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BVDV and Vaccination of the Young Animal

BVDV and Vaccination of the Young Animal. James A. Roth, DVM, PhD, DACVM Center for Food Security and Public Health College of Veterinary Medicine Iowa State University. Issues Regarding BVDV Vaccination in the Young Calf. Protection by maternal antibody Immunocompetence of the young calf

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BVDV and Vaccination of the Young Animal

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  1. BVDV and Vaccination of the Young Animal James A. Roth, DVM, PhD, DACVM Center for Food Security and Public Health College of Veterinary Medicine Iowa State University

  2. Issues Regarding BVDV Vaccination in the Young Calf • Protection by maternal antibody • Immunocompetence of the young calf • Induction of protective immune mechanisms by MLV vs. killed vaccines • Cross protection of antigenic variants • Interference by maternal antibody • Immunosuppression by MLV vaccine

  3. Pathogenic Mechanisms Defense Mechanisms

  4. Respiratory Virus Infection Pathogenic Mechanisms Defense Mechanisms

  5. Internal and external antigens surrounding the viral genetic material external antigens internal antigens non-protective protective non-protective Roitt, I., Brostoff, J., Male, D. Immunology. 1985

  6. Exogenous (MHCII) and Endogenous (MHCI) Pathways of Antigen Presentation

  7. Detection of Antigen Specific T Cell-Mediated Immunity to Bovine Respiratory Disease Viruses by Flow CytometryT Helper cells (CD4)Cytotoxic T cells (CD8)Gamma Delta T cells

  8. T T Cytokine Cell surface marker CD25 (IL-2R) T Cell Activation Unstimulated Stimulated Antigen stimulated

  9.  T CD 25 IL-4 CD 4 CD 8 T T IFN Multi-parameter Flow Cytometry Detects expression of CD25, intracellular IFNg, and IL-4 in T cell subsets. • PBMC are incubated with antigens for 4-5 days. • Cell surface markers and intracellular cytokines are stained with different fluorochromes and analyzed by flow cytometer.

  10. Monitoring T Cell Responses by CD25 and CytokineExpression Analysis • Immunize calves • Collect blood samples from vaccinated and control calves

  11. Monitoring T Cell Responses by CD25 and IFNg Expression Analysis • Isolate peripheral blood mononuclear cells (PBMC) • Incubate PBMC in vitro with antigens in microtiter plates • Stain cell phenotype markers and activation markers

  12. Induction of Antigen Specific T Cell Subset Activation to Bovine Respiratory Disease Viruses by MLV Vaccine Platt, R., W. Burdett, and J. A. Roth. 2006. Induction of antigen specific T cell subset activation to bovine respiratory disease viruses by a modified-live virus vaccine. Am J Vet Res, 67:1179-1184, 2006 Supported by Intervet

  13. Experimental Design • Vaccination with modified-live virus vaccine (BHV-1, BRSV, BVDV types 1 and 2, and PI3) (Vista vaccine – Intervet) • Week 0 • Blood collection for CMI assay • Weeks 0, 4, 5, 6, 8, 24, 25, 26, 27 • Challenge • BHV-1 Cooper strain on week 25 (2 ml of 108 TCID50/ml) • Nasal secretion collection for virus titration • Days 0-14 post-challenge

  14. CMI assay • In vitro stimulation of PBMC • Unstimulated • Mitogen stimulated • Live virus stimulated • BHV-1 (Bovishield) • BRSV (Bovishield) • BVDV type 1 (TGAN-NADC) • BVDV type 2 (890-NADC)

  15. Statistically significant (p<0.01) (p<0.05) BHV-1 Results CD25 Expression Index Pre-vaccination Week 0 Post-vaccination Weeks 4, 5, 6, 8 Pre- challenge Weeks 24, 25 Week 1 post-challenge Week 26 Week 2 post-challenge Week 27

  16. Statistically significant (p<0.01) (p<0.05) BVDV Type 1 Results CD25 Expression Index Pre-vaccination Week 0 Post-vaccination Weeks 4, 5, 6, 8 Pre- challenge Weeks 24, 25 Week 1 post-challenge Week 26 Week 2 post-challenge Week 27

  17. Statistically significant (p<0.01) (p<0.05) BVDV Type 2 Results CD25 Expression Index Pre-vaccination Week 0 Post-vaccination Weeks 4, 5, 6, 8 Pre- challenge Weeks 24, 25 Week 1 post-challenge Week 26 Week 2 post-challenge Week 27

