Search results for: influenza epidemic dynamics

Authors: Hsiang-Yu Yuan, Marc Baguelin, Kin O. Kwok, Nimalan Arinaminpathy, Edwin Leeuwen, Steven Riley

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Traditional syndromic surveillance for influenza has substantial public health value in characterizing epidemics. Because the relationship between syndromic incidence and the true infection events can vary from one population to another and from one year to another, recent studies rely on combining serological test results with syndromic data from traditional surveillance into epidemic models to make inference on epidemiological processes of influenza. However, despite the widespread availability of serological data, epidemic models have thus far not explicitly represented antibody titre levels and their correspondence with immunity. Most studies use dichotomized data with a threshold (Typically, a titre of 1:40 was used) to define individuals as likely recently infected and likely immune and further estimate the cumulative incidence. Underestimation of Influenza attack rate could be resulted from the dichotomized data. In order to improve the use of serosurveillance data, here, a refinement of the concept of the stratified immunity within an epidemic model for influenza transmission was proposed, such that all individual antibody titre levels were enumerated explicitly and mapped onto a variable scale of susceptibility in different age groups. Haemagglutination inhibition titres from 523 individuals and 465 individuals during pre- and post-pandemic phase of the 2009 pandemic in Hong Kong were collected. The model was fitted to serological data in age-structured population using Bayesian framework and was able to reproduce key features of the epidemics. The effects of age-specific antibody boosting and protection were explored in greater detail. RB was defined to be the effective reproductive number in the presence of stratified immunity and its temporal dynamics was compared to the traditional epidemic model using use dichotomized seropositivity data. Deviance Information Criterion (DIC) was used to measure the fitness of the model to serological data with different mechanisms of the serological response. The results demonstrated that the differential antibody response with age was present (ΔDIC = -7.0). The age-specific mixing patterns with children specific transmissibility, rather than pre-existing immunity, was most likely to explain the high serological attack rates in children and low serological attack rates in elderly (ΔDIC = -38.5). Our results suggested that the disease dynamics and herd immunity of a population could be described more accurately for influenza when the distribution of immunity was explicitly represented, rather than relying only on the dichotomous states 'susceptible' and 'immune' defined by the threshold titre (1:40) (ΔDIC = -11.5). During the outbreak, RB declined slowly from 1.22[1.16-1.28] in the first four months after 1st May. RB dropped rapidly below to 1 during September and October, which was consistent to the observed epidemic peak time in the late September. One of the most important challenges for infectious disease control is to monitor disease transmissibility in real time with statistics such as the effective reproduction number. Once early estimates of antibody boosting and protection are obtained, disease dynamics can be reconstructed, which are valuable for infectious disease prevention and control.

Keywords: effective reproductive number, epidemic model, influenza epidemic dynamics, stratified immunity

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Authors: Dina A. Amu, Fifi N. Afifah, Catur Rosidati, Tirton Nefianto

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Indonesia was shaken up by the avian influenza cases that had caused the country suffered losses of millions of dollars. The avian influenza case had even been suspected as a bioterrorism attack since it was an uncommon case in epidemiology. Furthermore, this avian influenza virus is a high pathogenic one and Indonesia has the highest case of fatality rate in the world. Bioterrorism threats or epidemic potential disease outbreaks currently does not exist in Tanjung Priok port yet. However, the surveillance system enhancement on epidemic potential diseases should be taken as a prevention, especially because Indonesia is currently facing the ASEAN Economic Society (AES). Therefore, this research evaluates the health surveillance system which is organized by Control, Quarantine and Surveillance Department, Health Office of Tanjung Priok Port. This study uses qualitative-evaluative method which utilizes Urgency Seriousness Growth (USG) method to determine priority issues and Root Cause analysis to determine the cause of prior problem. The result of this research shows that the implementation of epidemic potential disease surveillance in Tanjung Priok port has not done in the best possible way. It is because the lack of time allocation and the succinctness of the check list of ship's environmental health inspection. Therefore, Health Ministry of Indonesia should recruit more employees at the health office of Tanjung Priok port, hold a simulation of ship's inspection and simplify the list for ship's environmental health inspection.

Keywords: surveillance, epidemic potential disease, port health, bioterrorism

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Authors: Nazish Badar

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Background: To investigate the epidemic resurgent wave of influenza A (H1N1) pdm09 infections during 2015-16 Influenza season(Nov,15 –Feb,16) we compared epidemiological features of influenza A (H1N1) pdm09 associated hospitalizations and deaths during this period in Pakistan. Methods: Respiratory samples were tested using CDC Real-Time RT-PCR protocols. Demographic and epidemiological data was analyzed using SPSS. Risk ratio was calculated between age groups to compare patients that were hospitalized and died due to influenza A (H1N1) pdm09 during this period. Results: A total of 1970 specimens were analyzed; influenza virus was detected in 494(25%) samples, including 458(93%) Influenza type A and 36(7%) influenza type B viruses. Amongst influenza A viruses, 351(77%) A(H1N1) pdm09 and 107(23%) were A/H3N2. Influenza A(H1N1)pdm09 peaked in January 2016 when 250(54%) of tested patients were positive. The resurgent waves increased hospitalizations due to pdmH1N1 as compared to the rest part of the year. Overall 267(76%) A(H1N1) pdm09 cases were hospitalized. Adults ≥18 years showed the highest relative risk of hospitalization (1.2). Median interval of hospitalization and symptom onset was five days for all age groups. During this period, a total of 34 laboratory-confirmed deaths associated with pandemic influenza A (H1N1) were reported out of 1970 cases, the case fatality rate was 1.72%. the male to female ratio was 2:1in reported deaths. The majority of the deaths during that period occurred in adults ≥18 years of age. Overall median age of the death cases was 42.8 years with underlying medical conditions. The median number of days between symptom onset was two days. The diagnosis upon admission in influenza-associated fatal cases was pneumonia (53%). Acute Respiratory Distress Syndrome 9 (26%), eight out of which (88%) required mechanical ventilation. Conclusions: The present resurgence of pandemic virus cannot be attributed to a single factor. The prolong cold and dry weather, possibility of drift in virus and absence of annual flu vaccination may have played an integrated role in resurfacing of pandemic virus.

Keywords: influenza A (H1N1)pdm 09, resurgence, epidemiology, Pakistan

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Authors: Sadia Arshad, Ayesha Sohail, Sana Javed, Khadija Maqbool, Salma Kanwal

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The dynamical study of the carriers play an essential role in the evolution and global transmission of infectious diseases and will be discussed in this study. To make this approach novel, we will consider the fractional order model which is generalization of integer order derivative to an arbitrary number. Since the integration involved is non local therefore this property of fractional operator is very useful to study epidemic model for infectious diseases. An extended numerical method (ODE solver) is implemented on the model equations and we will present the simulations of the model for different values of fractional order to study the effect of carriers on transmission dynamics. Global dynamics of fractional model are established by using the reproduction number.

