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      El factor de transferencia como inductor de la expresión de RNAm de IFN-γ e IL-2 en pollos vacunados contra influenza aviar Translated title: Transfer factor acting as IFN-γ and IL-2 mRNA expression inductor in chicken vaccinated against avian influenza

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          Abstract

          La influenza aviar es una enfermedad de gran importancia económica para la industria avícola. En México sólo se ha reportado la cepa H5N2 de baja patogenicidad y ésta se controla mediante la vacunación con virus inactivado. Esta vacuna en emulsión reduce la presencia de signos, pero no la eliminación viral. Desde hace más de 50 años se ha informado acerca de la eficacia del Factor de Transferencia (FT) como inmunomodulador en casos clínicos humanos y en menor cantidad en modelos animales. El objetivo de este trabajo fue el de establecer la dosis que produce un mayor porcentaje de expresión del RNAm de dos citocinas: IL-2 y de IFN-γ. Se diseñó un experimento para evidenciar la expresión del RNAm de estas dos citocinas en pollos previamente inoculados con FT específico para influenza aviar. En la primera fase se aplicaron 0,1, 1, y 10 unidades de FT a diferentes grupos de pollos, posteriormente se realizó la PCR a partir de tejido esplénico. En la segunda fase se aplicó el FT junto con la vacuna a tres nuevos grupos de pollos. Del experimento 1 solamente IL-2 tuvo un porcentaje mayor de positivos (58,33%) con 1 unidad (P < 0,05). En cambio, en el experimento 2, con 1 unidad se obtuvo 75% de positivos para IL-2 (P < 0,05) y 100% para IFN-γ (P < 0,01). De estos resultados su puede concluir que al aplicar una unidad de FT (equivalente a 7,3 μg de proteína) al inicio del experimento y 10 días después otra unidad de FT junto con la vacuna inactivada de IA se indujo la expresión del RNAm de IFN-γ e IL-2.

          Translated abstract

          Avian influenza is a disease of paramount economical importance for the poultry industry. In Mexico, only low pathogenicity H5N2 strain has been reported and it is controlled through inactivated-virus inoculation. This emulsified vaccine reduces clinical signs indeed, but not viral shedding. Over the last 50 years Transfer Factor (TF) has shown to be an efficient immunomodulator and has been used successfully in human clinical cases, and less commonly in animal models. The aim of this work was to establish an avian influenza-specific TF dose able to produce the highest percentage of mRNA expression of the following cytokines: IL-2 and IFN-γ. An experiment to show the mRNA expression of these cytokines in chicken previously inoculated with avian influenza-specific TF was set up. In the first experiment 0.1, 1 and 10 TF units were inoculated into 3 different groups of chickens; PCR for cytokines from splenic tissue was performed. For the second experiment, a second TF inoculation in combination with the vaccine was carried out using 3 new groups of chicken. Experiment 1: Only IL-2 expression was achieved in 58.33% of chickens using 1 TF unit (P < 0.05). In experiment 2, 75% of chickens showed IL-2 with 1 TF unit (P < 0.05) and all of them (100%) expressed IFN-γ (P < 0.01). From these results it can be concluded that IFN-γ and IL-2 expression can be induced by the inoculation of 1 TF unit (equivalent to 7.3 μg of protein) at the beginning of the experiment procedure and after a second inoculation of TF (10 days after) together with the inactivated virus vaccine.

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          Most cited references39

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          Antiviral actions of interferons.

          C Samuel (2001)
          Tremendous progress has been made in understanding the molecular basis of the antiviral actions of interferons (IFNs), as well as strategies evolved by viruses to antagonize the actions of IFNs. Furthermore, advances made while elucidating the IFN system have contributed significantly to our understanding in multiple areas of virology and molecular cell biology, ranging from pathways of signal transduction to the biochemical mechanisms of transcriptional and translational control to the molecular basis of viral pathogenesis. IFNs are approved therapeutics and have moved from the basic research laboratory to the clinic. Among the IFN-induced proteins important in the antiviral actions of IFNs are the RNA-dependent protein kinase (PKR), the 2',5'-oligoadenylate synthetase (OAS) and RNase L, and the Mx protein GTPases. Double-stranded RNA plays a central role in modulating protein phosphorylation and RNA degradation catalyzed by the IFN-inducible PKR kinase and the 2'-5'-oligoadenylate-dependent RNase L, respectively, and also in RNA editing by the IFN-inducible RNA-specific adenosine deaminase (ADAR1). IFN also induces a form of inducible nitric oxide synthase (iNOS2) and the major histocompatibility complex class I and II proteins, all of which play important roles in immune response to infections. Several additional genes whose expression profiles are altered in response to IFN treatment and virus infection have been identified by microarray analyses. The availability of cDNA and genomic clones for many of the components of the IFN system, including IFN-alpha, IFN-beta, and IFN-gamma, their receptors, Jak and Stat and IRF signal transduction components, and proteins such as PKR, 2',5'-OAS, Mx, and ADAR, whose expression is regulated by IFNs, has permitted the generation of mutant proteins, cells that overexpress different forms of the proteins, and animals in which their expression has been disrupted by targeted gene disruption. The use of these IFN system reagents, both in cell culture and in whole animals, continues to provide important contributions to our understanding of the virus-host interaction and cellular antiviral response.
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            How cells respond to interferons.

