Experimental Human Influenza Research Articles

Clinical and virological responses to a broad-spectrum human monoclonal antibody in an influenza virus challenge study

Influenza A infections cause significant seasonal morbidity and mortality as well as periodic pandemic infections. Currently, no approved therapies exist for patients hospitalized with influenza. The efficacy of VIS410, a broadly neutralizing human immunoglobulin IgG1 monoclonal antibody engineered to bind to the stem region of group 1 and 2 influenza A hemagglutinins, was explored in experimental human influenza infection. Healthy volunteers were inoculated with influenza A/California/07/2009 (H1N1) and received a single dose of VIS410 or placebo 24 h later. Subjects were monitored for symptoms, viral shedding, and safety, including cytokine measurements. The primary efficacy endpoint was the area under the curve (AUC) of viral load (VL) in the VIS410 group versus placebo. VIS410 treatment was associated with a 76% reduction in median VL AUC as measured by qRT-PCR (p = 0.024). Similar VIS410 antiviral activity was observed by virus culture, with a 91% reduction in median VL AUC by TCID50 (p = 0.019) compared to placebo-treated volunteers. Influenza symptoms were generally mild or moderate, with a trend toward faster resolution in VIS410-treated subjects. Treatment with VIS410 was generally safe, with an increase in gastrointestinal events that were largely mitigated by pre-treatment with oral diphenhydramine (50 mg) in combination with 600 mg of ibuprofen. Transient elevation of specific cytokines (IL-8 and TNFα) were associated with gastrointestinal adverse events. Treatment with VIS410 did not interfere with the endogenous immune response to influenza A. These data indicate that VIS410 may provide therapeutic benefit in influenza A infection. Trial registrationClinicaTtrials.gov Identification NCT02468115; https://clinicaltrials.gov/ct2/show/NCT02468115?term=NCT02468115&rank=1).

Symptoms associated with influenza vaccination and experimental human pneumococcal colonisation of the nasopharynx

BackgroundNasopharyngeal colonisation by S. pneumoniae is a prerequisite for invasive pneumococcal infections. Influenza co-infection leads to increased susceptibility to secondary pneumonia and mortality during influenza epidemics. Increased bacterial load and impaired immune responses to pneumococcus caused by influenza play a role in this increased susceptibility. Using an Experimental Human Challenge Model and influenza vaccines, we examined symptoms experienced by healthy adults during nasal co-infection with S. pneumoniae and live attenuated influenza virus. MethodsRandomised, blinded administration of Live Attenuated Influenza Vaccine (LAIV) or Tetravalent Inactivated Influenza Vaccine (TIV) either preceded bacterial inoculation or followed it, separated by a 3-day interval. The presence and density of S. pneumoniae was determined from nasal washes. Participants completed a symptom questionnaire from the first intervention until 6 days post second intervention. ResultsThe timing and type of influenza vaccination and presence of S. pneumoniae in the nasopharynx significantly affected symptom reporting. In the study where influenza vaccination preceded bacterial inoculation: nasal symptoms were less common in the LAIV group than the TIV group (OR 0.57, p < 0.01); with colonisation status only affecting the TIV group where more symptoms were reported by colonised participants compared to non-colonised participants following inoculation (n = 12/23 [52.17%] vs n = 13/38 [34.21%], respectively; p < 0.05). In the study where influenza vaccination followed bacterial inoculation: no difference was seen in the symptoms reported between the LAIV and TIV groups following inoculation and subsequent vaccination; and symptoms were unaffected by colonisation status. ConclusionSymptoms experienced during live viral vaccination and bacterial co-infection in the nasopharynx are directly affected by the precedence of the pathogen acquisition. Symptoms were directly affected by nasal pneumococcal colonisation but only when TIV was given prior to bacterial exposure.

