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Natsumi Omura, Tomoki Yoshikawa, Hikaru Fujii, Miho Shibamura, Takuya Inagaki, Hirofumi Kato, Kazutaka Egawa, Shizuko Harada, Souichi Yamada, Haruko Takeyama, Masayuki Saijo
A novel system was developed for generating a highly-attenuated vaccinia virus LC16m8 (m8, third generation smallpox vaccine) that expresses foreign genes. The innovations in this system are its excisable selection marker, specificity of the integration site of a gene of interest, and easy identification of clones with the fluorescent signal. Using this system, recombinant m8s, which expressed either herpes simplex virus 2 (HSV-2) glycoprotein B (gB)-, gD-, or both gB and gD (gB+gD) were developed, and their efficacy was evaluated...
April 27, 2018: Japanese Journal of Infectious Diseases
Tomoki Yoshikawa, Hikaru Fujii, Akiko Okutani, Miho Shibamura, Natsumi Omura, Kazutaka Egawa, Hirofumi Kato, Takuya Inagaki, Shizuko Harada, Souichi Yamada, Shigeru Morikawa, Masayuki Saijo
LC16m8 (m8), a highly attenuated vaccinia virus (VAC) strain, was developed as a smallpox vaccine, and its safety and immunogenicity have been confirmed. Here, we aimed to develop a system that recovers infectious m8 from a bacterial artificial chromosome (BAC) that retains the full-length viral genomic DNA (m8-BAC system). The infectious virus was successfully recovered from a VAC-BAC plasmid, named pLC16m8-BAC. Furthermore, the bacterial replicon-free virus was generated by intramolecular homologous recombination and was successfully recovered from a modified VAC-BAC plasmid, named pLC16m8...
2018: PloS One
Itoe Iizuka, Yasushi Ami, Yuriko Suzaki, Noriyo Nagata, Shuetsu Fukushi, Momoko Ogata, Shigeru Morikawa, Hideki Hasegawa, Masashi Mizuguchi, Ichiro Kurane, Masayuki Saijo
Monkeypox virus (MPXV) causes human monkeypox (human MPX), which is a similar disease with smallpox in humans. A previous study showed that a single vaccination of monkeys with LC16m8, a highly attenuated smallpox vaccine, protected them from MPX at 4-5 weeks after vaccination. In the present study, we evaluated the long-term efficacy of a single vaccination with LC16m8 in a nonhuman primate model of MPXV infection. The monkeys were inoculated with LC16m8, Lister (parental strain of LC16m8), or mock and then challenged with MPXV via the subcutaneous route at 6 and 12 months after vaccination, which we compared with either Lister or mock vaccination...
December 22, 2016: Japanese Journal of Infectious Diseases
Yasumasa Nishiyama, Tatsuya Fujii, Yasuhiro Kanatani, Yasuhiko Shinmura, Hiroyuki Yokote, So Hashizume
BACKGROUND: In Japan, production of smallpox vaccine LC16m8 (named LC16-KAKETSUKEN) was restarted and was determined to be maintained as a national stockpile in March 2002. OBJECTIVE: To conduct a post-marketing surveillance study of the vaccination of freeze-dried live attenuated smallpox vaccine prepared in cell culture LC16-KAKETSUKEN using attenuated vaccinia strain LC16m8. The study complied with Good Clinical Practice, focusing on a comparison between primary vaccinees and re-vaccinees...
November 9, 2015: Vaccine
Akiko Eto, Tomoya Saito, Hiroyuki Yokote, Ichiro Kurane, Yasuhiro Kanatani
LC16m8 is a live, attenuated, cell-cultured smallpox vaccine that was developed and licensed in Japan in the 1970s, but was not used in the campaign to eradicate smallpox. In the early 2000s, the potential threat of bioterrorism led to reconsideration of the need for a smallpox vaccine. Subsequently, LC16m8 production was restarted in Japan in 2002, requiring re-evaluation of its safety and efficacy. Approximately 50,000 children in the 1970s and about 3500 healthy adults in the 2000s were vaccinated with LC16m8 in Japan, and 153 adults have been vaccinated with LC16m8 or Dryvax in phase I/II clinical trials in the USA...
