya viruses, and JEV has recently invaded continental Australia for the first time. These events amply illustrate the potential threat of JEV to US health security. Susceptible indigenous mosquito vectors, birds, feral and domestic pigs, and possibly bats, constitute the receptive ecological ingredients for the spread of JEV in the US. Fortunately, unlike the other virus invaders mentioned above, an inactivated whole virus JE vaccine (IXIARO®) has been approved by the US Food and Drug Administration for human use in advance of a public health emergency, but there is no veterinary vaccine. This paper describes the risks and potential consequences of the introduction of JEV into the US, the need to integrate planning for such an event in public health policy, and the requirement for additional countermeasures, including antiviral drugs and an improved single dose vaccine that elicits durable immunity in both humans and livestock.

Shift in dominant genotypes of Japanese encephalitis virus and its impact on current vaccination strategies.

Japanese encephalitis (JE) is a zoonotic ailment from the Japanese encephalitis virus (JEV). JEV belongs to the flavivirus genus and is categorized into a solitary serotype consisting of five genetically diverse genotypes (I, II, III, IV, and V). The JEV genotype III (GIII) was the prevailing strain responsible for multiple outbreaks in countries endemic to JEV until 1990. In recent years, significant improvements have occurred in the epidemiology of JE, encompassing the geographical expansion of the epidemic zone and the displacement of prevailing genotypes. The dominant genotype of the JEV has undergone a progressive shift from GIII to GI due to variations in its adaptability within avian populations. From 2021 to 2022, Australia encountered an epidemic of viral encephalitis resulting from infection with the GIV JEV pathogen. The current human viral encephalitis caused by GIV JEV is the initial outbreak since its initial discovery in Indonesia during the late 1970s. Furthermore, following a time frame of 50 years, the detection and isolation of GV JEV have been reported in Culex mosquitoes across China and South Korea. Evidence suggests that the prevalence of GIV and GV JEV epidemic regions may be on the rise, posing a significant threat to public safety and the sustainable growth of animal husbandry. The global approach to preventing and managing JE predominantly revolves around utilizing the GIII strain vaccine for vaccination purposes. Nevertheless, research has demonstrated that the antibodies generated by the GIII strain vaccine exhibit limited capacity to neutralize the GI and GV strains. Consequently, these antibodies cannot protect against JEV challenge caused by animal GI and GV strains. The limited cross-protective and neutralizing effects observed between various genotypes may be attributed to the low homology of the E protein with other genotypes. In addition, due to the GIV JEV outbreak in Australia, further experiments are needed to evaluate the protective efficiency of the current GIII based JE vaccine against GIV JEV. The alteration of the prevailing genotype of JEV and the subsequent enlargement of the geographical extent of the epidemic have presented novel obstacles in JE prevention and control. This paper examines the emerging features of the JE epidemic in recent years and the associated problems concerning prevention and control.

New Vaccine Introductions in WHO African Region between 2000 and 2022.

Significant progress has been made in vaccine development worldwide. This study examined the WHO African Region's vaccine introduction trends from 2000 to 2022, excluding COVID-19 vaccines. We extracted data on vaccine introductions from the WHO/UNICEF joint reporting form for 17 vaccines. We examined the frequency and percentages of vaccine introductions from 2000 to 2022, as well as between two specific time periods (2000-2010 and 2011-2022). We analysed Gavi eligible and ineligible countries separately and used a Chi-squared test to determine if vaccine introductions differed significantly. Three vaccines have been introduced in all 47 countries within the region: hepatitis B (HepB), Haemophilus influenzae type b (Hib), and inactivated polio vaccine (IPV). Between 2011 and 2022, HepB, Hib, IPV, the second dose of measles-containing vaccine (MCV2), and pneumococcal conjugate vaccine (PCV) were the five most frequently introduced vaccines. Hepatitis A vaccine has only been introduced in Mauritius, while Japanese encephalitis vaccine has not been introduced in any African country. Between 2000-2010 and 2011-2022, a statistically significant rise in the number of vaccine introductions was noted (p < 0.001) with a significant positive association between Gavi eligibility and vaccine introductions (p < 0.001). Significant progress has been made in the introduction of new vaccines between 2000 and 2022 in the WHO African Region, with notable introductions between 2011 and 2022. Commitments from countries, and establishing the infrastructure required for effective implementation, remain crucial.

