Efficacy of single intravenous injection of peramivir against influenza B virus infection in

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "Efficacy of single intravenous injection of peramivir against influenza B virus infection in"

Transcription

1 AAC Accepts, published online ahead of print on 15 August 2011 Antimicrob. Agents Chemother. doi: /aac Copyright 2011, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved. Kitano et al., Page Efficacy of single intravenous injection of peramivir against influenza B virus infection in ferrets and cynomolgus macaques Mitsutaka Kitano, 1,3 Yasushi Itoh, 1 Makoto Kodama, 3 Hirohito Ishigaki, 1 Misako Nakayama, 1 Hideaki Ishida, 1 Kaoru Baba, 3 Takahiro Noda, 3 Kenji Sato 4, Yoichiro Nihashi 4, Takushi Kanazu 4, Ryu Yoshida, 3 Ryuzo Torii, 2 Akihiko Sato, 3 * Kazumasa Ogasawara 1 Department of Pathology, Shiga University of Medical Science, Otsu, Shiga , 1 Research Center for Animal Life Science, Shiga University of Medical Science, Otsu, Shiga , 2 Infectious Diseases, Medicinal Research Laboratories, Shionogi & Co., Ltd., Settsu, Osaka, , 3 Drug Metabolism & Pharmacokinetics, Drug Developmental Research Laboratories, Shionogi & Co., Ltd., Toyonaka, Osaka, , Japan Running title: In vivo effects of peramivir against influenza B virus infection *Corresponding author Infectious Diseases, Medicinal Research Laboratories Shionogi & Co., Ltd. 5-1 Mishima 2-chome, Settsu, Osaka , Japan Tel: +81 (6) , Fax: +81 (6) ,

2 Kitano et al., Page ABSTRACT We evaluated the efficacy of single intravenous dose peramivir for treatment of influenza B virus infection in ferrets and cynomolgus macaques in the present study. A single dose of peramivir (30 and 60 mg/kg) given to ferrets on 1 day post-infection with influenza B virus significantly reduced virus titers in nasal washes and total virus shedding as measured by areas under the curve (AUC) by 2 log 10 compared with control group. Furthermore, nasal virus titers on day 2 post infection and AUCs of the body temperature raise of the ferrets singly injected with peramivir (30 and 60 mg/kg) were lower than those of ferrets orally administered with oseltamivir phosphate (30 and 60 mg/kg/day twice daily for 3 days). In macaques infected with influenza B virus, viral titers in nasal swab fluid on days 2 and 3 post infection and body temperature after single injection with peramivir (30 mg/kg) were lower than those after oral administration with oseltamivir phosphate (30 mg/kg/day for 5 days). Two animal models used in the present study demonstrated that inhibition of viral replication at the early time point after infection was critical in reduction of AUC of virus titers and IL-6 production, resulting in amelioration of symptoms. Our results shown in animal models suggest the early treatment with a single intravenous injection of peramivir is significantly effective for the treatment of influenza B infection. 2

3 Kitano et al., Page INTRODUCTION Peramivir is a neuraminidase (NA) inhibitor for intravenous administration and it has been introduced into clinical practice for adult and child patients in Japan. Peramivir has potent activity against various influenza A and B viruses including highly pathogenic avian influenza (HPAI) viruses and pandemic (H1N1) 2009 influenza virus in vitro and in vivo mouse models (2, 6, 7, 10). Phase 2 clinical trials of intravenous peramivir in Japanese patients with seasonal influenza demonstrated the efficacy and safety of a single intravenous treatment (24). On the basis of these results, the United States Food and Drug Administration issued an emergency use authorization for intravenous peramivir exclusively for severe pandemic H1N1 hospitalized patients (4). Influenza B virus causes seasonal influenza infection and indistinguishable clinical symptoms from those caused by influenza A virus though pandemic influenza A virus infection attracts more attention than does influenza B virus infection (26, 29, 30). Influenza B virus was isolated more frequently and for a longer period after antiviral therapy than influenza A virus (21). Furthermore, influenza B virus epidemics did not diminish after the pandemic of 2009 H1N1 influenza A virus (34). Therefore, treatment for influenza B virus infection is important as well as for influenza A virus infection. NA inhibitors are major antiviral agents of influenza B virus infection because available M2 inhibitors, amantadine and rimantadine, are not active in influenza B viruses since the amino acid sequences of the B/M2 protein differ from those of the A/M2 protein (31). Nonetheless, some studies have claimed that an NA inhibitor oseltamivir was less effective in treating influenza B virus than influenza A virus with regard to duration of fever, irrespective of patient age or the timing of administration of the first dosing (21, 32). The 3

4 Kitano et al., Page difference would be explained by higher 50% inhibitory concentration (IC 50 ) values of oseltamivir against NA activity of influenza B viruses than those of influenza A viruses (33). In addition, influenza B viruses with low susceptibility to NA inhibitors were isolated from immunocompromised children treated with oseltamivir or zanamivir (11, 12). Therefore, further development and evaluation of clinical efficacy of antiviral drugs against influenza B virus are required. Problems in the drug development are that the appearance of influenza B virus is variable season by season compared with influenza A virus that consistently appears almost every year (20) and that preexisting immunity against influenza B virus in humans may make evaluation of drug effects complicated. To solve the problems of shortage of human patients infected with influenza B virus and of preexisting immunity in clinical trials, studies using immunologically naïve animals against influenza B virus would offer the essential experimental approach for testing efficacy of antiviral drugs. Although mice are naturally resistant to most seasonal human influenza viruses unless the viruses are adapted, ferrets are susceptible to human influenza virus, including various influenza A and B viruses, without adaptation (14, 16, 17, 19, 28). Human influenza viruses infect upper respiratory tracts in ferrets and cause clinical symptoms, including fever, nasal congestion, anorexia and sneezing. Therefore, ferrets are often used for examining efficacy of antiviral agents in treatment and prevention of influenza infection (9, 25, 35). On the other hand, nonhuman primates are also susceptible to unadapted human influenza viruses (17). Since clinical symptoms by infection with influenza B viruses in cynomolgus macaque closely reflects the signs of diseases observed in humans (23), we investigated the therapeutic efficacy of a new NA inhibitor peramivir against influenza B virus infection using not only ferrets but also 4

5 Kitano et al., Page cynomolgus macaques. In the present study, efficacy of single intravenous administration of peramivir was compared with those of multiple oral administration of oseltamivir phosphate in ferret and macaque models of influenza B virus infection. We demonstrated that a single intravenous injection of peramivir was more effective on reduction of viral titers and amelioration of symptoms in ferrets and cynomolgus macaques after influenza B virus infection than administration with oseltamivir phosphate. These results suggest that peramivir has potential as a treatment against influenza B virus infection. Downloaded from on May 13, 2018 by guest 5

6 Kitano et al., Page MATERIALS AND METHODS Compounds. Peramivir was synthesized by BioCryst Pharmaceuticals (Birmingham, AL). Oseltamivir phosphate was purchased from Sequoia Research Products (Oxford, UK), and oseltamivir carboxylic acid was purchased from Toronto Research Chemicals (Ontario, Canada). Viruses and cells. Influenza B viruses, B/SendaiH/1051/2007 and B/Kadoma/1/2005, were kindly provided by Sendai Medical Center and Osaka Prefectural Institute of Public Health, respectively. Madin-Darby canine kidney (MDCK) cells were obtained from the American Type Culture Collection (Manassas, VA, USA) and were grown in minimum essential medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (Invitrogen), 100 µg/ml kanamycin sulfate (Invitrogen) in a humidified atmosphere of 5% CO 2 at 37 o C. Ferrets. Approximately nine- to eleven-month-old female ferrets (Japan SLC Inc., Shizuoka, Japan) were used. All ferret studies were conducted under applicable laws and guidelines and after approval from the Shionogi Animal Care and Use Committee. Under anesthesia at least one week before virus inoculation, a data logger (DS1921H-F5, Maxim Integrated Products, Inc., Sunnyvale, CA) was implanted in the peritoneal cavity of each ferret to monitor body temperature every 15 min. The absence of influenza B-specific antibody in sera was confirmed before experiments using the hemagglutination inhibition (HI) test. Cynomolgus macaques. Approximately two- to five-year-old female cynomolgus macaques from the Philippines (Ina Research Inc., Ina, Japan) were used for challenge experiments. Macaque studies were conducted under applicable laws and guidelines and after approval from the Shiga University of Medical Sciences Animal Experiment Committee and Biosafety Committee. Under anesthesia at least one week before virus inoculation, a telemetry probe 6

7 Kitano et al., Page (TA10CTA-D70, Data Sciences International, St. Paul, MN) was implanted in the peritoneal cavity of each macaque to monitor body temperature every 15 min. The absence of influenza B-specific antibody in sera was confirmed before experiments using the HI test. Individual macaques are distinguished by identification numbers. The macaques used in this study did not carry herpes B virus, hepatitis E virus, Mycobacterium tuberculosis, Shigella spp., Salmonella spp., or Entamoeba histolytica. NA inhibition assay. Whole viruses inactivated by NP-40 were used as a source of NA activity. 2 -(4-methylumbelliferyl)-α-D-N-acethylneuraminic acid (MUNANA, Sigma-Aldrich, St. Louis, MO) was used as substrate. The virus, a compound and MUNANA (final concentration 10 mm) were mixed in reaction buffer, and incubated at 37 o C for 30 min. The fluorometric intensity of 4-methylumbelliferon released from MUNANA was measured, the percent inhibition at each concentration of drug was determined, and the 50% inhibitory concentration (IC 50 ) was determined. The results are reported as the average of three experiments. Pharmacokinetic (PK) analysis. Female ferrets and cynomolgus macaques (three animals per group) were given peramivir (30 mg/kg) intravenously or oseltamivir phosphate (30 mg/kg) orally. Blood samples were collected from h to 24 h after dosing and centrifuged to obtain plasma. The concentration of peramivir and oseltamivir carboxylate in plasma was determined by liquid chromatography-tandem mass spectrometry (LC/MS/MS). PK parameters were calculated and modeling performed using WinNonlin software ver. 4.0 (Pharsight Corp., Mountain View, CA). Antiviral study in a ferret model. Under anesthesia, ferrets were inoculated intranasally with B/Kadoma/1/2005 (300 tissue culture infective dose (TCID 50 )) in 200 µl of PBS. Four and six 7

