The Experts below are selected from a list of 240 Experts worldwide ranked by ideXlab platform

Seiji Hongo - One of the best experts on this subject based on the ideXlab platform.

  • EffeCt of Phosphorylation of CM2 Protein on Influenza C Virus RepliCation.
    Journal of virology, 2017
    Co-Authors: Takanari Goto, Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Yoshitaka Shimotai, Ri Sho, Seiji Hongo
    Abstract:

    CM2 is the seCond membrane protein of the Influenza C Virus and has been demonstrated to play a role in the unCoating and genome paCkaging proCesses in Influenza C Virus repliCation. Although the effeCts of N-linked glyCosylation, disulfide-linked oligomerization, and palmitoylation of CM2 on Virus repliCation have been analyzed, the effeCt of the phosphorylation of CM2 on Virus repliCation remains to be determined. In this study, a phosphorylation site(s) at residue 78 and/or 103 of CM2 was replaCed with an alanine residue(s), and the effeCts of the loss of phosphorylation on Influenza C Virus repliCation were analyzed. No signifiCant differenCes were observed in the paCkaging of the reporter gene between Influenza C Virus-like partiCles (VLPs) produCed from 293T Cells expressing wild-type CM2 and those from the Cells expressing the CM2 mutants laCking the phosphorylation site(s). Reporter gene expression in HMV-II Cells infeCted with VLPs Containing the CM2 mutants was inhibited in Comparison with that in Cells infeCted with wild-type VLPs. The Virus produCtion of the reCombinant Influenza C Virus possessing CM2 mutants Containing a serine-to-alanine Change at residue 78 was signifiCantly lower than that of wild-type reCombinant Influenza C Virus. Furthermore, the Virus growth of the reCombinant Viruses possessing CM2 with a serine-to-aspartiC aCid Change at position 78, to mimiC Constitutive phosphorylation, was virtually identiCal to that of the wild-type Virus. These results suggest that phosphorylation of CM2 plays a role in effiCient Virus repliCation, probably through the addition of a negative Charge to the Ser78 phosphorylation site.IMPORTANCE It is well-known that many host and viral proteins are posttranslationally modified by phosphorylation, whiCh plays a role in the funCtions of these proteins. In Influenza A and B Viruses, phosphorylation of viral proteins NP, M1, NS1, and the nuClear export protein (NEP), whiCh are not integrated into the membranes, affeCts the funCtions of these proteins, thereby affeCting Virus repliCation. However, it was reported that phosphorylation of the Influenza A Virus M2 ion Channel protein, whiCh is integrated into the membrane, has no effeCt on Virus repliCation in vitro or in vivo We previously demonstrated that the Influenza C Virus CM2 ion Channel protein is modified by N-glyCosylation, oligomerization, palmitoylation, and phosphorylation and have analyzed the effeCts of these modifiCations, exCept phosphorylation, on Virus repliCation. This is the first report demonstrating that phosphorylation of the Influenza C Virus CM2 ion Channel protein, unlike that of the Influenza A Virus M2 protein, plays a role in Virus repliCation.

  • IntrinsiC Temperature Sensitivity of Influenza C Virus Hemagglutinin-Esterase-Fusion Protein
    Journal of virology, 2012
    Co-Authors: Emi Takashita, Yasushi Muraki, Kanetsu Sugawara, Seiji Hongo, Hidekazu Nishimura, Hironobu Asao, Koji Suzuki, Takashi Tsuji, Yoshihiro Ohara, Yoshihiro Kawaoka
    Abstract:

    Influenza C Virus repliCates more effiCiently at 33°C than at 37°C. To determine whether hemagglutinin-esterase-fusion protein (HEF), a surfaCe glyCoprotein of Influenza C Virus, is a restriCting faCtor for this temperature sensitivity, we analyzed the biologiCal and bioChemiCal properties of HEF at 33°C and 37°C. We found that HEF exhibits intrinsiC temperature sensitivities for surfaCe expression and fusion aCtivity.

  • DeteCtion and quantifiCation of Influenza C Virus in pediatriC respiratory speCimens by real-time PCR and Comparison with infeCtious viral Counts
    Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2012
    Co-Authors: Yoko Matsuzaki, Kanetsu Sugawara, Chieko Abiko, Tatsuya Ikeda, Katsumi Mizuta, Yoshitaka Shimotai, Yoko Aoki, Seiji Hongo
    Abstract:

    AbstraCt BaCkground The epidemiologiCal and CliniCal impaCts of Influenza C Virus infeCtion may have been underestimated by Conventional viral Culture sCreening alone. ObjeCtive To evaluate a newly developed real-time polymerase Chain reaCtion (PCR) assay as a tool for diagnosing Influenza C Virus infeCtion. Study design The primers and probe for real-time PCR were designed to amplify the Conserved region of the nuCleoprotein gene based on the aligned sequenCes of nine isolates from 1967 to 2010. Respiratory speCimens from Children ColleCted between January 2010 and August 2010 were examined for the presenCe of Influenza C Virus by Cell Culture and real-time PCR. SpeCimens that were positive for the Virus using real-time PCR were further examined using an infeCtivity assay with embryonated hen's eggs. Results Of the 1203 speCimens examined, 34 (2.8%) tested positive for the Influenza C Virus by Cell Culture and 51 (4.2%) tested positive by real-time PCR. The mean viral load and infeCtivity titer in speCimens that tested positive using Cell Culture were 3.97 × 108 Copies/ml and 5.43 × 105 EID50/ml, respeCtively, and those in speCimens that were negative using Cell Culture were 2.18 × 106 Copies/ml and 3.67 × 102 EID50/ml, respeCtively. In the CliniCal speCimens with viral loads less than 105 Copies/ml, it was not possible to isolate the Virus using embryonated hen's eggs. The Copy number-to-EID50 ratio of the CliniCal speCimens was muCh higher, ranging from 32 to 278,000, than those of Culture fluid, ranging from 2.3 to 13.5. ConClusion The real-time PCR assay desCribed here Can be used as a sensitive method for diagnosing Influenza C Virus infeCtion.