November 9, 2015: Vaccine
Hiroyuki Yokote, Yasuhiko Shinmura, Tomomi Kanehara, Shinichi Maruno, Masahiko Kuranaga, Hajime Matsui, So Hashizume
BACKGROUND: Attenuated vaccinia virus strain, LC16m8, defective in the B5R envelope protein gene, is used as a stockpile smallpox vaccine strain in Japan against bioterrorism: the defect in the B5R gene mainly contributes to its highly attenuated properties. METHODS: The protective activity of LC16m8 vaccine against challenge with a lethal dose of vaccinia Western Reserve strain was assessed in wild-type and immunodeficient mice lacking CD4, MHC class I, MHC class II or MHC class I and II antigens...
November 9, 2015: Vaccine
Hiroyuki Yokote, Yasuhiko Shinmura
No abstract text is available yet for this article.
May 2015: Clinical and Vaccine Immunology: CVI
Clement A Meseda, Jordan Kuhn, Vajini Atukorale, Joseph Campbell, Jerry P Weir
The vaccinia virus complement control protein (VCP) is a secreted viral protein that binds the C3b and C4b complement components and inhibits the classic and alternative complement pathways. Previously, we reported that an attenuated smallpox vaccine, LC16m8, which was derived from the Lister strain of vaccinia virus (VV-Lister), expressed a glycosylated form of VCP, whereas published sequence data at that time indicated that the VV-Lister VCP has no motif for N-linked glycosylation. We were interested in determining whether the glycosylation of VCP impairs its biological activity, possibly contributing to the attenuation of LC16m8, and the likely origin of the glycosylated VCP...
September 2014: Clinical and Vaccine Immunology: CVI
Hiroyuki Yokote, Yasuhiko Shinmura, Tomomi Kanehara, Shinichi Maruno, Masahiko Kuranaga, Hajime Matsui, So Hashizume
Freeze-dried live attenuated smallpox vaccine LC16m8 prepared in cell culture has been the sole smallpox vaccine licensed in Japan since 1975 and was recently recommended as a WHO stockpile vaccine. We evaluated the safety of recently remanufactured lots of LC16m8 using a series of immunodeficient mouse models. These models included suckling mice, severe combined immunodeficiency disease (SCID) mice, and wild-type mice treated with cyclosporine. LC16m8 showed extremely low virulence in each of the three mouse models compared with that of its parental strains, Lister and LC16mO...
September 2014: Clinical and Vaccine Immunology: CVI
Takashi Ohashi, Takafumi Nakamura, Minoru Kidokoro, Xianfeng Zhang, Hisatoshi Shida
Adult T cell leukemia (ATL) is a malignant lymphoproliferative disease caused by human T cell leukemia virus type I (HTLV-I). To develop an effective therapy against the disease, we have examined the oncolytic ability of an attenuated vaccinia virus (VV), LC16m8Δ (m8Δ), and an HTLV-I Tax-specific cytotoxic T lymphocyte (CTL) line, 4O1/C8, against an HTLV-I-infected rat T cell line, FPM1. Our results demonstrated that m8Δ was able to replicate in and lyse tumorigenic FPM1 cells but was incompetent to injure 4O1/C8 cells, suggesting the preferential cytolytic activity toward tumor cells...
2014: BioMed Research International
Mao Isshiki, Xianfeng Zhang, Hirotaka Sato, Takashi Ohashi, Makoto Inoue, Hisatoshi Shida
Previously, we developed a vaccination regimen that involves priming with recombinant vaccinia virus LC16m8Δ (rm8Δ) strain followed by boosting with a Sendai virus-containing vector. This protocol induced both humoral and cellular immune responses against the HIV-1 envelope protein. The current study aims to optimize this regimen by comparing the immunogenicity and safety of two rm8Δ strains that express HIV-1 Env under the control of a moderate promoter, p7.5, or a strong promoter, pSFJ1-10. m8Δ-p7.5-JRCSFenv synthesized less gp160 but showed significantly higher growth potential than m8Δ-pSFJ-JRCSFenv...
