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Detection of sponta...
Detection of spontaneous and antigen-induced human interleukin-4 responses in vitro : Comparison of ELISPOT, a novel ELISA and real-time RT-PCR
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- Ekerfelt, Christina, 1957- (author)
- Östergötlands Läns Landsting,Linköpings universitet,Hälsouniversitetet,Klinisk immunologi,Klinisk immunologi och transfusionsmedicin
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- Ernerudh, Jan, 1952- (author)
- Östergötlands Läns Landsting,Linköpings universitet,Hälsouniversitetet,Klinisk immunologi,Klinisk immunologi och transfusionsmedicin
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- Jenmalm, Maria, 1971- (author)
- Linköpings universitet,Hälsouniversitetet,Pediatrik
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(creator_code:org_t)
- 2002
- 2002
- English.
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In: JIM - Journal of Immunological Methods. - 0022-1759 .- 1872-7905. ; 260:1-2, s. 55-67
- Related links:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
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- 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.
Keyword
- MEDICINE
- MEDICIN
Publication and Content Type
- ref (subject category)
- art (subject category)
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