Abstract
Crohn´s disease is a chronic inflammatory pathology of the gastrointestinal tract. The cause is unknown. It is characterized by a granulomatous inflammation that could affect any portion of the digest tract, frequently discontinuous and with tendency of fistula formation. Crohn´s disease is common in Europe, North America and Australia. It is less common in India, tropical Africa and South America, and rare in Japan. From 1950 the incidence and prevalence are increasing in several regions of the world. The changes in the incidence profile are not clear. Crohn's disease can occur in any age, it is commonly seen in young adults between fifteen and thirty five years. Male and female are equally affected. There is no correlation with occupational activity or social class. Familiar incidences vary from six to fifteen per cent, genetically correlated with chromosome 16. The inflammation is transmural and consists mainly of lymphocytes, histiocytes and plasma cells. Granuloma is seen in sixty five per cent of patients, it is common in the cases with rectal involvement, less common in the ileum manifestation. The mucosa structure is well preserved and, the cuboidal epithelium is usually seen. The mucosa superficial of the intestine is exposed to a diverse food antigens and bacteria agent from the intestinal flora. The immune physiological process toward the intestinal antigens does not trigger an injury to the own organism, it is mediated by secretion of immunoglobulin A and adaptation of immune system from the stimulus. The cells of the innate immune system, e.g.: macrophages and monocytes are able to organize a rapid process toward a tissue injury, e.g.: against an infectious agent, where there is secretion of pro-inflammatory cytokines as Interleukin 1 (IL1), IL6, IL8, IL12, Tumor Necrosis Factor alpha (TNFα). The group of cytokines afterwards leads to the development of adaptation of the immunity mediated by T and B lymphocytes. ObjectiveCharacterize the effect of the Human Recombinant Granulocyte/Macrophage Colony Stimulating Factor and Anti TNF alpha taken individually, and in combination. Compare the quantitatively the mononuclear cells and cytokines from chronic inflammatory intestinal injury in mice C57/BL6 WT chemical induced and also in IL-10 know-out mice, and after each group treated. MethodsMice C57/BL6 WT and C57/BL6 IL10-/- specific pathogens free will obtained from the biotherium of the Instituto de Biociências da Universidade de São Paulo. The mice will be kept within cages in the Gastroenterology Medical Investigation Laboratory of the Universidade de São Paulo. Mononuclear cells identification in the lamina propria and in the intestinal mucosa: CD4+ and CD8+ cells, NK cells, dendritic cells S100+ using the immunohistochemistry method with streptavidin, biotin and peroxidase. Induction of intestinal inflammation in mice C57/BL6 WT: The mice will be submitted to anesthesia with ether. The inflammation will be induced by instillation of 0.5 mg of TNBS reagent dissolved in equal quantity of ethanol 50%. A volume of 100 µL of the mixture of TNBS and ethanol will be slowly infused by rectal with a catheter of 3.5 F attached to a syringe of one milliliter. The cytokines quantity will be done from biopsies of the caecum and colon using immunohistochemistry. The results will be evaluated before and after the treatment with GM-CSF.Cytokines quantification in paraffined samples in the caecum and colon:Gamma-Interferon, Transforming growth factor-beta1, Tumoral necrosis factor-alpha. The quantification of cytokines will be done by chromogenic reaction.Interleukin 10 and 12 will be done by immunohistochemistry method. (AU)