Keywords: Fractional differential equation, Numerical simulations, epidemic model, transmission dynamics

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Authors: Aneta Nitsch-Osuch, Katarzyna Zycinska, Ewa Gyrczuk, Agnieszka Topczewska-Cabanek, Kazimierz Wardyn

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Introduction: Influenza is an important clinical and epidemiological problem and should be considered as a possible nosocomial infection. The aim of the study was to determine the influenza vaccine coverage rates among Polish nurses and to find out drivers and barriers for influenza vaccination among this group of health care workers (HCWs). Material and methods: The self- fulfilled survey with 26 questions about the knowledge, perception, and influenza coverage rates was distributed among 461 nurses. Results: Only 15% of nurses were vaccinated against influenza in the consecutive seasons. The majority (75%) of the regularly vaccinated nurses were ambulatory careworkers. The difference between the number of vaccinated hospitals and ambulatory care nurses was statistically significant (p < 0.05). The main motivating factors for an influenza vaccination were: a fear of the illness and its complications (97%) and a free of charge vaccine available at the workplace (87%). Ambulatory care nurses more often declared that they were vaccinated mainly to protect themselves while hospital care nurses more often declared the will to protect their patients, these differences in the perception and attitudes to an influenza vaccination among hospital and ambulatory care nurses were statistically significant (p < 0.05). The main barriers for an influenza vaccination among the nursing staff were: a lack of reimbursement of the vaccine (95%), a lack of insufficient knowledge about the effectiveness, and safety of the influenza vaccine (54%). The ambulatory care nurses more often found influenza vaccination as the ethical duty compared to hospital care nurses (p < 0.05). Conclusions: The influenza vaccine coverage rates among the Polish nurses are low and must be improved in the future. More educational activities dedicated to HCWs may result in the increased awareness of influenza vaccination benefits for both medical professionals and patients.

Keywords: influenza, vaccination, nurses, ambulatory careworkers

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Authors: Henry Memczak, Marc Hovestaedt, Bernhard Ay, Sandra Saenger, Thorsten Wolff, Frank F. Bier

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The influenza viruses cause flu epidemics every year and serious pandemics in larger time intervals. The only cost-effective protection against influenza is vaccination. Due to rapid mutation continuously new subtypes appear, what requires annual reimmunization. For a correct vaccination recommendation, the circulating influenza strains had to be detected promptly and exactly and characterized due to their antigenic properties. During the flu season 2016/17, a wrong vaccination recommendation has been given because of the great time interval between identification of the relevant influenza vaccine strains and outbreak of the flu epidemic during the following winter. Due to such recurring incidents of vaccine mismatches, there is a great need to speed up the process chain from identifying the right vaccine strains to their administration. The monitoring of subtypes as part of this process chain is carried out by national reference laboratories within the WHO Global Influenza Surveillance and Response System (GISRS). To this end, thousands of viruses from patient samples (e.g., throat smears) are isolated and analyzed each year. Currently, this analysis involves complex and time-intensive (several weeks) animal experiments to produce specific hyperimmune sera in ferrets, which are necessary for the determination of the antigen profiles of circulating virus strains. These tests also bear difficulties in standardization and reproducibility, which restricts the significance of the results. To replace this test a peptide-based assay for influenza virus subtyping from corresponding virus samples was developed. The differentiation of the viruses takes place by a set of specifically designed peptidic recognition molecules which interact differently with the different influenza virus subtypes. The differentiation of influenza subtypes is performed by pattern recognition guided by machine learning algorithms, without any animal experiments. Synthetic peptides are immobilized in multiplex format on various platforms (e.g., 96-well microtiter plate, microarray). Afterwards, the viruses are incubated and analyzed comparing different signaling mechanisms and a variety of assay conditions. Differentiation of a range of influenza subtypes, including H1N1, H3N2, H5N1, as well as fine differentiation of single strains within these subtypes is possible using the peptide-based subtyping platform. Thereby, the platform could be capable of replacing the current antigenic characterization of influenza strains using ferret hyperimmune sera.

Keywords: antigenic characterization, influenza-binding peptides, influenza subtyping, influenza surveillance

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Authors: Lei Zhou, Dan Li, Ruiqi Ren, Chao Li, Yali Wang, Daxin Ni, Zijian Feng, Timothy M. Uyeki, Qun Li

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Since the first human infections with avian influenza A(H7N9) virus were identified in early 2013, 1533 cases of laboratory-confirmed A(H7N9) virus infections were reported and confirmed as of September 13, 2017. The fifth epidemic was defined as starting from September 1, 2016, and the number of A(H7N9) cases has surged since the end of December in 2016. On February 18, 2017, the A(H7N9) cases who were infected with highly pathogenic avian influenza (HPAI) virus was reported from Southern China. The HPAI A(H7N9) cases were identified and then an investigation and analyses were conducted to assess whether disease severity in humans has changed with HPAI A(H7N9) compared with low pathogenic avian influenza (LPAI) A(H7N9) virus infection. Methods: All confirmed cases with A(H7N9) virus infections reported throughout mainland China from September 1, 2016, to September 13, 2017, were included. Cases' information was extracted from field investigation reports and the notifiable infectious surveillance system to describe the demographic, clinical, and epidemiologic characteristics. Descriptive statistics were used to compare HPAI A(H7N9) cases with all LPAI A(H7N9) cases reported during the fifth epidemic. Results: A total of 27 cases of HPAI A(H7N9) virus were identified infection from five provinces, including Guangxi (44%), Guangdong (33%), Hunan (15%), Hebei (4%) and Shangxi (4%). The median age of cases of HPAI A(H7N9) virus infection was 60 years (range, 15 to 80) and most of them were male (59%) and lived in rural areas (78%). All 27 cases had live poultry related exposures within 10 days before their illness onset. In comparison with LPAI A(H7N9) case-patients, HPAI A(H7N9) case-patients were significantly more likely to live in rural areas (78% vs. 51%; p = 0.006), have exposure to the sick or dead poultry (56% vs. 19%; p = 0.000), and be hospitalized earlier (median 3 vs. 4 days; p = 0.007). No significant differences were observed in median age, sex, prevalence of underlying chronic medical conditions, median time from illness onset to first medical service seeking, starting antiviral treatment, and diagnosis. Although the median time from illness onset to death (9 vs. 13 days) was shorter and the overall case-fatality proportion (48% vs. 38%) was higher for HPAI A(H7N9) case-patients than for LPAI A(H7N9) case-patients, these differences were not statistically significant. Conclusions: Our findings indicate that HPAI A(H7N9) virus infection was associated with exposure to sick and dead poultry in rural areas when visited live poultry market or in the backyard. In the fifth epidemic in mainland China, HPAI A (H7N9) case-patients were hospitalized earlier than LPAI A(H7N9) case-patients. Although the difference was not statistically significant, the mortality of HPAI A (H7N9) case-patients was obviously higher than that of LPAI A(H7N9) case-patients, indicating a potential severity change of HPAI A(H7N9) virus infection.

