Assessment of the cross-protective capability of recombinant capsid proteins derived from pig, rat, and avian hepatitis E viruses (HEV) against challenge with a genotype 3 HEV in pigs

Hepatitis E virus (HEV), the causative agent of hepatitis E, is primarily transmitted via the fecal-oral route through contaminated water supplies, although many sporadic cases of hepatitis E are transmitted zoonotically via direct contact with infected animals or consumption of contaminated animal meats. Genotypes 3 and 4 HEV are zoonotic and infect humans and other animal species, whereas genotypes 1 and 2 HEV are restricted to humans. There exists a single serotype of HEV, although the cross-protective ability among the animal HEV strains is unknown. Thus, in this study we expressed and characterized N-terminal truncated ORF2 capsid antigens derived from swine, rat, and avian HEV strains and evaluated their cross-protective ability in a pig challenge model. Thirty, specific-pathogen-free, pigs were divided into 5 groups of 6 pigs each, and each group of pigs were vaccinated with 200 µg of swine HEV, rat HEV, or avian HEV ORF2 antigen or PBS buffer (2 groups) as positive and negative control groups. After a booster dose immunization at 2 weeks post-vaccination, the vaccinated animals all seroconverted to IgG anti-HEV. At 4 weeks post-vaccination, the animals were intravenously challenged with a genotype 3 mammalian HEV, and necropsied at 4 weeks post-challenge. Viremia, fecal virus shedding, and liver histological lesions were compared to assess the protective and cross-protective abilities of these antigens against HEV challenge in pigs. The results indicated that pigs vaccinated with truncated recombinant capsid antigens derived from three animal strains of HEV induced a strong IgG anti-HEV response in vaccinated pigs, but these antigens confer only partial cross-protection against a genotype 3 mammalian HEV. The results have important implications for the efficacy of current vaccines and for future vaccine development, especially against the novel zoonotic animal strains of HEV

FUNCTIONAL CHARACTERIZATION OF THE INTERACTION OF HEPATITIS E VIRUS ORF3 PRODUCT WITH THE CYTOSKELETON

Hepatitis E virus (HEV) causes several outbreaks of hepatitis in humans. Many aspects of HEV pathogenesis are not well understood. The HEV ORF3 product (henceforth known as vp13) is a multifunctional protein essential for infection of animals. To better understand the vp13 functions, this study was performed. We observed that vp13 protein was associated with the microtubules (MT) in transfected cells. Mutational studies revealed that both hydrophobic domains at the N-terminal region of vp13 are required for the vp13-MT interaction. Our studies also showed that HEV vp13 protein increased the stability of the MT, activated the apoptotic pathway, and, increased the levels of tumor suppressor gene p53 and its downstream effector p21Cip/WAF1 in the transfected cells. However, no noticeable effect on cell survival was observed. These results indicated that HEV vp13 protein may act as a viral regulatory protein

Interferense of Host Innate Immune Response by Hepatitis E Virus

The host antiviral innate immunity mainly relies on host pattern recognition receptors (PRR) and downstream interferon (IFN) signaling. Host PRR for RNA viruses include Toll-like receptors (TLR) and Retinoic acid-inducible gene I (RIG-I) like receptors (RLR). Activation of both TLR and RLR pathways can eventually lead to the secretion of type I IFNs, which can modulate both innate and adaptive immune responses against viral pathogens, including hepatitis E virus (HEV). HEV causes acute hepatitis in humans and has been responsible for several outbreaks of hepatitis across the world. Currently, no commercial vaccine is available for the prevention of HEV infection in any country except China. HEV biology and pathogenesis as well as its responses to host innate immunity are poorly understood, though other hepatitis viruses, including the hepatitis A, B and C viruses, have been much better studied. In this study, how HEV interferes with IFN induction and IFN-activated signaling had been examined. Results showed that the protein encoded by HEV ORF1 can inhibit type I IFN synthesis and downstream JAK/STAT signaling pathway. However, the HEV ORF3 product is able to enhance RIG-I-mediated signaling to a certain extent. These data suggest that HEV proteins interfere with the host innate immune response and may exert the diverse roles depending on the stage and/or context of infection. These studies contribute to a better understanding of HEV pathogenesis and may facilitate a strategy development for the prevention and control of HEV infection

Studies on feline calicivirus with particular reference to persistence.

