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Updated: Jul 12, 2025
Determination of Immune Cell Identity and Purity Using Epigenetic-Based Quantitative PCR
Published on: February 19, 2020
Rapid qPCR-based quantitative immune cell phenotyping in mouse tissues
Jinghao Huang1, Richard Demmler1, Mariam Mohamed Abdou1
1Division of Molecular and Experimental Surgery, Translational Research Center, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.
Insights
Researchers developed a rapid RT-qPCR method to quantify immune cells in mouse tissues, offering a cost-effective alternative to FACS and IHC. This technique accurately measures T cells, B cells, and macrophages, aiding disease research.
Area of Science:
- Immunology
- Pathogenesis Research
- Molecular Biology
Background:
- The immune microenvironment is crucial for disease regulation.
- Characterizing immune cell infiltrates is vital for understanding pathogenesis and diagnostics.
- Current methods like FACS and IHC are costly, labor-intensive, and antibody-dependent.
Purpose of the Study:
- To develop a rapid, cost-effective RT-qPCR approach for quantifying immune cell populations in mouse tissues.
- To provide a complementary method to existing techniques for immune cell assessment in experimental models.
Main Methods:
- Development of a robust reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay.
- Quantification of specific immune cell populations including T cells (helper T cells, cytotoxic T cells, Th1 cells), B cells, and macrophages.
- Validation of RT-qPCR results against gold-standard techniques: immunohistochemistry (IHC) and fluorescence-activated cell sorting (FACS).
Main Results:
- The developed RT-qPCR approach accurately quantifies major immune cell types in mouse tissues.
- Results showed high concordance with established methods (IHC and FACS).
- The method is rapid and suitable for experimental mouse models where human-specific qPCR assays are unavailable.
Conclusions:
- A validated, rapid RT-qPCR method for immune cell quantification in mouse tissues has been established.
- This approach offers a valuable, cost-effective alternative for immune cell profiling in research and diagnostics.
- The technique facilitates deeper understanding of immune cell roles in disease pathogenesis using mouse models.
Abstract:
The immune microenvironment plays an important role in the regulation of diseases. The characterization of the cellular composition of immune cell infiltrates in diseases and respective models is a major task in pathogenesis research and diagnostics. For the assessment of immune cell populations in tissues, fluorescence-activated cell sorting (FACS) or immunohistochemistry (IHC) are the two most common techniques presently applied, but they are cost intensive, laborious, and sometimes limited by the availability of suitable antibodies. Complementary rapid qPCR-based approaches exist for the human situation but are lacking for experimental mouse models. Accordingly, we developed a robust, rapid RT-qPCR-based approach to determine and quantify the abundance of prominent immune cell populations such as T cells, helper T (Th) cells, cytotoxic T cells, Th1 cells, B cells, and macrophages in mouse tissues. The results were independently validated by the gold standards IHC and FACS in corresponding tissues and showed high concordance.
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