Analysis of the V beta specificity of superantigen activation with a rapid and sensitive method using RT PCR and an automatic DNA analyser.
Polymerase chain reaction (PCR) by the specific amplification of a DNA target sequence has been shown to permit analysis of T cell receptor usage. The complete repertoire is amplified using oligonucleotide primers specific for each of the known V alpha or V beta regions of the T cell receptor. One of the methods currently used to appreciate the relative quantity of different V chains of the TCR is by coamplifying in the same reaction tube the variable region of one chain together with the constant region of the other chain. We have optimised PCR conditions and analysed PCR products on an automatic DNA analyser facilitating the quantification of the amplified products, avoiding the use of radioisotopes, and allowing the determination of the sizes of CDR3 regions, thus giving new information on the modification of the T cell repertoire. This method was used to analyse the precise V beta specificity of the T cell activation with the superantigen SEB.
Cottrez
F
F
Unité mixte INSERM U167-CNRS 624, Institut Pasteur, Lille, France.
Auriault
C
C
Capron
A
A
Groux
H
H
D001322:Autoanalysis / Q000295:instrumentation; D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D018076:DNA, Complementary; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology*; D018089:Superantigens / Q000276:immunology*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta; D018089:Superantigens; D004247:DNA