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Article ; Online: High-throughput isolation of SARS-CoV-2 nucleocapsid antibodies for improved antigen detection.

Fujisawa, Mizuki / Adachi, Yu / Onodera, Taishi / Shiwa-Sudo, Nozomi / Iwata-Yoshikawa, Naoko / Nagata, Noriyo / Suzuki, Tadaki / Takeoka, Shinji / Takahashi, Yoshimasa

Biochemical and biophysical research communications

2023 Volume 673, Page(s) 114–120

Abstract: SARS-CoV-2 nucleocapsid protein (NP) is the main target for COVID-19-diagnostic PCR and antigen rapid diagnostic tests (Ag-RDTs). Ag-RDTs are more convenient than PCR tests for point-of-care testing or self-testing to identify the SARS-CoV-2 antigen. The ...

Abstract	SARS-CoV-2 nucleocapsid protein (NP) is the main target for COVID-19-diagnostic PCR and antigen rapid diagnostic tests (Ag-RDTs). Ag-RDTs are more convenient than PCR tests for point-of-care testing or self-testing to identify the SARS-CoV-2 antigen. The sensitivity and specificity of this method depends mainly on the affinity and specificity of NP-binding antibodies; therefore, antigen-antibody binding is key elements for the Ag-RDTs. Here, we applied the high-throughput antibody isolation platform that has been utilized to isolate therapeutic antibodies against rare epitopes. Two NP antibodies were identified to recognize non-overlapping epitopes with high affinity. One antibody specifically binds to SARS-CoV-2 NP, and the other rapidly and tightly binds to SARS-CoV-2 NP with cross-reactivity to SARS-CoV NP. Furthermore, these antibodies were compatible with a sandwich enzyme-linked immunosorbent assay that exhibited enhanced sensitivity for NP detection compared to the previously isolated NP antibodies. Thus, the NP antibody pair is applicable to more sensitive and specific Ag-RDTs, highlighting the utility of a high-throughput antibody isolation platform for diagnostics development.
MeSH term(s)	Humans ; SARS-CoV-2 ; COVID-19/diagnosis ; Nucleocapsid ; Antibodies, Viral ; Epitopes ; Sensitivity and Specificity
Chemical Substances	Antibodies, Viral ; Epitopes
Language	English
Publishing date	2023-06-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	205723-2
ISSN	1090-2104 ; 0006-291X ; 0006-291X
ISSN (online)	1090-2104 ; 0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2023.06.067
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Article ; Online: Prophylactic Vaccine Targeting TLR3 on Dendritic Cells Ameliorates Eosinophilic Pneumonia in a Mouse SARS-CoV Infection Model.

Iwata-Yoshikawa, Naoko / Nagata, Noriyo / Takaki, Hiromi / Matsumoto, Misako / Suzuki, Tadaki / Hasegawa, Hideki / Seya, Tsukasa

ImmunoHorizons

2022 Volume 6, Issue 4, Page(s) 275–282

Abstract: Putative subcomponent vaccines of severe acute respiratory syndrome coronavirus spike protein and ARNAX (TLR3-specific adjuvant for priming dendritic cells) were examined and compared with spike protein + Alum in a mouse BALB/c model. Survival, body ... ...

Abstract	Putative subcomponent vaccines of severe acute respiratory syndrome coronavirus spike protein and ARNAX (TLR3-specific adjuvant for priming dendritic cells) were examined and compared with spike protein + Alum in a mouse BALB/c model. Survival, body weight, virus-neutralizing Ab titer in the blood, and viral titer in the lung were evaluated for prognosis markers. The infiltration degrees of eosinophils in the lung were histopathologically monitored at 10 d postinfection. The results were: (1) adjuvant was essential in vaccines to achieve a complete recovery from infection, (2) ARNAX displayed optimal body weight recovery compared with Alum, (3) ARNAX was optimal for the amelioration of eosinophilic pneumonia, and (4) the eosinophil infiltration score was not associated with the neutralizing Ab titer in the blood or viral titer in the lung. Although the pathological link between the TLR3 vaccine and lung eosinophil infiltration remains unclear, severe acute respiratory syndrome-mediated eosinophilic pneumonia can be blocked by the prior induction of dendritic cell priming by ARNAX.
MeSH term(s)	Adjuvants, Immunologic/pharmacology ; Animals ; Body Weight ; Dendritic Cells ; Disease Models, Animal ; Mice ; Pulmonary Eosinophilia/prevention & control ; SARS Virus ; Toll-Like Receptor 3 ; Viral Vaccines/pharmacology
Chemical Substances	Adjuvants, Immunologic ; TLR3 protein, mouse ; Toll-Like Receptor 3 ; Viral Vaccines
Language	English
Publishing date	2022-04-27
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2573-7732
ISSN (online)	2573-7732
DOI	10.4049/immunohorizons.2200020
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Article ; Online: Author Correction: Immune response and protective efficacy of the SARS-CoV-2 recombinant spike protein vaccine S-268019-b in mice.

