Non Susceptibility of Neonatal and Adult Rats against the Middle East Respiratory Syndrome Coronavirus

Iwata‐Yoshikawa, Naoko; Fukushi, Shuetsu; Fukuma, Aiko; Suzuki, Tadaki; Takeda, Makoto; Tashiro, Masato; Hasegawa, Hideki; Nagata, Noriyo

doi:10.7883/yoken.jjid.2015.589

Cited by 3 publications

(9 citation statements)

References 17 publications

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“…When generating and validating the cELISA, three positive control sera and 42 negative control sera were used. The three positive controls were from two rabbits that had been immunized with recombinant MERS-CoV S and a rat that had been infected with MERS-CoV (Fukuma et al, 2015;Iwata-Yoshikawa et al, 2016). The 42 negative controls were sera from ten mice that had been immunized with recombinant SARS-CoV S protein (Ishii et al, 2009), two rabbits immunized with recombinant MERS-CoV N protein (Fukuma et al, 2015), a rabbit immunized with recombinant SARS-CoV S protein (Fukuma et al, 2015), a rabbit immunized with UV-inactivated SARS-CoV particles (Fukushi et al, 2005), four mock-infected rats, 12 non-immunized mice, and 12 healthy human donors.…”

Section: Serum Samplesmentioning

confidence: 99%

“…MERS-CoV was cultured in BSL-3 laboratory of National Institute of Infectious Diseases as described previously (Iwata-Yoshikawa et al, 2016). Vero cells were inoculated with MERS-CoV at a multiplicity of infection (MOI) of 1.0 per cell.…”

Section: Preparation Of Mers-cov Antigen For the Celisamentioning

confidence: 99%

“…Vero cells were inoculated with MERS-CoV at a multiplicity of infection (MOI) of 1.0 per cell. After 26 h, the cells were lysed with PBS containing 1% NP40 to extract the viral antigens from infected cells (Iwata-Yoshikawa et al, 2016). After centrifugation at 8000g for 10 min, the supernatant was collected and used as the MERS-CoV antigen in the cELISA.…”

Section: Preparation Of Mers-cov Antigen For the Celisamentioning

confidence: 99%

“…Hybridomas were produced by fusing Sp2/O-Ag14 cells with the splenic cells from the mice that were obtained 3 days after the last immunization. The culture supernatants of the hybridoma cells were screened for the presence of antibodies against MERS-CoV antigen by ELISA as described previously (Iwata-Yoshikawa et al, 2016). To exclude hybridomas that produced antibodies that were cross-reactive with other coronaviruses, the culture supernatants were also subjected to ELISA using antigens prepared from cells infected with human coronavirus 229E, NL63, and OC43 (kindly provided by Dr. Shutoku Matsuyama, NIID).…”

Section: Production and Isotyping Of Mabsmentioning

confidence: 99%

“…To exclude hybridomas that produced antibodies that were cross-reactive with other coronaviruses, the culture supernatants were also subjected to ELISA using antigens prepared from cells infected with human coronavirus 229E, NL63, and OC43 (kindly provided by Dr. Shutoku Matsuyama, NIID). This ELISA was also performed as described previously (Iwata-Yoshikawa et al, 2016). The MAb-producing hybridoma cells that reacted specifically with MERS-CoV were selected and were further cloned by using the limiting dilution method.…”

Section: Production and Isotyping Of Mabsmentioning

confidence: 99%

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Characterization of novel monoclonal antibodies against the MERS-coronavirus spike protein and their application in species-independent antibody detection by competitive ELISA

Fukushi

Fukuma

Kurosu

et al. 2018

Journal of Virological Methods

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Since discovering the Middle East respiratory syndrome coronavirus (MERS-CoV) as a causative agent of severe respiratory illness in the Middle East in 2012, serological testing has been conducted to assess antibody responses in patients and to investigate the zoonotic reservoir of the virus. Although the virus neutralization test is the gold standard assay for MERS diagnosis and for investigating the zoonotic reservoir, it uses live virus and so must be performed in high containment laboratories. Competitive ELISA (cELISA), in which a labeled monoclonal antibody (MAb) competes with test serum antibodies for target epitopes, may be a suitable alternative because it detects antibodies in a species-independent manner. In this study, novel MAbs against the spike protein of MERS-CoV were produced and characterized. One of these MAbs was used to develop a cELISA. The cELISA detected MERS-CoV-specific antibodies in sera from MERS-CoV-infected rats and rabbits immunized with the spike protein of MERS-CoV. The MAb-based cELISA was validated using sera from Ethiopian dromedary camels. Relative to the neutralization test, the cELISA detected MERS-CoV-specific antibodies in 66 Ethiopian dromedary camels with a sensitivity and specificity of 98% and 100%, respectively. The cELISA and neutralization test results correlated well (Pearson's correlation coefficients=0.71-0.76, depending on the cELISA serum dilution). This cELISA may be useful for MERS epidemiological investigations on MERS-CoV infection.

