Generation and Characterization of DNA Vaccines Targeting the Nucleocapsid Protein of Severe Acute Respiratory Syndrome Coronavirus

Kim, Tae Woo; Lee, Jin Hyup; Hung, Chien Fu; Peng, Shiwen; Roden, Richard B.S.; Wang, Mei Cheng; Viscidi, Raphael P.; Tsai, Ya Chea; He, Liangmei; Chen, Pei‐Jer; Boyd, David A.; Wu, T. C.

doi:10.1128/jvi.78.9.4638-4645.2004

Cited by 173 publications

(174 citation statements)

References 37 publications

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Section: Introductionmentioning

confidence: 99%

“…N protein has been shown to generate CoV-specific CD8 + T cells (Boots et al, 1991;Seo et al, 1997; Stohlman et al, 1993 Stohlman et al, , 1995; it also provides protection in animals in response to infection by animal CoV (Collisson et al, 2000;Seo et al, 1997). In addition, vaccination with SARS N protein decreased replication of a vaccinia virus expressing SARS N protein in mice (Kim et al, 2004).The SARS Accelerated Vaccine Initiative (SAVI) (Finlay et al, 2004) described the first head-to-head comparison of a WKV SARS-CoV vaccine and a combination of attenuated Ad expressing either S or N glycoproteins (Ad S/N) for their ability to protect against live SARS-CoV challenge in a mouse model (See et al, 2006). In the mouse model, the WKV vaccine, in the presence or absence of alum adjuvant, provided protection against live SARS-CoV challenge by the induction of high levels of neutralizing antibodies and reduced SARS-CoV load in the respiratory tract compared with mock-vaccinated mice.…”

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confidence: 99%

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Severe acute respiratory syndrome vaccine efficacy in ferrets: whole killed virus and adenovirus-vectored vaccines

See

Petric

Lawrence

et al. 2008

Journal of General Virology

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Although the 2003 severe acute respiratory syndrome (SARS) outbreak was controlled, repeated transmission of SARS coronavirus (CoV) over several years makes the development of a SARS vaccine desirable. We performed a comparative evaluation of two SARS vaccines for their ability to protect against live SARS-CoV intranasal challenge in ferrets. Both the whole killed SARS-CoV vaccine (with and without alum) and adenovirus-based vectors encoding the nucleocapsid (N) and spike (S) protein induced neutralizing antibody responses and reduced viral replication and shedding in the upper respiratory tract and progression of virus to the lower respiratory tract. The vaccines also diminished haemorrhage in the thymus and reduced the severity and extent of pneumonia and damage to lung epithelium. However, despite high neutralizing antibody titres, protection was incomplete for all vaccine preparations and administration routes. Our data suggest that a combination of vaccine strategies may be required for effective protection from this pathogen. The ferret may be a good model for SARS-CoV infection because it is the only model that replicates the fever seen in human patients, as well as replicating other SARS disease features including infection by the respiratory route, clinical signs, viral replication in upper and lower respiratory tract and lung damage. INTRODUCTIONSevere acute respiratory syndrome (SARS) caused 8098 reported cases and 774 deaths in 26 countries (WHO, 2004a) in a single autumn-to-spring period from 2002 to 2003, and had significant effects on the global economy. Serological evidence suggests zoonotic transmission of SARS coronavirus (CoV) into the human population for several years before this outbreak (Zheng et al., 2004); transmission to humans has continued, resulting in at least four independent non-laboratory associated cases in (Che et al., 2006Fleck, 2004; Guan et al., 2005; WHO, 2004b). The aetiological agent of SARS has been identified as a novel human CoV by sequencing of its genome (Marra et al., 2003;Rota et al., 2003) and by experimental infection of macaques to fulfil Koch's postulates (Fouchier et al., 2003). It is of particular concern as a zoonosis because it can replicate in a large number of animals including cats, pigs, ferrets, foxes, monkeys and rats (Chen et al., 2005; et al., 1996;Olsen, 1993). Although antibodies to CoV N proteins have no virus-neutralizing activity, there is evidence that the protein may provide protection in vivo by induction of cell-mediated immunity, although it has also been suggested to induce eosinophilic infiltrates resulting in immunopathology (Deming et al., 2006;Enjuanes et al., 1995; Stohlman et al., 1995;Wesseling et al., 1993). N protein has been shown to generate CoV-specific CD8 + T cells (Boots et al., 1991;Seo et al., 1997; Stohlman et al., 1993 Stohlman et al., , 1995; it also provides protection in animals in response to infection by animal CoV (Collisson et al., 2000;Seo et al., 1997). In addition, vaccination with SARS N protein decre...