The molecular evolution of feline calicivirus (FCV) was studied in cell culture and in persistently infected cats. Sequence analysis of the 5' hypervariable region of the FCV capsid (5'HVR; located at the 5' end of variable region E), a region known to contain linear neutralising B cell epitopes, showed FCV existed as a quasispecies which evolved at the nucleotide and amino acid level during persistent infection. Quasispecies heterogeneity tended to decrease the course of persistence. Sequential isolates from a cat showed marked antigenic variation during the course of persistent infection. Sequential passage of FCV in cell culture was also associated with sequence evolution of the 5'HVR. However, these isolates showed no change in antigenicity suggesting that individual substitutions observed in viruses from cats, but not in viruses from cell culture, may be responsible for changes in antigenicity. Alternatively, the observed antigenic changes may be associated with mutations elsewhere in the genome. In order to identify regions of the FCV capsid protein containing liner B-cell epitopes, two approaches were used. Firstly, an expression library containing random, short (100-300bp) fragments of an FCV capsid gene was constructed. This library was screened using polyclonal antisera from a cat that had been challenged experimentally with FCV to identify immunoreactive clones containing B-cell epitopes. Initial screening identified five clones that reacted positively to feline antisera in immunoblots. FCV derived sequence from these clones all mapped to the 5'HVR, suggesting this region contains the immunodominant linear epitopes of the capsid. The second approach used to identify B-cell epitopes was to map more accurately the epitope of a neutralising monoclonal antibody (IG9) which had already been shown to lie in a 37 amino acid region of the 5'HVR (Milton et al. (1992), Journal of General Virology 73, 2435-2439). Replication of plaque purified IG9-sensitive parent virus in sub-neutralising concentrations of IG9 led to the generation of a neutralisation resistant escape mutant. Sequence analysis of this mutant and the parent virus revealed a single non-synonymous nucleotide substitution within the 5'HVR, suggesting this residue is critical to the correct formation of the IG9 epitope

Evaluation of nano-systems as vaccine platforms against infectious diseases

Τα εμβόλια αποτελούν μια από τις πιο σημαντικές σελίδες της ιστορίας της ανθρώπινης υγείας παρέχοντας προστασία εναντίον πολλών λοιμωδών νοσημάτων. Παρά τα αναμφισβήτητα πλεονεκτήματα των κλασικών εμβολίων, αυτά έχουν και σοβαρούς περιορισμούς όπως είναι η χαμηλή ανοσογονικότητα και η μικρής διάρκειας δραστικότητα, η χρονοβόρα διαδικασία ανάπτυξης και παρασκευής, το υψηλό κόστος καθώς και οι περιορισμένοι τρόποι χορήγησης και αποθήκευσης των εμβολίων αυτών. Τα παραπάνω οδήγησαν στην ανάπτυξη του κλάδου των νανο-εμβολίων, που περιλαμβάνει συστήματα εμβολίων μεγέθους 1-1.000 nm. Αυτές οι πλατφόρμες λειτουργούν είτε ως συστήματα μεταφοράς αντιγόνων είτε ως ανοσοενισχυτικά και οδηγούν σε ενεργοποίηση του έμφυτου και του επίκτητου ανοσοποιητικού συστήματος. Τα νανο-εμβόλια μπορούν να ταξινομηθούν με βάση τα φυσικοχημικά και μορφολογικά χαρακτηριστικά τους, σχηματίζοντας πλατφόρμες με μοναδικές ιδιότητες. Τα λιποσώματα, τα όμοια με ιό σωματίδια (VLPs), τα νανο-γαλακτώματα, τα πολυμερικά και ανόργανα νανο-σωματίδια όπως και οι ιικοί φορείς είναι οι κύριες κατηγορίες νανο-εμβολίων που αυτήν τη στιγμή βρίσκονται σε φάσεις προ-κλινικών και κλινικών δοκιμών. Οι νανο-διαμορφώσεις επιτρέπουν την παρασκευή καινοτόμων εμβολίων, όπως τα εμβόλια τρίτης γενιάς, αλλά θα πρέπει να μελετώνται σχολαστικά ώστε να μειωθούν οι πιθανοί κίνδυνοι από τη χρήση τους στη κλινική πράξη. Οι Οργανισμοί Φαρμάκων είναι υπεύθυνοι για τη σύνταξη νομοθετικών πλαισίων για τα νανο-εμβόλια. Με αυτόν τον τρόπο, τόσο οι φαρμακευτικές εταιρείες όσο και οι πληθυσμιακές κοινότητες θα ωφεληθούν μειώνοντας τον κίνδυνο μιας επερχόμενης πανδημίας ή άλλων λοιμωδών νοσημάτων.Vaccines are one of the most important pages of human health by providing protection against various infections. Although the indisputable benefits of classic vaccines, they have some serious limitations such as low immunogenicity and short-term effectiveness, time-consuming developing and/or manufacturing processes, high cost, and limited variety of administration routes and storage. The above led to the development of nanovaccinology, a science area which concludes vaccine platforms of 1-1000 nm. These platforms function either as antigen delivery systems or as immune-stimulators that induce both innate and adaptive immune responses. Nanovaccines can be classified by their physicochemical and morphological characteristics and, thus, platforms with unique properties are able to form. Liposomes, virus-like particles, nano-emulsions, polymeric or inorganic nanoparticles as well as viral vectors are the main categories that are currently on the market or in clinical and pre-clinical phases. Nano-formulations allow the manufacture of innovative vaccines, such as the third generation vaccines, but they shall be carefully studied to minimize probable dangers from their use in clinical practice. Medicine Agencies are responsible for composing a legislative framework about nanovaccinology. In this way, both pharmaceutical companies and human populations will be benefited by eliminating the danger of upcoming pandemics or other infectious diseases

