Detection of spontaneous and antigen-induced human interleukin-4 responses in vitro: Comparison of ELISPOT, a novel ELISA and real-time RT-PCR
2002 (English)In: JIM - Journal of Immunological Methods, ISSN 0022-1759, Vol. 260, no 1-2, 55-67 p.Article in journal (Refereed) Published
Interleukin-4 (IL-4) is an important T-helper cell type 2 (Th2) cytokine in man, driving Th2 polarisation and exerting the most antagonistic effects to the Th1 cytokine interferon-? (IFN-?). Nevertheless, few data on spontaneous and antigen-specific secretion of IL-4 in man are available, mainly due to difficulties in the detection of IL-4. In this study, we compared three assays that can detect antigen-induced IL-4 responses, ELISPOT, ELISA after blocking the IL-4 receptor during cell culture, and real-time reverse transcription polymerase chain reaction (RT-PCR). Spontaneous, antigen- and allergen-induced responses were analysed in peripheral blood mononuclear cells from three groups with different secretion patterns for IL-4: atopic individuals, nonatopic individuals and pregnant women. ELISPOT displayed the highest sensitivity and was the only assay that could detect spontaneous secretion of IL-4 in all analysed samples. The IL-4 receptor blocking ELISA was considered best for the detection of in vitro antigen- and allergen-induced responses, since the results obtained from the ELISPOT and real-time RT-PCR displayed lower specificity, possibly because of seemingly aberrant IL-4 responses in the group of pregnant women. The real-time RT-PCR for detection of IL-4 mRNA proved to be sensitive, but expression of IL-4 mRNA was not correlated with the secretion of IL-4.
Place, publisher, year, edition, pages
2002. Vol. 260, no 1-2, 55-67 p.
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:liu:diva-26419DOI: 10.1016/S0022-1759(01)00520-8Local ID: 10961OAI: oai:DiVA.org:liu-26419DiVA: diva2:246968