  • Palmitoylation of CM2 is dispensable to Influenza C Virus repliCation.
    Virus research, 2011
    Co-Authors: Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Seiji Hongo, Takatoshi Furukawa, Takako Okuwa, Toshiki Himeda, Yoshiro Ohara
    Abstract:

    CM2 is the seCond membrane protein of Influenza C Virus. The signifiCanCe of the posttranslational modifiCations of CM2 remains to be Clarified in the Context of viral repliCation, although the positions of the modified amino aCids on CM2 have been determined. In the present study, using reverse genetiCs we generated rCM2-C65A, a reCombinant Influenza C Virus laCking CM2 palmitoylation site, in whiCh Cysteine at residue 65 of CM2 was mutated to alanine, and examined viral growth and viral protein synthesis in the reCombinant-infeCted Cells. The rCM2-C65A Virus grew as effiCiently as did the parental Virus in Cultured HMV-II Cells as well as in embryonated ChiCken eggs. The synthesis and bioChemiCal features of HEF, NP, M1 and mutant CM2 in the rCM2-C65A-infeCted HMV-II Cells were similar to those in the parental Virus-infeCted Cells. Furthermore, membrane flotation analysis of the infeCted Cells revealed that equal amount of viral proteins was reCovered in the plasma membrane fraCtions of the rCM2-C65A-infeCted Cells to that in the parental Virus-infeCted Cells. These findings indiCate that defeCt in palmitoylation of CM2 does not affeCt transport and maturation of HEF, NP and M1 as well as CM2 in Virus-infeCted Cells, and palmitoylation of CM2 is dispensable to Influenza C Virus repliCation.

  • role of the Cm2 protein in the Influenza C Virus repliCation CyCle
    Journal of Virology, 2011
    Co-Authors: Takatoshi Furukawa, Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Emi Takashita, Yoshitaka Shimotai, Ri Sho, Takeshi Noda, Seiji Hongo
    Abstract:

    CM2 is the seCond membrane protein of Influenza C Virus. Although its bioChemiCal CharaCteristiCs, Coding strategy, and properties as an ion Channel have been extensively studied, the role(s) of CM2 in the Virus repliCation CyCle remains to be Clarified. In order to eluCidate this role, in the present study we generated CM2-defiCient Influenza C Virus-like partiCles (VLPs) and examined the VLP-produCing 293T Cells, VLPs, and VLP-infeCted HMV-II Cells. QuantifiCation of viral RNA (vRNA) in the VLPs by real-time PCR revealed that the CM2-defiCient VLPs Contain approximately one-third of the vRNA found in wild-type VLPs although no signifiCant differenCes were deteCted in the expression levels of viral Components in VLP-produCing Cells or in the number and morphology of the generated VLPs. This finding suggests that CM2 is involved in the genome paCkaging proCess into VLPs. Furthermore, HMV-II Cells infeCted with CM2-defiCient VLPs exhibited signifiCantly reduCed reporter gene expression. Although CM2-defiCient VLPs Could be internalized into HMV-II Cells as effiCiently as wild-type VLPs, a smaller amount of vRNA was deteCted in the nuClear fraCtion of CM2-defiCient VLP-infeCted Cells than in that of wild-type VLP-infeCted Cells, suggesting that the unCoating proCess of the CM2-defiCient VLPs in the infeCted Cells did not proCeed in an appropriate manner. Taken together, the data obtained in the present study indiCate that CM2 has a potential role in the genome paCkaging and unCoating proCesses of the Virus repliCation CyCle.

Yasushi Muraki - One of the best experts on this subject based on the ideXlab platform.

  • unique direCtional motility of Influenza C Virus Controlled by its filamentous morphology and short range motions
    Journal of Virology, 2017
    Co-Authors: Tatsuya Sakai, Yasushi Muraki, Hiroaki Takagi, Mineki Saito
    Abstract:

    Influenza Virus motility is based on Cooperation between two viral spike proteins, hemagglutinin (HA) and neuraminidase (NA), and is a major determinant of Virus infeCtivity. To transloCate a Virus partiCle on the Cell surfaCe, HA moleCules exChange viral reCeptors and NA moleCules aCCelerate the reCeptor exChange of HA. This type of Virus motility was reCently identified in Influenza A Virus (IAV). To determine if other Influenza Virus types have a similar reCeptor exChange meChanism-driven motility, we investigated Influenza C Virus (ICV) motility on a reCeptor-fixed glass surfaCe. This system exCludes reCeptor mobility, whiCh makes it more desirable than a Cell surfaCe for demonstrating Virus motility by reCeptor exChange. Like IAV, ICV was observed to move aCross the reCeptor-fixed surfaCe. However, in Contrast to the random movement of IAV, a filamentous ICV strain, Ann Arbor/1/50 (AA), moved in a straight line, in a direCted manner, and at a Constant rate, whereas a spheriCal ICV strain, Taylor/1233/47 (Taylor), moved randomly, similar to IAV. The AA and Taylor Viruses eaCh moved with a Combination of gradual (Crawling) and rapid (gliding) motions, but the distanCes of Crawling and gliding for the AA Virus were shorter than those of the Taylor Virus. Our findings indiCate that like IAV, ICV also has a motility that is driven by the reCeptor exChange meChanism. However, Compared with IAV movement, filamentous ICV movement is highly regulated in both direCtion and speed. Control of ICV movement is based on its speCifiC motility employing short Crawling and gliding motions as well as its own filamentous morphology.IMPORTANCE Influenza Virus enters into a host Cell for infeCtion via Cellular endoCytosis. Human Influenza Virus infeCts epithelial Cells of the respiratory traCt, the surfaCes of whiCh are hidden by abundant Cilia that are inaCtive in endoCytosis. An open question is the manner by whiCh the Virus migrates to endoCytosis-aCtive domains. In analyzing individual Virus behaviors through single-Virus traCking, we identified a novel funCtion of the hemagglutinin and esterase of Influenza C Virus (ICV) as the motility maChinery. Hemagglutinin iteratively exChanges a viral reCeptor, Causing Virus movement. Esterase degrades the reCeptors along the trajeCtory traveled by the Virus and prevents the Virus from moving baCkward, Causing direCtional movement. We propose that ICV has a unique motile maChinery direCtionally Controlled via hemagglutinin sensing the reCeptor density manipulated by esterase.

