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Detection of spontaneous and antigen-induced human interleukin-4 responses in vitro: Comparison of ELISPOT, a novel ELISA and real-time RT-PCR
Linköping University, Faculty of Health Sciences. Linköping University, Department of Molecular and Clinical Medicine, Clinical Immunology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Immunology and Transfusion Medicine.ORCID iD: 0000-0002-3993-9985
Linköping University, Faculty of Health Sciences. Linköping University, Department of Molecular and Clinical Medicine, Clinical Immunology. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Immunology and Transfusion Medicine.
Linköping University, Faculty of Health Sciences. Linköping University, Department of Molecular and Clinical Medicine, Pediatrics.
2002 (English)In: JIM - Journal of Immunological Methods, ISSN 0022-1759, Vol. 260, no 1-2, 55-67 p.Article in journal (Refereed) Published
Abstract [en]

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.
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Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-26419DOI: 10.1016/S0022-1759(01)00520-8Local ID: 10961OAI: oai:DiVA.org:liu-26419DiVA: diva2:246968
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2013-08-29

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Ekerfelt, ChristinaErnerudh, JanJenmalm, Maria

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