Homma, Tomoyuki / Nagata, Noriyo / Hashimoto, Masayuki / Iwata-Yoshikawa, Naoko / Seki, Naomi M / Shiwa-Sudo, Nozomi / Ainai, Akira / Dohi, Keiji / Nikaido, Eiji / Mukai, Akiko / Ukai, Yuuta / Nakagawa, Takayuki / Shimo, Yusuke / Maeda, Hiroki / Shirai, Seiki / Aoki, Miwa / Sonoyama, Takuhiro / Sato, Mamoru / Fumoto, Masataka /

Nagira, Morio / Nakata, Fumihisa / Hashiguchi, Takao / Suzuki, Tadaki / Omoto, Shinya / Hasegawa, Hideki

Scientific reports

2024 Volume 14, Issue 1, Page(s) 2599

Language	English
Publishing date	2024-01-31
Publishing country	England
Document type	Published Erratum
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-024-52772-3
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Article: Lethal severe fever with thrombocytopenia syndrome virus infection causes systemic germinal centre failure and massive T cell apoptosis in cats.

Frontiers in microbiology

2024 Volume 14, Page(s) 1333946

Abstract: Introduction: Severe fever with thrombocytopenia syndrome (SFTS) is a fatal viral disease characterized by high fever, thrombocytopenia, leukopenia, and multi-organ haemorrhage. Disruption of the humoral immune response and decreased lymphocyte numbers ... ...

Abstract	Introduction: Severe fever with thrombocytopenia syndrome (SFTS) is a fatal viral disease characterized by high fever, thrombocytopenia, leukopenia, and multi-organ haemorrhage. Disruption of the humoral immune response and decreased lymphocyte numbers are thought to contribute to the disease severity. These findings have been obtained through the analysis of peripheral blood leukocytes in human patients, whereas analysis of lymph nodes has been limited. Thus, in this study, we characterized the germinal centre response and apoptosis in the lymph nodes of cats with fatal SFTS, because SFTS in cats well mimics the pathology of human SFTS. Methods: Lymph node tissue sections collected during necropsy from seven fatal SFTS patients and five non-SFTS cases were used for histopathological analysis. Additionally, lymph node tissue sections collected from cats with experimental infection of SFTS virus (SFTSV) were also analysed. Results: In the lymphoid follicles of cats with SFTS, a drastic decrease in Bcl6- and Ki67-positive germinal centre B cells was observed. Together, the number of T cells in the follicles was also decreased in SFTS cases. In the paracortex, a marked increase in cleaved-caspase3 positivity was observed in T cells. These changes were independent of the number of local SFTS virus-positive cell. Furthermore, the analysis of cats with experimental SFTSV infection revealed that the intrafollicular Bcl6- and CD3-positive cell numbers in cats with low anti-SFTSV antibody production were significantly lower than those in cats with high anti-SFTSV antibody production. Discussion: These results suggest that dysfunction of the humoral response in severe SFTS was caused by the loss of germinal centre formation and massive apoptosis of T cells in the lymph nodes due to systemically circulating viruses.
Language	English
Publishing date	2024-01-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2023.1333946
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Article ; Online: Impact of Reinfection with SARS-CoV-2 Omicron Variants in Previously Infected Hamsters.