8 Kitano et al., Page ferrets were used into treated groups and an untreated control group, respectively. Six groups were compared in the study using ferrets as followings: (1) Peramivir (30 mg/kg) in saline was administered once intravenously one day after infection (P30x1, abbreviations were described in Fig. 2). (2) Peramivir (60 mg/kg) in saline was administered once intravenously one day after infection (P60x1). (3) Peramivir (30 mg/kg/day) in saline was administered once a day for 3 days from day 1 to day 3 after infection (P30x3). (4) Oseltamivir phosphate (30 mg/kg/day) in 0.5% methylcellulose solution (MC) was administered orally twice a day for 3 days from day 1 to day 3 (O30x3). (5) Oseltamivir phosphate (60 mg/kg/day) in 0.5% MC was administered orally twice a day for 3 days from day 1 to day 3 (O60x3). The first administration of peramivir and oseltamivir phosphate was performed on day 1 post infection (pi). (6) Control animals inoculated with virus but not treated with any reagents were orally administered with 0.5% MC twice a day from day 1 to day 3. To monitor virus replication in nasal cavities, nasal washes were collected from infected ferrets on days 1 to 4 pi. Collected samples were stored at a temperature below -80 o C until use. For virus titration, serial dilutions of nasal washes were inoculated onto confluent MDCK cells in 96-well plates. After 1-h incubation, the suspension was removed, and the cells were cultured in MEM including 0.5% BSA (Sigma-Aldrich) and 3 µg/ml trypsin. The plates were incubated at 37 o C in 5% CO 2 for 3 days. The presence of cytopathic effects (CPE) was determined under a microscope and viral titers were calculated as log 10 TCID 50 /ml. When no CPE was observed using undiluted viral solution, it was defined as the undetectable level that was considered to be lower than 1.4 log 10 TCID 50 /ml. Inflammatory cells in nasal washes were counted microscopically in a hemocytometer. The protein concentration in cell-free nasal washes was 8

9 Kitano et al., Page measured by using a protein reagent (Wako Pure Chemicals, Osaka, Japan). Body temperature was expressed by calculating the average temperatures during nighttime (10 pm 8 am) for avoiding the effect of anesthesia and was compared with that before virus inoculation. Antiviral study in a cynomolgus model. Under anesthesia, cynomolgus macaques were inoculated intranasally with B/SendaiH/1051/2007 (2 x 10 5 TCID 50 ) in 1 ml of PBS. Three macaques were used in each group. Four groups were compared in the macaque study as followings: (1) Peramivir (30 mg/kg) in saline was administered intravenously once immediately after virus inoculation (P30-d0, abbreviations were described in Fig. 5). (2) Peramivir (30 mg/kg) in saline was administered intravenously on day 1 pi (P30-d1). (3) Oseltamivir phosphate (30 mg/kg/day) in 0.5% MC was administered orally once a day for 5 days from day 0 to day 4 (O30x5). The first administration of oseltamivir phosphate was performed immediately after virus inoculation. (4) Control animals inoculated with virus but not treated with any reagents were orally received with 0.5% MC from day 0 to day 4. On day 0 before virus inoculation and days 1 to 12 after virus inoculation, the macaques were anesthetized and then nasal swab samples were collected with two cotton sticks (TE8201, Eiken Chemical, Ltd., Tokyo, Japan) and the sticks were subsequently immersed in 1 ml of PBS containing 0.1% BSA and Penicillin-Streptomycin. Blood samples were collected for measuring anti-ha antibody responses in sera. Collected samples were stored at a temperature below 80 ºC until use. Virus titers in nasal swab fluid were determined using MDCK cells as described above. Levels of inflammatory cytokines and chemokines (interleukin (IL)-6, tumor-necrotizing factor (TNF)-α and monocyte chemotactic protein (MCP)-1) in the nasal swab fluid were assessed using the Cytometric Bead Assay (Becton, Dickinson and Company, Franklin Lakes, NJ) 9

10 Kitano et al., Page according to the manufacturer's instructions. Results of assays were analyzed using FCAP Array software (Becton, Dickinson and Company). Body temperature was expressed by calculating the averages of the highest and lowest temperatures during 1 day and the body temperature after the virus inoculation was compared with that before the virus inoculation. Hemagglutination inhibition (HI) assay. Sera were treated with receptor destroying enzyme (RDEII, Denka Seiken, Tokyo, Japan). Serially diluted sera were mixed with 4 HA units of virus antigen for 1 h at room temperature. The mixture was then incubated with 0.5% chicken red blood cells for 30 min at room temperature. The HI titers were expressed as reciprocals of the highest dilution of serum samples that completely inhibited hemagglutination. Statistical analysis. Virus titers, cytokine levels and body temperature of each animal were calculated as the area under the curve (AUC) by the trapezoidal method. Differences of AUC in virus titers, cytokine levels and body temperature change were analyzed by the Dunnett s multiple comparison method. The Dunnett s test was also carried out for comparison in the virus titers, body temperature, body weight and the number of inflammatory cells on each day. The efficacy of peramivir was compared with that of oseltamivir by using Student's t-test. Statistical analysis was performed using the statistical analysis software, SAS version 9.2 for Windows (SAS Institute, Cary, NC). P values below 0.05 were considered as statistically significant. Sequence analysis of NA genes. Viral RNA was isolated directly from nasal swab fluid of infected macaques or nasal wash fluid of infected ferrets by using the RNeasy Mini Kit (Qiagen, Duesseldorf, Germany). The NA region of influenza virus was amplified by PCR using OneStep RT-PCR kit (Qiagen) and specific primers. The primers for amplification of the NA included a forward primer, 5 -ATGCTACCTTCAACTGTACAAAC -3 or 10

11 Kitano et al., Page CCTAAGAACACAAGAAAGTGCC -3 and a reverse primer 5 - GCCAGCAATAAAACCATAGAATG-3 or 5 - CACAGGTGTTGATATGGCTCTGTAA-3. The amplified DNA was sequenced in Applied Biosystems 3730xl DNA Analyzer, by TAKARA sequencing service. The sequences of the NA region derived from isolated viruses were compared with that of inoculation viruses, and amino acid substitutions were analyzed. Downloaded from on May 13, 2018 by guest 11

12 Kitano et al., Page RESULTS Sensitivity of influenza B virus NA to NA inhibitors in vitro. Initially, we examined the sensitivity of B/Kadoma/1/2005 and B/SendaiH/1051/2007 neuraminidases to peramivir and oseltamivir carboxylate in vitro (Table1). Since virus sensitivity to NA inhibitors in vivo is not always correlated with that analyzed in cell culture, we determined enzyme 50 % inhibitory concentrations (IC 50 ) (27). IC 50 of peramivir against NA activity of B/Kadoma/1/2005 and B/SendaiH/1051/2007 were 1.78 ± 0.08 nm and 4.26 ± 0.17 nm, respectively. On the other hand, IC 50 values of oseltamivir carboxylate against NA activity of B/Kadoma/1/2005 and B/SendaiH/1051/2007 were 5.93 ± 0.51 nm and 13.1 ± 0.50 nm, respectively. These values were comparable to previous results that IC 50 s of peramivir against NA activity of other strains of influenza B virus were between 0.60 and 10.8 nm and IC 50 s of oseltamivir carboxylate were 5.0 to 24.3 nm (3). Regarding these two influenza B viruses, the inhibitory activity of peramivir was more potent than that of oseltamivir carboxylate (p < 0.001). Pharmacokinetics of peramivir and oseltamivir carboxylate in ferrets and cynomolgus macaques. Pharmacokinetic analysis of peramivir in ferrets and cynomolgus macaques showed rapid uptake into the circulation following intravenous injection (Fig.1). After administration of 30 mg/kg, peramivir concentrations on average was 126 µg/ml and 147 µg/ml at h, then decreased to 0.14 µg/ml and 0.76 µg/ml at 8h in ferrets and macaques, respectively. C max were calculated as 194 µg/ml in ferrets and 160 µg/ml in macaques (Table 2). Area under the curve (AUC) of plasma concentration of peramivir after injection with 30 mg/kg in the macaques (234 µg hr/ml) 12

13 Kitano et al., Page was higher than that observed in ferrets injected with 30 mg/kg peramivir (89.1 µg hr/ml). On the other hand, mean times to maximum concentration (T max ) of oseltamivir carboxylate were 2.67 h and 3 h in ferrets and macaques, respectively. AUC of plasma concentration of oseltamivir carboxylate after injection with 30 mg/kg in ferrets (23.7 µg hr/ml) was almost equivalent as that in macaques (25.4 µg hr/ml). We have confirmed low toxicity of peramivir in cynomolgus macaques. When peramivir was administered daily by intravenous infusion to macaques at a dose of 720 mg/kg/day for 30 days, which was the maximum feasible dose, no toxic findings were evident in the clinical observations, body weights, food consumption, hematology, urinalysis, organ weights or histopathology (data not shown). Efficacy of peramivir against influenza B virus infection in ferrets. To evaluate the efficacy of peramivir and oseltamivir against influenza B virus in vivo, peramivir or oseltamivir phosphate was administered on day 1 pi into ferrets infected intranasally with B/Kadoma/1/2005 (300 TCID 50 /ferret) of which titer in nasal samples of ferrets was highest among various clinical isolates examined (data not shown) and that induced fever in ferrets, thereafter virus titers in nasal washes were examined. No viruses were detected in nasal washes of 6 control ferrets on day 1 pi, the maximum virus titer in the control group was observed on day 3 pi and then the virus titer was declined (Fig. 2A). According to these results in control ferrets, we focused the analysis on days 2, 3 and 4 pi to examine the effects of peramivir and oseltamivir on virus replication in the early phase after infection. On day 2 pi, virus titers in all groups treated with peramivir were significantly reduced compared with that in the control group (p < 0.01), but no 13

14 Kitano et al., Page significant difference on days 3 and 4 pi was observed in the group treated with peramivir (30 mg/kg) once (P30x1, see abbreviations in Fig. 2). On day 3 pi, significant reduction in virus titers was also observed in the groups P60x1 and P30x3 compared with control. Significant reduction in virus titers on day 2 pi was observed in the group treated with oseltamivir (60 mg/kg/day) (O60x3, also see abbreviations in Fig. 2) compared with control (p < 0.05) but not in O30x3. In addition, on day 3 pi, virus titers in two groups treated with oseltamivir phosphate were significantly lower than that in the control group (p < 0.05). In comparison of efficacy between peramivir and oseltamivir, the virus titer on day 2 pi in the group P30x1 was significantly lower than that in the group O30x3 (p < 0.05). To examine the effects of peramivir against virus replication during infection, the AUCs of the viral titers from days 2 to 4 pi were calculated. Virus titer AUCs of the groups treated with peramivir and oseltamivir phosphate were significantly reduced in a dose-dependent manner (Fig. 2B). In addition, the virus titer AUC in the group P60x1 was comparable to those in the groups P30x3 and O60x3. These results indicated that a single intravenous injection of peramivir had potent antiviral activity in the early phase after virus inoculation and that the efficacy of a single intravenous injection of peramivir against influenza B virus was comparable to that of oseltamivir phosphate administered for 3 days in the ferret model. To monitor the emergence of resistant variants after peramivir treatment and during oseltamivir phosphate treatment, we determined NA gene sequences of virus obtained from nasal wash fluid on day 4 pi. An isolated virus in the group O30x3 showed a mutation resulting in an amino acid change at position 31 (L to I) of the NA gene. This mutation is located around transmembrane domain and not associated with reported resistance or decreased susceptibility to 14

