Kristiyan Kanev1, Patrick Roelli1,3, Ming Wu1, Christine Wurmser2, Mauro Delorenzi3, Michael Pfaffl1, Dietmar Zehn1
1) Division of Animal Physiology and Immunology, School of Life Sciences Weihenstephan, Technical University of Munich, 85354 Freising, Germany;
2) Division of Animal Breeding, School of Life Sciences Weihenstephan, Technical University of Munich, 85354 Freising, Germany;
3) BCF, Swiss Institute of Bioinformatics, University of Lausanne 1015 Lausanne, Switzerland
Cytotoxic T-cells (CTLs) are major players in the protective immunity against viral infections and cancer. Given this central role, CTLs are of key interest for the development of novel and improvement of existing immunotherapies. This requires deeper understanding of the factors and molecular programs guiding their differentiation and functionality. This can be obtained through Single-cell RNA sequencing, which allows for unbiased discovery of single-cell resolved gene expression networks, which can be further used to define new cell subpopulations in an unbiased way. To allow for comprehensive single-cell resolved analysis of CTLs, as well as to deal with their minute amounts of messenger RNA, we have systematically optimized a droplet-based method providing breadth of cellular profiling and a plate based method providing depth of cellular profiling. We have achieved significant improvement in their sensitivity, providing tools for the establishment of an atlas of single-cell defined CTL cell states which will contribute to development of better conceptual understanding and new therapeutic opportunities.
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