The reemerging and outbreak of genotypes 4 and 5 of Japanese encephalitis virus.

The Japanese encephalitis virus (JEV) is classified into five distinct genotypes, with genotypes 1 and 3 historically showing higher activity. These genotypes are the primary agents of viral encephalitis in the Asian continent. Genotypes 4 and 5 have remained silent in low-latitude tropical regions since their discovery. From 2009, the hidden genotype 5 suddenly emerged simultaneously in mosquitoes from the Tibetan region of China and those from South Korea in East Asia. The detection of genotype 5 of JEV in these mosquitoes was associated with cases of viral encephalitis in the local population. Similarly, in 2022, the long-silent genotype 4 of JEV emerged in Australia, resulting in a local outbreak of viral encephalitis that primarily affected adults and caused fatalities. The emergence and outbreaks of genotypes 4 and 5 of JEV present new challenges for the prevention and control of Japanese encephalitis (JE). This study not only analyzes the recent emergence of these new genotypes but also discusses their implications in the development of JE vaccines and laboratory tests for newly emerging JEV infections.

Live and Inactivated Japanese Encephalitis Vaccine Interchangeability in Travelers: Review of Evidence and Recommendation to Travelers

Live and Inactivated Japanese Encephalitis Vaccine Interchangeability in Travelers: Review of Evidence and Recommendation to Travelers

Japanese Encephalitis Vaccine Acceptance and Strategies for Travelers: Insights from a Scoping Review and Practitioners in Endemic Countries.

Japanese encephalitis (JE) remains the cause of vaccine-preventable encephalitis in individuals living in endemic areas and international travelers. Although rare, the disease's high fatality rate emphasizes the need for effective immunization. This review aims to provide updated data on the JE burden between 2017 and 2023, vaccine acceptance, and vaccine strategies for travelers. We prospectively identified studies, using MEDLINE and PubMed, published through 2023. JE incidence has decreased in local populations and remains low among travelers from non-endemic countries. The local JE risk cannot be utilized to determine traveler risk. Adult travelers naïve to JEV infection or immunization may be at potentially higher risk. The JE vaccine acceptance rates among international travelers visiting JE endemic areas range from 0.2% to 28.5%. The cost of the vaccine and low risk perception could be barriers to JE vaccination. For travelers, an accelerated two-dose regimen of inactivated Vero cell JE vaccine (JE-VC) or a single dosage of live attenuated JE vaccine (JE-LV) may be an option. In conclusion, the JE burden among residents and travelers is lower, but the risk is not negligible. Practitioners should prioritize sharing knowledge, increasing awareness, and promoting vaccinations and preventive measures to reduce tourists' risk of JE along their journey.

Cost-effectiveness analysis of Japanese Encephalitis (JE) vaccination program in Bali Province, Indonesia

ObjectiveThe incidence of Japanese Encephalitis (JE) in Bali Province remains high, and is one among the highest in Indonesia. The Indonesian Government initiated the JE vaccination campaign followed by a JE vaccine introduction program in Bali Province in 2018. The JE vaccination program then has been fully integrated into the provincial routine immunization program since 2019. We conducted a retrospective economic analysis of JE vaccination program in Bali Province, Indonesia; considering multiple vaccination strategies. MethodsWe conducted a cost-effectiveness analysis using a decision analytic model comparing two vaccination strategies with no vaccination from the societal and government perspectives. These vaccination strategies were: (1) JE vaccination campaign and introduction program, and (2) a routine JE vaccination program. We compared costs and outcomes for three hypothetical cohorts of 100,000 children followed from birth to the age of 10 years, with impacts measured throughout the child’s life-time. We measured the economic consequences as costs per case, per death, and per disability-adjusted life year (DALY) averted. ResultsA routine JE immunization program was the most cost-effective strategy with a cost per DALYs averted of US$ 212.59 and US$ 94.09 from the government and societal perspectives respectively. In contrast, costs per DALYs averted through the JE vaccination campaign and introduction strategy was US$ 1,473.53 and US$ 1,224.20 from the government and societal perspectives respectively. ConclusionsBoth JE vaccination strategies are cost-effective but they are not cost-saving when compared to no immunization program.

Epidemiology and risk factors of Japanese encephalitis in Taiwan, 2010-2022.