Shiwa-Sudo, Nozomi / Sakai, Yusuke / Iwata-Yoshikawa, Naoko / Watanabe, Shinji / Yamada, Souichi / Kuroda, Yudai / Yamamoto, Tsukasa / Shirakura, Masayuki / Fujisaki, Seiichiro / Miyazaki, Kaya / Miura, Hideka / Nagata, Shiho / Fukushi, Shuetsu / Maeda, Ken / Hasegawa, Hideki / Suzuki, Tadaki / Nagata, Noriyo

Journal of virology

2023 Volume 97, Issue 1, Page(s) e0136622

Abstract: The diversity of SARS-CoV-2 mutations raises the possibility of reinfection of individuals previously infected with earlier variants, and this risk is further increased by the emergence of the B.1.1.529 Omicron variant. In this study, we used ... ...

Abstract	The diversity of SARS-CoV-2 mutations raises the possibility of reinfection of individuals previously infected with earlier variants, and this risk is further increased by the emergence of the B.1.1.529 Omicron variant. In this study, we used an
MeSH term(s)	Animals ; Cricetinae ; COVID-19 ; Mesocricetus ; Reinfection ; RNA, Viral ; SARS-CoV-2/genetics
Chemical Substances	RNA, Viral
Language	English
Publishing date	2023-01-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/jvi.01366-22
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Article ; Online: A lethal mouse model for evaluating vaccine-associated enhanced respiratory disease during SARS-CoV-2 infection.

Iwata-Yoshikawa, Naoko / Shiwa, Nozomi / Sekizuka, Tsuyoshi / Sano, Kaori / Ainai, Akira / Hemmi, Takuya / Kataoka, Michiyo / Kuroda, Makoto / Hasegawa, Hideki / Suzuki, Tadaki / Nagata, Noriyo

Science advances

2022 Volume 8, Issue 1, Page(s) eabh3827

Abstract: One safety concern during severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine development has been the vaccine-associated enhanced disease, which is characterized by eosinophilic immunopathology and T helper cell type 2 ( ... ...

Abstract	One safety concern during severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine development has been the vaccine-associated enhanced disease, which is characterized by eosinophilic immunopathology and T helper cell type 2 (T
Language	English
Publishing date	2022-01-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	2810933-8
ISSN	2375-2548 ; 2375-2548
ISSN (online)	2375-2548
ISSN	2375-2548
DOI	10.1126/sciadv.abh3827
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Article ; Online: TMPRSS2 Contributes to Virus Spread and Immunopathology in the Airways of Murine Models after Coronavirus Infection.

Iwata-Yoshikawa, Naoko / Okamura, Tadashi / Shimizu, Yukiko / Hasegawa, Hideki / Takeda, Makoto / Nagata, Noriyo

Journal of virology

2019 Volume 93, Issue 6

Abstract: Transmembrane serine protease TMPRSS2 activates the spike protein of highly pathogenic human coronaviruses such as severe acute respiratory syndrome-related coronavirus (SARS-CoV) and Middle East respiratory syndrome-related coronavirus (MERS-CoV). ...

Abstract	Transmembrane serine protease TMPRSS2 activates the spike protein of highly pathogenic human coronaviruses such as severe acute respiratory syndrome-related coronavirus (SARS-CoV) and Middle East respiratory syndrome-related coronavirus (MERS-CoV).
MeSH term(s)	Animals ; Cell Line ; Chlorocebus aethiops ; Coronavirus Infections/immunology ; Coronavirus Infections/metabolism ; Coronavirus Infections/virology ; Disease Models, Animal ; Female ; Humans ; Lung/immunology ; Lung/metabolism ; Lung/virology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mice, Transgenic ; Middle East Respiratory Syndrome Coronavirus/immunology ; Poly I-C/metabolism ; SARS Virus ; Serine Endopeptidases/metabolism ; Severe Acute Respiratory Syndrome/immunology ; Severe Acute Respiratory Syndrome/metabolism ; Severe Acute Respiratory Syndrome/virology ; Spike Glycoprotein, Coronavirus/metabolism ; Toll-Like Receptor 3/metabolism ; Vero Cells
Chemical Substances	Spike Glycoprotein, Coronavirus ; Toll-Like Receptor 3 ; Serine Endopeptidases (EC 3.4.21.-) ; TMPRSS2 protein, mouse (EC 3.4.21.-) ; Poly I-C (O84C90HH2L)
Keywords	covid19
Language	English
Publishing date	2019-03-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/JVI.01815-18
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Article ; Online: Results from a preclinical study in rodents and a Phase 1/2, randomized, double-blind, placebo-controlled, parallel-group study of COVID-19 vaccine S-268019-a in Japanese adults