Keywords: Avian influenza A (H7N9) virus, highly pathogenic avian influenza (HPAI), case-patients, poultry

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Authors: Maria Ines Azambuja

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Graphic displays of Age-Period-Cohort (APC) mortality trends generally uses data aggregated within 5 or 10-year intervals. Technology allows one to increase the amount of processed data. Displaying occurrences by 1-year intervals is a logic first step in the direction of attaining higher quality landscapes of variations in temporal occurrences. Method: 1) Comparison of UK mortality trends plotted by 10-, 5- and 1-year intervals; 2) Comparison of UK and US mortality trends (period X age and cohort X age) displayed by 1-year intervals. Source: Mortality data (period, 1x1, males, 1933-1912) uploaded from the Human Mortality Database to Excel files, where Period X Age and Cohort X Age graphics were produced. The choice of transforming age-specific trends from calendar to birth-cohort years (cohort = period – age) (instead of using cohort 1x1 data available at the HMD resource) was taken to facilitate the comparison of age-specific trends when looking across calendar-years and birth-cohorts. Yearly live births, males, 1933 to 1912 (UK) were uploaded from the HFD. Influenza references are from the literature. Results: 1) The use of 1-year intervals unveiled previously unsuspected period, cohort and interacting period x cohort effects upon all-causes mortality. 2) The UK and US figures showed variations associated with particular calendar years (1936, 1940, 1951, 1957-68, 72) and, most surprisingly, with particular birth-cohorts (1889-90 in the US, and 1900, 1918-19, 1940-41 and 1946-47, in both countries. Also, the figures showed ups and downs in age-specific trends initiated at particular birth-cohorts (1900, 1918-19 and 1947-48) or a particular calendar-year (1968, 1972, 1977-78 in the US), variations at times restricted to just a range of ages (cohort x period interacting effects). Importantly, most of the identified “scars” (period and cohort) correlates with the record of occurrences of Influenza A epidemics since the late 19th Century. Conclusions: The use of 1-year intervals to describe APC mortality trends both increases the amount of information available, thus enhancing the opportunities for patterns’ recognition, and increases our capability of interpreting those patterns by describing trends across smaller intervals of time (period or birth-cohort). The US and the UK mortality landscapes share many but not all 'scars' and distortions suggested here to be associated with influenza epidemics. Different size-effects of wars are evident, both in mortality and in fertility. But it would also be realistic to suppose that the preponderant influenza A viruses circulating in UK and US at the beginning of the 20th Century might be different and the difference to have intergenerational long-term consequences. Compared with the live births trend (UK data), birth-cohort scars clearly depend on birth-cohort sizes relatives to neighbor ones, which, if causally associated with influenza, would result from influenza-related fetal outcomes/selection. Fetal selection could introduce continuing modifications on population patterns of immune-inflammatory phenotypes that might give rise to 'epidemic constitutions' favoring the occurrence of particular diseases. Comparative analysis of mortality landscapes may help us to straight our record of past circulation of Influenza viruses and document associations between influenza recycling and fertility changes.

Keywords: age-period-cohort trends, epidemic constitution, fertility, influenza, mortality

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Authors: Nadia Nisar

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Background: In the temperate regions Influenza activities occur sporadically all year round with peaks coinciding during cold months. Meteorological and environmental conditions play significant role in the transmission of influenza globally. In this study, we assessed the relationship between meteorological parameters and influenza activity in two geographical areas of Pakistan. Methods: Influenza data were collected from Islamabad (north) and Multan (south) regions of national influenza surveillance system during 2010-2015. Meteorological database was obtained from National Climatic Data Center (Pakistan). Logistic regression model with a stepwise approach was used to explore the relationship between meteorological parameters with influenza peaks. In statistical model, we used the weekly proportion of laboratory-confirmed influenza positive samples to represent Influenza activity with metrological parameters as the covariates (temperature, humidity and precipitation). We also evaluate the link between environmental conditions associated with seasonal influenza epidemics: 'cold-dry' and 'humid-rainy'. Results: We found that temperature and humidity was positively associated with influenza in north and south both locations (OR = 0.927 (0.88-0.97)) & (OR = 0.1.078 (1.027-1.132)) and (OR = 1.023 (1.008-1.037)) & (OR = 0.978 (0.964-0.992)) respectively, whilst precipitation was negatively associated with influenza (OR = 1.054 (1.039-1.070)) & (OR = 0.949 (0.935-0.963)). In both regions, temperature and humidity had the highest contribution to the model as compared to the precipitation. We revealed that the p-value for all of climate parameters is <0.05 by Independent-sample t-test. These results demonstrate that there were significant relationships between climate factors and influenza infection with correlation coefficients: 0.52-0.90. The total contribution of these three climatic variables accounted for 89.04%. The reported number of influenza cases increased sharply during the cold-dry season (i.e., winter) when humidity and temperature are at minimal levels. Conclusion: Our findings showed that measures of temperature, humidity and cold-dry season (winter) can be used as indicators to forecast influenza infections. Therefore integrating meteorological parameters for influenza forecasting in the surveillance system may benefit the public health efforts in reducing the burden of seasonal influenza. More studies are necessary to understand the role of these parameters in the viral transmission and host susceptibility process.

Keywords: influenza, climate, metrological, environmental

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Authors: Sasiporn Rattanasupha, Settapat Chinviriyasit

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The treatment function adopts a continuous and differentiable function which can describe the effect of delayed treatment when the number of infected individuals increases and the medical condition is limited. In this paper, the SEIR epidemic model with treatment function is studied to investigate the dynamics of the model due to the effect of treatment. It is assumed that the treatment rate is proportional to the number of infective patients. The stability of the model is analyzed. The model is simulated to illustrate the analytical results and to investigate the effects of treatment on the spread of infection.