Department of Veterinary and Biomedical Sciences: 2006 Annual Report

Department of Veterinary and Biomedical Sciences Personnel . 9-16 Faculty .. 9 VBMS Researchers, Postdoctoral and Senior Research Associates . 10 VBMS Adjunct and Courtesy Faculty .... 11 Emeriti Faculty .. 11 VBMS Faculty and Staff by Function and Unit .. 9-16 VBMS Honors, Awards and Recognitions, 2006 17-19 VBMS Committee Assignments, 2006/2007 . 20-22 Faculty Profiles . 23-45 Raul G. Barletta, BS, MS, PhD .. 23 Bruce W. Brodersen, BS, DVM, MS, PhD .... 24 Michael P. Carlson, BS, MS, PhD ... 25 Subash Das, BSV c, MVSc, PhD .... 26 Alan R. Doster, DVM, MS, PhD, ACVP . 27 Gerald E. Duhamel, BS, DMV, PhD, ACVP .. 28 M.RohanFernando,BS,MSc,PhD,MPhil ... 29 Dicky Dee Griffm, BS, DVM, MS ... 30 Clin ton]. ] ones, BA, PhD . 31 Clayton L. Kelling, BS, MS, PhD, DVI\u27v1 .. 32 Marjorie F. Lou, BS, MS, PhD .. 33 David S. McVey\ PhD, DVM .. 34 Rodeny A. Moxley, DVM, PhD . 35 Fernando A. Osorio, MV, MS, PhD, ACVM .. 36 Asit K. Pattnaik, BS, MS, PhD .. 37 Douglas G. Rogers, BS, DVM, MS, PhD . 38 Gary P. Rupp, DVM, MS .. 39 John A. Schmiti, DVI\u27vl, PhD, .. A.CVP ... 40 David R. Smith, BS, DVM, PhD, ACVPM, ABVP . 41 Greg A. Somerville, PhD, MS, BS ... 42 David]. Steffen, BS, DVM, PhD, ABVP . 43 Arden R. Wohlers, BS, DVM ... 44 Y. \u27\u27Joe\u27\u27 Zhou, BSc, PhD .. 45 VBMS Researchers, Postdoctoral Research Associates and Senior Research Associates Profiles .... 46-47 Academic Appointment Summary .... 48 Veterinary and Biomedical Sciences Trend, 11-Year Report .. 49 VBMS Teaching Program - Courses, 2006 .. 50-51 Veterinary and Biomedical Sciences Department Courses . 50-51 Enrollment VBMS Courses .... 52 Spring, Summer and Fall Semesters 2006 .. 52 Undergraduate Enrollment/Degrees Obtained .... 53 Nebraska Residents Enrolled in KSU/CVM, Academic Year 2006 (05-2005/04-2006) .... 54 Nebraska Residents that Graduated from Kansas State University, May 2006 ... 55 UNL Students Attending Veterinary Colleges Other Than Kansas State University or Iowa State ... 56 Graduate Students (MS) ... 57 Graduate Students (PhD) ... 57-59 Graduate Degrees Obtaind in 2006 .. 60 Veterinary and Biomedical Science - Seminars .. 61-64 US Meat Animal Research Center (MARC) In-House Seminars ... 64 Great Plains Veterinary Educational Center (GPVEC) .. 65-72 Overview - Gary P. Rupp, Professor and Director .... 65-68 Beef Cattle Production Management Series - 1993-2005 . 69 Beef Cattle Production Management Series Participant Location .. 70 UNL - GPVEC Student Enrollment - 1990-2007 .. 71 2007-2008 Electives .. 72 VBMS Research Program .... 73-99 Faculty Research Interests .... 73-74 VBMS Agricultural Research Division (ARD) Research Projects .. 75-77 Research Projects - Progress Summaries ... 79-99 International Activities, 2006 ... 100-101 Department of Veterinary and Biomedical Sciences Veterinary Extension Program, 2006 . . . 102-107 Topics/Titles of Extension Program Emphases .. 102 D. Dee Griffin .. 102 David R. Smith .. 102 Extension Faculty Programs .... 103-107 D. Dee Griffin .. 103-105 David R. Smith .. 105-107 Nebraska Veterinary and Diagnostic Center (NVDC), 2006 .... 