            Interferons play key roles in mediating antiviral and antigrowth responses and in modulating immune response. The main signaling pathways are rapid and direct. They involve tyrosine phosphorylation and activation of signal transducers and activators of transcription factors by Janus tyrosine kinases at the cell membrane, followed by release of signal transducers and activators of transcription and their migration to the nucleus, where they induce the expression of the many gene products that determine the responses. Ancillary pathways are also activated by the interferons, but their effects on cell physiology are less clear. The Janus kinases and signal transducers and activators of transcription, and many of the interferon-induced proteins, play important alternative roles in cells, raising interesting questions as to how the responses to the interferons intersect with more general aspects of cellular physiology and how the specificity of cytokine responses is maintained.
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              Cloning and characterization of chicken IL-10 and its role in the immune response to Eimeria maxima.

              We isolated the full-length chicken IL-10 (chIL-10) cDNA from an expressed sequence tag library derived from RNA from cecal tonsils of Eimeria tenella-infected chickens. It encodes a 178-aa polypeptide, with a predicted 162-aa mature peptide. Chicken IL-10 has 45 and 42% aa identity with human and murine IL-10, respectively. The structures of the chIL-10 gene and its promoter were determined by direct sequencing of a bacterial artificial chromosome containing chIL-10. The chIL-10 gene structure is similar to (five exons, four introns), but more compact than, that of its mammalian orthologues. The promoter is more similar to that of Fugu IL-10 than human IL-10. Chicken IL-10 mRNA expression was identified mainly in the bursa of Fabricius and cecal tonsils, with low levels of expression also seen in thymus, liver, and lung. Expression was also detected in PHA-activated thymocytes and LPS-stimulated monocyte-derived macrophages, with high expression in an LPS-stimulated macrophage cell line. Recombinant chIL-10 was produced and bioactivity demonstrated through IL-10-induced inhibition of IFN-gamma synthesis by mitogen-activated lymphocytes. We measured the expression of mRNA for chIL-10 and other signature cytokines in gut and spleen of resistant (line C.B12) and susceptible (line 15I) chickens during the course of an E. maxima infection. Susceptible chickens showed higher levels of chIL-10 mRNA expression in the spleen, both constitutively and after infection, and in the small intestine after infection than did resistant chickens. These data indicate a potential role for chIL-10 in changing the Th bias during infection with an intracellular protozoan, thereby contributing to susceptibility of line 15I chickens.
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                Author and article information

                Journal
                amv
                Archivos de medicina veterinaria
                Arch. med. vet.
                Facultad de Ciencias Veterinarias, Universidad Austral de Chile (Valdivia, , Chile )
                0301-732X
                2010
                : 42
                : 1
                : 67-71
                Affiliations
                [01] México D.F orgnameUniversidad Nacional Autónoma de México orgdiv1Facultad de Medicina Veterinaria y Zootecnia orgdiv2Departamento de Microbiología e Inmunología México
                [02] México D.F orgnameIPN orgdiv1Escuela Nacional de Ciencias Biológicas orgdiv2Departamento de Inmunología México
                Article
                S0301-732X2010000100009 S0301-732X(10)04200109
                151c6d2c-6ea5-47ef-ab82-1a5a3a77617d

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

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                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 42, Pages: 5
                Product

                SciELO Chile

                Categories
                ARTICULOS ORIGINALES

                PCR,dyalizable leukocyte extracts,cytokines,cellular immune response,extractos dializables de leucocitos,citocinas,respuesta inmune celular

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