15 Kitano et al., Page NA inhibitors (1, 8). In the other ferrets, no NA amino acid changes were detected in viruses from the nasal wash fluid (data not shown). We examined the effects of peramivir and oseltamivir on body temperature in ferrets after inoculation with B/Kadoma/1/2005. Higher body temperature than that before the inoculation was observed for 4 days in the control group (Fig. 3A). On day 2 pi, the groups P60x1 and P30x3 showed significantly lower body temperature than did the control group (p < 0.05). The group P30x1 showed slightly but not significant reduction in body temperature on day 2 pi. In contrast, two groups administered with oseltamivir phosphate showed no apparent reduction in body temperature. AUCs of the body temperature change of the groups P30x1 and P60x1 were lower than those of O30x3 and O60x3, respectively (p < 0.05 and p < 0.01) (Fig. 3B). To assess the effects of peramivir and oseltamivir on inflammation in nasal cavities of infected ferrets, the protein concentration and the number of inflammatory cells in nasal wash fluid were measured. The protein concentration and the number of inflammatory cells on day 3 pi were reduced in all groups treated with peramivir and the group O60x3 but not in the group O30x3 (Fig. 4A and B). In addition, body weights were measured as one of the clinical signs. A significant difference in the average percent body weight (weight on day 4 / weight on day 0) between the group P30x3 (102.0%) and the untreated control group (97.4%) was observed (p < 0.05) (Fig. 4C). While nasal signs (nasal discharge, sneezing and mouth breathing) induced by influenza B infection in the control group were relatively mild, ferrets treated with peramivir or oseltamivir phosphate exhibited little or no nasal sign (data not shown) Efficacy of a single intravenous injection of peramivir against influenza B virus in 15

16 Kitano et al., Page cynomolgus macaques. We have reported that cynomolgus macaques were susceptible to influenza B virus that replicated in the upper respiratory tracts. Especially, B/SendaiH/1051/2007 caused fever with loss of appetite and body weight more clearly than did B/Kadoma /1/2005 (23). Therefore, cynomolgus macaques were intranasally inoculated with B/SendaiH/1051/2007 (2 x 10 5 TCID 50 ) and 30 mg/kg of peramivir was administered intravenously once immediately after virus inoculation (P30-d0, see abbreviations in Fig. 5) or 24 h after virus inoculation (P30-d1). Thirty mg/kg/day of oseltamivir phosphate was administered orally once a day for 5 days as a comparison (O30x5). When the virus titers in nasal swab fluid were measured from day 1 to 12 pi, the maximum virus titer in the control group was observed on day 3 pi and then the virus titer declined after day 4 pi (Fig. 5A). On days 2 and 3 pi, virus titers in all groups treated with peramivir were significantly reduced compared with those in the control group (p < 0.01). On days 5 and 7 pi, significant reduction in virus titers was also observed in the group P30-d1. In contrast, no apparent reduction of virus titers was observed in the early phase after virus inoculation in the group O30x5 and the virus titer on day 7 pi in the group O30x5 was significantly lower than that of the control group (p < 0.05). In comparison between peramivir and oseltamivir on day 2 pi, virus titers of P30-d0 and P30-d1 were significantly lower than that of O30x5 (p < 0.05 and p < 0.01, respectively). On day3 pi, the virus titer of P30-d1 was also significantly lower than that of O30x5 (p < 0.05). When the statistical analysis was performed on the basis of the AUCs of the viral titers from day 1 to 7 pi, the virus titer AUCs of the groups treated with peramivir were significantly lower than that of the control group (p < 0.05), while that treated with oseltamivir was not (Fig. 5B). Furthermore, the virus titer AUC of P30-d1 was significantly lower than that of O30x5 (p < 16

17 Kitano et al., Page ). We determined NA gene sequences of virus obtained from nasal swab fluid on day 5 pi. Viruses isolated from macaques treated with peramivir and oseltamivir showed no mutation in NA amino acid sequences (data not shown). Body temperatures were compared among the treated and untreated groups. Higher body temperatures than those before the virus inoculation were observed in the untreated control group for 1 to 4 days (Fig. 6). Body temperature of macaques treated with peramivir was substantially lower than those of macaques in the control group, whereas no significant suppression in body temperature was observed in all treated macaques using statistical analysis on the basis of the AUC of body temperature change (data not shown). However, the averages of maximal temperature rise in the groups P30-d0 (mean ± SD = 0.33 ± 0.15 o C) and P30-d1 (0.43 ± 0.31 o C) after infection were significantly lower than that in the control (1.13 ± 0.21 o C) (p < 0.05). In contrast, no significant suppression of maximal temperature raise was observed in the group O30x5 (0.60 ± 0.26 o C) compared with the control group. To examine the effects of peramivir and oseltamivir on immune responses in infected macaques, production of inflammatory cytokines and chemokines in nasal swab fluid was measured (Fig. 7A). Levels of cytokine and chemokine secretion were increased after infection. However, the kinetics of cytokine and chemokine production varied in individual macaques. Therefore, levels of cytokines and chemokines were expressed as AUC from day 1 to 12 and statistical analysis was conducted on the basis of the AUC (Fig. 7A). Levels of IL-6 production in groups P30-d0, P30-d1 and O30x5 were significantly lower than those in the control group. On the other hand, no significant effect on TNF-α and MCP-1 production was observed in treated groups compared with the control group. 17

18 Kitano et al., Page To serologically confirm influenza B virus infection and to compare the effects of the reagents on production of anti-ha antibodies, sera were collected on indicated days after virus inoculation. HI activities against B/SendaiH/1051/2007 were observed in sera of groups treated with peramivir or oseltamivir phosphate on day 8 pi and increased until day 14 pi (Fig. 7B). These results indicate that peramivir and oseltamivir did not inhibit the production of serum anti-ha antibodies against B/SendaiH/1051/2007 in cynomolgus macaques. Downloaded from on May 13, 2018 by guest 18

19 Kitano et al., Page DISCUSSION Influenza B virus causes seasonal influenza even after newly emerging H1N1 influenza A virus infection has become pandemic in 2009 and the number of Russian H1N1 virus cases decreased (34). Since influenza B virus infection causes significant morbidity as influenza A virus does (26, 29, 30), antiviral agents against influenza B virus are required. The efficacy of peramivir against influenza B virus in vitro was more potent than that of oseltamivir and zanamivir (5, 15). Therefore, in the present study, we examined the efficacy of peramivir against influenza B virus in vivo using ferrets and macaques that were susceptible to infection with a number of unadapted human influenza isolates and showed clinical symptoms. Since absence of HI activity in sera was confirmed before experiments and animals were maintained under the specific-pathogen-free condition in the present study, use of immunologically naïve animals against influenza B virus might support the interpretation that change of virus titers in the early stage of infection was mainly caused by antiviral agents but not by memory immune responses against influenza B virus, which might exist in many humans. In addition, animal models enable to examine effective time points of initiation of treatments after virus exposure since it might be difficult to know when patients were infected with virus in clinical studies. These points might be critical if differences of effects among antiviral agents would be modest. In ferrets infected with human influenza B virus, viral titers and virus titer AUC were decreased with single intravenous administration of peramivir and repeated oral administration of oseltamivir phosphate compared with those of control (Fig. 2). However, single administration of 60 mg/kg of peramivir attenuated fever by about 1 o C, while no change in fever was observed in the groups treated with oseltamivir phosphate (Fig. 3). The difference of virus titers in between 19

20 Kitano et al., Page ferrets treated with peramivir and ferrets treated with oseltamivir phosphate was observed on day 2 pi. This suggested that viral propagation in an early phase after virus infection contributed to severity of symptoms including fever and that suppression of virus replication in the early phase during infection was crucial to ameliorate symptoms in the ferret model. To further evaluate the therapeutic efficacy of peramivir against influenza B virus, we next compared the inhibitory effects of single intravenous administration of peramivir with those of repeated oral administration of oseltamivir phosphate using cynomolgus macaques. Virus titers, virus titer AUC, body temperature and the IL-6 AUC in nasal swabs of macaques treated with peramivir were lower than those of the control group (Figs. 5 to 7). These findings were concordant with our previous study that the levels of IL-6 in the nasal swabs were correlated with both virus replication and symptoms (23). These results observed in macaques and ferrets suggest that virus propagation in the nasal cavity in the early phase of infection is related to body temperature during infection. On the other hand, the virus titer on day 7 pi and the IL-6 AUC in nasal swabs of macaques treated with oseltamivir phosphate were significantly lower than those of the control group but body temperature and virus titer AUC were not. It was thought that reduction of virus titers in the late phase might cause a low IL-6 level on days 7 to 8 pi resulting in a decrease in the IL-6 AUC (data not shown). However, it did not affect suppression of fever in macaques treated with oseltamivir phosphate since the virus titer in the early phase was not suppressed. These results also support importance of suppression on virus replication in the early phase of infection to reduce symptoms of influenza and significance of the early treatment with antiviral agents. Although oseltamivir phosphate has been administered twice a day in humans, we 20

21 Kitano et al., Page administered oseltamivir phosphate once a day for 5 days in the present macaque study for the following reasons. Firstly, since anesthesia was necessary for drug administration and it decreased body temperature, anesthesia twice a day would make evaluation of macaque s body temperature change and feeding of food to macaques difficult. Secondly, there was no significant difference in duration of virus shedding and total symptom scores after treatment with oseltamivir phosphate between once and twice a day in the previous study although a group treated with oseltamivir once a daily showed a higher virus titer AUC than did a group treated twice daily (13). Thirdly, we confirmed that the C max of oseltamivir carboxylate in plasma of macaques was 6440 ± 3180 ng/ml after oral administration of 30 mg/kg dose of oseltamivir phosphate, which was 1,570 times higher than enzyme inhibition IC 50 value (4.1 ng/ml = 13.1 nm) against B/SendaiH/1051/2007 (Tables 1 and 2). In addition, the plasma concentration of oseltamivir carboxylate after 24 h (36 ± 8.0 ng/ml) was still 9 times higher than the IC 50 value (4.1 ng/ml) (Fig. 1 and Table 1). Therefore, one dose administration of oseltamivir in the macaque model was expected to show pharmacological effects from the aspect of pharmacokinetics. Furthermore, we did not detect mutations in NA genes in nasal samples from macaques after treatment with oseltamivir once daily as well as after treatment with peramivir though substitution of an amino acid was detected in a nasal wash sample of a ferret treated with oseltamivir twice a daily. Nonetheless, low effectiveness of oseltamivir against influenza B virus was demonstrated by not only virus titer in nasal swab fluid but also symptom in the macaque model. These results partially coincide with clinical studies that showed the longer duration of virus shedding and fever in patients treated with oseltamivir phosphate in influenza B virus infection than in influenza A virus infection (21, 22, 32). On the other hand, in the macaque model, intravenous 21