Keywords: basic reproduction number, local stability, SEIR epidemic model, treatment function

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Authors: Ines Cherif, Ghassen Kharroubi, Leila Bouabid, Adel Gharbi, Aicha Boukthir, Margaret Mccarron, Nissaf Ben Alaya, Afif Ben Salah, Jihene Bettaieb

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Healthcare workers' flu vaccination prevents influenza disease among both patients and caregivers. We aimed in this study to assess influenza vaccine (IV) coverage in 2018-2019 among Tunisian physicians and to determine factors associated with IV receipt. A cross sectional study was carried out in Tunisian primary and secondary health care facilities in the 2018-2019 influenza season. Physicians with direct patient contact were recruited according to a self-weighted multistage sampling. Data were collected through a face to face questionnaire containing questions on knowledge, attitudes, and practices regarding IV. Bivariate analysis was used in order to determine factors associated with IV receipt. A total of 167 physicians were included in the study with a mean age of 48.2 ± 7.7 years and a sex-ratio (M: F) of 0.37. Among participants, 15.1% (95% CI: [9.7%-20.3%]) were vaccinated against influenza in the 2018-2019 influenza season. Bivariate analysis revealed that previous flu immunization in the four years preceding the 2018-2019 influenza season (OR=32.3; p < 10-3), belief that vaccinating healthcare workers may reduce work absenteeism (OR=4.7, p=0.028), belief that flu vaccine should be mandatory to healthcare workers (OR=3.3, p=0.01) and high confidence towards IV efficacy in preventing influenza among caregivers (OR= 4.5, p=0.01) were associated with a higher IV receipt in 2018-2019 among physicians. Less than one fifth of Tunisian physicians were vaccinated against influenza in 2018-2019. Higher vaccine uptake was related to a higher belief in vaccine efficacy in preventing influenza disease among both patients and caregivers. This underscores the need for periodic educational campaigns to raise physicians' awareness about IV efficacy. The switch to an IV mandatory policy should also be considered.

Keywords: influenza vaccine, physicians, Tunisia, vaccination uptake

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Authors: N. G. Klivleyeva, T. I. Glebova, G. V. Lukmanova, S. B. Bayseit, S. Z. Taubaeva, M. K. Kalkozhaeva

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The role of viral diseases in the general infectious disease incidence increases every year and requires special attention to the problem of interpreting the etiology of infectious agents. Influenza and acute respiratory viral infections are one of the most pressing public health issues. In the period 2012-2014, collection of 419 nasal swabs and 150 blood sera has been carried out in the patient care institutions of the various Kazakhstan regions from patients with symptoms of ARVI and pneumonia. Primary identification of biosamples for the presence of influenza viral antigens in enzyme immunoassay on nitrocellulose membrane gave positive results in 125 swabs (29.8%). Biosample screening in immunofluorescence test revealed the presence of influenza viral antigens against A/H1 in 63 samples (15.0%), A/H3 – in 70 samples (16.7%) and type B – in 9 samples (2.1%). As a result of primary infection, and successive passages in chick embryos and MDCK cell cultures, 38 HAAg were isolated from 419 samples with a clear cytopathic effect and hemagglutination titre in MDCK cell culture within 1:2-1:4, in CE - 1:8-1:256. The infectivity of isolates in chicken embryos were 3.5-6.5 lg EID50/0.2, in MDCK cell culture – 2.5-6.5 lg PFU/ml. Identification of 28 isolates was carried out in inhibition reactions of hemagglutinating activity and neuraminidase activity, showed their belonging to the influenza virus: 26 strains to A/H1N1, one - to A/H3N2, and one - to type B. Serological examination of blood sera for the presence of specific antibodies being an indirect evidence of the performed isolation and contributing to the timely interpretation of the disease etiology in the epidemics takes an important place in the comprehensive study of influenza viruses circulating among people. Serological analyzes were carried out in HAI assay using a kit consisting of 12 reference strains obtained from the WHO centre for reference and research on Influenza (CDC, Atlanta, USA) and three Kazakhstan (A/Almaty/347/09 (H1N1v), A/Almaty/462/11 (H3N2) and B/Almaty/414/10) human influenza viruses that are stored in the laboratory collection. The results of serological analysis of 150 blood sera showed that antihaemagglutinins against the A/H3N2 virus serosubtype were found in 46 samples (49.4%) out of 93 sera collected in 2012-2013. The antibody titres were within 1:160-1:320. 19 sera (20.4%) were seropositive against influenza A/H1N1 virus, the antibodies were observed in titres of 1:20-1:40. Six sera (6.4%) were positive against the influenza A/H1N1+A/H3N2 virus (mixed infection); the antibodies were recorded in titres of 1:20-1:40. Antihaemagglutinins against influenza type B virus were detected only in five sera (5.4%). The results of analysis of 57 sera collected in 2014 showed that antihaemagglutinins against A/H3N2 virus subtype were detected in 32 blood sera (56.1%) in titres of 1:160-1:640. Ten sera (17.5%) were seropositive against A/H1N1 virus; antihaemagglutinins against influenza type B virus were not detected. Therefore, virological and serological studies have shown that in Kazakhstan, as well as in the world, the influenza viruses A/H1N1, A/H3N2 and influenza B viruses were actively circulating during the epidemic seasons in 2012-2014.

Keywords: influenza, MDCK cell, serological analysis, virus

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Authors: Haile Berihulay, Chenglong Luo

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Avian influenza, commonly known as bird flu, is a highly contagious viral disease primarily affecting poultry and wild birds, with significant implications for both animal health and public safety. The influenza virus (IV) is notorious for its ability to mutate and infect multiple species, including humans, leading to severe consequences. Avian influenza poses considerable pandemic risks due to the potential evolution of low pathogenic avian influenza (LPAI) into highly pathogenic avian influenza (HPAI), which can cause rapid outbreaks in domestic flocks. While AVI viruses typically do not replicate well in humans, strains such as H5N1 and H7N9 have crossed the species barrier, raising alarm over human infections. The recent documentation of human transmission of the H5N8 strain from birds underscores the ongoing threat posed by avian influenza. This review necessitates a thorough discussion about the genetic foundation of viral pathogens, identifying key candidate genes linked to disease resilience, and discussing powerful tools. This review can help researchers to comprehensively overview the disease severity and combat related to AIV, which causes significant economic impact and set effective control strategies to mitigate the risks associated with avian influenza outbreaks.

Keywords: Avian, candidate genes, chicken, molecular, pathogen, virus

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Authors: Marina Kljaković-Gašpić Batinjan, Tea Petrović, Frano Vučković, Irzal Hadžibegović, Barbara Radovani, Ivana Jurin, Lovorka Đerek, Eva Huljev, Alemka Markotić, Ivica Lukšić, Irena Trbojević-Akmačić, Gordan Lauc, Ivan Gudelj, Rok Čivljak

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Immunoglobulin G has essential role in defense against infectious diseases, but its role cannot be fully recognized without understanding of changes in its N-glycans attached to the Fc domain. We analyzed and compared total IgG glycome in plasma samples of patients with influenza, patients with COVID-19 and healthy controls. We found similarities in IgG glycosylation changes in COVID-19 survivors and influenza patients that could be the consequence of adequate immune response to enveloped viruses, while observed changes in deceased COVID-19 patients may indicate its deviation.

