Detection of CLCF1 protein expression by flow cytometry.
Véronique Laplante1, Marine Rousseau2, Félix Lombard-Vadnais3
1Département de pharmacologie et physiologie, Université de Montréal, Montréal, QC, H3T 1J4, Canada.
Scientific Reports
|June 10, 2024
Summary
Researchers developed a flow cytometry method to detect Cardiotrophin-like cytokine factor 1 (CLCF1) protein in immune cells. This tool aids in studying CLCF1
Area of Science:
- Immunology
- Cytokine Biology
- Flow Cytometry
Background:
- Cardiotrophin-like cytokine factor 1 (CLCF1), an IL-6 family member, exhibits neurotrophic and immune-modulating roles.
- CLCF1 mRNA is found in lymphoid organs and linked to T helper 17 cell differentiation.
- Protein-level expression data for CLCF1 in immune cells is limited.
Purpose of the Study:
- To develop a reliable method for detecting CLCF1 protein expression in immune cells.
- To characterize CLCF1 expression in mouse immune cells using flow cytometry.
- To investigate the functional impact of a CLCF1-specific monoclonal antibody.
Main Methods:
- Development of a monoclonal antibody (mAb) against CLCF1.
- Detection of human and mouse CLCF1 protein via flow cytometry.
- Utilizing CLCF1-expressing Ba/F3 cells and conditional knock-out mouse models for validation.
Main Results:
- Successfully detected CLCF1 protein in transduced Ba/F3 cells and mouse splenic T cells, including in vitro differentiated Th1 cells.
- The anti-CLCF1 mAb demonstrated inhibitory activity against CLCF1 in vitro, binding to a critical epitope.
- Specificity was confirmed using lymphocytes from Clcf1-deficient mice.
Conclusions:
- Flow cytometry with a specific mAb is a viable method for detecting CLCF1 protein expression.
- This technique enables the study of CLCF1 expression in various immune cell types.
- The developed method will advance research into CLCF1's role in immune responses.
