A highly attenuated vaccinia virus strain LC16m8-based vaccine for severe fever with thrombocytopenia syndrome

Severe fever with thrombocytopenia syndrome (SFTS) caused by a species Dabie bandavirus (formerly SFTS virus [SFTSV]) is an emerging hemorrhagic infectious disease with a high case-fatality rate. One of the best strategies for preventing SFTS is to develop a vaccine, which is expected to induce both humoral and cellular immunity. We applied a highly attenuated but still immunogenic vaccinia virus strain LC16m8 (m8) as a recombinant vaccine for SFTS. Recombinant m8s expressing SFTSV nucleoprotein (m8-N), envelope glycoprotein precursor (m8-GPC), and both N and GPC (m8-N+GPC) in the infected cells were generated. Both m8-GPC- and m8-N+GPC-infected cells were confirmed to produce SFTSV-like-particles (VLP) in vitro, and the N was incorporated in the VLP produced by the infection of cells with m8-N+GPC. Specific antibodies to SFTSV were induced in mice inoculated with each of the recombinant m8s, and the mice were fully protected from lethal challenge with SFTSV at both 103 TCID50 and 105 TCID50. In mice that had been immunized with vaccinia virus strain Lister in advance of m8-based SFTSV vaccine inoculation, protective immunity against the SFTSV challenge was also conferred. The pathological analysis revealed that mice immunized with m8-GPC or m8-N+GPC did not show any histopathological changes without any viral antigen-positive cells, whereas the control mice showed focal necrosis with inflammatory infiltration with SFTSV antigen-positive cells in tissues after SFTSV challenge. The passive serum transfer experiments revealed that sera collected from mice inoculated with m8-GPC or m8-N+GPC but not with m8-N conferred protective immunity against lethal SFTSV challenge in naïve mice. On the other hand, the depletion of CD8-positive cells in vivo did not abrogate the protective immunity conferred by m8-based SFTSV vaccines. Based on these results, the recombinant m8-GPC and m8-N+GPC were considered promising vaccine candidates for SFTS.

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A highly attenuated vaccinia virus strain LC16m8-based vaccine for severe fever with thrombocytopenia syndrome

10.1101/2020.08.06.239368 ◽

2020 ◽

Author(s):

Tomoki Yoshikawa ◽

Satoshi Taniguchi ◽

Hirofumi Kato ◽

Naoko Iwata-Yoshikawa ◽

Hideki Tani ◽

...

Keyword(s):

Vaccinia Virus ◽

Virus Strain ◽

Protective Immunity ◽

Case Fatality Rate ◽

Case Fatality ◽

Hemorrhagic Fever ◽

Recombinant Vaccine ◽

Infected Cells ◽

Vaccinia Virus Strain ◽

Severe Fever

AbstractSevere fever with thrombocytopenia syndrome (SFTS) caused by Dabie bandavirus (formerly SFTS virus [SFTSV]) is an emerging hemorrhagic infectious disease with a high case-fatality rate. One of the best strategies for preventing SFTS is to develop a vaccine, which is expected to induce both humoral and cellular immunity. We applied a highly attenuated but still immunogenic vaccinia virus strain LC16m8 (m8) as a recombinant vaccine for SFTS. Recombinant m8s expressing SFTSV nucleoprotein (m8-N), envelope glycoprotein precursor (m8-GPC), and both N and GPC (m8-N+GPC) in the infected cells were generated. Both m8-GPC- and m8-N+GPC-infected cells were confirmed to produce SFTSV-like-particles (VLP) in vitro, and the N was incorporated in the VLP produced by the infection of cells with m8-N+GPC. Specific antibodies to SFTSV were induced in mice inoculated with each of the recombinant m8s, and the mice were fully protected from lethal challenge with SFTSV at both 103 TCID50 and 105 TCID50. In mice that had been immunized with vaccinia virus strain Lister in advance of m8-based SFTSV vaccine inoculation, protective immunity against the SFTSV challenge was also conferred. The pathological analysis revealed that mice immunized with m8-GPC or m8-N+GPC did not show any histopathological changes without any viral antigen-positive cells, whereas the control mice showed focal necrosis with inflammatory infiltration with SFTSV antigen-positive cells in tissues after SFTSV challenge. The passive serum transfer experiments revealed that sera collected from mice inoculated with m8-GPC or m8-N+GPC but not with m8-N conferred protective immunity against lethal SFTSV challenge in naïve mice. On the other hand, the depletion of CD8-positive cells in vivo did not abrogate the protective immunity conferred by m8-based SFTSV vaccines. Based on these results, the recombinant m8-GPC and m8-N+GPC were considered promising vaccine candidates for SFTS.Author SummarySevere fever with thrombocytopenia syndrome (SFTS) is an emerging viral hemorrhagic fever with a high case-fatality rate (approximately 5% to >40%). Indigenous SFTS has been reported in China, Japan, South Korea, and Vietnam. Thus, the development of an effective vaccine for SFTS is urgently needed. Vaccinia virus (VAC) was previously used as a vaccine for smallpox. Unfortunately, after these strains, the so-called second generation of VAC used during the eradication campaign was associated with severe adverse events, and the third generation of VAC strains such as LC16m8 (m8) and modified vaccinia Ankara (MVA) was established. m8 is confirmed to be highly attenuated while still maintaining immunogenicity. m8 is licensed for use in healthy people in Japan. At the present time, approximately 100,000 people have undergone vaccination with m8 without experiencing any severe postvaccine complications. At present, third-generation VAC strains are attractive for a recombinant vaccine vector, especially for viral hemorrhagic infectious diseases, such as Ebola virus disease, Lassa fever, Crimean-Congo hemorrhagic fever, and SFTS. We investigated the practicality of an m8-based recombinant vaccine for SFTS as well as other promising recombinant VAC-based vaccines for viral hemorrhagic infectious diseases.