22 Kitano et al., Page administration of 30 mg/kg dose of peramivir can achieve C max of up to 160 µg/ml, which is 100,000 times higher than the IC 50 values (1.4 ng/ml = 4.26 nm) against B/SendaiH/1051/2007 (Fig. 1, Tables 1 and 2). The concentration of peramivir in plasma dropped quickly after injection but the level of peramivir at 24 h postdosing (52.4 ng/ml) was still 37 times higher than the IC 50 value (1.4 ng/ml). Therefore, the high concentration of peramivir in plasma during 24 h after injection is thought to be effective to reduce virus replication in the early phase of infection. In our previous study, HI and neutralization activities in sera of macaque were detected 6 to 8 days after infection and coincided with decreasing virus titers in swab samples (23). In the present study, HI activities against B/SendaiH/1051/2007 were observed in sera of macaques treated with peramivir or oseltamivir phosphate after day 8 pi at the comparable level of sera from untreated macaques. These results indicate that peramivir and oseltamivir did not inhibit production of anti-ha antibodies against B/SendaiH/1051/2007 in cynomolgus macaques. It is thought that the titers of anti-ha antibodies against B/SendaiH/1051/2007 in treated cynomolgus macaques may be sufficient to protect from challenge with the same virus strain since macaques possessing similar or lower HI titers of sera after vaccination with an H7 vaccine were protected from infection with a different H7 strain in the previous experiment (18). In the present study, we demonstrated that peramivir injected once intravenously had beneficial effects on viral titers and symptoms of ferrets and cynomolgus macaques with influenza B virus infection. The effects observed with peramivir treatment seem to be superior to those observed with once daily oseltamivir phosphate treatment in cynomolgus macaque when oseltamivir was given once daily. Therefore, single intravenous administration of peramivir could be an alternative to oseltamivir to treat patients with acute influenza B virus infection. 22

23 Kitano et al., Page ACKNOWLEDGEMENTS We would like to thank Dr. Hidekazu Nishimura for providing B/SendaiH/1051/2007, Dr. Tetsuo Kase for providing B/Kadoma/1/2005, Drs. Hideaki Tsuchiya, Norio Okahara and Takahiro Nakagawa for animal care, Drs. Shinya Omoto and Shinobu Kawauchi-Miki for valuable discussions and Hiroko Iwasaki and Mayumi Kakui for help on enzyme assay. All work reported here was financially supported by Shionogi Co., Ltd. REFERENCES 1. Air, G. M., W. G. Laver, M. Luo, S. J. Stray, G. Legrone, and R. G. Webster Antigenic, sequence, and crystal variation in influenza B neuraminidase. Virology 177: Babu, Y. S., P. Chand, S. Bantia, P. Kotian, A. Dehghani, Y. El-Kattan, T. H. Lin, T. L. Hutchison, A. J. Elliott, C. D. Parker, S. L. Ananth, L. L. Horn, G. W. Laver, and J. A. Montgomery BCX-1812 (RWJ ): discovery of a novel, highly potent, orally active, and selective influenza neuraminidase inhibitor through structure-based drug design. J Med Chem 43: Bantia, S., C. D. Parker, S. L. Ananth, L. L. Horn, K. Andries, P. Chand, P. L. Kotian, A. Dehghani, Y. El-Kattan, T. Lin, T. L. Hutchison, J. A. Montgomery, D. L. Kellog, and Y. S. Babu Comparison of the anti-influenza virus activity of RWJ with those of oseltamivir and zanamivir. Antimicrob Agents Chemother 45:

24 Kitano et al., Page Birnkrant, D., and E. Cox The Emergency Use Authorization of peramivir for treatment of 2009 H1N1 influenza. N Engl J Med 361: Boivin, G., and N. Goyette Susceptibility of recent Canadian influenza A and B virus isolates to different neuraminidase inhibitors. Antiviral Res 54: Boltz, D. A., N. A. Ilyushina, C. S. Arnold, Y. S. Babu, R. G. Webster, and E. A. Govorkova Intramuscularly administered neuraminidase inhibitor peramivir is effective against lethal H5N1 influenza virus in mice. Antiviral Res 80: Chand, P., Y. S. Babu, S. Bantia, N. Chu, L. B. Cole, P. L. Kotian, W. G. Laver, J. A. Montgomery, V. P. Pathak, S. L. Petty, D. P. Shrout, D. A. Walsh, and G. M. Walsh Design and synthesis of benzoic acid derivatives as influenza neuraminidase inhibitors using structure-based drug design. J Med Chem 40: Collins, P. J., L. F. Haire, Y. P. Lin, J. Liu, R. J. Russell, P. A. Walker, J. J. Skehel, S. R. Martin, A. J. Hay, and S. J. Gamblin Crystal structures of oseltamivir-resistant influenza virus neuraminidase mutants. Nature 453: Govorkova, E. A., N. A. Ilyushina, D. A. Boltz, A. Douglas, N. Yilmaz, and R. G. Webster Efficacy of oseltamivir therapy in ferrets inoculated with different clades of H5N1 influenza virus. Antimicrob Agents Chemother 51: Govorkova, E. A., I. A. Leneva, O. G. Goloubeva, K. Bush, and R. G. Webster Comparison of efficacies of RWJ , zanamivir, and oseltamivir against H5N1, H9N2, and other avian influenza viruses. Antimicrob Agents Chemother 45: Gubareva, L. V., M. N. Matrosovich, M. K. Brenner, R. C. Bethell, and R. G. Webster Evidence for zanamivir resistance in an immunocompromised child 24

25 Kitano et al., Page infected with influenza B virus. J Infect Dis 178: Hatakeyama, S., N. Sugaya, M. Ito, M. Yamazaki, M. Ichikawa, K. Kimura, M. Kiso, H. Shimizu, C. Kawakami, K. Koike, K. Mitamura, and Y. Kawaoka Emergence of influenza B viruses with reduced sensitivity to neuraminidase inhibitors. JAMA 297: Hayden, F. G., J. J. Treanor, R. S. Fritz, M. Lobo, R. F. Betts, M. Miller, N. Kinnersley, R. G. Mills, P. Ward, and S. E. Straus Use of the oral neuraminidase inhibitor oseltamivir in experimental human influenza: randomized controlled trials for prevention and treatment. JAMA 282: Herlocher, M. L., S. Elias, R. Truscon, S. Harrison, D. Mindell, C. Simon, and A. S. Monto Ferrets as a transmission model for influenza: sequence changes in HA1 of type A (H3N2) virus. J Infect Dis 184: Hurt, A. C., P. Iannello, K. Jachno, N. Komadina, A. W. Hampson, I. G. Barr, and J. L. McKimm-Breschkin Neuraminidase inhibitor-resistant and -sensitive influenza B viruses isolated from an untreated human patient. Antimicrob Agents Chemother 50: Hurt, A. C., S. S. Nor'e, J. M. McCaw, H. R. Fryer, J. Mosse, A. R. McLean, and I. G. Barr Assessing the viral fitness of oseltamivir-resistant influenza viruses in ferrets, using a competitive-mixtures model. J Virol 84: Itoh, Y., K. Shinya, M. Kiso, T. Watanabe, Y. Sakoda, M. Hatta, Y. Muramoto, D. Tamura, Y. Sakai-Tagawa, T. Noda, S. Sakabe, M. Imai, Y. Hatta, S. Watanabe, C. Li, S. Yamada, K. Fujii, S. Murakami, H. Imai, S. Kakugawa, M. Ito, R. Takano, K. 25

26 Kitano et al., Page Iwatsuki-Horimoto, M. Shimojima, T. Horimoto, H. Goto, K. Takahashi, A. Makino, H. Ishigaki, M. Nakayama, M. Okamatsu, K. Takahashi, D. Warshauer, P. A. Shult, R. Saito, H. Suzuki, Y. Furuta, M. Yamashita, K. Mitamura, K. Nakano, M. Nakamura, R. Brockman-Schneider, H. Mitamura, M. Yamazaki, N. Sugaya, M. Suresh, M. Ozawa, G. Neumann, J. Gern, H. Kida, K. Ogasawara, and Y. Kawaoka In vitro and in vivo characterization of new swine-origin H1N1 influenza viruses. Nature 460: Itoh, Y., H. Ozaki, H. Ishigaki, Y. Sakoda, T. Nagata, K. Soda, N. Isoda, T. Miyake, H. Ishida, K. Okamoto, M. Nakayama, H. Tsuchiya, R. Torii, H. Kida, and K. Ogasawara Subcutaneous inoculation of a whole virus particle vaccine prepared from a non-pathogenic virus library induces protective immunity against H7N7 highly pathogenic avian influenza virus in cynomolgus macaques. Vaccine 28: Jackson, S., N. Van Hoeven, L. M. Chen, T. R. Maines, N. J. Cox, J. M. Katz, and R. O. Donis Reassortment between avian H5N1 and human H3N2 influenza viruses in ferrets: a public health risk assessment. J Virol 83: Kawai, N., H. Ikematsu, N. Iwaki, N. Hirotsu, and S. Kashiwagi Prevalence of influenza B during the season in Japan. Clin Infect Dis 43: Kawai, N., H. Ikematsu, N. Iwaki, T. Kawashima, T. Maeda, S. Mitsuoka, K. Kondou, I. Satoh, K. Miyachi, S. Yamaga, T. Shigematsu, N. Hirotsu, and S. Kashiwagi Longer virus shedding in influenza B than in influenza A among outpatients treated with oseltamivir. J Infect 55: Kawai, N., H. Ikematsu, N. Iwaki, T. Maeda, I. Satoh, N. Hirotsu, and S. Kashiwagi. 26