Sonoyama, Takuhiro / Iwata, Satoshi / Shinkai, Masaharu / Iwata-Yoshikawa, Naoko / Shiwa-Sudo, Nozomi / Hemmi, Takuya / Ainai, Akira / Nagata, Noriyo / Matsunaga, Nobuaki / Tada, Yukio / Homma, Tomoyuki / Omoto, Shinya / Yokokawa Shibata, Risa / Igarashi, Kenji / Suzuki, Tadaki / Hasegawa, Hideki / Ariyasu, Mari

Vaccine. 2023 Mar., v. 41, no. 11 p.1834-1847

2023

Abstract: In early 2020, developing vaccines was an urgent need for preventing COVID-19 from a contingency perspective. S-268019-a is a recombinant protein-based vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), comprising a modified ... ...

Abstract	In early 2020, developing vaccines was an urgent need for preventing COVID-19 from a contingency perspective. S-268019-a is a recombinant protein-based vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), comprising a modified recombinant spike protein antigen adjuvanted with agatolimod sodium, a Toll-like receptor-9 agonist. In the preclinical phase, it was administered intramuscularly twice at a 2-week interval in 7-week-old mice. Immunogenicity was assessed, and the mice were challenged intranasally with mouse-adapted SARS-CoV-2 at 2 and 8 weeks, respectively, after the second immunization. After confirming the preclinical effect, a Phase 1/2, randomized, parallel-group clinical study was conducted in healthy adults (aged 20-64 years). All participants received 2 intramuscular injections at various combinations of the antigen and the adjuvant (S-910823/agatolimod sodium, in μg: 12.5/250, 25/250, 50/250, 25/500, 50/500, 100/500, 10/500, 100/100, 200/1000) or placebo (saline) in an equivalent volume at a 3-week interval and were followed up until Day 50 in this interim analysis. In the preclinical studies, S-268019-a was safe and elicited robust immunoglobulin G (IgG) and neutralizing antibody responses in mice. When challenged with SARS-CoV-2, all S-268019-a-treated mice survived and maintained weight until 10 days, whereas all placebo- or adjuvant-treated (without antigen) mice died within 6 days. In the Phase 1/2 trial, although S-268019-a was well tolerated in adult participants, was safe up to Day 50, and elicited robust anti-spike protein IgG antibodies, it did not elicit sufficient neutralizing antibody levels. The S-268019-a vaccine was not sufficiently immunogenic in Japanese adults despite robust immunogenicity and efficacy in mice. Our results exemplify the innate challenges in translating preclinical data in animals to clinical trials, and highlight the need for continued research to overcome such barriers. (jRCT2051200092)
Keywords	COVID-19 infection ; Severe acute respiratory syndrome coronavirus 2 ; Toll-like receptor 9 ; adjuvants ; adults ; agonists ; antibodies ; antigens ; immunization ; immunogenicity ; immunoglobulin G ; placebos ; sodium ; vaccines ; COVID-19 vaccine ; Preclinical study ; Clinical trial ; Recombinant spike protein ; Safety
Language	English
Dates of publication	2023-03
Size	p. 1834-1847.
Publishing place	Elsevier Ltd
Document type	Article ; Online
Note	Use and reproduction
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2022.12.025
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Article ; Online: Results from a preclinical study in rodents and a Phase 1/2, randomized, double-blind, placebo-controlled, parallel-group study of COVID-19 vaccine S-268019-a in Japanese adults.

Vaccine

2022 Volume 41, Issue 11, Page(s) 1834–1847

Abstract: Background: In early 2020, developing vaccines was an urgent need for preventing COVID-19 from a contingency perspective.: Methods: S-268019-a is a recombinant protein-based vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), ...