Assessment of Wearable Sensors During Experimental Human Influenza Infection (Sigma Plus)

Assessment of Wearable Sensors During Experimental Human Influenza Infection (Sigma Plus)

Commentary

Commentary

Using experimental human influenza infections to validate a viral dynamic model and the implications for prediction

The aim of this work was to use experimental infection data of human influenza to assess a simple viral dynamics model in epithelial cells and better understand the underlying complex factors governing the infection process. The developed study model expands on previous reports of a target cell-limited model with delayed virus production. Data from 10 published experimental infection studies of human influenza was used to validate the model. Our results elucidate, mechanistically, the associations between epithelial cells, human immune responses, and viral titres and were supported by the experimental infection data. We report that the maximum total number of free virions following infection is 10(3)-fold higher than the initial introduced titre. Our results indicated that the infection rates of unprotected epithelial cells probably play an important role in affecting viral dynamics. By simulating an advanced model of viral dynamics and applying it to experimental infection data of human influenza, we obtained important estimates of the infection rate. This work provides epidemiologically meaningful results, meriting further efforts to understand the causes and consequences of influenza A infection.

Assessing the Exacerbations Risk of Influenza-Associated Chronic Occupational Asthma

The purpose of this article was to conduct a risk‐based study based on a linkage of experimental human influenza infections and fluctuation analysis of airway function to assess whether influenza viral infection was risk factor for exacerbations of chronic occupational asthma. Here we provided a comprehensive probabilistic analysis aimed at quantifying influenza‐associated exacerbations risk for occupational asthmatics, based on a combination of published distributions of viral shedding and symptoms scores and lung respiratory system properties characterized by long‐range peak expiratory flow (PEF) dynamics. Using a coupled detrended fluctuation analysis‐experimental human influenza approach, we estimated the conditional probability of moderate or severe lung airway obstruction and hence the exacerbations risk of influenza‐associated occupational asthma in individuals. The long‐range correlation exponent (α) was used as a predictor of future exacerbations risk of influenza‐associated asthma. For our illustrative distribution of PEF fluctuations and influenza‐induced asthma exacerbations risk relations, we found that the probability of exacerbations risk can be limited to below 50% by keeping α to below 0.53. This study also found that limiting wheeze scores to 0.56 yields a 75% probability of influenza‐associated asthma exacerbations risk and a limit of 0.34 yields a 50% probability that may give a representative estimate of the distribution of chronic respiratory system properties. This study implicates that influenza viral infection is an important risk factor for exacerbations of chronic occupational asthma.

Understanding influenza virus-specific epidemiological properties by analysis of experimental human infections

This study aimed to estimate the natural history and transmission parameters based on experimental viral shedding and symptom dynamics in order to understand the key epidemiological factors that characterize influenza (sub)type epidemics. A simple statistical algorithm was developed by combining a well-defined mathematical scheme of epidemiological determinants and experimental human influenza infection. Here we showed that (i) the observed viral shedding dynamics mapped successfully the estimated time-profile of infectiousness and (ii) the profile of asymptomatic probability was obtained based on observed temporal variation of symptom scores. Our derived estimates permitted evaluation of relationships between various model-derived and data-based estimations, allowing evaluation of trends proposed previously but not tested fully. As well as providing insights into the dynamics of viral shedding and symptom scores, a more profound understanding of influenza epidemiological parameters and determinants could enhance the viral kinetic studies of influenza during infection in the respiratory tracts of experimentally infected individuals.

Efficacy and Tolerability of the Oral Neuraminidase Inhibitor Peramivir in Experimental Human Influenza: Randomized, Controlled Trials for Prophylaxis and Treatment