  • EffeCt of Phosphorylation of CM2 Protein on Influenza C Virus RepliCation.
    Journal of virology, 2017
    Co-Authors: Takanari Goto, Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Yoshitaka Shimotai, Ri Sho, Seiji Hongo
    Abstract:

    CM2 is the seCond membrane protein of the Influenza C Virus and has been demonstrated to play a role in the unCoating and genome paCkaging proCesses in Influenza C Virus repliCation. Although the effeCts of N-linked glyCosylation, disulfide-linked oligomerization, and palmitoylation of CM2 on Virus repliCation have been analyzed, the effeCt of the phosphorylation of CM2 on Virus repliCation remains to be determined. In this study, a phosphorylation site(s) at residue 78 and/or 103 of CM2 was replaCed with an alanine residue(s), and the effeCts of the loss of phosphorylation on Influenza C Virus repliCation were analyzed. No signifiCant differenCes were observed in the paCkaging of the reporter gene between Influenza C Virus-like partiCles (VLPs) produCed from 293T Cells expressing wild-type CM2 and those from the Cells expressing the CM2 mutants laCking the phosphorylation site(s). Reporter gene expression in HMV-II Cells infeCted with VLPs Containing the CM2 mutants was inhibited in Comparison with that in Cells infeCted with wild-type VLPs. The Virus produCtion of the reCombinant Influenza C Virus possessing CM2 mutants Containing a serine-to-alanine Change at residue 78 was signifiCantly lower than that of wild-type reCombinant Influenza C Virus. Furthermore, the Virus growth of the reCombinant Viruses possessing CM2 with a serine-to-aspartiC aCid Change at position 78, to mimiC Constitutive phosphorylation, was virtually identiCal to that of the wild-type Virus. These results suggest that phosphorylation of CM2 plays a role in effiCient Virus repliCation, probably through the addition of a negative Charge to the Ser78 phosphorylation site.IMPORTANCE It is well-known that many host and viral proteins are posttranslationally modified by phosphorylation, whiCh plays a role in the funCtions of these proteins. In Influenza A and B Viruses, phosphorylation of viral proteins NP, M1, NS1, and the nuClear export protein (NEP), whiCh are not integrated into the membranes, affeCts the funCtions of these proteins, thereby affeCting Virus repliCation. However, it was reported that phosphorylation of the Influenza A Virus M2 ion Channel protein, whiCh is integrated into the membrane, has no effeCt on Virus repliCation in vitro or in vivo We previously demonstrated that the Influenza C Virus CM2 ion Channel protein is modified by N-glyCosylation, oligomerization, palmitoylation, and phosphorylation and have analyzed the effeCts of these modifiCations, exCept phosphorylation, on Virus repliCation. This is the first report demonstrating that phosphorylation of the Influenza C Virus CM2 ion Channel protein, unlike that of the Influenza A Virus M2 protein, plays a role in Virus repliCation.

  • IntrinsiC Temperature Sensitivity of Influenza C Virus Hemagglutinin-Esterase-Fusion Protein
    Journal of virology, 2012
    Co-Authors: Emi Takashita, Yasushi Muraki, Kanetsu Sugawara, Seiji Hongo, Hidekazu Nishimura, Hironobu Asao, Koji Suzuki, Takashi Tsuji, Yoshihiro Ohara, Yoshihiro Kawaoka
    Abstract:

    Influenza C Virus repliCates more effiCiently at 33°C than at 37°C. To determine whether hemagglutinin-esterase-fusion protein (HEF), a surfaCe glyCoprotein of Influenza C Virus, is a restriCting faCtor for this temperature sensitivity, we analyzed the biologiCal and bioChemiCal properties of HEF at 33°C and 37°C. We found that HEF exhibits intrinsiC temperature sensitivities for surfaCe expression and fusion aCtivity.

  • GlyCosylation of CM2 is important for effiCient repliCation of Influenza C Virus.
    Virology, 2012
    Co-Authors: Takako Okuwa, Yasushi Muraki, Toshiki Himeda, Yoshiro Ohara
    Abstract:

    CM2 is the seCond membrane protein of Influenza C Virus and possesses a Conserved motif for N-glyCosylation. To investigate the role(s) of CM2 glyCosylation in the Virus repliCation, we generated rN11A, a reCombinant Influenza C Virus laCking the glyCosylation site. The rN11A Virus grew less effiCiently than the wild-type (WT) Virus, although the bioChemiCal CharaCteristiCs of the mutant CM2 were similar to those of authentiC CM2. The amount of the genome (GFP-vRNA) in the CM2-N11A-Virus-like partiCles (VLPs) was 13% of that found in WT-VLPs. The inComing GFP-vRNA was less effiCiently transported to the nuCleus in CM2-N11A-VLP-infeCted Cells than WT-VLP-infeCted Cells, leading to the reduCed reporter gene expression in CM2-N11A-VLP-infeCted Cells. Thus the glyCosylation of CM2 is required for effiCient repliCation of Influenza C Virus, and the obtained findings Confirmed and extended the previous observation that CM2 is involved in the genome paCkaging and unCoating proCesses.

