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Chapter 11.14 : Quantitation of Human Interleukin 4, Interleukin 6, and Gamma Interferon

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Abstract:

Interleukin 4 (IL-4), IL-6, and gamma interferon (IFN-γ) are cytokines with many cellular effects, including mediation of the immune response to infection. Among other biological activities, IL-4 and IL-6 stimulate B cells and IFN-c has antiviral activities ( ). There is an increasing interest in using cytokine levels to monitor disease progression ( ). There are several commercially prepared kits available to quantify cytokines by ELISA. This procedure is written for use with kits manufactured by Pierce Endogen. If IL-4, IL-6, or IFN-c is present in the clinical specimen, it will bind to an ELISA plate that is precoated with antibody specific for that cytokine. After the patient specimen is incubated with a biotinylated detection antibody, the plate is washed and then streptavidin conjugated to horseradish peroxidase (HRP) is added. After another incubation, tetramethylbenzidine (TMB) substrate is added to the plate. The enzyme-substrate reaction produces a color change that can be measured spectrophotometrically. The unknown values can be determined from a standard curve in which the color intensity is directly proportional to the standard concentration.

Citation: Garcia L. 2010. Quantitation of Human Interleukin 4, Interleukin 6, and Gamma Interferon, p 232-237. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.14
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Figures

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Figure 11.14-1

Plate plan for Endogen IL-6 ELISA with 10 samples

Citation: Garcia L. 2010. Quantitation of Human Interleukin 4, Interleukin 6, and Gamma Interferon, p 232-237. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.14
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References

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1. Katial, R. K.,, D. Sachanandani,, C. Pinney,, and M. M. Lieberman. 1998. Cytokine production in cell culture by peripheral blood mononuclear cells from immunocompetent hosts. Clin. Diagn. Lab. Immunol. 5:7881.
2. Lee, B. N.,, J. G. Lu,, M. W. Kline,, M. Paul,, M. Doyle,, C. Kozinetz,, W. T. Shearer,, and J. M. Reuben. 1996. Type 1 and type 2 cytokine profiles in children exposed to or infected with vertically transmitted human immunodeficiency virus. Clin. Diagn. Lab. Immunol. 3:493499.
3.Pierce Endogen. 2006. Package insert for human interleukin 4 ELISA. Pierce Endogen, Rockford, IL.
4.Pierce Endogen. 2007. Package insert for human interleukin 6 ELISA. Pierce Endogen, Rockford, IL.
5.Pierce Endogen. 2006. Package insert for human interferon gamma ELISA. Pierce Endogen, Rockford, IL.
6. Remick, D. G., 2002. Protein analysis and bioassays of cytokines and cytokine receptors, p. 320337. In N. R. Rose,, R. G. Hamilton,, and B. Detrick (ed.), Manual of Clinical Laboratory Immunology, 6th ed. ASM Press, Washington, DC.
7. Wallis, R. S.,, H. M. Lederman,, J. Spritzler,, J. L. Devers,, D. Georges,, A. Weinberg,, S. Stehn,, M. M. Lederman, and the ACTG Inducible Cytokines Focus Group. 1998. Measurement of induced cytokines in AIDS clinical trials using whole blood: a preliminary report. Clin. Diagn. Lab. Immunol. 5:556560.
8. Aziz, N.,, P. Nishanian,, and J. L. Fahey. 1998. Levels of cytokines and immune activation markers in plasma in human immunodeficiency virus infection: quality control procedures. Clin. Diagn. Lab. Immunol. 5:755761.
1.. Pierce Endogen. 2006. Package insert for human interleukin 4 ELISA. Pierce Endogen, Rockford, IL.
2.. Pierce Endogen. 2007. Package insert for human interleukin 6 ELISA. Pierce Endogen, Rockford, IL.
3.. Pierce Endogen. 2006. Package insert for human interferon gamma ELISA. Pierce Endogen, Rockford, IL.

Tables

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Table 11.14-1

Pierce Endogen observed cytokine values in body fluids

The number of subjects evaluated is in parentheses (3–5).

Citation: Garcia L. 2010. Quantitation of Human Interleukin 4, Interleukin 6, and Gamma Interferon, p 232-237. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.14

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