  18. Statistically significant (p<0.01) (p<0.05) BHV-1 Results D %IFNg + Pre-vaccination Week 0 Post-vaccination Weeks 4, 5, 6, 8 Pre- challenge Weeks 24, 25 Week 1 post-challenge Week 26 Week 2 post-challenge Week 27

  19. Statistically significant (p<0.01) (p<0.05) BVDV Type 1 Results D %IFNg + Pre-vaccination Week 0 Post-vaccination Weeks 4, 5, 6, 8 Pre- challenge Weeks 24, 25 Week 1 post-challenge Week 26 Week 2 post-challenge Week 27

  20. Statistically significant (p<0.01) (p<0.05) BVDV Type 2 Results D %IFNg + Pre-vaccination Week 0 Post-vaccination Weeks 4, 5, 6, 8 Pre- challenge Weeks 24, 25 Week 1 post-challenge Week 26 Week 2 post-challenge Week 27

  21. Efficacy of Killed BVDV Vaccines for Induction of T Cell Mediated Immunity? Humoral and T cell-mediated immune responses to bivalent killed bovine viral diarrhea virus vaccine in beef cattle. Ratree Platt, Christopher Coutu, Todd Meinert, James A. Roth. Vet Immunol Immunopathol. 122:8-15, 2008. Supported by Pfizer Animal Health

  22. Materials and Methods Animals and vaccines • Two groups of 10, 9-12 month old, BVDV seronegative, castrated male crossbred beef cattle were used. • Gr. 1 served as negative mock-vaccinated control. • Group 2 was vaccinated subcutaneously twice, 3 weeks apart, with 2 ml of modified live BHV-1, PI3, and BRSV diluted in diluent containing killed BVDV type 1 (strain 5960) and type 2 (strain 53637) in an adjuvant containing Quil A, Amphigen, and cholesterol

  23. Materials and Methods Cell-mediated immunity responses • Anticoagulated blood samples were collected on vaccination day and days 28, 35, 56 and 70 post-vaccination. • Specific T cell responses assays for BVDV types 1 and 2 were tested for • The up-regulation of CD25 by multi-parameter flow cytometry • The expression of IFNg in cultured cells supernatants by indirect ELISA.

  24. Net increase of %CD25+ area under the curve analysis results ( p<0.05)

  25. Net increase of IFNg area under the curve analysis results ( p<0.05)

  26. Induction of T Lymphocytes Specific for Bovine Viral Diarrhea Virus in Calves with Maternal Antibody Janice Endsley1, Julia Ridpath2, John Neill2, Matt Sandbulte1, James Roth1 1Iowa State University, 2USDA ARS National Animal Disease Center Viral Immunology 17:13-23, 2004

  27. Hypothesis Calves infected with Bovine Viral Diarrhea virus in the presence of passive antibody will develop CD4, CD8, and  T cells specific for BVDV, without a detectable antibody response and will be protected from subsequent challenge.

  28. Experimental Design • Pooled colostrum from BVDV hyper-immunized cows was fed to 12 calves. • Six of 12 calves were inoculated with BVDV type 2 (strain 1373) at 2 to 5 weeks of age. • Three calves received no colostrum and no BVDV inoculation. • Three calves received no colostrum and were challenged at 2 to 5 weeks of age (all died). • All surviving calves were challenged with BVDV type 2 (strain 1373) at 8 to 9 months of age.

  29. 107 106 105 104 103 Temp (F) 102 101 100 0 2 4 6 8 10 12 14 16 18 20 Day Post Inoculation First inoculation Colostrum No Colostrum Temperature (After 1st Inoculation)

  30. 4 3.5 3 2.5 SVN Titer (log10) 2 1.5 1 0.5 0 1 2 3 4 5 6 7 8 9 10 Month after first BVDV inoculation Colostrum Colostrum, BVDV Neutralizing Antibody Responses to BVDV Type 2

  31. 30 25 P = 0.07 20 Expression Index P = 0.04 15 10 5 0 0-10 wks 11-20 wks 21-32 wks Weeks after first BVDV inoculation Colostrum, BVDV Colostrum Activation of CD4 T Cells by BVDV Type 2