  • Role of the CM2 Protein in Influenza C Virus RepliCation: Analyses of ReCombinant Viruses possessing CM2 Mutants
    Metabolomics, 2012
    Co-Authors: Yasushi Muraki, Takako Okuwa, Toshiki Himeda, Yoshiro Ohara
    Abstract:

    CM2 is the seCond membrane protein of Influenza C Virus. In the present study, to investigate the role(s) of CM2 in the Virus repliCation, we generated reCombinant Influenza C Viruses, rC65A and rN11A, whiCh laCk the palmitoylatioin and glyCosylation sites of CM2, respeCtively. The rC65A Virus grew as effiCiently as the reCombinant wild-type (rWT) Virus did, whereas the rN11A grew less effiCiently than the rWT Virus. To study the differenCe more preCisely, we generated Influenza C Virus-like partiCles (VLPs) laCking the CM2 glyCosylation site (N11A-VLPs) and examined the VLPs and VLP-infeCted Cells. As a result, the N11A-VLPs Contain approximately 13% of the Virus RNA found in wildtype VLPs (WT-VLPs), and N11A-VLP-infeCted HMV-II Cells exhibited reduCed reporter gene expression Compared with that of WT-VLP-infeCted Cells (WT : N11A = 1:0.1). Thus, supportive evidenCe was obtained that CM2 is involved in paCkaging and unCoating proCesses and that observed growth differenCe between rWT and rN11A Can be attributed to the differenCe in the CM2 funCtion.

Kanetsu Sugawara - One of the best experts on this subject based on the ideXlab platform.

  • EffeCt of Phosphorylation of CM2 Protein on Influenza C Virus RepliCation.
    Journal of virology, 2017
    Co-Authors: Takanari Goto, Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Yoshitaka Shimotai, Ri Sho, Seiji Hongo
    Abstract:

    CM2 is the seCond membrane protein of the Influenza C Virus and has been demonstrated to play a role in the unCoating and genome paCkaging proCesses in Influenza C Virus repliCation. Although the effeCts of N-linked glyCosylation, disulfide-linked oligomerization, and palmitoylation of CM2 on Virus repliCation have been analyzed, the effeCt of the phosphorylation of CM2 on Virus repliCation remains to be determined. In this study, a phosphorylation site(s) at residue 78 and/or 103 of CM2 was replaCed with an alanine residue(s), and the effeCts of the loss of phosphorylation on Influenza C Virus repliCation were analyzed. No signifiCant differenCes were observed in the paCkaging of the reporter gene between Influenza C Virus-like partiCles (VLPs) produCed from 293T Cells expressing wild-type CM2 and those from the Cells expressing the CM2 mutants laCking the phosphorylation site(s). Reporter gene expression in HMV-II Cells infeCted with VLPs Containing the CM2 mutants was inhibited in Comparison with that in Cells infeCted with wild-type VLPs. The Virus produCtion of the reCombinant Influenza C Virus possessing CM2 mutants Containing a serine-to-alanine Change at residue 78 was signifiCantly lower than that of wild-type reCombinant Influenza C Virus. Furthermore, the Virus growth of the reCombinant Viruses possessing CM2 with a serine-to-aspartiC aCid Change at position 78, to mimiC Constitutive phosphorylation, was virtually identiCal to that of the wild-type Virus. These results suggest that phosphorylation of CM2 plays a role in effiCient Virus repliCation, probably through the addition of a negative Charge to the Ser78 phosphorylation site.IMPORTANCE It is well-known that many host and viral proteins are posttranslationally modified by phosphorylation, whiCh plays a role in the funCtions of these proteins. In Influenza A and B Viruses, phosphorylation of viral proteins NP, M1, NS1, and the nuClear export protein (NEP), whiCh are not integrated into the membranes, affeCts the funCtions of these proteins, thereby affeCting Virus repliCation. However, it was reported that phosphorylation of the Influenza A Virus M2 ion Channel protein, whiCh is integrated into the membrane, has no effeCt on Virus repliCation in vitro or in vivo We previously demonstrated that the Influenza C Virus CM2 ion Channel protein is modified by N-glyCosylation, oligomerization, palmitoylation, and phosphorylation and have analyzed the effeCts of these modifiCations, exCept phosphorylation, on Virus repliCation. This is the first report demonstrating that phosphorylation of the Influenza C Virus CM2 ion Channel protein, unlike that of the Influenza A Virus M2 protein, plays a role in Virus repliCation.

  • epidemiologiCal information regarding the periodiC epidemiCs of Influenza C Virus in japan 1996 2013 and the seroprevalenCe of antibodies to different antigeniC groups
    Journal of Clinical Virology, 2014
    Co-Authors: Yoko Matsuzaki, Kanetsu Sugawara, Chieko Abiko, Tatsuya Ikeda, Katsumi Mizuta, Noriko Katsushima, Yoko Aoki, Fumio Katsushima, Yuriko Katsushima, Tsutomu Itagaki
    Abstract:

    AbstraCt BaCkground Although Influenza C Virus is widely distributed throughout the world, epidemiologiCal information, based on long-term surveillanCe, has not yet been aCquired. ObjeCtives To Clarify the epidemiologiCal features of Influenza C Virus infeCtion, and to examine whether the prevalenCe of the antibodies against the Influenza C Virus is assoCiated with the epidemiCs. Study design Between 1996 and 2013, 36,973 respiratory speCimens were ColleCted from two pediatriC outpatient CliniCs in Yamagata, Japan. The speCimens were examined for the presenCe of Influenza C Virus using Cell Culture methods. Isolated Viruses were antigeniCally analyzed. The differenCes in seropositivity, with respeCt to the different antigeniC groups, were examined using serum samples ColleCted in 2001 and 2011 by a hemagglutination inhibition assay. Results Influenza C Viruses were isolated from 190 speCimens during an 18-year period. Most Influenza C Viruses were isolated from winter to early summer in even-numbered years, and the frequenCy of Virus isolation per year ranged from 0.43% to 1.73%. An antigeniC analysis revealed that the dominant antigeniC groups were the C/Yamagata/26/81 from 1996 to 2000, the C/Kanagawa/1/76 in 2002 and 2004, and the C/Sao Paulo/378/82 from 2006 to 2012. When Compared to the other antigeniC groups, the seroprevalenCe of the C/Sao Paulo/378/82 group was lower in 2001 for individuals older than 5 years and was higher in 2011 in individuals younger than 40 years. ConClusions The results from our study suggest that epidemiCs of Influenza C Virus infeCtion periodiCally oCCur and the replaCement of the dominant antigeniC group may be Caused by immune seleCtion within older Children and/or adults in the Community.

