Conclusions and Recommendations for Further Work
7.1 Thesis aims
Componentes
Members
Tafalla Piñeiro, Carolina Investigador OPIs Cuesta Peñafiel, Alberto Ramón y Cajal
Chaves Pozo, Elena Juan de la Cierva Montero Calle, Jana Becaria INIA
Se produjeron las quimioquinas IL-8 y CK-6 recombinantes y se estudió su capacidad atra- yente e inmunomoduladora utilizando la línea celular establecida de macrófagos-monocitos de trucha arcoiris RTS11.
Hemos estudiado la presencia del virus de la necrosis pancreática infecciosa (IPNV), nodavirus y VHSV en un total aproximado de 1.000 mues- tras de peces tanto salvajes como cultivados de la región de Murcia. Se ha detectado la presen- cia de IPNV en algunas muestras aisladas, sin que en ningún caso se observaran síntomas clínicos.
We have characterized the efficacy of alternative promoters that could be used in fish antiviral DNA vaccines in the future instead of currently used human cytomegalovirus promoter that so many biosecurity problems creates.
We have studied the immune effects caused by viral hemorrhagic septicemia virus (VHSV) in sea bream. In this work, we have demonstrated that the most important immune effects provoked by the virus affect cytotoxic cellular reactions as NK function, and oxidative responses. We have performed a study to determine which factors affect the expression of the interleukin 8 receptor (IL-8R).VHSV in vivo produced a significant inhibition in the levels of expression of IL- 8R. This could be considered as a viral suppressive effect which allows it to elude the immune response. DNA vaccination only provoked an increase in the levels of IL-8R expression in the muscle. We could also demonstrate that not only IL-8, but also CK-6 activates the expression of IL-8R. We have used the human defensin HNP1 as a model, to study the antiviral and immunomodulating capacity of defensins in trout, demonstrating that HNP1 has an antiviral activity against VHSV and is immunostimula- tory for trout.
We have produced in bacterial systems recombinant IL-8 and CK-6 and studied the chemoattractant and immunomodulatory capacity using the established rainbow trout monocyte-macrophage cell line, RTS-11. We have studied the presence of infectious pancre- atic necrosis virus (IPNV), nodavirus and VHSV in more than 1,000 wild and cultured fish in the Murcia region during 2008. During this year, we
have detected the presence of IPNV in some fish, without ever having clinical symptoms.
Artículos científicos
Scientific papers
Esteban MA, Meseguer J, Tafalla C, Cuesta A.
(2008).“NK-like and oxidative burst activities are the main early cellular innate immune responses activated after virus inoculation in reservoir fish”.
Fish & Shellfish Immunolology 25, 433-8.
Montero J,Estepa A,Coll J,Tafalla C.(2008).“Regu- lation of rainbow trout (Oncorhynchus mykiss) inter- leukin-8 receptor (IL-8R) gene transcription in response to viral hemorrhagic septicemia virus (VHSV), DNA vaccination and chemokines”. Fish &
Shellfish Immunolology 25, 271-80.
Montero J, Coll J, Sevilla N, Cuesta A, Bols NC, Tafalla C.(2008). “Interleukin 8 and CK-6 che- mokines specifically attract rainbow trout (Oncorhynchus mykiss) RTS11 monocyte- macrophage cells and have variable effects on their immune functions”. Developmental and
Comparative Immunolology 32, 1374-84.
Falco A, Brocal I, Pérez L, Coll JM, Estepa A, Tafalla C.(2008).“In vivo modulation of the rain- bow trout (Oncorhynchus mykiss) immune re- sponse by the human alpha defensin 1, HNP1”.
Fish & Shellfish Immunolology 24,102-12.
Tafalla C, Sánchez E, Lorenzen N, DeWitte-Orr SJ, Bols NC.(2008).“Effects of viral hemorrhagic septicemia virus (VHSV) on the rainbow trout (Oncorhynchus mykiss) monocyte cell line RTS- 11”. Molecular Immunolology 45,1439-48.
Cuesta A, Esteban MA, Meseguer J.(2008). “The expression profile of TLR9 mRNA and CpG ODNs immunostimulatory actions in the teleost gilthead seabream points to a major role of lymphocytes”. Cell and Molecular Life Science 65,
2091-2104.
Cuesta A, Salinas I, Esteban MA, Meseguer J.
(2008). “Unmethylated CpG motifs mimicking bacterial DNA triggers the local and systemic teleost innate immune parameters and expres- sion of immune-relevant genes in gilthead sea- bream”. Fish Shellfish Immunology 25, 617-624.
Objetivos
Objectives
El objetivo general del grupo es avanzar en el conocimiento de la biología y la patogénesis de los priones mediante el desarrollo de modelos basados en ratones transgénicos y cultivos celu- lares que expresan diferentes secuencias de la
PrP. Para ello, estamos siguiendo 4 líneas de tra- bajo estrechamente relacionadas:
• Desarrollo de bioensayos de alta sensibilidad para la detección de priones y el diagnóstico de las enfermedades producidas por priones.
• Estudio de los elementos moleculares que determinan la barrera de transmisión de las diferentes cepas de priones.
• Estudio de los elementos moleculares implicados en la replicación y en la patogé- nesis de los priones.
• Discriminación de las diferentes cepas de priones en especies ganaderas y su capaci- dad de transmisión a humanos.
Palabras clave:
PrP, prion, barrera de transmisión, cepas de priones, replicación.The general objective of the group is to advance in the knowledge of the biology and pathogenesis of prions by using models based on transgenic mice and cell culture that express different sequences of the cellular prion protein (PrPC). Four lines of work are being developed. Development of bioassay of high sensitivity for prion detection.
Study of the molecular elements determining species barrier for the different prion strains. Study of the ele- ments involved in prion replication and pathogenesis. Discrimination of the different prion strains in the different species and their capability of transmission to human.
Key words:
PrP, prion, prion transmision barrier, prion strains, replication.Proyectos en convocatorias
competitivas
Competitive funding projects
AGL2005-03066 (2005-2008). Desarrollo de nuevos bioensayos celulares susceptibles a la replicación de los priones.
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