Abstract	Background: In early 2020, developing vaccines was an urgent need for preventing COVID-19 from a contingency perspective. Methods: S-268019-a is a recombinant protein-based vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), comprising a modified recombinant spike protein antigen adjuvanted with agatolimod sodium, a Toll-like receptor-9 agonist. In the preclinical phase, it was administered intramuscularly twice at a 2-week interval in 7-week-old mice. Immunogenicity was assessed, and the mice were challenged intranasally with mouse-adapted SARS-CoV-2 at 2 and 8 weeks, respectively, after the second immunization. After confirming the preclinical effect, a Phase 1/2, randomized, parallel-group clinical study was conducted in healthy adults (aged 20-64 years). All participants received 2 intramuscular injections at various combinations of the antigen and the adjuvant (S-910823/agatolimod sodium, in μg: 12.5/250, 25/250, 50/250, 25/500, 50/500, 100/500, 10/500, 100/100, 200/1000) or placebo (saline) in an equivalent volume at a 3-week interval and were followed up until Day 50 in this interim analysis. Results: In the preclinical studies, S-268019-a was safe and elicited robust immunoglobulin G (IgG) and neutralizing antibody responses in mice. When challenged with SARS-CoV-2, all S-268019-a-treated mice survived and maintained weight until 10 days, whereas all placebo- or adjuvant-treated (without antigen) mice died within 6 days. In the Phase 1/2 trial, although S-268019-a was well tolerated in adult participants, was safe up to Day 50, and elicited robust anti-spike protein IgG antibodies, it did not elicit sufficient neutralizing antibody levels. Conclusions: The S-268019-a vaccine was not sufficiently immunogenic in Japanese adults despite robust immunogenicity and efficacy in mice. Our results exemplify the innate challenges in translating preclinical data in animals to clinical trials, and highlight the need for continued research to overcome such barriers. (jRCT2051200092).
MeSH term(s)	Animals ; Humans ; Mice ; Adjuvants, Immunologic ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19/prevention & control ; COVID-19 Vaccines/immunology ; Double-Blind Method ; East Asian People ; Immunogenicity, Vaccine ; Immunoglobulin G ; SARS-CoV-2 ; Sodium ; Vaccines, Synthetic/immunology
Chemical Substances	Adjuvants, Immunologic ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 Vaccines ; Immunoglobulin G ; Sodium (9NEZ333N27) ; Vaccines, Synthetic
Language	English
Publishing date	2022-12-16
Publishing country	Netherlands
Document type	Randomized Controlled Trial ; Clinical Trial, Phase I ; Clinical Trial, Phase II ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2022.12.025
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Article ; Online: Saturation time of exposure interval for cross-neutralization response to SARS-CoV-2: Implications for vaccine dose interval.

iScience

2023 Volume 26, Issue 5, Page(s) 106694

Abstract: Evaluating the serum cross-neutralization responses after breakthrough infection with various SARS-CoV-2 variants provides valuable insight for developing variant-proof COVID-19 booster vaccines. However, fairly comparing the impact of breakthrough ... ...

Abstract	Evaluating the serum cross-neutralization responses after breakthrough infection with various SARS-CoV-2 variants provides valuable insight for developing variant-proof COVID-19 booster vaccines. However, fairly comparing the impact of breakthrough infections with distinct epidemic timing on cross-neutralization responses, influenced by the exposure interval between vaccination and infection, is challenging. To compare the impact of pre-Omicron to Omicron breakthrough infection, we estimated the effects on cross-neutralizing responses by the exposure interval using Bayesian hierarchical modeling. The saturation time required to generate saturated cross-neutralization responses differed by variant, with variants more antigenically distant from the ancestral strain requiring longer intervals of 2-4 months. The breadths of saturated cross-neutralization responses to Omicron lineages were comparable in pre-Omicron and Omicron breakthrough infections. Our results highlight the importance of vaccine dosage intervals of 4 months or longer, regardless of the antigenicity of the exposed antigen, to maximize the breadth of serum cross-neutralization covering SARS-CoV-2 Omicron lineages.
Language	English
Publishing date	2023-04-19
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2023.106694
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