  • EpidemiologiCal information regarding the periodiC epidemiCs of Influenza C Virus in Japan (1996-2013) and the seroprevalenCe of antibodies to different antigeniC groups.
    Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2014
    Co-Authors: Yoko Matsuzaki, Kanetsu Sugawara, Chieko Abiko, Tatsuya Ikeda, Katsumi Mizuta, Noriko Katsushima, Yoko Aoki, Fumio Katsushima, Yuriko Katsushima, Tsutomu Itagaki
    Abstract:

    Although Influenza C Virus is widely distributed throughout the world, epidemiologiCal information, based on long-term surveillanCe, has not yet been aCquired. To Clarify the epidemiologiCal features of Influenza C Virus infeCtion, and to examine whether the prevalenCe of the antibodies against the Influenza C Virus is assoCiated with the epidemiCs. Between 1996 and 2013, 36,973 respiratory speCimens were ColleCted from two pediatriC outpatient CliniCs in Yamagata, Japan. The speCimens were examined for the presenCe of Influenza C Virus using Cell Culture methods. Isolated Viruses were antigeniCally analyzed. The differenCes in seropositivity, with respeCt to the different antigeniC groups, were examined using serum samples ColleCted in 2001 and 2011 by a hemagglutination inhibition assay. Influenza C Viruses were isolated from 190 speCimens during an 18-year period. Most Influenza C Viruses were isolated from winter to early summer in even-numbered years, and the frequenCy of Virus isolation per year ranged from 0.43% to 1.73%. An antigeniC analysis revealed that the dominant antigeniC groups were the C/Yamagata/26/81 from 1996 to 2000, the C/Kanagawa/1/76 in 2002 and 2004, and the C/Sao Paulo/378/82 from 2006 to 2012. When Compared to the other antigeniC groups, the seroprevalenCe of the C/Sao Paulo/378/82 group was lower in 2001 for individuals older than 5 years and was higher in 2011 in individuals younger than 40 years. The results from our study suggest that epidemiCs of Influenza C Virus infeCtion periodiCally oCCur and the replaCement of the dominant antigeniC group may be Caused by immune seleCtion within older Children and/or adults in the Community. Copyright © 2014 Elsevier B.V. All rights reserved.

  • IntrinsiC Temperature Sensitivity of Influenza C Virus Hemagglutinin-Esterase-Fusion Protein
    Journal of virology, 2012
    Co-Authors: Emi Takashita, Yasushi Muraki, Kanetsu Sugawara, Seiji Hongo, Hidekazu Nishimura, Hironobu Asao, Koji Suzuki, Takashi Tsuji, Yoshihiro Ohara, Yoshihiro Kawaoka
    Abstract:

    Influenza C Virus repliCates more effiCiently at 33°C than at 37°C. To determine whether hemagglutinin-esterase-fusion protein (HEF), a surfaCe glyCoprotein of Influenza C Virus, is a restriCting faCtor for this temperature sensitivity, we analyzed the biologiCal and bioChemiCal properties of HEF at 33°C and 37°C. We found that HEF exhibits intrinsiC temperature sensitivities for surfaCe expression and fusion aCtivity.

  • DeteCtion and quantifiCation of Influenza C Virus in pediatriC respiratory speCimens by real-time PCR and Comparison with infeCtious viral Counts
    Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2012
    Co-Authors: Yoko Matsuzaki, Kanetsu Sugawara, Chieko Abiko, Tatsuya Ikeda, Katsumi Mizuta, Yoshitaka Shimotai, Yoko Aoki, Seiji Hongo
    Abstract:

    AbstraCt BaCkground The epidemiologiCal and CliniCal impaCts of Influenza C Virus infeCtion may have been underestimated by Conventional viral Culture sCreening alone. ObjeCtive To evaluate a newly developed real-time polymerase Chain reaCtion (PCR) assay as a tool for diagnosing Influenza C Virus infeCtion. Study design The primers and probe for real-time PCR were designed to amplify the Conserved region of the nuCleoprotein gene based on the aligned sequenCes of nine isolates from 1967 to 2010. Respiratory speCimens from Children ColleCted between January 2010 and August 2010 were examined for the presenCe of Influenza C Virus by Cell Culture and real-time PCR. SpeCimens that were positive for the Virus using real-time PCR were further examined using an infeCtivity assay with embryonated hen's eggs. Results Of the 1203 speCimens examined, 34 (2.8%) tested positive for the Influenza C Virus by Cell Culture and 51 (4.2%) tested positive by real-time PCR. The mean viral load and infeCtivity titer in speCimens that tested positive using Cell Culture were 3.97 × 108 Copies/ml and 5.43 × 105 EID50/ml, respeCtively, and those in speCimens that were negative using Cell Culture were 2.18 × 106 Copies/ml and 3.67 × 102 EID50/ml, respeCtively. In the CliniCal speCimens with viral loads less than 105 Copies/ml, it was not possible to isolate the Virus using embryonated hen's eggs. The Copy number-to-EID50 ratio of the CliniCal speCimens was muCh higher, ranging from 32 to 278,000, than those of Culture fluid, ranging from 2.3 to 13.5. ConClusion The real-time PCR assay desCribed here Can be used as a sensitive method for diagnosing Influenza C Virus infeCtion.

Yoko Matsuzaki - One of the best experts on this subject based on the ideXlab platform.

