Evaluación inmunológica en pacientes con Fibromialgia Tipo 1

Zusammenfassung

OBJECTIVES 1. Determine the role of autoimmune mechanisms in the pathogenesis of fibromyalgia. To evaluate the presence of anti-serotonin antibodies, anti-ganglioside antibodies and other autoantibodies in peripheral blood of patients with fibromyalgia and healthy controls, as well as to analyze the variation of these antibodies according to the clinical status of patients. 2. Evaluate the presence of imbalances between pro-inflammatory and anti-inflammatory cytokines and to study the association of cytokine genetic polymorphisms with the susceptibility and clinical manifestations of fibromyalgia. 3. To analyze the polymorphism of KIR and HLA class I and class II genes in fibromyalgia patients and healthy controls in order to investigate their potential association with the susceptibility and clinical manifestations of the disease. METHODOLOGY The determination of anti-serotonin IgG and IgM antibodies was performed bysandwich ELISA, and the determination of anti-Ganglioside antibodies was performed by dot-blot. Serum levels of serotonin were determined by high performance liquid chromatography (HPLC). Serum cytokine determination was performed using Luminex technology, with the exception of TGF-?1 that was performed by sandwich ELISA. The cytokine gene polymorphisms were determined by allelic discrimination assay, with specific Taqman probes and alternatively by Sanger sequencing. The study of the IL-4 gene VNTR polymorphism was performed by conventional PCR using the specific primers to amplify the region of interest. Typing of the HLA-A, B , C, and KIR genes was performed using PCR-SSO and PCR-SSP techniques. CONCLUSIONS: 1. Serotonin levels are decreased in the serum of FM patients but doesn't appear to be associated with the severity of clinical manifestations. 2. The prevalence and serum levels of IgG and IgM anti-serotonin antibodies are increased in patients with FM compared to the healthy population. 3. The prevalence of IgG and IgM anti-serotonin antibodies is higher in FM patients with moderate or severe anxiety levels than in patients with mild anxiety levels, suggesting an association of these antibodies with more severe forms of the disease. 4. Serum levels of the studied pro-inflammatory and anti-inflammatory cytokines are similar in patients with FM and in healthy population. 5. SNPs of the studied pro-inflammatory and anti-inflammatory cytokine genes doesn't appear to be associated with susceptibility to FM development. 6. The AA genotype of the IL-1? gene SNP rs16944 is associated with a severe or very severe manifestation of the disease according to the FIQ test in patients with FM, by contrast, the presence of GA genotype in this SNP is associated with a milder manifestation of the disease. 7. The AA genotype of the IL-1? gene SNP rs1143634 is associated with a moderate or severe manifestation of FM according to the CGI test. 8. The 3R/3R genotype of the IL-4 gene VNTR region is associated with a worse quality of life in patients with FM., by contrast the 2R/3R genotype is associated with a better quality of life. 9. The GG genotype of the TGF-? gene SNP rs1800469 associated with a better quality of life in patients with FM, by contrast the AG genotype is associated with severe anxiety levels in these patients. 10. KIR and HLA genes polymorphism is not associated with the susceptibility and clinical manifestations of FM. 11. This work confirms that FM is conditioned by genetic factors, but also may be related to immunological factors such as autoimmune phenomena or a deregulation of inflammatory mechanisms, which may be relevant for an adequate clinical assessment of patients as well as for the design personalized treatments