  32. 30 25 P = 0.008 Expression Index 20 P = 0.10 15 10 5 0 0-10 wks 11-20 wks 21-32 wks Weeks after first BVDV inoculation Colostrum, BVDV Colostrum Activation of CD8 T Cells by BVDV Type 2

  33. 30 25 P = 0.04 P = 0.006 P = 0.04 20 Expression Index 15 10 5 0 0-10 wks 11-20 wks 21-32 wks Weeks after first BVDV inoculation Colostrum, BVDV Colostrum Activation of gd T Cells by BVDV Type 2

  34. IFNg Production (ELISA) IFNg ng/ml

  35. 107 Colostrum + Virus 106 Colostrum No Colostrum 105 104 Temp (F) 103 102 101 100 0 2 4 6 8 10 12 14 16 18 20 Days post challenge Challenge Temperature (After Challenge)

  36. Virus Isolation from Buffy Coats (After Challenge) Days post challenge

  37. Mean SN Titers to BVDV 1373 After Challenge Colostrum + Virus Colostrum No Colostrum

  38. Effect of maternal antibody on T cell and antibody responses to BVDV type 2 modified live virus vaccine and virulent challenge Ratree Platt1, Philip W. Widel2, Lyle D. Kesl3, James A. Roth1 1 Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, IA 50011, USA 2 Boehringer Ingelheim Vetmedica, Inc., St. Joseph, MO 64507, USA 3 Veterinary Resources, Inc., Ames, IA 50010, USA Supported by Boehringer Ingelheim Vetmedica, Inc

  39. Materials and Methods • Animals • Three age groups of bottle-fed Holstein calves, (1-2 week, 4-5 week, and 7-8 week old), 12 calves in each group • Eight calves were vaccinated and 4 calves served as non-vaccinated controls • Vaccine • MLV vaccine (Express™5) • BVDV type 2 challenge • Virulent strain 1373

  40. Challenge Experimental design Vaccination wk0 wk3 wk6 wk9 wk11 wk12 wk13 wk14 SVN test CMI assay Clinical signs, rectal temperature, and body weight gain observation

  41. Cell-mediated immunity assay • Multi-parameter flow cytometry (6 fluorochromes) using BVDV type 2 (strain 890) as recall antigen • Detected IL-2 receptor (CD25) and intracellular IFN and IL-4 expressions in major T cell subsets (CD4+, CD8+, TCR+) CD25 expression index = (%CD25+)(MFI) of stimulated cells (%CD25+)(MFI) of unstimulated cells %IFN+ = %IFN+ of stimulated cells - %IFN+ of unstimulated cells %IL-4+ = % IL-4+ of stimulated cells - % IL-4+ of unstimulated cells

  42. Protection from virulent challenge • Clinical signs included depression, anorexia, nasal and ocular discharges, oral lesion, respiratory signs, cough, and diarrhea • Rectal temperature • Body weight gain

  43. Challenge Mean log2 BVDV type 2 SVN titer Levels not connected by same letter are significantly different within same group. * Statistically different (p<0.05) * Statistically different (p<0.01) from control group of the same age

  44. CD25 Expression Index Levels not connected by same letter are significantly different within same group. * Statistically different (p<0.05) * Statistically different (p<0.01) from control group of the same age

  45.  % IFN+ * Statistically different (p<0.05) * Statistically different (p<0.01) from control group of the same age

  46. D % IL-4+ * Statistically different (p<0.05) * Statistically different (p<0.01) from control group of the same age

  47. Mean cumulative clinical score Challenge Levels not connected by same letter are significantly different within same group. * Statistically different (p<0.05) * Statistically different (p<0.01) from control group of the same age

  48. Mean rectal temperature Challenge Levels not connected by same letter are significantly different within same group. * Statistically different (p<0.05) * Statistically different (p<0.01) from control group of the same age

  49. Challenge Mean log2 BVDV type 2 SVN titer Levels not connected by same letter are significantly different within same group. * Statistically different (p<0.05) * Statistically different (p<0.01) from control group of the same age

  50. Response to BHV1, BRSV, and PI3 • No antibody response in any age group of calves • In contrast to BVDV1 and BVDV2 there was no evidence of T cell responsiveness to BHV1, BRSV, or PI3 in any age group of calves • Since the calves were not challenged with BHV1, BRSV, or PI3, the influence of vaccination on resistance to challenge is not known

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