Detection of epsilon class switching and IgE synthesis in human B cells.
Methods Mol Biol
; 315: 319-29, 2006.
Article
en En
| MEDLINE
| ID: mdl-16110166
ABSTRACT
We observed that mast cells, as other cells expressing the CD40 ligand CD154, can trigger IgE synthesis in B cells in the presence of interleukin (IL)-4. Numerous complementary techniques can be used to follow the succession of molecular events leading to IgE synthesis. This chapter will illustrate how human B cells (naïve or memory) can be purified, stored, and cultivated in medium that is permissive for IgE synthesis and stimulated with IL-4 or IL-13 and CD40 activation, the latter being induced by soluble CD154, anti-CD40 antibodies, or CD154-expressing cells. All these molecules are expressed by mast cells. The quantification of the epsilon-sterile transcript synthesis by polymerase chain reaction or Northern blot, the epsilon excision circles produced during immunoglobulin heavy chain locus rearrangement by polymerase chain reaction, and the IgE production by enzyme-linked immunosorbent assay will be described.
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Banco de datos:
MEDLINE
Asunto principal:
Inmunoglobulina E
/
Linfocitos B
/
Cadenas epsilon de Inmunoglobulina
/
Cambio de Clase de Inmunoglobulina
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2006
Tipo del documento:
Article
País de afiliación:
Francia