Taiwan introduced a two-dose inactivated Japanese encephalitis (JE) mouse brain-derived (JE-MB) vaccine into routine childhood immunization in 1968, with booster vaccination implemented in 1974 and 1983. In 2017, JE-MB vaccine was replaced by a two-dose live-attenuated chimeric vaccine (JE-CV). After implementation of JE vaccination programs, JE cases have shifted from children to adults. In this study, we described the JE epidemiology and identify high-risk groups to further inform vaccine policy. We extracted data from Taiwan's notifiable disease surveillance database, vital statistics, and employment statistics from 2010 to 2022. Diagnosis of JE was confirmed by JE seroconversion, a four-fold increase in virus-specific antibodies, a positive JE viral nucleic-acid test, or JE virus isolation. From 2010 to 2022, a total of 313 cases of JE were diagnosed, resulting in an overall incidence rate of 0.10 cases per 100,000 person-years and a mortality rate of 0.006 per 100,000 population per year. Among these patients, 64% were male, and the median age was 51 years (range 0-82). Compared with people born in or after 1976 (vaccinated with four doses of JE-MB vaccine or two doses of JE-CV), those born in or before 1962 (unvaccinated) and those born during 1963-1975 (vaccinated with two or three doses of JE-MB vaccine) had a 4.2-fold (95% confidence interval [CI] 3.0-5.7) and 5.9-fold (95% CI 4.3-8.1) higher risk of JE, respectively. The relative risk of working in agriculture, forestry, fishing, or animal husbandry, compared to other occupations, was 5.0 (95% CI 3.5-7.0). In Taiwan, individuals born before 1976 and those employed in agriculture, forestry, fishing, or animal husbandry had a higher risk of JE. We recommend JE vaccination for people in these high-risk groups who have not been fully vaccinated or have an unknown vaccination history.

A decision support tool for risk-benefit analysis of Japanese encephalitis vaccine in travellers.

During pre-travel consultations, clinicians and travellers face the challenge of weighing the risks verus benefits of Japanese encephalitis (JE) vaccination due to the high cost of the vaccine, low incidence in travellers (~1 in 1million), but potentially severe consequences (~30% case-fatality rate). Personalised JE risk assessment based on the travellers' demographics and travel itinerary is challenging using standard risk matrices. We developed an interactive digital tool to estimate risks of JE infection and severe health outcomes under different scenarios to facilitate shared decision-making between clinicians and travellers. A Bayesian network (conditional probability) model risk-benefit analysis of JE vaccine in travellers was developed. The model considers travellers' characteristics (age, sex, co-morbidities), itinerary (destination, departure date, duration, setting of planned activities) and vaccination status to estimate the risks of JE infection, the development of symptomatic disease (meningitis, encephalitis), clinical outcomes (hospital admission, chronic neurological complications, death) and adverse events following immunization. In low-risk travellers (e.g. to urban areas for <1month), the risk of developing JE and dying is low (<1per million) irrespective of the destination; thus, the potential impact of JE vaccination in reducing the risk of clinical outcomes is limited. In high-risk travellers (e.g. to rural areas in high JE incidence destinations for >2months), the risk of developing symptomatic disease and mortality is estimated at 9.5and 1.4per million, respectively. JE vaccination in this group would significantly reduce the risk of symptomatic disease and mortality (by ~80%) to 1.9and 0.3per million, respectively. The JE tool may assist decision-making by travellers and clinicians and could increase JE vaccine uptake. The tool will be updated as additional evidence becomes available. Future work needs to evaluate the usability of the tool. The interactive, scenario-based, personalised JE vaccine risk-benefit tool is freely available on www.VaxiCal.com.

Arboviral vaccines for use in pregnant travelers

Pregnant women traveling abroad can be exposed to a variety of arboviruses, primarily spread by mosquitoes or ticks. Some arboviral infections can be of particular concern for pregnant women or their fetuses. Vaccination is one preventive measure that can reduce the risk for infection. Several arboviral vaccines have been licensed for many years and can be used to prevent infection in travelers, namely Japanese encephalitis, yellow fever, and tick-borne encephalitis vaccines. Recommendations on use of these vaccines in pregnancy vary. Other arboviral vaccines have been licensed but are not indicated for use in pregnant travelers (e.g., dengue vaccines) or are in development (e.g., chikungunya, Zika vaccines). This review describes arboviral vaccines for travelers, focusing on women who are pregnant and those planning travel during pregnancy.

