Molecular cloning and expression of equine interleukin 2

Vandergrifft, Evie V.; Horohov, David W.

doi:10.1016/0165-2427(93)90070-k

Cited by 16 publications

(3 citation statements)

References 22 publications

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“…Additional wells containing sham infected allantoic fluid were included as negative controls. A duplicate set of plates containing 10 u/ml recombinant equine IL-2 in each well was also included (Vandergrifft and Horohov 1993). Plates were incubated at 39°C and 5% CO 2 for 5 days at which time they were pulsed with 0.5 µCi 3 H-thymidine for 4 h. Cultures were then harvested onto glass fibre filter paper and counted in a liquid scintillation counter.…”

Section: Equine Influenza Virus-specific Lymphoproliferationmentioning

confidence: 99%

Exercise alters the immune response to equine influenza virus and increases susceptibility to infection

Folsom

Littlefield-Chabaud

French

et al. 2001

Equine Veterinary Journal

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Summary Equine influenza virus remains a major health concern for the equine industry in spite of ongoing vaccination programmes. Previous work has shown that the immune system of horses can be affected by strenuous exercise. The possible adverse consequence of exercise‐induced alterations in lymphocyte responses measured in vitro was unknown. Here we demonstrate that subjecting vaccinated ponies to a 5 day strenuous exercise programme results in a significant suppression of their T cell‐mediated immune response to equine influenza virus as measured by decreased lymphoproliferation and gamma interferon production measured in vitro. These same ponies also demonstrated increased susceptibility to influenza disease following a challenge exposure to the same strain of virus. Rested ponies that had received the same vaccine and challenge were completely protected from disease. Our results demonstrate that exercise‐induced suppression of the equine immune response to influenza virus can be associated with an increased susceptibility to disease.

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Section: Equine Influenza Virus-specific Lymphoproliferationmentioning

confidence: 99%

Exercise alters the immune response to equine influenza virus and increases susceptibility to infection

Folsom

Littlefield-Chabaud

French

et al. 2001

Equine Veterinary Journal

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“…A dual sandwich ELISA was employed for quantification using commercial antibodies based on known equine specific protein sequences for IL‐1β , IL‐1RA , IL‐2 , IL‐4 , IL‐6 , IL‐10 , IL‐15, TNF‐α , IFN‐γ . Equine antigen‐specific polyclonal antibodies (Abs) , were used as capture Abs, their biotinylated conjugates served as detection Abs, and recombinant equine proteins , served as calibrators using methodology developed in our laboratory .…”

Section: Methodsmentioning

confidence: 99%

Inflammatory mediators are potential biomarkers for extracorporeal shockwave therapy in horses

Chen

Stefanovski

Haughan

et al. 2019

Equine Veterinary Journal

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Summary Background Extracorporeal shockwave therapy (ESWT) can potentially mask painful injuries in equine athletes. Tests to detect whether a horse has received ESWT prior to competition are needed. Extracorporeal shockwave therapy is known to affect inflammatory mediators in other species, and if these mediators are altered in the horse, these could serve as biomarkers of ESWT. Objectives To test the hypothesis that a single application of ESWT will alter the circulating protein concentrations of 10 inflammatory mediators in horse plasma. Study design Prospective repeated measures experimental study. Methods Eleven healthy horses were administered a single dose of ESWT on the dorsal surface of proximal MCIII. Blood samples were collected at −168, −144, −120, −96, −72, −70, −68, −66, −48, −24, −6, −4, −2, 0 h before and 2, 4, 6, 24, 48, 72, 96, 168, 336 and 504 h after ESWT. Plasma concentrations of interleukin 1 beta (IL‐1β), IL‐1 receptor antagonist (IL‐1RA), IL‐2, IL‐4, IL‐6, IL‐10, IL‐15, interferon gamma (IFN‐γ), soluble toll‐like receptor 2 (sTLR2) and tumour necrosis factor alpha (TNF‐α) were measured to assess the effects of ESWT on these mediators. Results Baseline concentrations of inflammatory mediators did not change substantially during the week prior to ESWT. Plasma concentrations of five inflammatory factors changed following ESWT. IL‐1β and IL‐6 were significantly down‐regulated (P<0.01), while TNF‐α, IL‐1RA and TLR2 were significantly up‐regulated (P<0.01). The remaining cytokines were not significantly affected by ESWT. Main limitations This study was performed in a small number of sedentary, healthy pasture‐kept horses using a single dose of ESWT applied to a single location. Additional studies are necessary to determine the effect of ESWT on inflammatory mediators in athletic horses undergoing treatment for musculoskeletal injuries. Conclusions Plasma concentrations of TNF‐α, IL‐1β, IL‐1RA, IL‐6 and TLR2 were significantly affected by ESWT, and deserve further investigation as possible biomarkers of ESWT.

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“…The plasmid pEQU-SPORT2 [63] was used as the source of equine IL-2 cDNA, and the plasmid IGHG1 [64] was used as the source of equine Cγ1 cDNA. The primers used to amplify IL-2 cDNA added a 5′ BamHI site for insertion into the multiple cloning site of the VR-1055 eukaryotic expression vector (Vical, San Diego, CA), a Kozak consensus for optimum eukaryotic expression, and a 3′ Ssp I site for ligation to Cγ1.…”

Section: Methodsmentioning

confidence: 99%

Viral load and clinical disease enhancement associated with a lentivirus cytotoxic T lymphocyte vaccine regimen

Mealey

Leib

Littke

et al. 2009

Vaccine

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Effective DNA-based vaccines against lentiviruses will likely induce CTL against conserved viral proteins. Equine infectious anemia virus (EIAV) infects horses worldwide, and serves as a useful model for lentiviral immune control. Although attenuated live EIAV vaccines have induced protective immune responses, DNA-based vaccines have not. In particular, DNA-based vaccines have had limited success in inducing CTL responses against intracellular pathogens in the horse. We hypothesized that priming with a codon-optimized plasmid encoding EIAV Gag p15/p26 with co-administration of a plasmid encoding an equine IL-2/IgG fusion protein as a molecular adjuvant, followed by boosting with a vaccinia vector expressing Gag p15/p26, would induce protective Gag-specific CTL responses. Although the regimen induced Gag-specific CTL in four of seven vaccinated horses, CTL were not detected until after the vaccinia boost, and protective effects were not observed in EIAV challenged vaccinates. Unexpectedly, vaccinates had significantly higher viral loads and more severe clinical disease, associated with the presence of vaccine-induced CTL. It was concluded that 1.) further optimization of the timing and route of DNA immunization was needed for efficient CTL priming in vivo, 2.) co-administration of the IL-2/IgG plasmid did not enhance CTL priming by the Gag p15/p26 plasmid, 3.) vaccinia vectors are useful for lentivirus-specific CTL induction in the horse, 4.) Gag-specific CTL alone are either insufficient or a more robust Gag-specific CTL response is needed to limit EIAV viremia and clinical disease, and 5.) CTL-inducing vaccines lacking envelope immunogens can result in lentiviral disease enhancement. Although the mechanisms for enhancement associated with this vaccine regimen remain to be elucidated, these results have important implications for development of lentivirus T cell vaccines.

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Molecular cloning and expression of equine interleukin 2

Cited by 16 publications

References 22 publications

Exercise alters the immune response to equine influenza virus and increases susceptibility to infection

Exercise alters the immune response to equine influenza virus and increases susceptibility to infection

Inflammatory mediators are potential biomarkers for extracorporeal shockwave therapy in horses

Viral load and clinical disease enhancement associated with a lentivirus cytotoxic T lymphocyte vaccine regimen

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