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Efficient production of human norovirus-specific IgY in egg yolks by vaccination of hens with a recombinant vesicular stomatitis virus expressing VP1 protein

Zhu, Yang
http://rave.ohiolink.edu/etdc/view?acc_num=osu1397750987

Abstract Details

2014, Master of Science, Ohio State University, Food Science and Technology.
Human norovirus (NoV) is responsible for more than 95% of outbreaks of acute non-bacterial gastroenteritis worldwide and over 50% of foodborne illnesses in the US. Despite significant health, social, and economical burden it causes, no vaccine or antiviral drug available for this virus. This is due to the fact that human NoV cannot be grown in cell culture and lacks a small animal model for pathogenesis study. Recent epidemiological studies showed that severe clinical outcomes including death are often associated with high-risk populations such as infants, children, the elderly, and immunocompromised individuals. There is an urgent need to develop an effective therapeutic agent for human NoV. Chicken immunoglobulin Y (IgY)-based passive immunization has been shown to be an effective strategy to prevent and treat many enteric viral diseases such as rotavirus. In the present study, we developed an efficient approach to generate a high titer of human NoV-specific IgY in chicken yolks using recombinant vesicular stomatitis virus (rVSV-VP1) expressing human NoV capsid protein (VP1) as an antigen. We first demonstrated that rVSV-VP1 replicated efficiently in a chicken cell line and VP1 protein can be highly expressed by VSV vector. Subsequently, White Leghorn chickens were immunized with recombinant rVSV-VP1 by intramuscular route or combination of intramuscular and nasal drop route. After immunization, eggs were collected daily and IgYs were purified from each egg. It was found that the purified IgY strongly reacted with human NoV virus-like particles (VLPs) by both enzyme linked immunosorbant assay (ELISA) and Western blot. In addition, hens vaccinated by intramuscular route triggered significantly higher human NoV-specific IgY than by the combination of intramuscular and nasal drop route. At week 4 post-vaccination, human NoV-specific IgY reached 4.8 mg per yolk in intramuscular vaccinated group, which was approximately 3 times more than that in the combined vaccination group. . Next, we determined whether human NoV-specific IgY has potential antiviral activity. Since human NoV is uncultivable, there is no standard virus-antibody neutralizing assay available. However, it is known that human NoV utilizes histo-blood group antigens (HBGAs) as functional receptors for attachment and subsequent viral entry. Importantly, we found that the purified human NoV-specific IgY efficiently blocked the binding of human NoV VLPs to all three types (A, B and O) of HBGA receptors using a saliva-based HBGA blocking assay. This result suggests that human NoV-specific IgY may have antiviral activity since blockage of viral receptor binding will likely inhibit viral attachment and entry. In order to utilize human NoV-specific IgYs as immunological supplements in food products, the thermal and pH stability of IgY was investigated. It was found that the receptor blocking activity of IgY remained stable at temperature below 70 oC and at pH ranging from 4 to 9.
Jianrong Li (Advisor)
Xin Li (Committee Member)
Melvin Pascall (Committee Member)
123 p.
Food Science
Human norovirus, IgY, Passive immunization

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Citations

  • Zhu, Y. (2014). Efficient production of human norovirus-specific IgY in egg yolks by vaccination of hens with a recombinant vesicular stomatitis virus expressing VP1 protein [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1397750987

    APA Style (7th edition)

  • Zhu, Yang. Efficient production of human norovirus-specific IgY in egg yolks by vaccination of hens with a recombinant vesicular stomatitis virus expressing VP1 protein. 2014. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1397750987.

    MLA Style (8th edition)

  • Zhu, Yang. "Efficient production of human norovirus-specific IgY in egg yolks by vaccination of hens with a recombinant vesicular stomatitis virus expressing VP1 protein." Master's thesis, Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1397750987

    Chicago Manual of Style (17th edition)

osu1397750987
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This open access ETD is published by The Ohio State University and OhioLINK.