  • Longitudinal Course of Influenza C Virus antibody titers of healthy adults in Sendai, Japan.
    Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2020
    Co-Authors: Feng Liao, Hidekazu Nishimura, Hiroko Ito, Yunhui Zhang, Yoko Matsuzaki
    Abstract:

    Influenza C Virus Causes mild respiratory diseases in humans. Previous studies suggested that the predominant hemagglutinin-esterase gene lineage CirCulating in Children might be seleCted among the adult population, yet the prevalenCe of Influenza C Virus in adults has not been desCribed. To evaluate the frequenCy of Influenza C Virus infeCtion in adults. We performed hemagglutination inhibition assays of serum samples ColleCted at periodiC oCCupational mediCal CheCkups from employees of a hospital. A total of 679 serum samples were ColleCted from 57 subjeCts who partiCipated in biannual mediCal CheCkups between 2011 and 2016 as part of a longitudinal series. Titers of antibodies against the C/Kanagawa and C/Sao Paulo lineage Viruses were deteCted. Ten serum sample pairs from among the 57 subjeCts showed at least a four-fold inCrease in Influenza C antibody titers. Samples from three subjeCts exhibited antibody titer inCreases for both the C/Kanagawa and C/Sao Paulo lineages, four subjeCts showed an inCreased titer against the C/Sao Paulo lineage, and three subjeCts showed an inCreased titer against the C/Kanagawa lineage. Half of the antibody titer inCreases for the C/Kanagawa lineage were deteCted in May 2014, while the inCreases for the C/Sao Paulo lineage were deteCted from 2011 to 2016. The 5-year Influenza C Virus infeCtion rate was estimated at 17.5 %. There were antibodies that Cross-reaCted with the C/Sao Paulo and C/Kanagawa lineages. The results suggest that C/Sao Paulo was the main lineage in the adult population of this area, with CoCirCulation of the C/Kanagawa lineage. Copyright © 2020 Elsevier B.V. All rights reserved.

  • EffeCt of Phosphorylation of CM2 Protein on Influenza C Virus RepliCation.
    Journal of virology, 2017
    Co-Authors: Takanari Goto, Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Yoshitaka Shimotai, Ri Sho, Seiji Hongo
    Abstract:

    CM2 is the seCond membrane protein of the Influenza C Virus and has been demonstrated to play a role in the unCoating and genome paCkaging proCesses in Influenza C Virus repliCation. Although the effeCts of N-linked glyCosylation, disulfide-linked oligomerization, and palmitoylation of CM2 on Virus repliCation have been analyzed, the effeCt of the phosphorylation of CM2 on Virus repliCation remains to be determined. In this study, a phosphorylation site(s) at residue 78 and/or 103 of CM2 was replaCed with an alanine residue(s), and the effeCts of the loss of phosphorylation on Influenza C Virus repliCation were analyzed. No signifiCant differenCes were observed in the paCkaging of the reporter gene between Influenza C Virus-like partiCles (VLPs) produCed from 293T Cells expressing wild-type CM2 and those from the Cells expressing the CM2 mutants laCking the phosphorylation site(s). Reporter gene expression in HMV-II Cells infeCted with VLPs Containing the CM2 mutants was inhibited in Comparison with that in Cells infeCted with wild-type VLPs. The Virus produCtion of the reCombinant Influenza C Virus possessing CM2 mutants Containing a serine-to-alanine Change at residue 78 was signifiCantly lower than that of wild-type reCombinant Influenza C Virus. Furthermore, the Virus growth of the reCombinant Viruses possessing CM2 with a serine-to-aspartiC aCid Change at position 78, to mimiC Constitutive phosphorylation, was virtually identiCal to that of the wild-type Virus. These results suggest that phosphorylation of CM2 plays a role in effiCient Virus repliCation, probably through the addition of a negative Charge to the Ser78 phosphorylation site.IMPORTANCE It is well-known that many host and viral proteins are posttranslationally modified by phosphorylation, whiCh plays a role in the funCtions of these proteins. In Influenza A and B Viruses, phosphorylation of viral proteins NP, M1, NS1, and the nuClear export protein (NEP), whiCh are not integrated into the membranes, affeCts the funCtions of these proteins, thereby affeCting Virus repliCation. However, it was reported that phosphorylation of the Influenza A Virus M2 ion Channel protein, whiCh is integrated into the membrane, has no effeCt on Virus repliCation in vitro or in vivo We previously demonstrated that the Influenza C Virus CM2 ion Channel protein is modified by N-glyCosylation, oligomerization, palmitoylation, and phosphorylation and have analyzed the effeCts of these modifiCations, exCept phosphorylation, on Virus repliCation. This is the first report demonstrating that phosphorylation of the Influenza C Virus CM2 ion Channel protein, unlike that of the Influenza A Virus M2 protein, plays a role in Virus repliCation.

  • analyses of evolutionary CharaCteristiCs of the hemagglutinin esterase gene of Influenza C Virus during a period of 68 years reveals evolutionary patterns different from Influenza a and b Viruses
    Viruses, 2016
    Co-Authors: Yuki Furuse, Yoko Matsuzaki, Hidekazu Nishimura, Hitoshi Oshitani
    Abstract:

    InfeCtions with the Influenza C Virus Causing respiratory symptoms are Common, partiCularly among Children. SinCe isolation and deteCtion of the Virus are rarely performed, Compared with Influenza A and B Viruses, the small number of available sequenCes of the Virus makes it diffiCult to analyze its evolutionary dynamiCs. ReCently, we reported the full genome sequenCe of 102 strains of the Virus. Here, we exploited the data to eluCidate the evolutionary CharaCteristiCs and phylodynamiCs of the Virus Compared with Influenza A and B Viruses. Along with our data, we obtained publiC sequenCe data of the hemagglutinin-esterase gene of the Virus; the dataset Consists of 218 unique sequenCes of the Virus ColleCted from 14 Countries between 1947 and 2014. InformatiCs analyses revealed that (1) multiple lineages have been CirCulating globally; (2) there have been weak and infrequent seleCtive bottleneCks; (3) the evolutionary rate is low beCause of weak positive seleCtion and a low Capability to induCe mutations; and (4) there is no signifiCant positive seleCtion although a few mutations affeCting its antigeniCity have been induCed. The unique evolutionary dynamiCs of the Influenza C Virus must be shaped by multiple faCtors, inCluding virologiCal, immunologiCal, and epidemiologiCal CharaCteristiCs.

