AUTHOR=Traenkle Bjoern , Kaiser Philipp D. , Pezzana Stefania , Richardson Jennifer , Gramlich Marius , Wagner Teresa R. , Seyfried Dominik , Weldle Melissa , Holz Stefanie , Parfyonova Yana , Nueske Stefan , Scholz Armin M. , Zeck Anne , Jakobi Meike , Schneiderhan-Marra Nicole , Schaller Martin , Maurer Andreas , Gouttefangeas Cécile , Kneilling Manfred , Pichler Bernd J. , Sonanini Dominik , Rothbauer Ulrich TITLE=Single-Domain Antibodies for Targeting, Detection, and In Vivo Imaging of Human CD4+ Cells JOURNAL=Frontiers in Immunology VOLUME=12 YEAR=2021 URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2021.799910 DOI=10.3389/fimmu.2021.799910 ISSN=1664-3224 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.