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J Immunol Methods ; 305(2): 188-98, 2005 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-16157348

RESUMO

One of the challenges for immunomonitoring in clinical trials is to detect an antigen specific T cell-mediated immune response. In an attempt to define the most suitable assay, tetanus toxoid was used to compare the capacity of 4 different methods to detect cytokine responses, before and after recall vaccination, in peripheral blood mononuclear cells (PBMC) of 14 healthy volunteers. ELISA, ELISPOT, intracytoplasmic detection and real-time RT-PCR were chosen to measure IFN-gamma production before and after vaccination. As far as the detection of memory T cell status (before vaccination) was concerned, we found that ELISPOT was the most sensitive method to discriminate TT-induced from spontaneous responses. On the other hand, intracytoplasmic cytokine detection was the most efficient method to detect the restimulating effect of TT vaccination.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Interferon gama/biossíntese , Monitorização Imunológica/métodos , Toxoide Tetânico/imunologia , Vacinação , Adulto , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Memória Imunológica , Interferon gama/sangue , Interferon gama/genética , Interferon gama/imunologia , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/química , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Estatísticas não Paramétricas
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