27 Kitano et al., Page A comparison of the effectiveness of oseltamivir for the treatment of influenza A and influenza B: a Japanese multicenter study of the and influenza seasons. Clin Infect Dis 43: Kitano, M., Y. Itoh, M. Kodama, H. Ishigaki, M. Nakayama, T. Nagata, H. Ishida, H. Tsuchiya, R. Torii, K. Baba, R. Yoshida, A. Sato, and K. Ogasawara Establishment of a cynomolgus macaque model of influenza B virus infection. Virology 407: Kohno, S., H. Kida, M. Mizuguchi, and J. Shimada Efficacy and safety of intravenous peramivir for treatment of seasonal influenza virus infection. Antimicrob Agents Chemother 54: Kubo, S., T. Tomozawa, M. Kakuta, A. Tokumitsu, and M. Yamashita Laninamivir prodrug CS-8958, a long-acting neuraminidase inhibitor, shows superior anti-influenza virus activity after a single administration. Antimicrob Agents Chemother 54: Lee, N., P. K. Chan, D. S. Hui, T. H. Rainer, E. Wong, K. W. Choi, G. C. Lui, B. C. Wong, R. Y. Wong, W. Y. Lam, I. M. Chu, R. W. Lai, C. S. Cockram, and J. J. Sung Viral loads and duration of viral shedding in adult patients hospitalized with influenza. J Infect Dis 200: Matrosovich, M., T. Matrossovich, J. Carr, N. A. Roberts, and H.-D. Klenk Overexpression of the alpha-2,6-sialyltransferase in MDCK cells increases influenza virus sensitivity to neuraminidase inhibitors. J Virol 77: Munster, V. J., E. de Wit, J. M. van den Brand, S. Herfst, E. J. Schrauwen, T. M. 27

28 Kitano et al., Page Bestebroer, D. van de Vijver, C. A. Boucher, M. Koopmans, G. F. Rimmelzwaan, T. Kuiken, A. D. Osterhaus, and R. A. Fouchier Pathogenesis and transmission of swine-origin 2009 A(H1N1) influenza virus in ferrets. Science 325: Newland, J. G., J. R. Romero, M. Varman, C. Drake, A. Holst, T. Safranek, and K. Subbarao Encephalitis associated with influenza B virus infection in 2 children and a review of the literature. Clin Infect Dis 36:e Peltola, V., T. Ziegler, and O. Ruuskanen Influenza A and B virus infections in children. Clin Infect Dis 36: Pinto, L. H., and R. A. Lamb The M2 proton channels of influenza A and B viruses. J Biol Chem 281: Sugaya, N., K. Mitamura, M. Yamazaki, D. Tamura, M. Ichikawa, K. Kimura, C. Kawakami, M. Kiso, M. Ito, S. Hatakeyama, and Y. Kawaoka Lower clinical effectiveness of oseltamivir against influenza B contrasted with influenza A infection in children. Clin Infect Dis 44: Tashiro, M., J. L. McKimm-Breschkin, T. Saito, A. Klimov, C. Macken, M. Zambon, and F. G. Hayden Surveillance for neuraminidase-inhibitor-resistant influenza viruses in Japan, Antivir Ther 14: World Health Organization Influenza activity in the world, Global alert and response. (http://www.who.int/csr/disease/influenza/update/en/index.html). 35. Yun, N. E., N. S. Linde, M. A. Zacks, I. G. Barr, A. C. Hurt, J. N. Smith, N. Dziuba, M. R. Holbrook, L. Zhang, J. M. Kilpatrick, C. S. Arnold, and S. Paessler Injectable peramivir mitigates disease and promotes survival in ferrets and mice infected 28

29 Kitano et al., Page with the highly virulent influenza virus, A/Vietnam/1203/04 (H5N1). Virology 374:

In Vivo Influenza Virus-Inhibitory Effects of the Cyclopentane Neuraminidase Inhibitor RWJ

In Vivo Influenza Virus-Inhibitory Effects of the Cyclopentane Neuraminidase Inhibitor RWJ ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Mar. 2001, p. 749 757 Vol. 45, No. 3 0066-4804/01/$04.00 0 DOI: 10.1128/AAC.45.3.749 757.2001 Copyright 2001, American Society for Microbiology. All Rights Reserved.

More information

WHO biosafety risk assessment and guidelines for the production and quality control of human influenza pandemic vaccines: Update

WHO biosafety risk assessment and guidelines for the production and quality control of human influenza pandemic vaccines: Update WHO biosafety risk assessment and guidelines for the production and quality control of human influenza pandemic vaccines: Update 23 July 2009 Introduction This document updates guidance 1 from the World

More information

Evaluation of a new immunochromatographic assay for rapid identification of influenza A, B, and A(H1N1)2009 viruses

Evaluation of a new immunochromatographic assay for rapid identification of influenza A, B, and A(H1N1)2009 viruses J Infect Chemother (2013) 19:633 638 DOI 10.1007/s10156-012-0533-1 ORIGINAL ARTICLE Evaluation of a new immunochromatographic assay for rapid identification of influenza A, B, and A(H1N1)2009 viruses Keiko

More information

Recommended laboratory tests to identify influenza A/H5 virus in specimens from patients with an influenza-like illness

Recommended laboratory tests to identify influenza A/H5 virus in specimens from patients with an influenza-like illness World Health Organization Recommended laboratory tests to identify influenza A/H5 virus in specimens from patients with an influenza-like illness General information Highly pathogenic avian influenza (HPAI)

More information

Blocking Interhost Transmission of Influenza Virus by Vaccination in the Guinea Pig Model

Blocking Interhost Transmission of Influenza Virus by Vaccination in the Guinea Pig Model JOURNAL OF VIROLOGY, Apr. 2009, p. 2803 2818 Vol. 83, No. 7 0022-538X/09/$08.00 0 doi:10.1128/jvi.02424-08 Copyright 2009, American Society for Microbiology. All Rights Reserved. Blocking Interhost Transmission

More information

Application of Reverse Genetics to Influenza Vaccine Development

Application of Reverse Genetics to Influenza Vaccine Development NIAID Application of Reverse Genetics to Influenza Vaccine Development Kanta Subbarao Laboratory of Infectious Diseases NIAID, NIH Licensed Vaccines for Influenza Principle: Induction of a protective

More information

1918 Influenza; Influenza A, H1N1. Basic agent information. Section I- Infectious Agent. Section II- Dissemination

1918 Influenza; Influenza A, H1N1. Basic agent information. Section I- Infectious Agent. Section II- Dissemination 1918 Influenza; Influenza A, H1N1 Basic agent information Section I- Infectious Agent Risk Group: - RG3 Synonym or Cross reference: - Spanish Flu - 1918 Flu - El Grippe Characteristics: - SELECT AGENT

More information

Retrospective and Prospective Verification of the Cepheid Xpert Flu Assay

Retrospective and Prospective Verification of the Cepheid Xpert Flu Assay JCM Accepts, published online ahead of print on 20 July 2011 J. Clin. Microbiol. doi:10.1128/jcm.01162-11 Copyright 2011, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights

More information

Micro-plaque assay of influenza virus sensitivity to neuraminidase inhibitors

Micro-plaque assay of influenza virus sensitivity to neuraminidase inhibitors VIRGIL Antiviral Course 3-66 October 2006 Micro-plaque assay of influenza virus sensitivity to neuraminidase inhibitors Mikhail Matrosovich and Tatyana Matrosovich Influenza virus receptors: sialic acids

More information

Influenza virus.

Influenza virus. INFLUENZA VIRUS Adapté en partie des exposés de la Chaire Franqui 2003 "Antiviral drugs and Discoveries in Medicine" Prof. E. De Clercq, KU-Leuven http://www.md.ucl.ac.be/chaire-francqui/ Influenza virus

More information

ESwab challenges influenza virus propagation in cell cultures

ESwab challenges influenza virus propagation in cell cultures Rapid communications ESwab challenges influenza virus propagation in cell cultures R Trebbien (ratr@ssi.dk) 1, B Andersen 1, J Rønn 1, J McCauley 2, T Kølsen Fischer 1 1. National Influenza Center Denmark,

More information

In Vitro Generation of Neuraminidase Inhibitor Resistance in A(H5N1) Influenza Viruses

In Vitro Generation of Neuraminidase Inhibitor Resistance in A(H5N1) Influenza Viruses ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Oct. 2009, p. 4433 4440 Vol. 53, No. 10 0066-4804/09/$08.00 0 doi:10.1128/aac.00334-09 Copyright 2009, American Society for Microbiology. All Rights Reserved. In

More information

What is avian influenza (bird flu)? How does avian influenza spread among birds? Do avian influenza viruses infect humans?

What is avian influenza (bird flu)? How does avian influenza spread among birds? Do avian influenza viruses infect humans? Bird Flu FAQ Questions and Answers About Avian Influenza (Bird Flu) and Avian Influenza A (H5N1) Virus U.S. Centers for Disease Control, March 17, 2006 What is avian influenza (bird flu)? Avian influenza

More information

6.2 Pandemic influenza

6.2 Pandemic influenza 6.2 Pandemic influenza See Background Paper 6.2 (BP6_2Pandemic.pdf) Background and developments since 2004 The WHO estimates that annual influenza epidemics account for about 3 to 5 million cases of severe

More information

Influenza A 6/23/2010. Lisa Winston, MD UCSF / San Francisco General Hospital Divisions of Infectious Diseases and Hospital Medicine

Influenza A 6/23/2010. Lisa Winston, MD UCSF / San Francisco General Hospital Divisions of Infectious Diseases and Hospital Medicine Influenza Update in a Pandemic Year Nothing to disclose. Lisa Winston, MD UCSF / San Francisco General Hospital Divisions of Infectious Diseases and Hospital Medicine Influenza Biology Influenza Biology

More information

Public health relevant virological features of Influenza A(H7N9) causing human infection in China

Public health relevant virological features of Influenza A(H7N9) causing human infection in China Public health relevant virological features of Influenza A(H7N9) causing human infection in China Address requests about publications of the WHO Regional Office for Europe to: Publications WHO Regional

More information

Influenza. Gwen Clutario, Terry Chhour, Karen Lee

Influenza. Gwen Clutario, Terry Chhour, Karen Lee Influenza Gwen Clutario, Terry Chhour, Karen Lee Overview Commonly referred to as the flu Defined as a highly contagious viral infection where it starts at the upper respiratory tract and attacks the nose,

More information

(;[rowth Charaeteristies of Influenza Virus Type C in Avian Hosts

(;[rowth Charaeteristies of Influenza Virus Type C in Avian Hosts Archives of Virology 58, 349--353 (1978) Archives of Virology by Springer-Verlag 1978 (;[rowth Charaeteristies of Influena Virus Type C in Avian Hosts Brief Report By M ~R A~N D. AUSTIn, A. S. MONTO, and

More information

INFLUENZA VIRUS. Influenza virus

INFLUENZA VIRUS. Influenza virus IFLUEZA VIRUS Adapté en partie des exposés de la Chaire Franqui 2003 "Antiviral drugs and Discoveries in Medicine" Prof. E. De Clercq, KU-Leuven http://www.md.ucl.ac.be/chaire-francqui/ Influenza virus

More information

Role of Interferon in the Propagation of MM Virus in L Cells

Role of Interferon in the Propagation of MM Virus in L Cells APPLIED MICROBIOLOGY, Oct. 1969, p. 584-588 Copyright ( 1969 American Society for Microbiology Vol. 18, No. 4 Printed in U S A. Role of Interferon in the Propagation of MM Virus in L Cells DAVID J. GIRON

More information

Influenza virus.

