Abstract:
:Assessment of cytokine expression has become crucial to understand host responses to infections as well as autoimmunity. Several approaches including Northern blot, RNase protection assay and enzyme-linked immunosorbent assay have been used for this purpose, but they are time consuming, labour intense, and relatively large quantity of the samples is usually required. Recently, a technique termed real-time reverse transcriptase-polymerase chain reaction (RT-PCR) has been developed to determine genetic expression with great sensitivity and specificity; however, specialized instrumentation and costly reagents are usually needed. We aimed at using low-cost reagents for real-time PCR. This was achieved by adapting a conventional RT-PCR protocol to the quantitative real-time format, by the addition of the SYBR Green I reagent. We validated the approach by assessing the cytokine gene expression of murine splenocytes upon stimulation with phorbol 12-myristate 12-acetate (PMA)-ionomycin. The results using this technique were compared with those obtained with the well-established gene array method. We conclude that the use of the SYBR Green I reagent during real-time RT-PCR provides a highly specific and sensitive method to quantify cytokine expression with accuracy and no post-PCR manipulation.
journal_name
Scand J Immunoljournal_title
Scandinavian journal of immunologyauthors
Ramos-Payán R,Aguilar-Medina M,Estrada-Parra S,González-Y-Merchand JA,Favila-Castillo L,Monroy-Ostria A,Estrada-Garcia ICdoi
10.1046/j.1365-3083.2003.01250.xkeywords:
subject
Has Abstractpub_date
2003-05-01 00:00:00pages
439-45issue
5eissn
0300-9475issn
1365-3083pii
1250journal_volume
57pub_type
杂志文章abstract::Modulation of cytotoxic responses by viral immunoevasins plays an important role in the establishment of latent and persistent viral infections. Together with MHC class-I-restricted CD8T-lymphocytes, non-MHC-restricted natural killer (NK) and lymphokine-activated killer (LAK) cells participate in this anti-viral contr...
journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
doi:10.1111/j.1365-3083.1982.tb00719.x
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
doi:10.1046/j.1365-3083.2000.00685.x
更新日期:2000-03-01 00:00:00
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章,收录出版
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journal_title:Scandinavian journal of immunology
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pub_type: 杂志文章
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
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更新日期:2017-09-01 00:00:00
abstract::Chronic inflammatory diseases are now treated with a range of different biopharmaceuticals, often requiring lifelong parenteral administrations. This exposure to drugs is unnatural and can trigger the immune system and result in the formation of antidrug antibodies. Drug-specific antibodies will, if of sufficiently hi...
journal_title:Scandinavian journal of immunology
pub_type: 杂志文章,评审
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abstract::Mycobacterium tuberculosis (MTB) secretory proteins are generally considered important antigens for immune protection against tuberculosis (TB). An 8.3-kDa secretory antigen of MTB and Mycobacterium bovis bacillus Calmette-Guérin (BCG), called SA5K, was recently identified and cloned in our laboratory. In this report,...
journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
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journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
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journal_title:Scandinavian journal of immunology
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abstract::Immune complexes of horseradish peroxidase (HRP) and rabbit IgG antibodies to HRP were used to study the Fcgamma receptors in normal human placenta. Cryostat sections of placental tissue were incubated with the complexes, and the peroxidase activity was revealed histochemically. The bound complexes were localized to t...
journal_title:Scandinavian journal of immunology
pub_type: 杂志文章
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更新日期:1978-01-01 00:00:00