Oseltamivir is the only oral neuraminidase inhibitor currently available; we determined the tolerability and antiviral efficacy of oral peramivir for treatment and prophylaxis of experimental human influenza A and B. 288 susceptible, healthy volunteers (ages 18-45) were inoculated intranasally with A/Texas/36/ 91/H1N1 or B/Yamagata/16/88 virus in four randomized, double-blind, placebo-controlled trials. For treatment dosing was initiated at 24 h after inoculation with peramivir doses ranging from 100-800 mg/day for 5 days. For prophylaxis dosing was initiated 24 h before inoculation and continued for 4 days with peramivir doses ranging from 50-800 mg/day. The primary outcome measure for treatment was quantitative viral detection defined by the area under the curve (AUC) for nasal wash viral titres. For prophylaxis the primary outcome measure was the incidence of virus recovery. In influenza A treatment, peramivir 400 mg q24h and 200mg q12h, but not lower doses, resulted in significant reductions in viral titre AUC. In influenza B treatment, both 400 and 800/400 mg once daily dose groups reduced AUC values. In influenza A prophylaxis, the percentage of individuals with nasal viral shedding did not differ significantly in the placebo (58%), 50 mg (61%), 200 mg (37%) and 400 mg (31%) dose groups. In influenza B prophylaxis, shedding frequencies were similar in placebo (55%), 200 mg (41%), 400 mg (35%) and 800 mg (47%) dose groups. The drug was well tolerated in all four studies, with nausea and headache being the most common side effects. No drug-resistant variants were detected. Early treatment with peramivir was associated with significant antiviral effects in experimentally induced influenza in humans. Prophylaxis did not significantly reduce viral shedding. The relatively low blood peramivir concentrations observed may explain the lack of more robust antiviral effects, and parenteral dosing should be studied.

Oseltamivir: a review of its use in influenza.

Oseltamivir is a prodrug of oseltamivir carboxylate (Ro 64-0802, GS4071), a potent and selective inhibitor of the neuraminidase glycoprotein essential for replication of influenza A and B viruses. Studies in volunteers with experimental human influenza A or B showed that administration of oral oseltamivir 20 to 200 mg twice daily for 5 days reduced both the quantity and duration of viral shedding compared with placebo. Subsequent assessment of the drug at a dosage of 75 mg twice daily for 5 days in otherwise healthy adults with naturally acquired febrile influenza showed that oseltamivir reduced the duration of the disease by up to 1.5 days and the severity of illness by up to 38% compared with placebo when initiated within 36 hours of symptom onset (earlier initiation of therapy was associated with faster resolution). The incidence of secondary complications and the use of antibacterials were also reduced significantly in oseltamivir recipients. A liquid formulation of oseltamivir (2 mg/kg twice daily for 5 days) has been shown to be effective in the treatment of children with influenza, and data presented in abstracts suggest that the drug may also be of use in high-risk populations such as the elderly or those with chronic cardiac or respiratory disease. In addition to treatment efficacy, the drug has demonstrated efficacy when used for seasonal or household prophylaxis. Oral oseltamivir (75 mg once or twice daily for 6 weeks) during a period of local influenza activity significantly prevented the development of naturally acquired influenza by >70% compared with placebo in unvaccinated otherwise healthy adults. The drug also demonstrated efficacy when used adjunctively in previously vaccinated high-risk elderly patients (92% protective efficacy). Short term administration of oseltamivir (75 mg once daily for 7 days) may significantly reduce the risk of illness in household contacts of infected persons when administered within 48 hours of symptom onset in the infected person. Oseltamivir 75 mg twice daily for 5 days was well tolerated in clinical trials in healthy adults and high-risk patients, with nausea and vomiting being the most commonly reported events. Gastrointestinal events were mild and transient and both nausea and vomiting were less likely when oseltamivir was taken with food. Oseltamivir is a well tolerated orally active neuraminidase inhibitor which significantly reduces the duration of symptomatic illness and hastens the return to normal levels of activity when initiated promptly in patients with naturally acquired influenza. It therefore represents a useful therapeutic alternative to zanamivir (especially in patients who prefer oral administration or who have an underlying respiratory disorder) and the M2 inhibitors amantadine and rimantadine (because of its broader spectrum of anti-influenza activity and lower likelihood of resistance) in patients with influenza. In addition, although annual vaccination remains the best means of influenza prevention, there may be a place for oseltamivir in providing household prophylaxis or adjunctive prophylaxis in high-risk vaccinated patients during an outbreak of the disease or for use in patients in whom vaccination is unsuitable or ineffective.

