Single-Domain Antibodies for Targeting, Detection, and In Vivo Imaging of Human CD4+ Cells
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Published on: December 27, 2017
Bjoern Traenkle1, Philipp D Kaiser1, Stefania Pezzana2
1NMI Natural and Medical Sciences Institute at the University of Tübingen, Reutlingen, Germany.
Insights
Researchers developed novel nanobodies (Nbs) targeting CD4+ cells, crucial for immune responses. These nanobodies enable noninvasive imaging of immune cells, aiding immunotherapy monitoring in cancer and inflammatory diseases.
Area of Science:
- Immunology
- Molecular Imaging
- Biotechnology
Background:
- Immunotherapies require tools to track immune cells like CD4+ T cells.
- CD4+ cells play a vital role in regulating immune responses.
Purpose of the Study:
- To develop and validate novel nanobodies (Nbs) that specifically target human CD4+ cells.
- To create immune tracers for noninvasive molecular imaging of CD4+ cells.
- To assess the utility of CD4-Nbs for monitoring immune responses in disease.
Main Methods:
- Generation and characterization of single-domain antibodies (nanobodies) against human CD4.
- In vitro analysis of nanobody binding properties, epitope recognition, and effects on T-cell function.
- In vivo validation using optical imaging in a xenograft model and immunoPET/MR in a human CD4 knock-in mouse model.
Main Results:
- Novel CD4-specific nanobodies were generated without interfering with T-cell functions.
- High-affinity CD4-Nbs successfully visualized CD4+ cells in vivo via optical imaging.
- Radiolabeled CD4-Nb1 showed rapid accumulation in CD4+ T cell-rich tissues using immunoPET/MR.
Conclusions:
- CD4-Nbs are effective tools for noninvasive molecular imaging of CD4+ cells.
- These nanobodies can serve as versatile probes for patient stratification and immune response monitoring in immunotherapy.
- The developed CD4-Nbs hold potential for personalized immunotherapy in cancer and inflammatory diseases.
Abstract:
The advancement of new immunotherapies necessitates appropriate probes to monitor the presence and distribution of distinct immune cell populations. Considering the key role of CD4+ cells in regulating immunological processes, we generated novel single-domain antibodies [nanobodies (Nbs)] that specifically recognize human CD4. After in-depth analysis of their binding properties, recognized epitopes, and effects on T-cell proliferation, activation, and cytokine release, we selected CD4-specific Nbs that did not interfere with crucial T-cell processes in vitro and converted them into immune tracers for noninvasive molecular imaging. By optical imaging, we demonstrated the ability of a high-affinity CD4-Nb to specifically visualize CD4+ cells in vivo using a xenograft model. Furthermore, quantitative high-resolution immune positron emission tomography (immunoPET)/MR of a human CD4 knock-in mouse model showed rapid accumulation of 64Cu-radiolabeled CD4-Nb1 in CD4+ T cell-rich tissues. We propose that the CD4-Nbs presented here could serve as versatile probes for stratifying patients and monitoring individual immune responses during personalized immunotherapy in both cancer and inflammatory diseases.
