RESUMO
Southern blotting is a method whereby DNA fragments in the gel are denatured by soaking in an alkali solution, carried out of the gel, and transferred onto a membrane. After drying the membrane, the DNA is fixed irreversibly. The net result is a replica on the membrane of the DNA fragment pattern from the agarose gel. This technique is used to demonstrate B-cell clonality in blood and bone marrow down to the 1% level, though more reliably at the 5% level. The analysis is relatively nonselective and will detect novel rearrangements in relapse that were not seen at diagnosis. Modifications of the technique have been used to determine illegitimate switch recombinations and mutations of oncogenes.
Assuntos
Linfócitos B/imunologia , Linfócitos B/patologia , Southern Blotting/métodos , Rearranjo Gênico , Cadeias Pesadas de Imunoglobulinas/genética , DNA/genética , DNA/imunologia , DNA/isolamento & purificação , Humanos , Indicadores e Reagentes , Leucemia/genética , Leucemia/imunologia , Desnaturação de Ácido Nucleico , Hibridização de Ácido Nucleico/métodos , Mapeamento por RestriçãoRESUMO
Multiple myeloma (MM) is a malignant disease characterized by accumulation of morphologically recognizable plasma cells producing immunoglobulin (Ig) in the bone marrow. The occurrence of clonal T cells in MM, as defined by the presence of rearrangements in the T-cell receptor (TCR)-beta chains detected on Southern blotting, is associated with an improved prognosis. This review aims to describe the various ways in which we have demonstrated the presence of such T cell clones, and to describe the phenotype of these cells. Finally, the specificities of these clinically important CD8+ T cell populations will be discussed in the context of immunotherapy.