  • Influenza C Virus and Human MetapneumoVirus InfeCtions in Hospitalized Children With Lower Respiratory TraCt Illness.
    The Pediatric Infectious Disease Journal, 2015
    Co-Authors: Yukitoshi Shimizu, Chieko Abiko, Tatsuya Ikeda, Katsumi Mizuta, Yoko Matsuzaki
    Abstract:

    A 6-month prospeCtive study in a hospital setting deteCted Influenza C Virus and human metapneumoVirus in 10.0% (29/289) and 16.6% (48/289), respeCtively, of Children hospitalized with lower respiratory traCt illness. Influenza C Virus infeCtion had a similar rate of pneumonia (53.3% vs. 57.1%), signifiCantly lower frequenCy of wheezing (13.3% vs. 68.6%) and higher values of white blood Cell and C-reaCtive protein than human metapneumoVirus infeCtion.

  • epidemiologiCal information regarding the periodiC epidemiCs of Influenza C Virus in japan 1996 2013 and the seroprevalenCe of antibodies to different antigeniC groups
    Journal of Clinical Virology, 2014
    Co-Authors: Yoko Matsuzaki, Kanetsu Sugawara, Chieko Abiko, Tatsuya Ikeda, Katsumi Mizuta, Noriko Katsushima, Yoko Aoki, Fumio Katsushima, Yuriko Katsushima, Tsutomu Itagaki
    Abstract:

    AbstraCt BaCkground Although Influenza C Virus is widely distributed throughout the world, epidemiologiCal information, based on long-term surveillanCe, has not yet been aCquired. ObjeCtives To Clarify the epidemiologiCal features of Influenza C Virus infeCtion, and to examine whether the prevalenCe of the antibodies against the Influenza C Virus is assoCiated with the epidemiCs. Study design Between 1996 and 2013, 36,973 respiratory speCimens were ColleCted from two pediatriC outpatient CliniCs in Yamagata, Japan. The speCimens were examined for the presenCe of Influenza C Virus using Cell Culture methods. Isolated Viruses were antigeniCally analyzed. The differenCes in seropositivity, with respeCt to the different antigeniC groups, were examined using serum samples ColleCted in 2001 and 2011 by a hemagglutination inhibition assay. Results Influenza C Viruses were isolated from 190 speCimens during an 18-year period. Most Influenza C Viruses were isolated from winter to early summer in even-numbered years, and the frequenCy of Virus isolation per year ranged from 0.43% to 1.73%. An antigeniC analysis revealed that the dominant antigeniC groups were the C/Yamagata/26/81 from 1996 to 2000, the C/Kanagawa/1/76 in 2002 and 2004, and the C/Sao Paulo/378/82 from 2006 to 2012. When Compared to the other antigeniC groups, the seroprevalenCe of the C/Sao Paulo/378/82 group was lower in 2001 for individuals older than 5 years and was higher in 2011 in individuals younger than 40 years. ConClusions The results from our study suggest that epidemiCs of Influenza C Virus infeCtion periodiCally oCCur and the replaCement of the dominant antigeniC group may be Caused by immune seleCtion within older Children and/or adults in the Community.

Emi Takashita - One of the best experts on this subject based on the ideXlab platform.

  • IntrinsiC Temperature Sensitivity of Influenza C Virus Hemagglutinin-Esterase-Fusion Protein
    Journal of virology, 2012
    Co-Authors: Emi Takashita, Yasushi Muraki, Kanetsu Sugawara, Seiji Hongo, Hidekazu Nishimura, Hironobu Asao, Koji Suzuki, Takashi Tsuji, Yoshihiro Ohara, Yoshihiro Kawaoka
    Abstract:

    Influenza C Virus repliCates more effiCiently at 33°C than at 37°C. To determine whether hemagglutinin-esterase-fusion protein (HEF), a surfaCe glyCoprotein of Influenza C Virus, is a restriCting faCtor for this temperature sensitivity, we analyzed the biologiCal and bioChemiCal properties of HEF at 33°C and 37°C. We found that HEF exhibits intrinsiC temperature sensitivities for surfaCe expression and fusion aCtivity.

  • role of the Cm2 protein in the Influenza C Virus repliCation CyCle
    Journal of Virology, 2011
    Co-Authors: Takatoshi Furukawa, Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Emi Takashita, Yoshitaka Shimotai, Ri Sho, Takeshi Noda, Seiji Hongo
    Abstract:

    CM2 is the seCond membrane protein of Influenza C Virus. Although its bioChemiCal CharaCteristiCs, Coding strategy, and properties as an ion Channel have been extensively studied, the role(s) of CM2 in the Virus repliCation CyCle remains to be Clarified. In order to eluCidate this role, in the present study we generated CM2-defiCient Influenza C Virus-like partiCles (VLPs) and examined the VLP-produCing 293T Cells, VLPs, and VLP-infeCted HMV-II Cells. QuantifiCation of viral RNA (vRNA) in the VLPs by real-time PCR revealed that the CM2-defiCient VLPs Contain approximately one-third of the vRNA found in wild-type VLPs although no signifiCant differenCes were deteCted in the expression levels of viral Components in VLP-produCing Cells or in the number and morphology of the generated VLPs. This finding suggests that CM2 is involved in the genome paCkaging proCess into VLPs. Furthermore, HMV-II Cells infeCted with CM2-defiCient VLPs exhibited signifiCantly reduCed reporter gene expression. Although CM2-defiCient VLPs Could be internalized into HMV-II Cells as effiCiently as wild-type VLPs, a smaller amount of vRNA was deteCted in the nuClear fraCtion of CM2-defiCient VLP-infeCted Cells than in that of wild-type VLP-infeCted Cells, suggesting that the unCoating proCess of the CM2-defiCient VLPs in the infeCted Cells did not proCeed in an appropriate manner. Taken together, the data obtained in the present study indiCate that CM2 has a potential role in the genome paCkaging and unCoating proCesses of the Virus repliCation CyCle.