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Vaccinia Virus H3L Envelope Protein Is a Major Target of Neutralizing Antibodies in Humans and Elicits Protection against Lethal Challenge in Mice

Journal of Virology ◽

10.1128/jvi.79.18.11724-11733.2005 ◽

2005 ◽

Vol 79 (18) ◽

pp. 11724-11733 ◽

Cited By ~ 138

Author(s):

D. Huw Davies ◽

Megan M. McCausland ◽

Conrad Valdez ◽

Devan Huynh ◽

Jenny E. Hernandez ◽

...

Keyword(s):

Antibody Response ◽

Vaccinia Virus ◽

Virus Strain ◽

Envelope Protein ◽

Neutralizing Antibodies ◽

High Titer ◽

Lethal Challenge ◽

Vaccinia Virus Strain ◽

Major Target

ABSTRACT The smallpox vaccine is the prototypic vaccine, yet the viral targets critical for vaccine-mediated protection remain unclear in humans. We have produced protein microarrays of a near-complete vaccinia proteome and used them to determine the major antigen specificities of the human humoral immune response to the smallpox vaccine (Dryvax). H3L, an intracellular mature virion envelope protein, was consistently recognized by high-titer antibodies in the majority of human donors, particularly after secondary immunization. We then focused on examining H3L as a valuable human antibody target. Purified human anti-H3L antibodies exhibited substantial vaccinia virus-neutralizing activity in vitro (50% plaque reduction neutralization test [PRNT50] = 44 μg/ml). Mice also make an immunodominant antibody response to H3L after vaccination with vaccinia virus, as determined by vaccinia virus protein microarray. Mice were immunized with recombinant H3L protein to examine H3L-specific antibody responses in greater detail. H3L-immunized mice developed high-titer vaccinia virus-neutralizing antibodies (mean PRNT50 = 1:3,760). Importantly, H3L-immunized mice were subsequently protected against lethal intranasal challenges with 1 or 5 50% lethal doses (LD50) of pathogenic vaccinia virus strain WR, demonstrating the in vivo value of an anti-H3L response. To formally demonstrate that neutralizing anti-H3L antibodies are protective in vivo, we performed anti-H3L serum passive-transfer experiments. Mice receiving H3L-neutralizing antiserum were protected from a lethal challenge with 3 LD50 of vaccinia virus strain WR (5/10 versus 0/10; P < 0.02). Together, these data show that H3L is a major target of the human anti-poxvirus antibody response and is likely to be a key contributor to protection against poxvirus infection and disease.

