A recombinant fusion protein consisting of West Nile virus envelope domain III fused in-frame with equine CD40 ligand induces antiviral immune responses in horses

Liu, Shiliang A.; Haque, Muzammel; Stanfield, Brent; Andrews, Frank M.; Roy, Alma A.; Kousoulas, Konstantin G.

doi:10.1016/j.vetmic.2016.12.008

Cited by 6 publications

(2 citation statements)

References 37 publications

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“…In equine mesenchymal stem cells CD154 RNA expression was reduced after in vitro stimulation with TNF-α and IFN-γ for 72h ( 23 ). Recombinant equine CD154 was used as an experimental vaccine adjuvant of a West Nile Virus subunit vaccine for horses in vivo and improved the induction of neutralizing antibodies compared to the subunit vaccine without CD154 ( 24 ). Furthermore, CHO cell expressed recombinant ( r ) equine CD154 stimulated equine monocyte-derived macrophages to produce reactive oxygen species in vitro ( 20 ).…”

Section: Introductionmentioning

confidence: 99%

CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses

et al. 2022

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Despite the relevance of adaptive immunity against equine pathogens antigen-specific T cell responses of horses are not well characterized and the lack of insight into T cell responses hampers the understanding of the pathogeneses of important diseases. In this study we used tetanus toxoid (TT) as a well-defined antigen to characterize antigen-reactive T cells. Six healthy adult horses received a routine booster against tetanus with an immune stimulating complex (ISCOM)-based vaccine and were followed for 28 days. TT-specific serum antibodies were quantified by ELISA and increased in all horses by day 7 after vaccination. CD154 is an established indicator of antigen-reactive T helper cells in other species, but has not been characterized in horses. CD154 detection in equine PBMC by an anti-human CD154 antibody (clone 5C8) was confirmed by Western blots and then applied for flow cytometry. As a common indicator of equine T cell activation, cytokine induction was studied in parallel. T cells were analyzed by multicolor flow cytometry of PBMC after re-stimulation with TT in vitro. Reactive T helper (Th) cells were characterized by increased frequencies of CD4+CD154+ lymphocytes in in vitro TT-re-stimulated PBMC on day 14 after vaccination of the horses compared to pre-vaccination. The majority of all CD154+ cells after TT re-stimulation were CD4+ Th cells, but CD154 was also induced on CD4- cells albeit in lower frequencies. CD154+CD4+ Th cells were enriched in cytokine-expressing cells compared to CD154-CD4+ Th cells. Similar to the CD4+CD154+ frequencies, CD4+IL-4+, CD4+IFN-γ+ and CD4+TNF-α+ were increased after vaccination, but IL-4+ increased later than IFN-γ+ and CD4+TNF-α+, which already exceeded pre-vaccination frequencies on day 7. CD4+CD154+ frequencies correlated positively with those of CD4+IL-4+ (Th2) on day 14, and negatively with CD4+IFN-γ+ induction on day 7, but did not correlate with CD4+TNF-α+ frequencies or TT-specific antibody concentrations. CD154 appears to be a useful marker of antigen-reactive equine Th cells in combination with cytokine expression. The T cell analyses established here with TT can be applied to other antigens relevant for infections or allergies of horses and in horse models for translational research.

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Section: Introductionmentioning

confidence: 99%

CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses

et al. 2022

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“…Vaccines in development for horses include a recombinant vaccine that targets the WNV-EIII domain. In one study, this vaccine demonstrated immunogenicity when combined with an equine CD40 ligand (CD40L) [ 225 ]. Another promising equine vaccine candidate under development utilizes an avian paramyxovirus 2 (APMV-2)-vectored WNV construct [ 226 ].…”

Section: Vaccines and Therapeutics Against Wnvmentioning

confidence: 99%

Comparison of West Nile Virus Disease in Humans and Horses: Exploiting Similarities for Enhancing Syndromic Surveillance

Schwarz

Long

2023

Viruses

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West Nile virus (WNV) neuroinvasive disease threatens the health and well-being of horses and humans worldwide. Disease in horses and humans is remarkably similar. The occurrence of WNV disease in these mammalian hosts has geographic overlap with shared macroscale and microscale drivers of risk. Importantly, intrahost virus dynamics, the evolution of the antibody response, and clinicopathology are similar. The goal of this review is to provide a comparison of WNV infection in humans and horses and to identify similarities that can be exploited to enhance surveillance methods for the early detection of WNV neuroinvasive disease.

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Purification of animal immunoglobulin G (IgG) using peptoid affinity ligands

Reese

Bordelon²,

Odeh³

et al. 2020

Biotechnology Progress

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The availability of highly pure animal antibodies is critical in the production of diagnostic tools and biosensors. The peptoid PL16, previously isolated from an ensemble of peptoid variants of the IgG‐binding peptide HWRGWV, was utilized in this work as affinity ligand on WorkBeads resin for the purification of immunoglobulin G (IgG) from a variety of mammalian sources and chicken immunoglobulin Y (IgY). The chromatographic protocol initially optimized for murine serum and ascites was subsequently employed for processing rabbit, goat and sheep, donkey, llama, and chicken sera. The PL16‐WorkBeads resin proved able to recover all antibody targets with values of yield between 50 and 90%, and purity consistently above 90%. Notably, PL16 not only binds a broader spectrum of animal immunoglobulins than the reference ligands Protein A and G, but it also binds equally well with all their subclasses. Unlike the protein ligands, in fact, PL16 afforded excellent values of yield and purity of mammalian polyclonal IgG, namely murine (47 and 94%), rabbit (66.5 and 91.7%), caprine IgG (63 and 91–95%), donkey, and llama (93 and 97%), as well as chicken IgY (42 and 92%). Of notice, it is also the ability of PL16 to target monomeric IgG without binding aggregated IgG; when challenged with a mixture of monomeric and aggregated murine IgG, PL16 eluted <3% of fed aggregates, against 11–13% eluted by Protein A and G. Collectively, these results prove the potential of the proposed peptoid ligand for large‐scale purification of animal immunoglobulins.

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A recombinant fusion protein consisting of West Nile virus envelope domain III fused in-frame with equine CD40 ligand induces antiviral immune responses in horses

Cited by 6 publications

References 37 publications

CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses

CD154 Expression Indicates T Cell Activation Following Tetanus Toxoid Vaccination of Horses

Comparison of West Nile Virus Disease in Humans and Horses: Exploiting Similarities for Enhancing Syndromic Surveillance

Purification of animal immunoglobulin G (IgG) using peptoid affinity ligands

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