Keywords: COVID-19, glycosylation, immunoglobulin G, influenza, pneumonia, viral infection

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Authors: A. Ferasat, S. Rostampour Yasouri

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Seasonal flu is a highly contagious infection caused by influenza viruses. These viruses undergo genetic changes that result in new epidemics across the globe. Medical attention is crucial in severe cases, particularly for the elderly, frail, and those with chronic illnesses, as their immune systems are often weaker. The purpose of this study was to detect new subtypes of the influenza A virus rapidly using a specific RT-PCR method based on the HA gene (hemagglutinin). In the winter and spring of 2022_2023, 120 embryonated egg samples were cultured, suspected of seasonal influenza. RNA synthesis, followed by cDNA synthesis, was performed. Finally, the PCR technique was applied using a pair of specific primers designed based on the HA gene. The PCR product was identified after purification, and the nucleotide sequence of purified PCR products was compared with the sequences in the gene bank. The results showed a high similarity between the sequence of the positive samples isolated from the patients and the sequence of the new strains isolated in recent years. This RT-PCR technique is entirely specific in this study, enabling the detection and multiplication of influenza and its subspecies from clinical samples. The RT-PCR technique based on the HA gene, along with sequencing, is a fast, specific, and sensitive diagnostic method for those infected with influenza viruses and its new subtypes. Rapid molecular diagnosis of influenza is essential for suspected people to control and prevent the spread of the disease to others. It also prevents the occurrence of secondary (sometimes fatal) pneumonia that results from influenza and pathogenic bacteria. The critical role of rapid diagnosis of new strains of influenza is to prepare a drug vaccine against the latest viruses that did not exist in the community last year and are entirely new viruses.

Keywords: influenza, molecular diagnosis, patients, RT-PCR technique

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Authors: Vladimir Berezin, Aizhan Turmagambetova, Andrey Bogoyavlenskiy, Pavel Alexyuk, Madina Alexyuk, Irina Zaitceva, Nadezhda Sokolova

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Introduction: Influenza viruses could infect approximately 5% to 10% of the global human population annually, resulting in serious social and economic damage. Vaccination and etiotropic antiviral drugs are used for the prevention and treatment of influenza. Vaccination is important; however, antiviral drugs represent the second line of defense against new emerging influenza virus strains for which vaccines may be unsuccessful. However, the significant drawback of commercial synthetic anti-flu drugs is the appearance of drug-resistant influenza virus strains. Therefore, the search and development of new anti-flu drugs efficient against drug-resistant strains is an important medical problem for today. The aim of this work was a study of four phenolic acids of plant origin (Gallic, Syringic, Vanillic, and Protocatechuic acids) as a possible tool for treatment against influenza virus. Methods: Phenolic acids; gallic, syringic, vanillic, and protocatechuic have been prepared by extraction from plant tissues and purified using high-performance liquid chromatography fractionation. Avian influenza virus, strain A/Tern/South Africa/1/1961 (H5N3) and human epidemic influenza virus, strain A/Almaty/8/98 (H3N2) resistant to commercial anti-flu drugs (Rimantadine, Oseltamivir) were used for testing antiviral activity. Viruses were grown in the allantoic cavity of 10 days old chicken embryos. The chemotherapeutic index (CTI), determined as the ratio of an average toxic concentration of the tested compound (TC₅₀) to the average effective virus-inhibition concentration (EC₅₀), has been used as a criteria of specific antiviral action. Results: The results of study have shown that the structure of phenolic acids significantly affected their ability to suppress the reproduction of tested influenza virus strains. The highest antiviral activity among tested phenolic acids was detected for gallic acid, which contains three hydroxyl groups in the molecule at C3, C4, and C5 positions. Antiviral activity of gallic acid against A/H5N3 and A/H3N2 influenza virus strains was higher than antiviral activity of Oseltamivir and Rimantadine. gallic acid inhibited almost 100% of the infection activity of both tested viruses. Protocatechuic acid, which possesses 2 hydroxyl groups (C3 and C4) have shown weaker antiviral activity in comparison with gallic acid and inhibited less than 10% of virus infection activity. Syringic acid, which contains two hydroxyl groups (C3 and C5), was able to suppress up to 12% of infection activity. Substitution of two hydroxyl groups by methoxy groups resulted in the complete loss of antiviral activity. Vanillic acid, which is different from protocatechuic acid by replacing of C3 hydroxyl group to methoxy group, was able to suppress about 30% of infection activity of tested influenza viruses. Conclusion: For pronounced antiviral activity, the molecular of phenolic acid must have at least two hydroxyl groups. Replacement of hydroxyl groups to methoxy group leads to a reduction of antiviral properties. Gallic acid demonstrated high antiviral activity against influenza viruses, including Rimantadine and Oseltamivir resistant strains, and could be used as a potential candidate for the development of antiviral drug against influenza virus.

Keywords: antiviral activity, influenza virus, drug resistance, phenolic acids

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Authors: Ofosuhene O. Apenteng, Noor Azina Ismail

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Over the last several years, concern has developed over how to minimize the spread of HIV/AIDS epidemic in many countries. AIDS epidemic has tremendously stimulated the development of mathematical models of infectious diseases. The transmission dynamics of HIV infection that eventually developed AIDS has taken a pivotal role of much on building mathematical models. From the initial HIV and AIDS models introduced in the 80s, various improvements have been taken into account as how to model HIV/AIDS frameworks. In this paper, we present the impact of migration on the spread of HIV/AIDS. Epidemic model is considered by a system of nonlinear differential equations to supplement the statistical method approach. The model is calibrated using HIV incidence data from Malaysia between 1986 and 2011. Bayesian inference based on Markov Chain Monte Carlo is used to validate the model by fitting it to the data and to estimate the unknown parameters for the model. The results suggest that the migrants stay for a long time contributes to the spread of HIV. The model also indicates that susceptible individual becomes infected and moved to HIV compartment at a rate that is more significant than the removal rate from HIV compartment to AIDS compartment. The disease-free steady state is unstable since the basic reproduction number is 1.627309. This is a big concern and not a good indicator from the public heath point of view since the aim is to stabilize the epidemic at the disease equilibrium.