Influenza virus. INFLUENZA VIRUS Adapté en partie des exposés de la Chaire Franqui 2003 "Antiviral drugs and Discoveries in Medicine" Prof. E. De Clercq, KU-Leuven http://www.md.ucl.ac.be/chaire-francqui/ et de l'exposé

More information

Influenza A H1N1 (Swine Flu 2009) Hemagglutinin / HA ELISA Pair Set

Influenza A H1N1 (Swine Flu 2009) Hemagglutinin / HA ELISA Pair Set Influenza A H1N1 (Swine Flu 2009) Hemagglutinin / HA ELISA Pair Set Catalog Number : SEK001 To achieve the best assay results, this manual must be read carefully before using this product and the assay

More information

Influenza: The Threat of a Pandemic

Influenza: The Threat of a Pandemic April, 2009 Definitions Epidemic: An increase in disease above what you what would normally expect. Pandemic: A worldwide epidemic 2 What is Influenza? Also called Flu, it is a contagious respiratory illness

More information

Infant and Pediatric Influenza. Mike Czervinske RRT-NPS University of Kansas Medical Center

Infant and Pediatric Influenza. Mike Czervinske RRT-NPS University of Kansas Medical Center Infant and Pediatric Influenza Mike Czervinske RRT-NPS University of Kansas Medical Center Influenza Infants and Influenza Acute infection of the respiratory tract Nose Throat Possibly lungs Pathophysiology

More information

THIS ACTIVITY HAS EXPIRED. CME CREDIT IS NO LONGER AVAILABLE

THIS ACTIVITY HAS EXPIRED. CME CREDIT IS NO LONGER AVAILABLE THIS ACTIVITY HAS EXPIRED. CME CREDIT IS NO LONGER AVAILABLE The following content is provided for informational purposes only. PREVENTION AND CONTROL OF INFLUENZA Lisa McHugh, MPH Influenza can be a serious

More information

Understanding Flu Testing

Understanding Flu Testing TECHNICAL BULLETIN Vol. 1 No. 2, August 2005 Understanding Flu Testing 1 Flu Facts 2 Choosing a Rapid Flu A+B Test 4 Improved Medical Outcomes 5 Enhanced Economic Value 8 9 BD Directigen EZ Flu A+B Technology:

More information

2009 H1N1 Influenza ( Swine Flu ) Hemagglutinin ELISA kit

2009 H1N1 Influenza ( Swine Flu ) Hemagglutinin ELISA kit 2009 H1N1 Influenza ( Swine Flu ) Hemagglutinin ELISA kit Catalog Number : SEK001 To achieve the best assay results, this manual must be read carefully before using this product and the assay is run as

More information

Possible Modes of Transmission of Avian Viruses to People: Studies in Experimental Models

Possible Modes of Transmission of Avian Viruses to People: Studies in Experimental Models Possible Modes of Transmission of Avian Viruses to People: Studies in Experimental Models Jackie Katz and Terry Tumpey Influenza Division CDC, Atlanta, Georgia David E. Swayne USDA/Agricultural Research

More information

Influenza vaccines: present and future

Influenza vaccines: present and future PERSPECTIVE SERIES The future of vaccine design Peter Palese and Adolfo García-Sastre, Series Editors Influenza vaccines: present and future Peter Palese and Adolfo García-Sastre Department of Microbiology,

More information

IN VIVO STUDIES ON VIRAL VIRULENCE

IN VIVO STUDIES ON VIRAL VIRULENCE IN VIVO STUDIES ON VIRAL VIRULENCE M.Phil student: Emily TSUI Supervisor: Professor Paul K.S Chan Department of Microbiology, CUHK Date: 15th Dec, 2014 Viral Virulence Capacity of a virus to cause disease

More information

Anti-influenza (M2 and NA) Drugs

Anti-influenza (M2 and NA) Drugs Anti-influenza (M2 and NA) Drugs Alan J. Hay MRC National Institute for Medical Research, London Summer School on Influenza Siena 1-5 August 2011 Antiviral drugs licensed for use against influenza M2 (Flu

More information

HL-60 ATP assay for predicting rat oral toxicity study

HL-60 ATP assay for predicting rat oral toxicity study AATEX 14, Special Issue, 699-703 Proc. 6th World Congress on Alternatives & Animal Use in the Life Sciences August 21-25, 2007, Tokyo, Japan HL-60 ATP assay for predicting rat oral toxicity study Yumiko

More information

Isolation of Influenza C Virus from Pigs and Experimental Infection of Pigs with Influenza C Virus

Isolation of Influenza C Virus from Pigs and Experimental Infection of Pigs with Influenza C Virus J. gen. Virol. (1983), 64, 177-182. Printed in Great Britain 177 Key words: influenza C virus/antibodies/pigs Isolation of Influenza C Virus from Pigs and Experimental Infection of Pigs with Influenza

More information

In vitro modeling of the interaction between human epithelial cells and lymphocytes upon influenza infection

In vitro modeling of the interaction between human epithelial cells and lymphocytes upon influenza infection DOI:10.1111/irv.12394 www.influenzajournal.com Short Article In vitro modeling of the interaction between human epithelial cells and lymphocytes upon influenza infection Natalia A. Ilyushina, a Peter F.

More information

Active and Passive Immunization for Avian Influenza Virus Infections

Active and Passive Immunization for Avian Influenza Virus Infections NIAID Active and Passive Immunization for Avian Influenza Virus Infections Kanta Subbarao, MD, MPH Laboratory of Infectious Diseases NIAID, NIH Immortalizing H5 HA-Specific Memory B Cells Collection of

More information

Effects of the combination of favipiravir (T-705) and oseltamivir on influenza a virus infections in mice

Effects of the combination of favipiravir (T-705) and oseltamivir on influenza a virus infections in mice Utah State University From the SelectedWorks of John D. Morrey January 1, 2010 Effects of the combination of favipiravir (T-705) and oseltamivir on influenza a virus infections in mice D F Smee B L Hurst

More information

Rapid-VIDITEST Swine Flu

Rapid-VIDITEST Swine Flu Rapid-VIDITEST Swine Flu One Step Influenza type A Antigen Card test. Instruction manual Producer: VIDIA spol. s r.o., Nad Safinou II 365, 252 50 Vestec, Czech Republic, Tel.: +420 261 090 565, www.vidia.cz

More information

Animal Models for Influenza Viruses: Implications for Universal Vaccine Development

Animal Models for Influenza Viruses: Implications for Universal Vaccine Development Pathogens 2014, 3, 845-874; doi:10.3390/pathogens3040845 Review OPEN ACCESS pathogens ISSN 2076-0817 www.mdpi.com/journal/pathogens Animal Models for Influenza Viruses: Implications for Universal Vaccine

More information

Influenza: Ecology and Continuing Evolution

Influenza: Ecology and Continuing Evolution Influenza: Ecology and Continuing Evolution Robert G. Webster, PhD Division of Virology Department of Infectious Diseases St. Jude Children s s Research Hospital Influenza Virus Negative sense RNA virus

More information

Pandemic Planning Update. Anita L. Barkin, DrPH, MSN, CRNP ACHA Annual Meeting Orlando, Florida 2008

Pandemic Planning Update. Anita L. Barkin, DrPH, MSN, CRNP ACHA Annual Meeting Orlando, Florida 2008 Pandemic Planning Update Anita L. Barkin, DrPH, MSN, CRNP ACHA Annual Meeting Orlando, Florida 2008 Current Status of H5N1 383 human cases (5/29/08) 62% fatality rate Median age 18-20 previously healthy

More information

NEXT GENERATION SEQUENCING OPENS NEW VIEWS ON VIRUS EVOLUTION AND EPIDEMIOLOGY. 16th International WAVLD symposium, 10th OIE Seminar

NEXT GENERATION SEQUENCING OPENS NEW VIEWS ON VIRUS EVOLUTION AND EPIDEMIOLOGY. 16th International WAVLD symposium, 10th OIE Seminar NEXT GENERATION SEQUENCING OPENS NEW VIEWS ON VIRUS EVOLUTION AND EPIDEMIOLOGY S. Van Borm, I. Monne, D. King and T. Rosseel 16th International WAVLD symposium, 10th OIE Seminar 07.06.2013 Viral livestock

More information

Pandemic Preparedness Team Immunology and Pathogenesis Branch Influenza Division Centers for Disease Control and Prevention USA VERSION 1

Pandemic Preparedness Team Immunology and Pathogenesis Branch Influenza Division Centers for Disease Control and Prevention USA VERSION 1 MODIFIED HEMAGGLUTINATION INHIBITION (HI) ASSAY USING HORSE RBCS FOR SEROLOGIC DETECTION OF ANTIBODIES TO H7 SUBTYPE AVIAN INFLUENZA VIRUS IN HUMAN SERA Pandemic Preparedness Team Immunology and Pathogenesis

More information

EVALUATION OF A NOVEL ANTIVIRAL FOR INFLUENZA INFECTION IN THE FERRET MODEL. Allison Leigh Brichacek

EVALUATION OF A NOVEL ANTIVIRAL FOR INFLUENZA INFECTION IN THE FERRET MODEL. Allison Leigh Brichacek EVALUATION OF A NOVEL ANTIVIRAL FOR INFLUENZA INFECTION IN THE FERRET MODEL by Allison Leigh Brichacek B.S., University of Pittsburgh at Greensburg, 2012 Submitted to the Graduate Faculty of the Department

More information

Appendix B: Provincial Case Definitions for Reportable Diseases

Appendix B: Provincial Case Definitions for Reportable Diseases Infectious Diseases Protocol Appendix B: Provincial Case Definitions for Reportable Diseases Disease: Influenza Revised December 2014 Influenza 1.0 Provincial Reporting Confirmed cases of disease 2.0 Type

More information

Reverse genetic platform for inactivated and live-attenuated influenza vaccine

Reverse genetic platform for inactivated and live-attenuated influenza vaccine EXPERIMENTAL and MOLECULAR MEDICINE, Vol. 42, No. 2, 116-121, February 2010 Reverse genetic platform for inactivated and live-attenuated influenza vaccine Eun-Ju Jung, Kwang-Hee Lee and Baik Lin Seong

More information

Diagnostic Methods of HBV and HDV infections

Diagnostic Methods of HBV and HDV infections Diagnostic Methods of HBV and HDV infections Zohreh Sharifi,ph.D Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine Hepatitis B-laboratory diagnosis Detection

More information

Influenza virus.

