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Effect of the viral protein Npro on virulence of bovine viral diarrhea virus and induction of interferon type I in calves

Jamie N. Henningson DVM, PhD1, Christina L. Topliff DVM, PhD2, Laura H. V. Gil DVM, PhD3, Ruben O. Donis MV, PhD4, David J. Steffen DVM, PhD5, Bryan Charleston BVetMed, PhD6, Kent M. Eskridge PhD7, and Clayton L. Kelling DVM, PhD8
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  • 1 Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences and Natural Resources, University of Nebraska, Lincoln, NE 68583.
  • | 2 Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences and Natural Resources, University of Nebraska, Lincoln, NE 68583.
  • | 3 Fundação Osvaldo Cruz, Centro Aggeu Magalhães, Av. Professor Moraes Rego, s/n CEP-50670-420, Recife-PE, Brazil.
  • | 4 Influenza Division, National Center for Immunization and Respiratory Diseases, Coordinating Center for Infectious Diseases, CDC, 1600 Clifton Rd, Mail Stop G-16, Atlanta, GA 30333.
  • | 5 Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences and Natural Resources, University of Nebraska, Lincoln, NE 68583.
  • | 6 Pirbright Laboratory, Institute for Animal Health, Woking, Surrey, GU24 0NF, England.
  • | 7 Department of Statistics, College of Arts and Sciences, University of Nebraska, Lincoln, NE 68583.
  • | 8 Department of Veterinary and Biomedical Sciences, College of Agricultural Sciences and Natural Resources, University of Nebraska, Lincoln, NE 68583.

Abstract

Objective—To characterize the influence of the viral protein Npro on virulence of bovine viral diarrhea virus (BVDV) and on type I interferon responses in calves.

Animals—10 calves, 4 to 6 months of age.

Procedures—BVDV virulence and type I interferon responses of calves (n = 5) infected with a noncytopathic BVDV with a deleted Npro were compared with those of calves (5) infected with a noncytopathic BVDV with a functional Npro. Rectal temperatures, clinical signs, platelet counts, and total and differential WBC counts were evaluted daily. Histologic examinations and immunohistochemical analyses of tissues were conducted to assess lesions and distribution of viral antigens, respectively. Serum type I interferon concentrations were determined.

Results—Calves infected with Npro-deleted BVDV developed leukopenia and lymphopenia, without developing increased rectal temperatures or lymphoid depletion of target lymphoid organs. There was minimal antigen deposition in lymphoid organs. Calves infected with Npro BVDV developed increased rectal temperatures, leukopenia, lymphopenia, and lymphoid depletion with marked BVDV antigen deposition in lymphatic tissues. Interferon type I responses were detected in both groups of calves.

Conclusions and Clinical Relevance—Deletion of Npro resulted in attenuation of BVDV as evidenced by reduced virulence in calves, compared with BVDV with a functional Npro. Deletion of Npro did not affect induction of type I interferon. The Npro-deleted BVDV mutant may represent a safe noncytopathic virus candidate for vaccine development.

Abstract

Objective—To characterize the influence of the viral protein Npro on virulence of bovine viral diarrhea virus (BVDV) and on type I interferon responses in calves.

Animals—10 calves, 4 to 6 months of age.

Procedures—BVDV virulence and type I interferon responses of calves (n = 5) infected with a noncytopathic BVDV with a deleted Npro were compared with those of calves (5) infected with a noncytopathic BVDV with a functional Npro. Rectal temperatures, clinical signs, platelet counts, and total and differential WBC counts were evaluted daily. Histologic examinations and immunohistochemical analyses of tissues were conducted to assess lesions and distribution of viral antigens, respectively. Serum type I interferon concentrations were determined.

Results—Calves infected with Npro-deleted BVDV developed leukopenia and lymphopenia, without developing increased rectal temperatures or lymphoid depletion of target lymphoid organs. There was minimal antigen deposition in lymphoid organs. Calves infected with Npro BVDV developed increased rectal temperatures, leukopenia, lymphopenia, and lymphoid depletion with marked BVDV antigen deposition in lymphatic tissues. Interferon type I responses were detected in both groups of calves.

Conclusions and Clinical Relevance—Deletion of Npro resulted in attenuation of BVDV as evidenced by reduced virulence in calves, compared with BVDV with a functional Npro. Deletion of Npro did not affect induction of type I interferon. The Npro-deleted BVDV mutant may represent a safe noncytopathic virus candidate for vaccine development.

Contributor Notes

Supported in part by Animal Health funds, USDA NRI grant 2002-35-204-11619, and the Nebraska Veterinary Diagnostic Center, Lincoln, Neb.

Presented in part at the Conference of Research Workers in Animal Diseases, St Louis, December 2005.

This study is a contribution of the University of Nebraska Research Division.

Address correspondence to Dr. Kelling.