Keywords: epidemic model, HIV, MCMC, parameter estimation

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Authors: Shuafeng Yuan, Bojian Zheng

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The RNA-dependent RNA polymerase of influenza a virus comprises conserved and independently folded subdomains with defined functionalities. The N-terminal domain of the PA subunit (PAN) harbors the endonuclease function so that it can serve as a desired target for drug discovery. To identify a class of anti-influenza inhibitors that impedes PAN endonuclease activity, a screening approach that integrated the fluorescence resonance energy transfer based endonuclease inhibitor assay with the DNA gel-based endonuclease inhibitor assay was conducted, followed by the evaluation of antiviral efficacies and potential cytotoxicity of the primary hits in vitro and in vivo. A small-molecule compound ANA-0 was identified as a potent inhibitor against the replication of multiple subtypes of influenza A virus, including H1N1, H3N2, H5N1, H7N7, H7N9 and H9N2, in cell cultures. Combinational treatment of zanamivir and ANA-0 exerted synergistic anti-influenza effect in vitro. Intranasal administration of ANA-0 protected mice from lethal challenge and reduced lung viral loads in H1N1 virus infected BALB/c mice. Docking analyses predicted ANA-0 bound the endonuclease cavity of PAN by interacting with the metal-binding and catalytic residues. In summary, ANA-0 shows potential to be developed to novel anti-influenza agents.

Keywords: anti-influenza, novel compound, inhibition of endonuclease, PA

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Authors: Dileep K. Verma, Sunil K. Lal

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Influenza still remains one of the most challenging diseases posing significant threat to public health causing seasonal epidemics and pandemics. Previous studies suggest that influenza hemagglutinin is essential for viral attachment to host sialic acid receptors and concentrate in lipid rafts for efficient viral fusion. Studies also reported selective nature of Influenza virus to utilize rafts micro-domain for efficient virus assembly and budding. However, the detailed mechanism of Influenza A Virus (IAV) binding to host cell membrane and entry inside the host remains elusive. In the present study, we investigated if host membrane lipid rafts play any significant role in early life cycle events of influenza A virus. Role of host lipid rafts was studied using raft disruption method by extraction of cholesterol and Methyl-β-Cyclodextrin was used to remove membrane cholesterol. We observed co-localization of Influenza A Virus to lipid rafts by visualization of known lipid raft marker GM1 on host cell membrane. Co-localization suggest direct involvement of these micro-domain in initiation of IAV life cycle. We found significant reduction in influenza A virus adsorption in raft disrupted target host cells indicating poor binding and attachment in absence of coherent membrane rafts. Taken together, the results of present study provide evidence for critical involvement of host lipid rafts and its constituents in adsorption process of Influenza A Virus and suggests crucial involvement in other early events of IAV life cycle. The present study opens a new domain to study influenza virus-host interaction and to combat flu at the very early steps of viral life cycle.

Keywords: lipid raft, adsorption, cholesterol, methyl-β-cyclodextrin, GM1

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Authors: Ghazanfar Ali, Fahad N Almajhdi

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This study provides data on the viral diagnosis and molecular epidemiology of influenza A(H3N2) virus isolated in Riyadh, Saudi Arabia. Nasopharyngeal aspirates from 80 clinically infected patients in the peak of the 2010-2011 winter seasons were processed for viral diagnosis by RT-PCR. Sequencing of entire HA and NA genes of representative isolates and molecular epidemiological analysis were performed. A total of 06 patients were positive for influenza A, B and respiratory syncytial viruses by RT-PCR assays; out of these only one sample was positive for influenza A(H3N2) by RT-PCR. Phylogenetic analysis of the HA and NA gene sequences showed identities higher than 99-98.8 % in both genes. They were also similar to reference isolates in HA sequences (99 % identity) and in NA sequences (99 % identity). Amino acid sequences predicted for the HA gene were highly identical to reference strains. The NA amino acid substitutions identified did not include the oseltamivir-resistant H275Y substitution. Conclusion: Viral isolation and RT-PCR together were useful for diagnosis of the influenza A (H3N2) virus. Variations in HA and NA sequences are similar to those identified in worldwide reference isolates and no drug resistance was found.

Keywords: influenza A (H3N2), genetic characterization, viral isolation, RT-PCR, Saudi Arabia

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Authors: Ghassen Kharroubi, Ines Cherif, Leila Bouabid, Adel Gharbi, Aicha Boukthir, Margaret McCarron, Nissaf Ben Alaya, Afif Ben Salah, Jihene Bettaieb

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Influenza vaccination (IV) is recommended for elderly persons, especially those with underlying conditions. In countries where IV rates in the elderly remain unsatisfactory, exploring attitudes of older persons toward the flu vaccine could be useful to identify barriers and facilitators to IV. The aim of this study was to determine the reasons for IV acceptance or decline in the Tunisian elderly. A national cross-sectional study was conducted in 2019, among persons aged 60 years and over with chronic disease. Data were collected using a standard administered questionnaire. Of the 1191 older persons included, 19.4% received the influenza vaccine in the 2018-2019 flu season. The two main reasons that may lead to refusal of vaccination were concerns that the vaccine could cause side effects (71.5%) and a belief that the vaccine was ineffective (33.9%). The main reason that may lead to accepting vaccination was a doctor’s recommendation (41.1%). Doctors were by far the most trusted source for information regarding influenza vaccine (91.5%) followed by pharmacists (17.6%). Our results highlighted the important role that doctors could play in promoting IV among the Tunisian elderly. Physicians should correct misconceptions about adverse events and the efficiency of the vaccine. In fact, influenza vaccines are generally effective and safe among older persons.

Keywords: attitudes, influenza vaccination, older persons, Tunisia

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Authors: Attapon Kamlangdee, Brock Kingstad-Bakke, Tavis K. Anderson, Tony L. Goldberg, Jorge E. Osorio

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A critical failure in our preparedness for an influenza pandemic is the lack of a universal vaccine. Influenza virus strains diverge by 1 to 2% per year, and commercially available vaccines often do not elicit protection from one year to the next, necessitating frequent formulation changes. This represents a major challenge to the development of a cross-protective vaccine that can protect against circulating viral antigenic diversity. We have constructed a recombinant modified vaccinia virus Ankara (MVA) that expresses an H5N1 mosaic hemagglutinin (H5M) (MVA-H5M). This mosaic was generated in silico using 2,145 field-sourced H5N1 isolates. A single dose of MVA-H5M provided 100% protection in mice against clade 0, 1, and 2 avian influenza viruses and also protected against seasonal H1N1 virus (A/Puerto Rico/8/34). It also provided short-term (10 days) and long-term (6 months) protection post vaccination. Both neutralizing antibodies and antigen-specific CD4+and CD8+ T cells were still detected at 5 months post vaccination, suggesting that MVA-H5M provides long-lasting immunity.