Tinospora cordifolia Miers enhances the immune response in mice immunized with JEV-vaccine: A network pharmacology and experimental approach

BackgroundTinospora cordifolia Miers. (TC) (Giloya/Guduchi) is a native Indian herb, reported for its wide array of medicinal activities including immunomodulatory activity. However, the exact pharmacological mechanism of TC as an immunomodulatory agent remains unclear. Central to this, to the best of our knowledge, no study has explored the immunoadjuvant potential of TC in response to the Japanese encephalitis (JE) vaccines. PurposeThe study aims to explore the immunoadjuvant potential of TC ethanolic extract in response to the JE vaccine and illustrates its potential mechanism of immunomodulation using an integrated approach of network pharmacology and in-vivo experimental study. Study design and methodsInitially, the extract was prepared and the components of TC were identified through high-resolution liquid chromatography mass spectrometry (HR-LC/MS). The compounds were then screened for network pharmacology analysis. Next, the drug and disease targets were identified and the network was constructed using Cytoscape 3.7.2 to obtain different signalling pathways of TC in JEV. We then evaluated the immunoadjuvant potential of TC ethanolic extract in mice immunized with inactivated JE vaccine (SA-14–14–2 strain). BALB/c mice were supplemented with TC extract (30 and 100 mg/kg, i.g.), daily for 56 days, marked with immunization on 28th day of the study, by JE vaccine. Blood was collected for flow cytometry and haematological analysis (total and differential cell counts). The surface expression of immune-cell markers (CD3+, CD4+, CD19+, CD11c+, CD40+) were evaluated on day 0 (pre-immunization), day 14 and 28 post-immunization. Additionally, inflammatory cytokines (IFN-γ+/IL-17A+) were evaluated post-14 and 28 days of immunization. ResultsThe HR-LC/MS analysis identified the presence of glycosides, terpenoids, steroids and alkaloids in the TC extract. Through network analysis, 09 components and 166 targets were obtained, including pathways that involve toll-like receptor signalling, pattern-recognition receptor signalling, cytokine receptor and cytokine mediated signalling, etc. The in-vivo results showed that preconditioning with TC ethanolic extract significantly elevated the haematological variables (leucocyte count) as well as the surface expression of CD markers (B and T cell subsets) on day 0 (pre-immunization), day 14 and 28 post-immunization. Furthermore, preconditioning of TC demonstrated a dose-dependant augmentation of immune cells (CD3+, CD4+, CD19+, CD11c+) and inflammatory cytokines (IFN-γ+/IL-17A+) on day 14 and 28 post-immunization when compared to vaccine alone group. ConclusionResults showed that preconditioning with TC extract before immunization might play a potential role in enhancing the cell-mediated as well as humoral immunity. Altogether, the combinatorial approach of network pharmacology and in-vivo animal experimentation demonstrated the immunoadjuvant potential of TC in response to JEV vaccine.

Phylogeny and spatial distribution of Japanese encephalitis virus vector species in Cambodia.

In Southeast Asia, despite the use of Japanese encephalitis vaccines and vaccination coverage, Japanese encephalitis (JE) transmission is still a major public health issue. The main vectors of this virus are mosquitoes from the genus Culex, which diversity and density are important in Southeast Asia. The main vector species of Japanese encephalitis virus (JEV) in Cambodia belong to the Vishnui subgroup. However, their morphological identification solely based on the adult stage remains challenging, making their segregation and detection difficult. In order to identify and describe the distribution of the three main JEV vector species in Cambodia, namely Culex vishnui, Cx. pseudovishnui and Cx. tritaeniorhynchus, mosquito samplings were carried out throughout the country in different environments. Phylogenetic analysis of the cytochrome c oxidase subunit I (coI) gene using maximum-likelihood tree with ultrafast bootstrap and phylogeographic analysis were performed. The three main Culex species are phylogenetically separated, and represent two distinct clades, one with Cx. tritaeniorhynchus and the second with Cx. vishnui and Cx. pseudovishnui, the latter appearing as a subgroup of Cx. vishnui. The phylogeographic analysis shows a distribution of the Vishnui subgroup on the entire Cambodian territory with an overlapped distribution areas leading to a sympatric distribution of these species. The three JEV vector species are geographically well-defined with a strong presence of Cx. pseudovishnui in the forest. Combined with the presence of Cx. tritaeniorhynchus and Cx. vishnui in rural, peri-urban, and urban areas, the presence of JEV-competent vectors is widespread in Cambodia.