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Section: Serum Samplesmentioning

confidence: 99%

Section: Preparation Of Mers-cov Antigen For the Celisamentioning

confidence: 99%

Section: Preparation Of Mers-cov Antigen For the Celisamentioning

confidence: 99%

Section: Production and Isotyping Of Mabsmentioning

confidence: 99%

Section: Production and Isotyping Of Mabsmentioning

confidence: 99%

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Characterization of novel monoclonal antibodies against the MERS-coronavirus spike protein and their application in species-independent antibody detection by competitive ELISA

Fukushi

Fukuma

Kurosu

et al. 2018

Journal of Virological Methods

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Acute Respiratory Infection in Human Dipeptidyl Peptidase 4-Transgenic Mice Infected with Middle East Respiratory Syndrome Coronavirus

Iwata‐Yoshikawa

Okamura

Shimizu

et al. 2019

J Virol

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Middle East respiratory syndrome coronavirus (MERS-CoV) infection can manifest as a mild illness, acute respiratory distress, organ failure, or death. Several animal models have been established to study disease pathogenesis and to develop vaccines and therapeutic agents. Here, we developed transgenic (Tg) mice on a C57BL/6 background; these mice expressed human CD26/dipeptidyl peptidase 4 (hDPP4), a functional receptor for MERS-CoV, under the control of an endogenous hDPP4 promoter. We then characterized this mouse model of MERS-CoV. The expression profile of hDPP4 in these mice was almost equivalent to that in human tissues, including kidney and lung; however, hDPP4 was overexpressed in murine CD3positive cells within peripheral blood and lymphoid tissues. Intranasal inoculation of young and adult Tg mice with MERS-CoV led to infection of the lower respiratory tract and pathological evidence of acute multifocal interstitial pneumonia within 7 days, with only transient loss of body weight. However, the immunopathology in young and adult Tg mice was different. On day 5 or 7 postinoculation, lungs of adult Tg mice contained higher levels of proinflammatory cytokines and chemokines associated with migration of macrophages. These results suggest that the immunopathology of MERS-CoV infection in the Tg mouse is age dependent. The mouse model described here will increase our understanding of disease pathogenesis and host mediators that protect against MERS-CoV infection. IMPORTANCE Middle East respiratory syndrome coronavirus (MERS-CoV) infections are endemic in the Middle East and a threat to public health worldwide. Rodents are not susceptible to the virus because they do not express functional receptors; therefore, we generated a new animal model of MERS-CoV infection based on transgenic mice expressing human DPP4 (hDPP4). The pattern of hDPP4 expression in this model was similar to that in human tissues (except lymphoid tissue). In addition, MERS-CoV was limited to the respiratory tract. Here, we focused on host factors involved in immunopathology in MERS-CoV infection and clarified differences in antiviral immune responses between young and adult transgenic mice. This new small-animal model could contribute to more in-depth study of the pathology of MERS-CoV infection and aid development of suitable treatments. FIG 9 Histopathological changes in the lungs of human dipeptidyl peptidase 4 (hDPP4)-transgenic mice inoculated with MERS-CoV. Representative histopathological images of the lungs from 25-week-old Tg2 mice at 1, 3, 5, 7, 14, and 35 days post-MERS-CoV infection. Images in the left (A, D, G, J, M, and P) and right (C, F, I, L, O, and R) columns show time-dependent recruitment of inflammatory cells to the lung. Marked inflammatory cell infiltration was noted at 7 days postinoculation (dpi) in panels J and L. Images in the middle column (B, E, H, K, N, and Q) show immunohistochemical staining for MERS-CoV antigen (Ag). Scale bars: 100 m (left and middle columns), 50 m (right column), and 20 m (insets of middle...

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Host Determinants of MERS-CoV Transmission and Pathogenesis

Widagdo¹,

Ayudhya²,

Hundie³

et al. 2019

Viruses

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Middle East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic pathogen that causes respiratory infection in humans, ranging from asymptomatic to severe pneumonia. In dromedary camels, the virus only causes a mild infection but it spreads efficiently between animals. Differences in the behavior of the virus observed between individuals, as well as between humans and dromedary camels, highlight the role of host factors in MERS-CoV pathogenesis and transmission. One of these host factors, the MERS-CoV receptor dipeptidyl peptidase-4 (DPP4), may be a critical determinant because it is variably expressed in MERS-CoV-susceptible species as well as in humans. This could partially explain inter- and intraspecies differences in the tropism, pathogenesis, and transmissibility of MERS-CoV. In this review, we explore the role of DPP4 and other host factors in MERS-CoV transmission and pathogenesis—such as sialic acids, host proteases, and interferons. Further characterization of these host determinants may potentially offer novel insights to develop intervention strategies to tackle ongoing outbreaks.

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Non Susceptibility of Neonatal and Adult Rats against the Middle East Respiratory Syndrome Coronavirus

Cited by 3 publications

References 17 publications

Characterization of novel monoclonal antibodies against the MERS-coronavirus spike protein and their application in species-independent antibody detection by competitive ELISA

Characterization of novel monoclonal antibodies against the MERS-coronavirus spike protein and their application in species-independent antibody detection by competitive ELISA

Acute Respiratory Infection in Human Dipeptidyl Peptidase 4-Transgenic Mice Infected with Middle East Respiratory Syndrome Coronavirus

Host Determinants of MERS-CoV Transmission and Pathogenesis

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