Cytokine Profiles and Long-Term Virus-Specific Antibodies Following Immunization of CBA Mice with Equine Herpesvirus 1 and Viral Glycoprotein D

Abstract

Equine herpesvirus 1 (EHV-1)-specific antibody-secreting cells (ASC) isolated from the lung and spleen of mice at 12 months after immunization with attenuated EHV-1 KyA, heat-killed KyA, or recombinant viral glycoprotein D (rgD) assessed by ELISPOT showed a three- to fivefold increase in three immunoglobulin isotypes at 3 days post-challenge with pathogenic EHV-1 RacL11 as compared to control mice. ELISPOT assays demonstrated a high frequency of cells secreting proinflammatory tumor necrosis factor-alpha (TNF-alpha), interferon gamma (IFN-gamma), and interleukin 4 (IL-4) in the lungs in response to infection with KyA or RacL11 or immunization with rgD. Cytokine production elicited by EHV-1 KyA or RacL11 infection revealed similar frequencies of EHV-1-specific IFN-gamma and IL-4 spot forming cells in the mediastinal lymph nodes and spleen. However, KyA induced significantly greater amounts of IFN-gamma producing cells in the lungs than did RacL11. Intranasal immunization with KyA or rgD induced long-term immunity that provided protection against pathogenic EHV-1 challenge infection at 12 months post-immunization. Overall, the data indicate that immunization with infectious KyA or rgD induces significant levels of cytokines, virus-specific ASC in the lungs and spleen, and long-term virus specific B-cell responses.

Evaluation of immune responses following infection of ponies with an EHV-1 ORF1/2 deletion mutant

“…Similar to what has been reported in murine studies of EHV-1 infection [34], increases in cytokine/chemokine responses with inflammatory, antiviral (IL-1, TNF-α, (PBMCs) and IFN-α (nasal secretions)), and regulatory function (TGF-β (PBMCs) and IL-10 (nasal secretions)) were seen in the first 48 h following infection. With decreasing body temperatures, overall cytokine expression returned to baseline levels and cytokine mRNA, associated with cellular immune responses (IL-2 and Tbet) decreased markedly in Ab4 WT virus infected ponies.…”

Expression of the Full-Length Form of gp2 of Equine Herpesvirus 1 (EHV-1) Completely Restores Respiratory Virulence to the Attenuated EHV-1 Strain KyA in CBA Mice

“…Poly(A) ϩ RNA was isolated from total RNA using the FastTrack 2.0 kit (Invitrogen) according to the manufacturer's instructions. The resulting poly(A) ϩ RNA was quantitated and analyzed by RNase protection analyses (RPA), which were performed using the 32 P-based Ribo-Quant multiprobe RPA system (BD/Pharmingen) as previously described (15,32,41). Hybridized samples were electrophoresed in 5% acrylamide gels, transferred to filter paper, and exposed to a phosphor screen.…”

Molecular Characterization of the Equine Herpesvirus 1 Strains RacL11 and Kentucky D

“…All of these glycoproteins have been suggested to play some roles in the pathogenicity of EHV-1 [5,10,15,17,21,23,24]. This led us to believe that some specific sequence differences may be related to RacL11 and Kentucky D causing hepatitis.…”