February 7, 2014: Vaccine
Hirotaka Sato, Chen Jing, Mao Isshiki, Kazuhiro Matsuo, Minoru Kidokoro, Shiki Takamura, Xianfeng Zhang, Takashi Ohashi, Hisatoshi Shida
We compared the effect of the very strong pSFJ1-10 and moderately strong p7.5 promoters on the immunogenicity and pathogenicity of the replication-competent vaccinia virus (VV) LC16m8Δ (m8Δ) vector harboring the SIV gag gene in a vaccination regimen consisting of a recombinant BCG-SIVGag (rBCG-SIVGag) prime followed by a recombinant vaccinia boost. m8Δ/pSFJ/SIVGag synthesized more Gag protein than m8Δ/p7.5/SIVGag but replicated less efficiently in vitro. In addition, m8Δ/pSFJ/SIVGag was less pathogenic and elicited Gag-specific IFN-γ(+), CD107a(+), CD8(+) cells more efficiently than m8Δ/p7...
August 2, 2013: Vaccine
Xianfeng Zhang, Tomoyoshi Sobue, Mao Isshiki, Shun-ichi Makino, Makoto Inoue, Kazunori Kato, Tatsuo Shioda, Takashi Ohashi, Hirotaka Sato, Jun Komano, Hideji Hanabusa, Hisatoshi Shida
For protection from HIV-1 infection, a vaccine should elicit both humoral and cell-mediated immune responses. A novel vaccine regimen and adjuvant that induce high levels of HIV-1 Env-specific T cell and antibody (Ab) responses was developed in this study. The prime-boost regimen that used combinations of replication-competent vaccinia LC16m8Δ (m8Δ) and Sendai virus (SeV) vectors expressing HIV-1 Env efficiently produced both Env-specific CD8(+) T cells and anti-Env antibodies, including neutralizing antibodies (nAbs)...
2012: PloS One
Bidhan Ch Bera, K Shanmugasundaram, Sanjay Barua, Taruna Anand, T Riyesh, Rajesh K Vaid, Nitin Virmani, Manish Bansal, Brihaspati N Shukla, Praveen Malik, Raj K Singh
Buffalopox virus (BPXV), a close variant of vaccinia virus (VACV) has emerged as a zoonotic pathogen. The host tropism of poxviruses is governed by host-range genes. Among the host-range genes: E3L, K3L, and C7L are essential for virus replication by preventing interferon resistance, whereas B5R is essential for spread of the virus and evasion from the host's immune response as in VACV. We report sequence analysis of host-range genes: E3L, K3L, C7L, and membrane protein gene (B5R) of BPXVs from buffalo, cattle, and human from recent outbreaks in India-their phylogenetic relationship with reference strain (BP4) and other Orthopoxviruses...
December 2012: Virus Genes
Jeffrey S Kennedy, Marc Gurwith, Cornelia L Dekker, Sharon E Frey, Kathryn M Edwards, Julie Kenner, Michael Lock, Cyril Empig, Shigeru Morikawa, Masayuki Saijo, Hiroyuki Yokote, Kevin Karem, Inger Damon, Mark Perlroth, Richard N Greenberg
INTRODUCTION: LC16m8 is an attenuated cell culture-adapted Lister vaccinia smallpox vaccine missing the B5R protein and licensed for use in Japan. METHODS: We conducted a phase I/II clinical trial that compared the safety and immunogenicity of LC16m8 with Dryvax in vaccinia-naive participants. Adverse events were assessed, as were electrocardiography and laboratory testing for cardiotoxicity and viral culturing of the vaccination sites. Neutralization titers to vaccinia, monkeypox, and variola major were assessed and cell-mediated immune responses were measured by interferon (IFN)-γ enzyme-linked immunosorbent spot and lymphoproliferation assays...