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Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

Severe acute respiratory syndrome vaccine efficacy in ferrets: whole killed virus and adenovirus-vectored vaccines

See

Petric

Lawrence

et al. 2008

Journal of General Virology

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“…It is indeed one of the most abundant structural proteins of coronaviruses and can trigger vigorous humoral and cellular immune responses to the point that it has been considered for production of a SARS vaccine. 20 It is worth noting that the amino acid sequence and localization within the FCoV N protein of both epitopes targeted by the 2 mAbs used in this study remain unknown. Nevertheless, the CCV2-2-recognized epitope seems more frequent than the FIPV3-70-recognized epitope in FIPV.…”

Section: Discussionmentioning

confidence: 99%

Detection of Antigenic Heterogeneity in Feline Coronavirus Nucleocapsid in Feline Pyogranulomatous Meningoencephalitis

Poncelet¹,

Coppens²,

Peeters

et al. 2008

Vet Pathol

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Abstract.A new monoclonal antibody (mAb), CCV2-2, was compared with the widely used FIPV3-70 mAb, both directed against canine coronavirus (CCoV), as a diagnostic and research tool. Western blot showed that both anti-CCoV mAbs only reacted with a protein of 50 kD, a weight consistent with the feline coronavirus (FCoV) viral nucleocapsid. A competitive inhibition enzyme-linked immunosorbent assay showed that the 2 recognized epitopes are distinct. Preincubation of CCV2-2 mAb with FCoV antigen suppressed the immunostaining. Formalin-fixed, paraffin-embedded sections from brains of 15 cats with the dry form of feline infectious peritonitis (FIP) were examined by immunohistochemistry. Immunohistochemistry was performed with both anti-CCoV mAbs, either on consecutive or on the same sections. A myeloid-histiocytic marker, MAC 387, was also used to identify FIP virus-infected cells. In all regions where MAC 387-positive cells were present, positive staining with the CCV2-2 mAb was systematically detected, except at some levels in 1 cat. In contrast, none or only a few cells were positive for the FIPV3-70 mAb. Double immunostaining showed macrophages that were immunopositive for either CCV2-2 alone or alternatively for CCV2-2 and FIPV3-70 mAbs. This reveals the coexistence of 2 cohorts of phagocytes whose FIP viral contents differed by the presence or absence of the FIPV3-70-recognized epitope. These findings provide evidence for antigenic heterogeneity in coronavirus nucleocapsid protein in FIP lesions, a result that is in line with molecular observations. In addition, we provide for the first time morphologic depiction of viral variants distribution in these lesions.

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“…To prevent and control the re-emerging and transmission of SARS-CoV, except for inactivated vaccine experiments [32][33][34] , several different research reports on DNA vaccines of SARSCoV have been recommended, such as S gene recombinant DNA vaccine [35][36][37][38] , N gene recombinant DNA vaccine [39,40] , and a combination of S, M and N gene recombinant DNA vaccines [41,42] . These genes are delivered and expressed by different plasmids or viral vectors and elicit different levels of humoral and cellular immune responses.…”

Section: Discussionmentioning

confidence: 99%

Enhanced Induction of SARS-CoV Nucleocapsid Protein-Specific Immune Response Using DNA Vaccination followed by Adenovirus Boosting in BALB/c Mice

Yao

et al. 2006

Intervirology

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Objective: To investigate immunogenicity in the induction of humoral and cellular immune responses to genetic vaccines of the recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV)-N gene expressing the same protein plasmid, pcDNA3.1-N, and replication-defective adenoviral vector, rAd-N, in a pcDNA3.1-N prime-rAd-N boost regimen and the reverse sequence in a rAd-N prime-pcDNA3.1-N boost regimen. Method: After the mice had been immunized intramuscularly and/or intraperitoneally with pcDNA3.1-N and rAd-N in prime-triple boost immunization, humoral and cellular immune responses were detected. Results: After detection, different levels of anti-N humoral and cellular responses are shown compared to controls. The humoral immune response was induced more effectively by the DNA priming and recombinant adenovirus boosting regimen and the reverse sequence of heterogeneous combinations. There is a significant difference between heterogeneous and homologous vaccinations. However, the cytotoxic T lymphocyte (CTL) response was not significantly altered by the different prime-boost immunizations or the recombinant adenovirus of pcDNA3.1-N prime-rAd-N boost regimen alone, but lymphoproliferation and interferon-γ (IFN-γ) secretion were all enhanced by heterologous combination immunizations compared to homologous combinations. For the reverse sequence immunization regimen, lymphoproliferation, IFN-γ and CTL responses were all significantly weaker compared with pcDNA3.1-N prime-rAd-N boost regimen. Conclusion: Taken together, of all the combinations, the prime-triple boost immunization of pcDNA3.1-N/pcDNA3.1-N/pcDNA3.1-N/rAd-N can effectively induce SARS-CoV-N-specific and strong humoral and cellular immune responses in mice. The present results suggest that DNA immunization followed by recombinant adenovirus boosting could be used as a potential SARS-CoV vaccine in the induction of an enhanced humoral and cellular immune response.

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Generation and Characterization of DNA Vaccines Targeting the Nucleocapsid Protein of Severe Acute Respiratory Syndrome Coronavirus

Cited by 173 publications

References 37 publications

Severe acute respiratory syndrome vaccine efficacy in ferrets: whole killed virus and adenovirus-vectored vaccines

Severe acute respiratory syndrome vaccine efficacy in ferrets: whole killed virus and adenovirus-vectored vaccines

Detection of Antigenic Heterogeneity in Feline Coronavirus Nucleocapsid in Feline Pyogranulomatous Meningoencephalitis

Enhanced Induction of SARS-CoV Nucleocapsid Protein-Specific Immune Response Using DNA Vaccination followed by Adenovirus Boosting in BALB/c Mice

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