Full text
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| Author(s): |
Dilayla Kelly de Abreu
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD) |
| Defense date: | 2010-12-16 |
| Examining board members: |
Carlos Eduardo Ambrosio;
David Feder;
Janaína Munuera Monteiro
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| Advisor: | Carlos Eduardo Ambrosio |
| Abstract | |
Devised to test this in order to standardize the hematological, biochemical, electrophoretic (Serum protein) and quantification of CD4 lymphocyte cells, CD5, CD8 by flow cytometry of Golden Retriever dogs normal (group GR) and dogs dystrophic (GRMD groups). To this end, we considered the division of groups according to age of animals, from birth to adulthood, thus composing six groups, as Gr I, II, III and GRMD I, II, III. In this project we performed electrophoretic studies of dogs belonging to all groups, haematological and biochemical studies of dogs belonging to the groups GR II, III, II and III GRMD, lymphocyte immunophenotyping in groups III and GR GRMD III. The results obtained for trombometric, erythometric and dogs belonging to the groups and GR II GR III showed mean values within normal limits. With respect to dogs belonging to groups III GRMD, we observed that the erythrocyte is within the reference values. However, considering the WBC, the mean (3.79 / mm3) relating to the measurement of basophils were above normal values, ranging from 0 to 159/mm3. Moreover, considering the maximum, some animals belonging to the group in question had leukocytosis with neutrophilia without left shift, and thrombocytosis, monocytosis and lymphopenia at the time of collection. The biochemical serum of all affected dogs had mean values above the normal range for the determination of AST, ALT and CK. For the study of proteins, SDS-PAGE technique allowed the fractionation of twenty-three proteins whose molecular weights ranged from 16,000 to 260,000 daltons in all groups. These could be identified by name twelve fractions: IgA (PM 142,000 Da), C-reactive protein (PM122.000 Da), ceruloplasmin (PM 110,000 Da), phosphorylase (95,000 Da PM), transferrin (82,000 Da PM), hemopexin (PM 78 000 Da), albumin (66,000 Da PM), alpha1 antitrypsin (PM 62,000 Da), IgG heavy chain (55,000 Da PM), haptoglobin (PM 45,000 Da), glycoprotein (PM 40,000 Da) and IgG light chain (PM 23 000 Da). The other proteins were identified by their molecular weights. We could not identify by name the protein fractions of molecular weight 260,000 Da, 230,000 Da, 180,000 Da, 165,000 Da, 158,000 Da, 91,000 Da, 35,000 Da, 30,000 Da, 28,000 Da and 16,000 Da. Among these proteins was observed that the protein molecular weight of 91,000 Da was found only in groups GR I and GRMD I was not identified in other groups. Considering the changes found with relation to acute phase proteins, we noted the acute phase response in the face of muscle injuries that occur gradually in dystrophic dogs, especially in young dystrophic animals, which may infer that they had abnormal protein before tissue injury as reported in many inflammatory-related pathologies. Likewise, animals affected by muscular dystrophy, alterations immunoglobulin when compared to normal animals. With respect to lymphocyte immunophenotyping was made a histogram of the populations of CD4, CD5 and CD8 from the region of the gate of lymphocytes. All animals affected by muscular dystrophy showed significantly higher percentage with respect to CD4 and CD5 lymphocyte population compared to normal dogs. Thus, we can infer that the process of progressive muscular dystrophy, is associated with changes in cell populations that make up the immune system of patients, because due to the absence of dystrophin, the muscle is more susceptible to damage, and its occurrence , a release of cytokines that stimulate liver cells to secrete acute phase proteins and promote a co-stimulation of T lymphocytes Thus, we suggest that the findings in our study are consequences of the inflammatory response generated by muscle injury.We hope that this study, provide more information about the pathophysiolgy of the disease, and promote a greater understanding of the immune and inflammatory responde assessment in this study model. Moreover, the base values found could help in assessing possible responses in preclinical testing, aiding the understanding of the beneficial or adverse reactions to validate and explain the mechanism of action of a cell therapy, gene or drug treatments (AU) | |
| FAPESP's process: | 09/02958-1 - Standardization of blood profile, biochemical, serum protein and immunophenotype of lymphocytes in healthy Golden Retriever and affected by muscular dystrophy. |
| Grantee: | Dilayla Kelly de Abreu |
| Support Opportunities: | Scholarships in Brazil - Master |