108-118 Overview and Director\u27s Message, Dr. David J. Steffen, Associate Professor and Director .... 108-112 Accessions by Species by Month - January 2006 - December 2006) .. 113 Summary of Laboratory Procedures - January 2006 - December 2006) .... 114 Number of Accessions, Previous Five Years . 115 Number of Laboratory Procedures Conducted, Previous Five Years .. 115 Lag Time Report - Veterinary Diagnostic Center (January 1, 2006 - December 31, 2006) ... 116 Distribution of Accessions by State - NVDLS (January 2006 - December 2006) .... 117 Distribution of Accessions by County - NVDLS (January 2006 - December 2006) .... 118 VBMS Grants and Contracts Program, 2006 . 119-144 Active Grants and Contracts Funded in 2006 .. 119-121 Active Grants and Contracts Continued from Previous Years .... 122-126 Commodity. . . . .. 126-127 Industry ... 127 State .. 127 Revenues Generated . 128 Grant Proposals Submitted in 2006 ... 129-130 Teaching Grants . 131 Private Industry . 131 Grants Submitted, not Funded in 2006 ... 132 Five-Year Record of Grants and Contracts .... 133-142 Industry Grants . 142 Generated Revenues . 142 Intramural Grants ... 142 Travel Grants ... 142 U CARE Program Grants . 143 Patents Pending in 2006 ... 144 Publications by VBMS Faculty in 2006 .. 145-178 Refereed Publications, Year 2006 .. 145-148 Referred Journal Articles, in Press or Accepted in 2006 . 149 Articles Submitted to Refereed Journals in 2006 .. 150-153 Five-Year Record of Refereed Publications, 2001-2006 . 154-173 Non-Refereed Publications and Research Reports for 2006 . 174 Books and Book Chapters in 2006 .... .. 175 Other Publications-Public Press, Lay Journals, Etc., for 2006 .... 176 Extension Publications in 2006 . . 177 Computer Software, Other Publications or Media Developed in 2006 ... 178 Brodersen, Bruce W. . 178 Doster, Alan R.. .. 178 Zhou, You (Joe) ... 178 Presentations by VBMS Faculty in 2006 .. 179-191 Selected Committees, Editorial and Other Appointments, 2006 . 192-199 Barletta, Raul G. . 192 Brodersen, Bruce W. . 192 Carlson, Michael P. . . 192 Doster, Alan R. . 192 Duhamel, Gerald E. .. 193-194 Griffin, Dickey D. ... 194 Jones, ClintonJ .. 195 Kelling, Clayton L .... 195 Lou, Marjorie F. . 195-196 McVey, D. Scott . 196 Moxley, Rodeny A. ... 196-197 Osorio, Fernando A. . 197 Pattnaik, Asit K. . 197 Rupp, Gary P .... 197 Smith, David R. . 198 Somerville, Greg A. .. 198-199 Steffen, David J. . 199 Wohlers, Arden . 199 Articles Regarding the Department in 2006 .. 200 Departmental Budget Summaries, 2006 .. 201-202 Budget, Veterinary and Biomedical Sciences Department - Fiscal Year 2006 . 201 Summary of Other Income - Fiscal Year 2006 .... 201 Nebraska Veterinary Diagnostic Laboratory System Revolving Account Summary for Fiscal Year 2006/2007 .... 202 Nebraska Agricultural Statistics, 2005/2006 .. 203-205 Nebraska 2005/2006 Cash Receipts from Farm Marketings by Commodity .. 203 Nebraska - 2006 Rank in Agriculture .... 204-205 Appendix .... 206-214 The 47th George A. Young Swine Health and :rvfanagement Conference, South Sioux City, Nebraska .... 207-210 The 45th Annual Meeting of the North Central Conference of Veterinary Laboratory Diagnosticians, Holiday Inn Downtown, Lincoln .... 211-21