Oral Oseltamivir in Human Experimental Influenza B Infection

Oseltamivir is the prodrug of Ro64-0802 (GS4071), a potent and selective inhibitor of influenza A and B virus neuraminidases. Three randomized, double-blind, placebo-controlled, parallel-group studies evaluated oral oseltamivir for early treatment (75 or 150 mg twice daily for 5 days) or prevention (75 mg once or twice daily for 7 days) of experimental influenza B virus infection in healthy susceptible adults. Treatment study A (n=60) demonstrated similar trends to treatment study B (n=117), in which 75 mg doses of oseltamivir introduced 24 h after inoculation reduced median area under curve (AUC) virus titre (oseltamivir, 22.7; placebo, 131.1 log10 TCID50 x h/ml; P=0.002) and duration of viral shedding (oseltamivir, 23.9 h; placebo, 95.8 h; P=0.0005). In prevention study C (n=58), oseltamivir did not reduce infection rates (85 versus 84%) but significantly reduced median AUC virus titre (10.0 versus 66.9 log10 TCID50 x h/ml; P=0.03) and duration of viral shedding (36 versus 84 h; P=0.03) compared with placebo. Oseltamivir was well tolerated. No emergence of drug-resistant variants was detected by testing last-day isolates (n=112) in neuraminidase inhibition assays. These results indicate that oseltamivir has significant antiviral activity in experimental human influenza B virus infection when used for prophylaxis or early treatment.

Safety and Efficacy of Once Daily Intranasal Zanamivir in Preventing Experimental Human Influenza a Infection

Zanamivir, a potent inhibitor of influenza A and B virus neuraminidases, is protective against experimental human influenza when given intranasally twice daily. We conducted two studies to assess the pharmacokinetics and protective efficacy of a reduced frequency dosing regimen of topical zanamivir. In the first study, 36 uninfected volunteers received a single dose of zanamivir by intranasal spray (6.4 mg), intranasal drops (16 mg) or dry powder oral inhalation (10 mg). At 4 h, median nasal wash concentrations were 50-fold higher after intranasal dosing than after inhalation. Substantial levels (spray group, median 4,596 ng/ml; drop group, 1,239 ng/ml) were detected in nasal wash 48 h after intranasal dosing. In the double-blinded efficacy study, 47 sero-susceptible volunteers were randomized to receive either placebo or zanamivir intranasal spray (6.4 mg). Among the 43 subjects evaluated, decreases in viral shedding occurred in the group receiving one dose of zanamivir 4 h prior to inoculation, whereas no significant benefit was observed in those receiving a single dose 48 h prior to challenge. In the group given three daily doses, reductions were seen in viral shedding and infection. In the two regimens providing zanamivir 4 h prior to inoculation, significant reductions in nasal mucus weight were observed. Decreases in total symptom scores and the incidence of upper respiratory illness also occurred, but they did not reach statistical significance. The efficacy of a single dose of zanamivir given 4 h prior to inoculation supports the hypothesis that influenza virus neuraminidase is essential for initial virus spread through respiratory secretions. These findings indicate that once daily dosing of zanamivir is protective against experimental influenza A infection.

Local and systemic cytokine responses during experimental human influenza A virus infection. Relation to symptom formation and host defense.

To further understand the role of cytokine responses in symptom formation and host defenses in influenza infection, we determined the levels of IL-1beta, IL-2, IL-6, IL-8, IFN-alpha, TGF-beta, and TNF-alpha in nasal lavage fluid, plasma, and serum obtained serially from 19 volunteers experimentally infected with influenza A/Texas/36/91 (H1N1) and correlated these levels with various measures of infection and illness severity. We found that IL-6 and IFN-alpha levels in nasal lavage fluids peaked early (day 2) and correlated directly with viral titers, temperature, mucus production, and symptom scores. IL-6 elevations were also found in the circulation at this time point. In contrast, TNF-alpha responses peaked later (day 3 in plasma, day 4 in nasal fluids), when viral shedding and symptoms were subsiding. Similarly, IL-8 peaked late in the illness course (days 4-6) and correlated only with lower respiratory symptoms, which also occurred late. None of IL-1beta, IL-2, or TGF-beta levels increased significantly. These data implicate IL-6 and IFN-alpha as key factors both in symptom formation and host defense in influenza.