Keywords: modified vaccinia Ankara, MVA, H5N1, hemagglutinin, influenza vaccine

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Authors: N. W. Paul, M. Banerjee

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Discussing the nexus between global health policy and local practices, this article addresses the recent Ebola outbreak as a role model for narrative co-constructions of epidemic risk. We will demonstrate in how far a theory-driven and methodologically rooted analysis of narrativity can help to improve mechanisms of prevention and intervention whenever epidemic risk needs to be addressed locally in order to contribute to global health. Analyzing the narrative transformation of Ebola, we will also address issues of transcultural problem-solving and of normative questions at stake. In this regard, we seek to contribute to a better understanding of a key question of global health and justice as well as to the underlying ethical questions. By highlighting and analyzing the functions of narratives, this paper provides a translational approach to refine our practices by which we address epidemic risk, be it on the national, the transnational or the global scale.

Keywords: ebola, epidemic risk, medical ethics, medical humanities

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Authors: Hanjun Zhao

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Limited efficacy of current antivirals and antiviral-resistant mutations impair anti-influenza treatment. Here, we evaluated the in vitro and in vivo antiviral effect of three defective interfering genes (DIG-3) of influenza virus. Virus replication was significantly reduced in 293T and A549 cells transfected with DIG-3. Mice transfected with DIG-3 encoded by jetPEI-vector, as prophylaxis and therapeutics against A(H7N7) virus respectively, had significantly better survivals (80% and 50%) than control mice (0%). We further developed a dual-functional peptide TAT-P1, which delivers DIG-3 with high transfection efficiency and concomitantly exerts antiviral activity by preventing endosomal acidification. TAT-P1/DIG-3 was more effective than jetPEI/DIG-3 in treating A(H7N7) or A(H1N1)pdm09-infected mice and showed potent prophylactic protection on A(H7N7) or A(H1N1)pdm09-infected mice. The addition of P1 peptide, preventing endosomal acidification, could enhance the protection of TAT-P1/DIG-3 on A(H1N1)pdm09-infected mice. Dual-functional TAT-P1 with DIG-3 can effectively protect or treat mice infected by avian and seasonal influenza virus infection.

Keywords: antiviral peptide, dual-functional peptide, defective interfering genes, influenza virus

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Authors: Thanh Phuong Doan, Narueporn Sutanthavibul

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The influence of freezing cycle on influenza haemagglutinin (HA) conformational stability was investigated in terms of freezing rates and formulation compositions. The results showed that appropriate HA conformation could be evaluated using circular dichroism (CD) spectroscopy with HA concentration of greater than 0.09 mg/ml. The intermediate freezing rate of approximately 1.0oC/min preserved the original HA conformation better than at slow freezing rate (0.5oC/min) and rapid freezing rate (2.6oC/min). The changes in CD spectra of the secondary HA structure were more pronounced than those of the tertiary HA structure during the evaluation. Additionally, the formulations, which resulted in the highest conformational stability were found to have sucrose present in the composition. As opposed to when only glycine was used, the stability of HA conformation was poor.

Keywords: freezing, haemagglutinin, influenza, circular dichroism

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Authors: Shuofeng Yuan, Bojian Zheng

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Assembly of the heterotrimeric polymerase complex of influenza virus from the individual subunits PB1, PA, and PB2 is a prerequisite for viral replication, in which the interaction between the N-terminal of PB1 (PB1N) and the C terminal of PA (PAC) may be a desired target for antiviral development. In this study, we first compared the feasibility of high throughput screening by enzyme-linked immunosorbent assay (ELISA) and fluorescence polarization (FP) assay. Among the two, ELISA was demonstrated to own broader dynamic range so that it was used for screening inhibitors, which blocked PA and PB1 interaction. Several binding inhibitors of PAC-PB1N were identified and subsequently tested for the antiviral efficacy. Apparently, 3-(2-chlorophenyl)-6-ethyl-7-methyl[1,2,4]triazolo[4,3-a]pyrimidin-5-ol, designated ANA-1, was found to be a strong inhibitor of PAC-PB1N interaction and act as a potent antiviral agent against the infections of multiple subtypes of influenza A virus, including H1N1, H3N2, H5N1, H7N7, H7N9 and H9N2 subtypes, in cell cultures. Intranasal administration of ANA-1 protected mice from lethal challenge and reduced lung viral loads in H1N1 virus infected BALB/c mice. Docking analyses predicted that ANA-1 bound to an allosteric site of PAC, which would cause conformational changes thereby disrupting the PAC-PB1N interaction. Overall, our study has identified a novel compound with potential to be developed as an anti-influenza drug.

Keywords: influenza, antiviral, viral polymerase, compounds

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Authors: Masoud Soltanialvar, Ali Bagherpour

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Documented cases of human infection with H9N2 avian influenza viruses, first detected in 1999 in Hong Kong and China, indicate that these viruses can be directly transmitted from birds to humans. In this study, we characterized the mutation in the Hemagglutinin (HA) genes and proteins that correlates with a shift in affinity of the Hemagglutinin (HA) protein from the “avian” type sialic receptors to the “human” type in 10 Iranian isolates. We delineated the genomes and receptor binding profile of HA gene of some field isolates and established their phylogenetic relationship to the other Asian H9N2 sub lineages. A total of 1200 tissue samples collected from 40 farms located in various states of Iran during 2008 – 2010 as part of a program to monitor Avian Influenza Viruses (AIV) infection. To determine the genetic relationship of Iranian viruses, the Hemagglutinin (HA) genes from ten isolates were amplified and sequenced (by RT-PCR method). Nucleotide sequences (orf) of the (HA) genes were used for phylogenetic tree construction. Deduced amino acid sequences showed the presence of L226 (234 in H9 numbering) in all ten Iranian isolates which indicates a preference to binding of α (2–6) sialic acid receptors, so these Iranian H9N2 viruses have the potential to infect human beings. These isolates showed high degree of homology with 2 human H9N2 isolates A/HK/1073/99, A/HK/1074/99. Phylogenetic analysis of showed that all the HA genes of the Iranian H9N2 viruses fall into a single group within a G1-like sublineage which had contributed as donor of six internal genes to H5N1 highly pathogenic avian influenza. The results of this study indicated that all Iranian viruses have the potential to emerge as highly pathogenic influenza virus, and considering the homology of these isolates with human H9N2 strains, it seems that the potential of these avian influenza isolates to infect human should not be overlooked.

