Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCR
Only limited amounts of peripheral blood samples can be obtained from small children. Therefore, a polymerase chain reaction (PCR) aided analysis of cytokine gene expression by PBMC or T cells is a valuable tool. We present a combination of procedures to obtain an accurate estimation of the expressi...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
1995-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/S0962935195000329 |
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| author | H. Koning M. R. M. Baert R. de Groot H. J. Neijens H. F. J. Savelkoul |
| author_facet | H. Koning M. R. M. Baert R. de Groot H. J. Neijens H. F. J. Savelkoul |
| author_sort | H. Koning |
| collection | DOAJ |
| description | Only limited amounts of peripheral blood samples can be obtained from small children. Therefore, a polymerase chain reaction (PCR) aided analysis of cytokine gene expression by PBMC or T cells is a valuable tool. We present a combination of procedures to obtain an accurate estimation of the expression of the cytokines IL-4 and IFN-γ. This can be performed on T cells purified from blood samples of up to 5 ml in volume from children aged 0–4 years with allergic asthma and atopic dermatitis. This procedure includes multiple sampling of PCR products to determine the linear phase of the PCR; inter-experiment correction using a helper T-cell clone, expressing both IL-4 and IFN-γ; interpatient correction by comparing the expression of a housekeeping gene (HPRT); and finally the development of specific software to analyse densitometric data obtained by scanning photographs of agarose gels, separating PCR products. In this way it is possible to study cytokine gene expression from a very small amount of material. |
| format | Article |
| id | doaj-art-7d9d1f4c5ccf40178c1b16edc8ecf687 |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 1995-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-7d9d1f4c5ccf40178c1b16edc8ecf6872025-02-03T01:03:18ZengWileyMediators of Inflammation0962-93511466-18611995-01-014319620410.1155/S0962935195000329Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCRH. Koning0M. R. M. Baert1R. de Groot2H. J. Neijens3H. F. J. Savelkoul4Department of Immunology, Erasmus University, Rotterdam, The NetherlandsDepartment of Immunology, Erasmus University, Rotterdam, The NetherlandsDepartment of Pediatrics, Sophia Children's Hospital, Rotterdam, The NetherlandsDepartment of Pediatrics, Sophia Children's Hospital, Rotterdam, The NetherlandsDepartment of Immunology, Erasmus University, Rotterdam, The NetherlandsOnly limited amounts of peripheral blood samples can be obtained from small children. Therefore, a polymerase chain reaction (PCR) aided analysis of cytokine gene expression by PBMC or T cells is a valuable tool. We present a combination of procedures to obtain an accurate estimation of the expression of the cytokines IL-4 and IFN-γ. This can be performed on T cells purified from blood samples of up to 5 ml in volume from children aged 0–4 years with allergic asthma and atopic dermatitis. This procedure includes multiple sampling of PCR products to determine the linear phase of the PCR; inter-experiment correction using a helper T-cell clone, expressing both IL-4 and IFN-γ; interpatient correction by comparing the expression of a housekeeping gene (HPRT); and finally the development of specific software to analyse densitometric data obtained by scanning photographs of agarose gels, separating PCR products. In this way it is possible to study cytokine gene expression from a very small amount of material.http://dx.doi.org/10.1155/S0962935195000329 |
| spellingShingle | H. Koning M. R. M. Baert R. de Groot H. J. Neijens H. F. J. Savelkoul Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCR Mediators of Inflammation |
| title | Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCR |
| title_full | Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCR |
| title_fullStr | Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCR |
| title_full_unstemmed | Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCR |
| title_short | Analysis of cyt0kine gene expression in stimulated T cells of small children by semi-quantitative PCR |
| title_sort | analysis of cyt0kine gene expression in stimulated t cells of small children by semi quantitative pcr |
| url | http://dx.doi.org/10.1155/S0962935195000329 |
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