New approaches to influenza chemotherapy. Neuraminidase inhibitors.

Epidemic influenza continues to be associated with significant morbidity in the general population, and mortality in the elderly and other high risk patients. Although the case fatality rate averages less than 0.01%, tens of thousands of deaths occur each year. Control through immunisation programmes has not been possible due to incomplete protective efficacy and antigenic variations that occur frequently. Currently available anti-influenza medications (amantadine and rimantadine) have had limited success due to underutilisation, lack of activity against influenza B, the rapid development of viral resistance to the drugs, and adverse effects. A new class of antiviral agents designed to inhibit influenza neuraminidase, an important surface glycoprotein, is currently under active development for use in the prophylaxis and treatment of influenza A and B infections. Two of these compounds, zanamivir (GG167) and GS4104 (the ethyl ester prodrug of GS4071) have reached clinical trials. Most studies of zanamivir have involved topical administration by inhalation of dry powder aerosols and/or intranasal doses of aqueous solutions. These routes rapidly provide high local concentrations at the sites of delivery. GS4104 is administered orally, which allows for greater ease of administration, and probably more uniform distribution of the parent compound GS4071 in the respiratory tract. Both have shown potent inhibitory activity against influenza in animal models and experimental human influenza with excellent tolerability profiles. Zanamivir treatment has been shown to reduce the severity and duration of naturally occurring, uncomplicated influenza illness in adults. Clinical resistance to these drugs has not been recognised as a significant problem to date, although strains resistant to each agent have been produced in the laboratory. This class of agents shows considerable promise as a novel approach to prophylaxis and treatment of influenza infections. Ongoing studies should provide the data needed to allow the addition of 1 or more of the neuraminidase inhibitors to the clinician's anti-influenza armamentarium.

Safety and Efficacy of the Neuraminidase Inhibitor GG167 in Experimental Human Influenza

The current study evaluated whether intranasal administration of the sialic acid analog 4-guanidino-Neu5Ac2en (GG167), an inhibitor of influenza virus neuraminidase, was effective and safe in either preventing or treating experimental human influenza. Four randomized, double-blind, placebo-controlled trials involving three prophylaxis limbs, two early treatment limbs, and one delayed treatment limb were conducted. Isolation in individual rooms. Susceptible (serum hemagglutination-inhibition antibody titer < or = 1:8) adult volunteers (n = 166) were inoculated intranasally with 10(5) TCID50 influenza A/Texas/91 (H1N1) virus. GG167, 3.6 to 16 mg, was administered intranasally two or six times daily beginning 4 hours before inoculation (prophylaxis) or 1 or 2 days afterward (early or delayed treatment). Virological measures were frequency of infection based on viral shedding and/or seroconversion (prophylaxis) or quantitative viral shedding based on titers and duration of virus recovery (treatment). Clinical measures were the frequency of febrile illness and symptom severity scores. Intranasal GG167 was well tolerated for both prophylaxis and therapy. For all dose groups combined, GG167 prophylaxis was 82% effective in preventing laboratory evidence of infection and 95% effective in preventing febrile illness (P < .01 vs placebo). Early treatment with GG167 reduced peak viral titers by 2.0 log10, the median duration of viral shedding by 3 days, and the frequency of febrile illness by 85% (P < .05 for each comparison). Other measures of illness were reduced by approximately 50% to 70% in the GG167 dosing groups. Twice daily dosing was as effective as six times daily. Direct respiratory administration of the selective neuraminidase inhibitor GG167 appears safe and effective for both prevention and early treatment of experimental influenza. Influenza virus neuraminidase is important for viral replication in humans.