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Vaccinia virus strain differences in cell attachment and entry

Virology ◽

10.1016/j.virol.2009.04.012 ◽

2009 ◽

Vol 389 (1-2) ◽

pp. 132-140 ◽

Cited By ~ 66

Author(s):

Zain Bengali ◽

Alan C. Townsley ◽

Bernard Moss

Keyword(s):

Vaccinia Virus ◽

Virus Strain ◽

Cell Attachment ◽

Strain Differences ◽

Vaccinia Virus Strain

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Notes on Transient Host Range Selection for Engineering Vaccinia Virus Strain MVA

BioTechniques ◽

10.2144/02331dd01 ◽

2002 ◽

Vol 33 (1) ◽

pp. 186-188 ◽

Cited By ~ 2

Author(s):

David C. Tscharke ◽

Geoffrey L. Smith

Keyword(s):

Vaccinia Virus ◽

Host Range ◽

Virus Strain ◽

Selection For ◽

Vaccinia Virus Strain ◽

Range Selection

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Complete Genome Sequence of Vaccinia Virus Strain L-IVP

Genome Announcements ◽

10.1128/genomea.00372-16 ◽

2016 ◽

Vol 4 (3) ◽

Cited By ~ 2

Author(s):

Alexander N. Shvalov ◽

Galina F. Sivolobova ◽

Elena V. Kuligina ◽

Galina V. Kochneva

Keyword(s):

Vaccinia Virus ◽

Genome Sequence ◽

Virus Strain ◽

Complete Genome Sequence ◽

Complete Genome ◽

Sequence Similarity ◽

Consensus Sequence ◽

Live Vaccine ◽

Eradication Programme ◽

Vaccinia Virus Strain

Most of the live vaccine doses of vaccinia virus donated to the Intensified Smallpox Eradication Programme after 1971 were prepared using the L-IVP strain. A mixture of three clones of the L-IVP strain was sequenced using MySEQ. Consensus sequence similarity with the vaccinia virus Lister strain is 99.5%.

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The N-terminal 22 amino acids encoded by the gene specifying the major secreted protein of vaccinia virus, strain Lister, can function as a signal sequence to direct the export of a foreign protein

Virus Research ◽

10.1016/0168-1702(92)90013-y ◽

1992 ◽

Vol 26 (3) ◽

pp. 197-212 ◽

Cited By ~ 3

Author(s):

Arvind H. Patel ◽

John H. Subak-Sharpe ◽

Nigel D. Stow

Keyword(s):

Amino Acids ◽

Vaccinia Virus ◽

Virus Strain ◽

Signal Sequence ◽

Foreign Protein ◽

Secreted Protein ◽

Vaccinia Virus Strain

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DNA sequence of the gene encoding a major secreted protein of vaccinia virus, strain Lister

Journal of General Virology ◽

10.1099/0022-1317-71-9-2013 ◽

1990 ◽

Vol 71 (9) ◽

pp. 2013-2021 ◽

Cited By ~ 36

Author(s):

A. H. Patel ◽

D. F. Gaffney ◽

J. H. Subak-Sharpe ◽

N. D. Stow

Keyword(s):

Vaccinia Virus ◽

Dna Sequence ◽

Virus Strain ◽

Secreted Protein ◽

Gene Encoding ◽

Vaccinia Virus Strain

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Oncolytic cancer therapy with a vaccinia virus strain

Oncology Reports ◽

10.3892/or.2018.6801 ◽

2018 ◽

Cited By ~ 2

Author(s):

Lili Deng ◽

Jun Fan ◽

Yuedi Ding ◽

Jue Zhang ◽

Bin Zhou ◽

...

Keyword(s):

Cancer Therapy ◽

Vaccinia Virus ◽

Virus Strain ◽

Vaccinia Virus Strain

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Smallpox vaccines induce antibodies to the immunomodulatory, secreted vaccinia virus complement control protein

Journal of General Virology ◽

10.1099/vir.0.008474-0 ◽

2009 ◽

Vol 90 (11) ◽

pp. 2604-2608 ◽

Cited By ~ 8

Author(s):

Joan E. Adamo ◽

Clement A. Meseda ◽

Jerry P. Weir ◽

Michael J. Merchlinsky

Keyword(s):

Vaccinia Virus ◽

Humoral Response ◽

Viral Proteins ◽

Secreted Proteins ◽

Complement Protein ◽

Lethal Challenge ◽

Complement Control Protein ◽

Infected Cells ◽

Control Protein ◽

Primary Band

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. Vaccinia virus deleted in VCP, vVCPko, protected mice from a lethal intranasal challenge of vaccinia Western Reserve strain. Mice vaccinated with purified VCP demonstrated a strong humoral response, but were not protected against a moderate lethal challenge of vaccinia virus, suggesting that the humoral response against VCP is not critical for protection.

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