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Clinical and Diagnostic Laboratory Immunology, November 2003, p. 1123-1128, Vol. 10, No. 6
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.6.1123-1128.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Quantification of CCR5 mRNA in Human Lymphocytes and Macrophages by Real-Time Reverse Transcriptase PCR Assay

Jian-Ping Lai, Ji-Hong Yang, Steven D. Douglas,* Xu Wang, Eric Riedel, and Wen-Zhe Ho*

Divisions of Immunologic and Infectious Diseases, Joseph Stokes, Jr., Research Institute, The Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104

Received 21 February 2003/ Returned for modification 22 April 2003/ Accepted 23 July 2003

CCR5, a ß-chemokine receptor, plays an important role in human immunodeficiency virus (HIV) infection of human immune cells, as it is a primary coreceptor for HIV entry into macrophages. We have applied a newly developed real-time reverse transcriptase PCR (RT-PCR) assay for the quantification of CCR5 mRNA in human blood immune cells. The CCR5 real-time RT-PCR assay has a sensitivity of 100 mRNA copies, with a dynamic range of detection between 102 and 106 copies of the CCR5 mRNA transcripts per reaction. The assay is highly reproducible, with an intra-assay coefficient of variation of the threshold cycle of less than 2.07%. When used for quantification of CCR5 mRNA in human monocyte-derived macrophages (MDM) and peripheral blood lymphocytes (PBL), the assay has precision and reproducibility. MDM expressed higher levels of CCR5 mRNA than did PBL. Thus, this assay has the potential and a wide application for the investigation of the role of CCR5 in inflammatory diseases and viral infections, including HIV disease.

* Corresponding author. Mailing address for Wen-Zhe Ho: Division of Immunologic and Infectious Diseases, Children's Hospital of Philadelphia, 34th & Civic Center Blvd., Philadelphia, PA 19104. Phone: (215) 590-4462. Fax: (215) 590-2025. E-mail: Ho{at}email.chop.edu. Mailing address for Steven D. Douglas: Division of Immunologic and Infectious Diseases, Children’s Hospital of Philadelphia, 34th & Civic Center Blvd., Philadelphia, PA 19104. Phone: (215) 590-3561. Fax: (215) 590-3044. E-mail: Douglas{at}email.chop.edu.

Clinical and Diagnostic Laboratory Immunology, November 2003, p. 1123-1128, Vol. 10, No. 6
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.6.1123-1128.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Lai, J.-P., Lai, S., Tuluc, F., Tansky, M. F., Kilpatrick, L. E., Leeman, S. E., Douglas, S. D. (2008). Differences in the length of the carboxyl terminus mediate functional properties of neurokinin-1 receptor. Proc. Natl. Acad. Sci. USA 105: 12605-12610 [Abstract] [Full Text]  
  • Lai, J.-P., Douglas, S. D., Wang, Y.-J., Ho, W.-Z. (2005). Real-Time Reverse Transcription-PCR Quantitation of Substance P Receptor (NK-1R) mRNA. CVI 12: 537-541 [Abstract] [Full Text]  


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