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Updated: Aug 16, 2026
Isolation and Th17 Differentiation of Naïve CD4 T Lymphocytes
Published on: September 26, 2013
A single-step enrichment of Th2 lymphocytes using CCR4 microbeads
John Pappas1, Nancy Quan, Namit Ghildyal
1Department of Immunotherapeutics, Purdue Pharma L.P., Cranbury, NJ 08512, USA. papregister@att.net
Insights
Researchers developed a rapid method to isolate Th2 effector cells from human blood using CCR4 expression. This technique yields significantly more Th2 cells than previous methods, aiding immune response studies.
Area of Science:
- Immunology
- Cell Biology
Background:
- Monitoring human immune responses requires efficient isolation of effector T cells.
- Current methods for isolating Th2 cells from peripheral blood are often inefficient.
Purpose of the Study:
- To develop and evaluate a novel, one-step method for enriching Th2 cell populations from human peripheral blood.
- To compare the efficiency of CCR4-based isolation with the existing CRTh2-based method.
Main Methods:
- Isolation of Th2 cells from peripheral blood mononuclear cells (PBMCs) using magnetic cell sorting.
- Utilized anti-CCR4 or anti-CRTh2 antibodies for cell separation.
- Compared cell purity and yield between the two isolation methods.
Main Results:
- The CCR4-based method achieved a mean purity of >80% for CD4+ T cells.
- The CRTh2-based method yielded significantly lower purity (approximately 7%).
- CCR4 isolation provided 8- to 10-fold greater numbers of Th2 cells compared to CRTh2.
Conclusions:
- A rapid and efficient one-step method using CCR4 expression effectively enriches for Th2 effector cells from human peripheral blood.
- This method offers a substantial improvement in yield and purity over CRTh2-based isolation.
- Facilitates further research into human immune responses and related pathologies.
Abstract:
Monitoring the effector T cells in humans should provide important insights into the regulation of human immune responses and their pathologic consequences. To monitor immune responses it is desirable to be able to isolate greater numbers of effector T cells (Th1/Th2) from peripheral blood. Here, we describe a one step method to enrich for Th2 cell populations from human peripheral blood based on CCR4 expression on Th2 cells and compared it to the CRTh2-based isolation method. Th2 cells were isolated from PBMCs using anti-CCR4 or anti-CRTh2 antibodies and magnetic cell sorting. The mean purity of CD4+ T cells isolated using CCR4 approach from four independent donors was >80% whereas lymphocytes isolated using the CRTh2 approach gave poor yields (approximately 7%). Further, the CCR4 approach gave 8- to 10-fold greater numbers of Th2 cells, as compared to the CRTh2 method, isolated directly from peripheral blood. Upon activation with PMA and ionomycin, the purified CD4+CCR4+ T lymphocytes produced IL-4 and no IFN-gamma. This is a rapid and efficient method to enrich for Th2 effector cells from human peripheral blood.