  • Influenza C Virus NS1 Protein Upregulates the SpliCing of Viral mRNAs
    Journal of virology, 2009
    Co-Authors: Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Emi Takashita, Takatoshi Furukawa, Yoshihiko Kohno, Seiji Hongo
    Abstract:

    Pre-mRNAs of the Influenza A Virus M and NS genes are poorly spliCed in Virus-infeCted Cells. By Contrast, in Influenza C Virus-infeCted Cells, the predominant transCript from the M gene is spliCed mRNA. The present study was performed to investigate the meChanism by whiCh Influenza C Virus M gene-speCifiC mRNA (M mRNA) is readily spliCed. The ratio of M1 enCoded by a spliCed M mRNA to CM2 enCoded by an unspliCed M mRNA in Influenza C Virus-infeCted Cells was about 10 times larger than that in M gene-transfeCted Cells, suggesting that a viral protein(s) other than M gene translational produCts faCilitates viral mRNA spliCing. RNase proteCtion assays showed that the spliCing of M mRNA in infeCted Cells was muCh higher than that in M gene-transfeCted Cells. The unspliCed and spliCed mRNAs of the Influenza C Virus NS gene enCode two nonstruCtural (NS) proteins, NS1(C/NS1) and NS2(C/NS2), respeCtively. The introduCtion of premature translational termination into the NS gene, whiCh bloCked the synthesis of the C/NS1 and C/NS2 proteins, drastiCally reduCed the spliCing of NS mRNA, raising the possibility that C/NS1 or C/NS2 enhanCes viral mRNA spliCing. The spliCing of Influenza C Virus M mRNA was inCreased by Coexpression of C/NS1, whereas it was reduCed by Coexpression of the Influenza A Virus NS1 protein (A/NS1). The spliCing of Influenza A Virus M mRNA was also inCreased by Coexpression of C/NS1, though it was inhibited by that of A/NS1. These results suggest that Influenza C Virus NS1, but not A/NS1, Can upregulate viral mRNA spliCing.

  • IntraCellular loCalization of Influenza C Virus NS2 protein (NEP) in infeCted Cells and its inCorporation into virions
    Archives of Virology, 2009
    Co-Authors: Yoshihiko Kohno, Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Emi Takashita, Seiji Hongo
    Abstract:

    RNA segment 7 of Influenza C Virus enCodes two non-struCtural (NS) proteins, NS1 and NS2. The Influenza C Virus NS2 protein has been proposed to possess nuClear export aCtivity like that of Influenza A and B Virus NS2 proteins (NEP). In the present study, we investigated the kinetiCs and loCalization of the NS2 protein in Influenza C Virus-infeCted Cells, and analysed whether NS2 is present in virions. ImmunofluoresCent staining analysis of the infeCted Cells indiCated that NS2 was loCalized in the nuCleus immediately after synthesis and predominantly in the Cytoplasm in the later stages of infeCtion. ConfoCal miCrosCopy revealed that a part of the NS2 protein was ColoCalized with nuCleoprotein NP/vRNP in the Cytoplasm and on the Cell membrane in the late stages of infeCtion. The NS2 protein was deteCted in Influenza C virions purified by gradient Centrifugations and/or affinity Chromatography. Trypsin treatment demonstrated that the NS2 protein was present inside the viral envelope. Furthermore, glyCerol gradient analysis of detergent-solubilized virions revealed that the NS2 protein Cosedimented with vRNPs. These data suggest that the Influenza C Virus NS2 protein is inCorporated into virions, where it assoCiates with vRNP.

  • A Mutation on Influenza C Virus M1 Protein AffeCts Virion Morphology by Altering the Membrane Affinity of the Protein
    Journal of Virology, 2007
    Co-Authors: Yasushi Muraki, Kanetsu Sugawara, Yoko Matsuzaki, Emi Takashita, Toshio Murata, Seiji Hongo
    Abstract:

    Reverse genetiCs has been doCumented for Influenza A, B, and Thogoto Viruses belonging to the family Orthomyxoviridae. We report here the reverse genetiCs of Influenza C Virus, another member of this family. The seven viral RNA (vRNA) segments of C/Ann Arbor/1/50 were expressed in 293T Cells from Cloned CDNAs, together with nine Influenza C Virus proteins. At 48 h posttransfeCtion, the infeCtious titer of the Culture supernatant was determined to be 2.51 x 10(3) 50% egg infeCtious doses/ml, whiCh is lower than the number of Influenza C Virus-like partiCles (VLPs) (10(6)/ml) generated using the same system. By generating Influenza C VLPs Containing a given vRNA segment, we showed that eaCh of the vRNA segments was similarly synthesized in the plasmid-transfeCted Cells but that some segments were less effiCiently inCorporated into the VLPs. This finding leads us to speCulate that the differenCes in inCorporation effiCienCy into VLPs between segments might be a reason for the ineffiCient produCtion of infeCtious Viruses. SeCond, we generated a mutant reCombinant Virus, rMG96A, whiCh possesses an Ala-->Thr mutation at residue 24 of the M1 protein, a substitution demonstrated to be involved in the morphology (filamentous or spheriCal) of the Influenza C VLPs. As expeCted, rMG96A exhibited a spheriCal morphology, whereas reCombinant wild-type of C/Ann Arbor/1/50, rWT, exhibited a mainly filamentous morphology. Membrane flotation analysis of the Cells infeCted with rWT or rMG96A revealed a differenCe in the ratio of membrane-assoCiated M1 proteins, suggesting that the affinity of M1 protein to the Cell membrane is a determinant for virion morphology.