November 2011: Journal of Infectious Diseases
Benjamin F Johnson, Yasuhiro Kanatani, Tatsuya Fujii, Tomoya Saito, Hiroyuki Yokote, Geoffrey L Smith
In response to potential bioterrorism with smallpox, members of the Japanese Self-Defense Forces were vaccinated with vaccinia virus (VACV) strain LC16m8, an attenuated smallpox vaccine derived from VACV strain Lister. The serological response induced by LC16m8 to four virion-surface proteins and the intracellular mature virus (IMV) and extracellular enveloped virus (EEV) was investigated. LC16m8 induced antibody response against the IMV protein A27 and the EEV protein A56. LC16m8 also induced IMV-neutralizing antibodies, but unlike the VACV strain Lister, did not induce either EEV-neutralizing antibody or antibody to EEV protein B5, except after revaccination...
October 2011: Journal of General Virology
Shari N Gordon, Valentina Cecchinato, Vibeke Andresen, Jean-Michel Heraud, Anna Hryniewicz, Robyn Washington Parks, David Venzon, Hye-kyung Chung, Tatiana Karpova, James McNally, Peter Silvera, Keith A Reimann, Hajime Matsui, Tomomi Kanehara, Yasuhiko Shinmura, Hiroyuki Yokote, Genoveffa Franchini
The licensed smallpox vaccine, ACAM2000, is a cell culture derivative of Dryvax. Both ACAM2000 and Dryvax are administered by skin scarification and can cause progressive vaccinia, with skin lesions that disseminate to distal sites. We have investigated the immunologic basis of the containment of vaccinia in the skin with the goal to identify safer vaccines for smallpox. Macaques were depleted systemically of T or B cells and vaccinated with either Dryvax or an attenuated vaccinia vaccine, LC16m8. B cell depletion did not affect the size of skin lesions induced by either vaccine...
April 15, 2011: Journal of Infectious Diseases
Clement A Meseda, Anne E Mayer, Arunima Kumar, Alonzo D Garcia, Joseph Campbell, Paul Listrani, Jody Manischewitz, Lisa R King, Hana Golding, Michael Merchlinsky, Jerry P Weir
The immune response elicited by LC16m8, a candidate smallpox vaccine that was developed in Japan by cold selection during serial passage of the Lister vaccine virus in primary rabbit kidney cells, was compared to Dryvax in a mouse model. LC16m8 carries a mutation resulting in the truncation of the B5 protein, an important neutralizing target of the extracellular envelope form of vaccinia virus (EV). LC16m8 elicited a broad-spectrum immunoglobulin G (IgG) response that neutralized both EV and the intracellular mature form of vaccinia virus and provoked cell-mediated immune responses, including the activation of CD4+ and CD8+ cells, similarly to Dryvax...
September 2009: Clinical and Vaccine Immunology: CVI
Joan E Adamo, Clement A Meseda, Jerry P Weir, Michael J Merchlinsky
Vaccination with Dryvax elicits a broad humoral response against many viral proteins. Human vaccinia immune globulin was used to screen the secreted proteins from cells infected with Dryvax or the candidate smallpox vaccine LC16m8 to determine whether the protective humoral response included antibodies against secreted viral proteins. Many proteins were detected, with the primary band corresponding to a band of 28 or 30 kDa in cells infected with Dryvax or LC16m8, respectively. This was identified as the vaccinia virus complement protein (VCP), which migrated more slowly in LC16m8-infected cells due to post-translational glycosylation...
November 2009: Journal of General Virology
Tomoya Saito, Tatsuya Fujii, Yasuhiro Kanatani, Masayuki Saijo, Shigeru Morikawa, Hiroyuki Yokote, Tsutomu Takeuchi, Noriyuki Kuwabara
CONTEXT: The attenuated, tissue-cultured, third-generation smallpox vaccine LC16m8 was administered to vaccinia-naive infants in Japan during the 1970s without serious adverse events. It is a good candidate for use as part of a prevention plan for bioterrorism. OBJECTIVE: To assess the immunogenicity and frequency of adverse events of LC16m8 vaccine in unvaccinated and previously vaccinated adults. DESIGN, SETTING, AND PARTICIPANTS: Between 2002 and 2005 we vaccinated and revaccinated 1529 and 1692 adults, respectively, in the Japan Self-Defense Forces with LC16m8 vaccine, given intraepidermally using a bifurcated needle...
March 11, 2009: JAMA: the Journal of the American Medical Association
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