Domínguez, M. C.; Del Barco, D. García; Taylor, C.; Urquiza, D.; Rodríguez, D.; Suárez, J.; Vázquez, A.; Merino, N.; Cosme, K.; Padrón, G.
October 2002
Revista VacciMonitor (Vacunología y Temas Afines);Oct2002, Vol. 11 Issue 4, p1
Academic Journal
Recent advances in our understanding of cellular and molecular mechanisms of Rheumatoid Arthritis (RA) have highlighted a critical role for th1 cytokines. Adjuvant arthritis (AA) is an experimental model of RA that can be induced in susceptible inbred strains of rats upon immunization with heat-killed Mycobacterium tuberculosis (MT) in CFA. Here we used Lewis rats for the AA model. Our purpose is to validate this animal model towards assays of novel drugs against RA. One of the first step is to corroborate that affected rats have a pattern of th1 cytokines similar to the RA in humans. So that a real judgment for evaluating a drug, will be the inversion of cytokine pattern from th1 to th2 or tendency to the balance th1/th2. To validate this animal model on induction of the cytokine profile, groups of rat were sacrificed at 14, 21, 28 and 35 days after the induction of AA. Spleens homogenates were processed to extract the total RNA by the Guanidium-Phenol-Clorophorm method. Total RNA from each samples was analyzed by RT-PCR using specific oligonucleotides for IL-2, TNFα, IFN, IL-4, IL-6. TGFβ and the constitutive gene GAP. The imagines of electrophoretic runnings were scanned and analyzed by Molecular Analyst Software (BioRad). The results were expressed as relative levels of mRNA from each cytokine. After immunization with MT, the levels of IL-2, TNFα and IFN were significantly increased. These dates are similar to the AR patients. Our results offer a correct characterization of an animal model intended to be used in futures pre-clinical studies.


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