Influenza virus. INFLUENZA VIRUS Adapté en partie des exposés de la Chaire Franqui 2003 "Antiviral drugs and Discoveries in Medicine" Prof. E. De Clercq, KU-Leuven http://www.md.ucl.ac.be/chaire-francqui/ et de l'exposé

More information

Plaque Assay of Sendai Virus in Monolayers of a Clonal Line

Plaque Assay of Sendai Virus in Monolayers of a Clonal Line JOURNAL OF CUNICAL MICROBIOLOGY, Feb. 1976. p. 91-95 Copyright 1976 American Society for Microbiology Vol. 3, No. 2 Printed in U.SA. Plaque Assay of Sendai Virus in Monolayers of a Clonal Line of Porcine

More information

Influenza: Seasonal, Avian, and Otherwise

Influenza: Seasonal, Avian, and Otherwise Influenza: Seasonal, Avian, and Otherwise Lisa Winston, MD University of California, San Francisco San Francisco General Hospital Influenza biology Antiviral medications Seasonal influenza Vaccination

More information

The PB2-E627K Mutation Attenuates Viruses Containing the 2009 H1N1 Influenza Pandemic Polymerase

The PB2-E627K Mutation Attenuates Viruses Containing the 2009 H1N1 Influenza Pandemic Polymerase RESEARCH ARTICLE The PB2-E627K Mutation Attenuates Viruses Containing the 2009 H1N1 Influenza Pandemic Polymerase Brett W. Jagger, a * Matthew J. Memoli, a Zong-Mei Sheng, a Li Qi, a Rachel J. Hrabal,

More information

Antiviral Research xxx (2005) xxx xxx. Short communication

Antiviral Research xxx (2005) xxx xxx. Short communication Antiviral Research xxx (2005) xxx xxx 3 4 5 6 7 8 Short communication Sensitivity of influenza viruses to zanamivir and oseltamivir: A study performed on viruses circulating in France prior to the introduction

More information

Summary of neuraminidase amino acid substitutions associated with reduced inhibition by neuraminidase inhibitors (NAI)*

Summary of neuraminidase amino acid substitutions associated with reduced inhibition by neuraminidase inhibitors (NAI)* Summary of neuraminidase amino acid substitutions associated with reduced inhibition by neuraminidase inhibitors (NAI)* Susceptibility assessed by NA inhibition assays Source of Type/Subtype Amino acid

More information

MAJOR ARTICLE. Department of Virology, Erasmus Medical Centre and 2 ViroClinics Biosciences BV, Rotterdam, The Netherlands

MAJOR ARTICLE. Department of Virology, Erasmus Medical Centre and 2 ViroClinics Biosciences BV, Rotterdam, The Netherlands MAJOR ARTICLE Severity of Pneumonia Due to New H1N1 Influenza Virus in Ferrets Is Intermediate between That Due to Seasonal H1N1 Virus and Highly Pathogenic Avian Influenza H5N1 Virus Judith M. A. van

More information

Modification of the Ferret Model for Pneumonia From Seasonal Human Influenza A Virus Infection

Modification of the Ferret Model for Pneumonia From Seasonal Human Influenza A Virus Infection Modification of the Ferret Model for Pneumonia From Seasonal Human Influenza A Virus Infection Veterinary Pathology 49(3) 562-568 ª The Author(s) 2012 Reprints and permission: sagepub.com/journalspermissions.nav

More information

Yellow Fever Vaccine: Direct Challenge of Monkeys Given Graded Doses of 17D

Yellow Fever Vaccine: Direct Challenge of Monkeys Given Graded Doses of 17D AppuzD MmcoaioLOGy, Apr. 1973, p. 539-544. Copyright i 1973 American Society for Microbiology Vol. 25, No. 4 Printed in U.SA. Yellow Fever Vaccine: Direct Challenge of Monkeys Given Graded Doses of 17D

More information

PUBLIC HEALTH SIGNIFICANCE SEASONAL INFLUENZA AVIAN INFLUENZA SWINE INFLUENZA

PUBLIC HEALTH SIGNIFICANCE SEASONAL INFLUENZA AVIAN INFLUENZA SWINE INFLUENZA INFLUENZA DEFINITION Influenza is an acute highly infectious viral disease characterized by fever, general and respiratory tract catarrhal manifestations. Influenza has 3 Types Seasonal Influenza Avian

More information

A Live Attenuated H7N7 Candidate Vaccine Virus Induces Neutralizing Antibody That Confers Protection from Challenge in Mice, Ferrets, and Monkeys

A Live Attenuated H7N7 Candidate Vaccine Virus Induces Neutralizing Antibody That Confers Protection from Challenge in Mice, Ferrets, and Monkeys JOURNAL OF VIROLOGY, Nov. 2010, p. 11950 11960 Vol. 84, No. 22 0022-538X/10/$12.00 doi:10.1128/jvi.01305-10 Copyright 2010, American Society for Microbiology. All Rights Reserved. A Live Attenuated H7N7

More information

Meta Analysis: Blood Products for Spanish Influenza Pneumonia: A Future H5N1 Treatment?

Meta Analysis: Blood Products for Spanish Influenza Pneumonia: A Future H5N1 Treatment? Meta Analysis: Blood Products for Spanish Influenza Pneumonia: A Future H5N1 Treatment? Luke TC, Kilbane EM, Jackson JE, Hoffman SL. Annals of Internal Medicine. 2006 Oct 17;145(8):599-609. Annual Death

More information

Telephone Triage of Patients with Influenza

Telephone Triage of Patients with Influenza Telephone Triage of Patients with Influenza JONATHAN L. TEMTE, MD, PhD University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin Published ahead of print (American Family Physician)

More information

Heat-killed Lactobacillus casei

Heat-killed Lactobacillus casei Heat-killed Lactobacillus casei confers broad protection against influenza A virus primary infection and develops heterosubtypic immunity against future secondary infection Yu-Jin Jung, Young-Tae Lee,

More information

Patricia Fitzgerald-Bocarsly

Patricia Fitzgerald-Bocarsly FLU Patricia Fitzgerald-Bocarsly October 23, 2008 Orthomyxoviruses Orthomyxo virus (ortho = true or correct ) Negative-sense RNA virus (complementary to mrna) Five different genera Influenza A, B, C Thogotovirus

More information

NOVEL INFLUENZA A (H1N1) Swine Flu

NOVEL INFLUENZA A (H1N1) Swine Flu Introduction Definitions Influenza-like Illness Emergency Department Assessment Anitiviral Medication Oseltamivir (Tamiflu) Dosing Infection Control Issues Staff Exposure References Introduction This guideline

More information

H5N1 ( Avian Flu ) Hemagglutinin ELISA Pair Set

H5N1 ( Avian Flu ) Hemagglutinin ELISA Pair Set H5N1 ( Avian Flu ) Hemagglutinin ELISA Pair Set Catalog Number : SEK002 To achieve the best assay results, this manual must be read carefully before using this product and the assay is run as summarized

More information

Effect of Complement and Viral Filtration on the

Effect of Complement and Viral Filtration on the APPLIED MICROBIOLOGY, JUlY 1968, p. 1076-1080 Copyright @ 1968 American Society for Microbiology Vol. 16, No. 7 Printed in U.S.A. Effect of Complement and Viral Filtration on the Neutralization of Respiratory

More information

Phosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for use with the PrimePCR Reverse Transcription Control Assay

Phosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for use with the PrimePCR Reverse Transcription Control Assay Catalog # Description 172-5080 SingleShot Cell Lysis Kit, 100 x 50 µl reactions 172-5081 SingleShot Cell Lysis Kit, 500 x 50 µl reactions For research purposes only. Introduction The SingleShot Cell Lysis

More information

HAI and NAI as Correlates of Protection After Influenza Vaccination

HAI and NAI as Correlates of Protection After Influenza Vaccination HAI and NAI as Correlates of Protection After Influenza Vaccination Arnold S. Monto Thomas Francis Jr. Professor University of Michigan School of Public Health Ann Arbor, Michigan Having Correlates is

More information

however, and the present communication is concerned with some of

however, and the present communication is concerned with some of THE AGGLUTINATION OF HUMAN ERYTHROCYTES MODIFIED BY TREATMENT WITH NEWCASTLE DISEASE AND INFLUENZA VIRUS' ALFRED L. FLORMAN' Pediatric Service and Division of Bacteriology, The Mount Sinai Hospital, New

More information

UNIVERSAL INFLUENZA VIRUS VACCINES Adolfo García Sastre. Icahn School of Medicine at Mount Sinai, New York

UNIVERSAL INFLUENZA VIRUS VACCINES Adolfo García Sastre. Icahn School of Medicine at Mount Sinai, New York UNIVERSAL INFLUENZA VIRUS VACCINES Adolfo García Sastre Icahn School of Medicine at Mount Sinai, New York INFLUENZA VIRUSES PAx B EPIDEMIOLOGY OF HUMAN INFLUENZA VIRUSES A H1N1 H3N2 1968 H2N2 1957 H1N1

More information

MEETING THE STANDARDS: FDA MANDATORY RAPID INFLUENZA DETECTION TEST (RIDTs) RECLASSIFICATION

MEETING THE STANDARDS: FDA MANDATORY RAPID INFLUENZA DETECTION TEST (RIDTs) RECLASSIFICATION MEETING THE STANDARDS: FDA MANDATORY RAPID INFLUENZA DETECTION TEST (RIDTs) RECLASSIFICATION NOVEMBER 6, 2017 SALLY A. HOJVAT M.Sc., Ph.D. Retired as Director of FDA Division of Microbiology Devices, CDRH

More information

Amino acid sequence identity between the HA1 of influenza A (H3N2) viruses grown in mammalian and primary chick kidney cells

Amino acid sequence identity between the HA1 of influenza A (H3N2) viruses grown in mammalian and primary chick kidney cells Journal of General Virology (1992), 73, 1159-1165. Printed in Great Britain 1159 Amino acid sequence identity between the HA1 of influenza A (H3N2) viruses grown in mammalian and primary chick kidney cells

More information

Pharmacological management of viruses in obese patients

Pharmacological management of viruses in obese patients Cubist Pharmaceuticals The Shape of Cures to Come Pharmacological management of viruses in obese patients Dr. Dimitar Tonev, Medical Director UKINORD 1 Disclosures } The author is a pharmaceutical physician

More information

Influenza A virus subtype H5N1

Influenza A virus subtype H5N1 Influenza A virus subtype H5N1 Influenza A virus subtype H5N1, also known as A(H5N1) or simply H5N1, is a subtype of the Influenza A virus which can cause illness in humans and many other animal species.