Keywords: influenza virus, hemagglutinin, neuraminidase, Iran

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Authors: Nazish Badar

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In early 2009, Pandemic type A (H1N1) Influenza virus emerged globally. Since then, it has continued circulation causing considerable morbidity and mortality. The purpose of this study was to evaluate the evolutionary changes in Influenza A (H1N1) pdm09 viruses from 2009-15 and their relevance with the current vaccine viruses. Methods: Respiratory specimens were collected with influenza-like illness and Severe Acute Respiratory Illness. Samples were processed according to CDC protocol. Sequencing and phylogenetic analysis of Haemagglutinin (HA) and neuraminidase (NA) genes was carried out comparing representative isolates from Pakistan viruses. Results: Between Jan2009 - Feb 2016, 1870 (13.2%) samples were positive for influenza A out of 14086. During the pandemic period (2009–10), Influenza A/ H1N1pdm 09 was the dominant strain with 366 (45%) of total influenza positives. In the post-pandemic period (2011–2016), a total of 1066 (59.6%) cases were positive Influenza A/ H1N1pdm 09 with co-circulation of different Influenza A subtypes. Overall, the Pakistan A(H1N1) pdm09 viruses grouped in two genetic clades. Influenza A(H1N1)pdm09 viruses only ascribed to Clade 7 during the pandemic period whereas viruses belong to clade 7 (2011) and clade 6B (2015) during the post-pandemic years. Amino acid analysis of the HA gene revealed mutations at positions S220T, I338V and P100S specially associated with outbreaks in all the analyzed strains. Sequence analyses of post-pandemic A(H1N1)pdm09 viruses showed additional substitutions at antigenic sites; S179N,K180Q (SA), D185N, D239G (CA), S202A (SB) and at receptor binding sites; A13T, S200P when compared with pandemic period. Substitution at Genetic markers; A273T (69%), S200P/T (15%) and D239G (7.6%) associated with severity and E391K (69%) associated with virulence was identified in viruses isolated during 2015. Analysis of NA gene revealed outbreak markers; V106I (23%) among pandemic and N248D (100%) during post-pandemic Pakistan viruses. Additional N-Glycosylation site; HA S179N (23%), NA I23T(7.6%) and N44S (77%) in place of N386K(77%) were only found in post-pandemic viruses. All isolates showed histidine (H) at position 275 in NA indicating sensitivity to neuraminidase inhibitors. Conclusion: This study shows that the Influenza A(H1N1)pdm09 viruses from Pakistan clustered into two genetic clades, with co-circulation of some variants. Certain key substitutions in the receptor binding site and few changes indicative of virulence were also detected in post-pandemic strains. Therefore, it is imperative to continue monitoring of the viruses for early identification of potential variants of high virulence or emergence of drug-resistant variants.

Keywords: Influenza A (H1N1) pdm09, evolutionary analysis, post pandemic period, Pakistan

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Authors: Yun Wang, Ruoyu Wang, Tuo Chen, Guangxiu Liu, Guodong Chen, Wei Zhang

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Tuberculosis (TB) is a major public health problem in China. China has the world's second largest tuberculosis epidemic (after India). Xinjiang almost has the highest annual attendance rate of TB in China, and the province is also famous because of its severe dust storms. The epidemic timing starts in February and ends in July, and the dust storm mainly distribute throughout the spring and early summer, which strongly indicate a close linkage between causative agent of TB and dust storm events. However, mechanisms responsible for the observed patterns are still not clearly indentified. By comparing the information on cases of TB from Centers for Disease Control of China annual reports with dust storm atmosphere datasets, we constructed the relationship between the large scale annual occurrence of TB in Xinjiang, a Northwest province of China, and dust storm occurrence. Regional atmospheric indexes of dust storm based on surface wind speed show a clear link between population dynamics of the disease and the climate disaster: the onset of epidemics and the dust storm defined by the atmospheric index share the same mean year. This study is the first that provides a clear demonstration of connections that exist between TB epidemics and dust storm events in China. The development of this study will undoubtedly help early warning for tuberculosis epidemic onset in China and help nationwide and international public health institutions and policy makers to better control TB disease in Norwest China.

Keywords: dust storm, tuberculosis, Xinjiang province, epidemic

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Authors: Vladimir Berezin, Andrey Bogoyavlenskiy, Pavel Alexyuk, Madina Alexyuk, Aizhan Turmagambetova, Irina Zaitseva, Nadezhda Sokolova, Elmira Omirtaeva

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Introduction: Triterpene saponins of plant origin are one of the most promising candidates for elaboration of novel adjuvants. Due to the combination of immunostimulating activity and the capacity interact with amphipathic molecules with formation of highly immunogenic nanocomplexes, triterpene saponins could serve as a good adjuvant/delivery system for vaccine use. In the research presented adjuvants on the base of nanocomplexes contained triterpene saponins isolated from Glycyrrhiza glabra and Saponaria officinalis plants indigenous to Kazakhstan were elaborated for influenza vaccine use. Methods: Purified triterpene saponins 'Glabilox' and 'SO1' with low toxicity and high immunostimulatory activity were isolated from plants Glycyrrhiza glabra L. and Saponaria officinalis L. by high-performance liquid chromatography (HPLC) and identified using electrospray ionization mass spectrometry (ESI-MS). Influenza virus A/St-Petersburg/5/09 (H1N1) propagated in 9-days old chicken embryos was concentrated and purified by centrifugation in sucrose gradient. Nanocomplexes contained lipids, and triterpene saponins Glabilox or SO1 were prepared by dialysis technique. Immunostimulating activity of experimental vaccine preparations was studied in vaccination/challenge experiments in mice. Results: Humoral and cellular immune responses and protection against influenza virus infection were examined after single subcutaneous and intranasal immunization. Mice were immunized subunit influenza vaccine (HA+NA) or whole virus inactivated influenza vaccine in doses 3.0/5.0/10.0 µg antigen/animal mixed with adjuvant in dose 15.0 µg/animal. Sera were taken 14-21 days following single immunization and mice challenged by A/St-Petersburg/5/09 influenza virus in dose 100 EID₅₀. Study of experimental influenza vaccine preparations in animal immunization experiments has shown that subcutaneous and intranasal immunization with subunit influenza vaccine mixed with nanocomplexes contained Glabilox or SO1 saponins stimulated high levels of humoral immune response (IgM, IgA, IgG1, IgG2a, and IgG2b antibody) and cellular immune response (IL-2, IL-4, IL-10, and IFN-γ cytokines) and resulted 80-90% protection against lethal influenza infection. Also, single intranasal and single subcutaneous immunization with whole virus inactivated influenza vaccine mixed with nanoparticulated adjuvants stimulated high levels of humoral and cellular immune responses and provided 100% protection against lethal influenza infection. Conclusion: The results of study have shown that nanocomplexes contained purified triterpene saponins Glabilox and SO1 isolated from plants indigenous to Kazakhstan can stimulate a broad spectrum of humoral and cellular immune responses and induce protection against lethal influenza infection. Both elaborated adjuvants are promising for incorporation to influenza vaccine intended for subcutaneous and intranasal routes of immunization.

Keywords: influenza vaccine, adjuvants, triterpene saponins, immunostimulating activity

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