Safety and efficacy of the neuraminidase inhibitor GG167 in experimental human influenza

Safety and efficacy of the neuraminidase inhibitor GG167 in experimental human influenza

Comparative therapeutic effect of aerosolized and oral rimantadine HC1 in experimental human influenza A virus infection

Thirty-six adult volunteers were inoculated intranasally with 7.2 log 10 egg infectious doses 50% of an attenuated A/Khabarovsk/77/H1N1 virus. Twenty-four hours later volunteers were begun on both aerosol treatments (rimantadine HC1 25 mg in saline or saline, 10 min exposure twice daily) and oral medications (rimantadine HC1 50 mg or placebo every 6 h, three times daily) which were administered for 5 days. Virus-positive volunteers receiving placebo by both of the two routes had a peak in clinical illness scores on the second treatment day (mean score 5.3), which was not observed in either the aerosol rimantadine (0.6) or oral rimantadine (0.9) treated volunteers. On the second treatment day, the proportion of virus-positive volunteers with elevated axillary temperature measurements and the mean peak temperature measurement were also significantly reduced in both drug groups. No significant effects on the duration of virus shedding were noted. In experimental influenza A virus infection, characterized by mild clinical illness and short duration of virus shedding, low doses of aerosolized rimantadine had a therapeutic effect comparable to that found with larger doses of oral rimantadine.

Formation of antibody to matrix protein in experimental human influenza A virus infections.

Antibodies to type A influenza virus matrix protein (M) were assayed by single radial diffusion in 180 paired sera of volunteers challenged intranasally with live H3N2 viruses of varying degrees of virulence. Of these volunteers 20 had had severe clinical reactions (influenza-like); there had been 19 moderate reactions (lesser degrees of constitutional illness), and the remaining 141 reactions had been graded mild, very mild, or nil. Only 2 volunteers were shown to have antibodies to M in the pre-trial serum samples, and 11 developed anti-M rises after virus inoculation. Nine of the 11 had had severe reactions, and 2 had had moderate reactions. There was, therefore, a clear correlation between severity of clinical illness and anti-M antibody formation. In general, anti-M increases coincided with increases to the hemagglutinins and nucleoprotein, and with virus shedding. However, no anti-M antibody could be demonstrated in paired sera of 18 additional volunteers of whom 12 had developed severe reactions after the inoculation of virulent H0N1 and H1N1 influenza A viruses and of whom 12 had shown laboratory evidence of infection.

Cyclooctylamine in the prevention of experimental human influenza

The prophylactic effectiveness of intranasally given SK&amp;F 23880A (cyclooctylamine hydrochloride) against induced A₂/Hong Kong influenza was evaluated in double-blind trials on nine drug-treated and seven placebo-treated subjects. The topical SK&amp;F 23880A formulation was effective in preventing overt influenza in these susceptible subjects. The degree and duration of febrile reactions were substantially diminished. The drug exerted little effect in minimizing other signs and symptoms, however. Influenza viruses were recorded from all subjects but in apparently smaller quantities from the drug-treated men. Postchallenge serum antibody titers were significantly lower in the drug-treated group. Respiratory secretory antibody responses in this group were also reduced, but only slightly. Three drug-treated men complained of transient, mild nasopharyngeal irritation associated with the intranasally given medication.

Experimental human influenza infections studied by means of immunofluorescent staining of nasal smears.

Three volunteers, aged 20, 25 and 27 were experimentally infected with an amniotic culture fluid of fresh influenza B isolate. Two cases, which previously had low HAT antibody against the used virus, did not develop influenza B antibody, suggesting failure of the virus infection. All specimens examined throughout the course of infection by an immunofluorescent technique did not show accumulation of virus antigens. Only one case, which previously had no detectable antibody against influenza, B, developed typical signs and symptoms of influenza. At 48 hours after infection, fluorescent cells were detected by the immunofluorescent technique in the preparations from nasal scrubs. Fluorescent cells were successively found in the nasal scrub specimens up to 120 hours after the infection. Whereas, no fluorescent cells were found in the specimens obtained from nasal washings, throat scrubs, and throat washings throughout the course of infection. First discovery of fluorescent cells was 8 hours ahead the temperature rise and 48 hours ahead the first virus isolation.