More information

Influenza A (h5n1) virus causes severe disease in humans and poses

Influenza A (h5n1) virus causes severe disease in humans and poses The new england journal of medicine brief report Oseltamivir Resistance during Treatment of Influenza A (H5N1) Infection Menno D. de Jong, M.D., Ph.D., Tran Tan Thanh, M.Sc., Truong Huu Khanh, M.D., Vo

More information

Affinity of Doripenem and Comparators to Penicillin-Binding Proteins in Escherichia coli and ACCEPTED

Affinity of Doripenem and Comparators to Penicillin-Binding Proteins in Escherichia coli and ACCEPTED AAC Accepts, published online ahead of print on February 00 Antimicrob. Agents Chemother. doi:./aac.01-0 Copyright 00, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights

More information

ORIGINAL INVESTIGATION. Clinical Signs and Symptoms Predicting Influenza Infection

ORIGINAL INVESTIGATION. Clinical Signs and Symptoms Predicting Influenza Infection ORIGINAL INVESTIGATION Clinical Signs and Symptoms Predicting Influenza Infection Arnold S. Monto, MD; Stefan Gravenstein, MD; Michael Elliott, MD; Michael Colopy, PhD; Jo Schweinle, MD Background: New

More information

Neuraminidase Inhibitors for Influenza

Neuraminidase Inhibitors for Influenza review article drug therapy Neuraminidase Inhibitors for Influenza Anne Moscona, M.D. the impact of infection is felt globally each year when the disease develops in approximately 20 percent of the world

More information

Current Vaccines: Progress & Challenges. Influenza Vaccine what are the challenges?

Current Vaccines: Progress & Challenges. Influenza Vaccine what are the challenges? Current Vaccines: Progress & Challenges Influenza Vaccine what are the challenges? Professor John S. Tam The Hong Kong Polytechnic University Asia-Pacific Alliance for the Control of Influenza (APACI)

More information

Human infection with pandemic (H1N1) 2009 virus: updated interim WHO guidance on global surveillance

Human infection with pandemic (H1N1) 2009 virus: updated interim WHO guidance on global surveillance Human infection with pandemic (H1N1) 2009 virus: updated interim WHO guidance on global surveillance 10 July 2009 Background This document updates the interim WHO guidance on global surveillance of pandemic

More information

Clinical Development Challenges: Trial Designs and Endpoints

Clinical Development Challenges: Trial Designs and Endpoints Clinical Development Challenges: Trial Designs and Endpoints Menno de Jong Department of Medical Microbiology Academic Medical Center, University of Amsterdam ISIRV - Options IX for the Control of Influenza

More information

Comparison of Four Rapid Diagnostic Kits of Immunochromatography for Detection of Influenza A and Influenza B Viruses

Comparison of Four Rapid Diagnostic Kits of Immunochromatography for Detection of Influenza A and Influenza B Viruses Comparison of Four Rapid Diagnostic Kits of Immunochromatography for Detection of Influenza A and Influenza B Viruses Pornpicha Ying Zhao, Yao Wang, Min Zhong, Yingchun Xu and Anping Ni* Department of

More information

Practical guidance for national influenza centres establishing or implementing neuraminidase inhibitor susceptibility surveillance

Practical guidance for national influenza centres establishing or implementing neuraminidase inhibitor susceptibility surveillance Practical guidance for national influenza centres establishing or implementing neuraminidase inhibitor susceptibility surveillance WHO Library Cataloguing-in-Publication Data Practical guidance for national

More information

Attaching zanamivir to a polymer markedly enhances its activity against drug-resistant strains of influenza a virus

Attaching zanamivir to a polymer markedly enhances its activity against drug-resistant strains of influenza a virus Attaching zanamivir to a polymer markedly enhances its activity against drug-resistant strains of influenza a virus The MIT Faculty has made this article openly available. Please share how this access

More information

The humoral immune responses to IBV proteins.

The humoral immune responses to IBV proteins. The humoral immune responses to IBV proteins. E. Dan Heller and Rosa Meir The Hebrew University of Jerusalem, Israel COST FA1207 meeting WG2 + WG3, Budapest, Jan. 2015 1 IBV encodes four major structural

More information

WHO Technical Consultation on the severity of disease caused by the new influenza A (H1N1) virus infections

WHO Technical Consultation on the severity of disease caused by the new influenza A (H1N1) virus infections WHO Technical Consultation on the severity of disease caused by the new influenza A (H1N1) virus infections Original short summary posted 6 May 2009. Revised full report posted May 9 2009. On 5 May 2009

More information

UPDATE ON CANINE INFLUENZA IN TENNESSEE. Staci Cannon, DVM, MPH, DACVPM, DABVP (Shelter Medicine Practice)

UPDATE ON CANINE INFLUENZA IN TENNESSEE. Staci Cannon, DVM, MPH, DACVPM, DABVP (Shelter Medicine Practice) UPDATE ON CANINE INFLUENZA IN TENNESSEE Staci Cannon, DVM, MPH, DACVPM, DABVP (Shelter Medicine Practice) Headlines What is Canine Influenza? Highly contagious respiratory infection of dogs Caused by influenza

More information

Laboratory Diagnosis of Avian Influenza and Newcastle Disease

Laboratory Diagnosis of Avian Influenza and Newcastle Disease Laboratory Diagnosis of Avian Influenza and Newcastle Disease Dennis A. Senne dennis.a.senne@aphis.usda.gov (515) 239-7551 U. S. Department of Agriculture, Animal and Plant Health Inspection Service, Veterinary

More information

Restricted Infectivity of a Human-Lineage H3N2 Influenza A Virus in Pigs Is Hemagglutinin and Neuraminidase Gene Dependent

Restricted Infectivity of a Human-Lineage H3N2 Influenza A Virus in Pigs Is Hemagglutinin and Neuraminidase Gene Dependent JOURNAL OF CLINICAL MICROBIOLOGY, Feb. 2006, p. 297 301 Vol. 44, No. 2 0095-1137/06/$08.00 0 doi:10.1128/jcm.44.2.297 301.2006 Copyright 2006, American Society for Microbiology. All Rights Reserved. Restricted

More information

Swine H1N1 Influenza Human Pandemic Strain

Swine H1N1 Influenza Human Pandemic Strain TM Primerdesign Ltd Swine H1N1 Influenza Human Pandemic Strain M1 - global Influenza A & N1- specific for Swine H1N1 Influenza Human Pandemic Strain genesig Standard Kit 150 tests For general laboratory

More information

The 10 th Japan-Taiwan Symposium on Vaccine Preventable Diseases and Vector-Borne Diseases

The 10 th Japan-Taiwan Symposium on Vaccine Preventable Diseases and Vector-Borne Diseases The 10 th Japan-Taiwan Symposium on Vaccine Preventable Diseases and Vector-Borne Diseases September 12-13, 2013 the National Institute of Infectious Diseases, Tokyo, JAPAN September 12, Thursday Opening

More information

Justin Seroy, DO Infectious Disease

Justin Seroy, DO Infectious Disease Justin Seroy, DO Infectious Disease History The Virus Current statistics Presentation Treatment Vaccination Influenza viruses enveloped, ss- RNA virus (Types A, B, C) Causes acute, febrile illness Fever,

More information

Influenza A and B viruses infect 5 to 15% of the global population

Influenza A and B viruses infect 5 to 15% of the global population Comparative Study of Influenza Virus Replication in MDCK Cells and in Primary Cells Derived from Adenoids and Airway Epithelium Natalia A. Ilyushina, a Mine R. Ikizler, b Yoshihiro Kawaoka, c,d,e Larisa

More information

ADCC Assay Protocol Vikram Srivastava 1, Zheng Yang 1, Ivan Fan Ngai Hung 2, Jianqing Xu 3, Bojian Zheng 3 and Mei- Yun Zhang 3*

ADCC Assay Protocol Vikram Srivastava 1, Zheng Yang 1, Ivan Fan Ngai Hung 2, Jianqing Xu 3, Bojian Zheng 3 and Mei- Yun Zhang 3* ADCC Assay Protocol Vikram Srivastava 1, Zheng Yang 1, Ivan Fan Ngai Hung 2, Jianqing Xu 3, Bojian Zheng 3 and Mei- Yun Zhang 3* 1 Department of Microbiology, Li Ka Shing Faculty of Medicine, University

More information

Treatment of Influenza. Dr. YU Wai Cho

Treatment of Influenza. Dr. YU Wai Cho Treatment of Influenza Dr. YU Wai Cho Symptomatic Treatment Analgesics/ Antipyretics (avoid aspirin) Adequate fluids Rest Specific Drug Treatment Synthetic amines Amantadine Rimantadine Neuraminidase inhibitors

More information

Influenza A H1N1 HA ELISA Pair Set

Influenza A H1N1 HA ELISA Pair Set Influenza A H1N1 HA ELISA Pair Set for H1N1 ( A/Puerto Rico/8/1934 ) HA Catalog Number : SEK11684 To achieve the best assay results, this manual must be read carefully before using this product and the

More information

Swine Flu, Fiction or Reality

Swine Flu, Fiction or Reality Philadelphia University, Jordan From the SelectedWorks of Philadelphia University, Jordan 2009 Swine Flu, Fiction or Reality Philadelphia University, Philadelphia University Available at: https://works.bepress.com/philadelphia_university/98/

More information

Correlates of Protection for Flu vaccines and Assays Overview. by Simona Piccirella, PhD Chief Executive Officer

Correlates of Protection for Flu vaccines and Assays Overview. by Simona Piccirella, PhD Chief Executive Officer Correlates of Protection for Flu vaccines and Assays Overview by Simona Piccirella, PhD Chief Executive Officer Company Overview: VisMederi is an Italian private small enterprise established in 2009 and

More information

- They come in all sizes. -- General Structure is similar.

- They come in all sizes. -- General Structure is similar. - They come in all sizes. -- General Structure is similar. Centers for Disease Control (CDC) and Prevention. Influenza Prevention and Control. Influenza. Available at: http://www.cdc.gov/ncidod/diseases/flu/fluinfo.htm.

More information

Influenza Epidemiology,Treatment, and Prevention. Matt Zahn, MD Medical Director Epidemiology and Assessment Orange County Health Care Agency

Influenza Epidemiology,Treatment, and Prevention. Matt Zahn, MD Medical Director Epidemiology and Assessment Orange County Health Care Agency Influenza Epidemiology,Treatment, and Prevention Matt Zahn, MD Medical Director Epidemiology and Assessment Orange County Health Care Agency Orange County Population of 3,010,232 60.8% White 33.7% Hispanic

More information

Development of safe and immunogenic reassortant viruses with 5:3 genotype for live attenuated influenza vaccine

Development of safe and immunogenic reassortant viruses with 5:3 genotype for live attenuated influenza vaccine Development of safe and immunogenic reassortant viruses with 5:3 genotype for live attenuated influenza vaccine Irina Isakova-Sivak, PhD Institute of Experimental Medicine, Saint Petersburg, Russia The

More information

ECDC TECHNICAL REPORT

ECDC TECHNICAL REPORT TECHNICAL REPORT External quality assessment scheme for antiviral susceptibility detection in influenza viruses for the Community Network of Reference Laboratories for Human Influenza in Europe 2010/2011

More information