phaidra.vetmeduni.ac.at o:1806 DOI 10.3389/fimmu.2022.849922 Transcriptome Profiling of Porcine Naïve, Intermediate and Terminally Differentiated CD8+ T Cells The pig has the potential to become a leading research model for human diseases, pharmacological and transplantation studies. Since there are many similarities between humans and pigs, especially concerning anatomy, physiology and metabolism, there is necessity for a better understanding of the porcine immune system. In adaptive immunity, cytotoxic T lymphocytes (CTLs) are essential for host defense. However, most data on CTLs come from studies in mice, non-human primates and humans, while detailed information about porcine CD8+ CTLs is still sparse. Aim of this study was to analyze transcriptomes of three subsets of porcine CD8β+ T-cell subsets by using next-generation sequencing technology. Specifically, we described transcriptional profiles of subsets defined by their CD11a/CD27 expression pattern, postulated as naïve (CD8β+CD27+CD11alow), intermediate differentiated (CD8β+CD27dimCD11a+), and terminally differentiated cells (CD8β+CD27-CD11ahigh). Cells were analyzed in ex vivo condition as well as upon in vitro stimulation with concanavalin A (ConA) and PMA/ionomycin. Our analyses show that the highest number of differentially expressed genes was identified between naïve and terminally differentiated CD8+ T-cell subsets, underlining their difference in gene expression signature and respective differentiation stages. Moreover, genes related to early (IL7-R, CCR7, SELL, TCF7, LEF1, BACH2, SATB1, ZEB1 and BCL2) and late (KLRG1, TBX21, PRDM1, CX3CR1, ZEB2, ZNF683, BATF, EZH2 and ID2) stages of CD8+ T-cell differentiation were highly expressed in the naïve and terminally differentiated CD8+ T-cell subsets, respectively. Intermediate differentiated CD8+ T-cell subsets shared a more comparable gene expression profile associated with later stages of T-cell differentiation. Genes associated with cytolytic activity (GNLY, PRF1, GZMB, FASL, IFNG and TNF) were highly expressed in terminally and intermediate differentiated CD8+ T-cell subsets, while naïve CD8+ T cells lacked expression even after in vitro stimulation. Overall, PMA/ionomycin stimulation induced much stronger upregulation of genes compared to stimulation with ConA. Taken together, we provided comprehensive results showing transcriptional profiles of three differentiation stages of porcine CD8+ T-cell subsets. In addition, our study provides a powerful toolbox for the identification of candidate markers to characterize porcine immune cell subsets in more detail. eng Cytotoxic-T; Effector; Memory; Expression; Receptor; Subsets; Virus; Lymphocytes; Infection; Responses 2023-07-07T07:49:15.124Z LOMv1.0 Author BEGIN:VCARD VERSION:3.0 N:Lagumdzic;Emil; FN:Emil Lagumdzic X-ORCID:https://orcid.org/0000-0002-0347-7125 END:VCARD BEGIN:VCARD VERSION:3.0 N:Mair;Kerstin H.; FN:Kerstin H. Mair X-ORCID:https://orcid.org/0000-0002-3538-8303 END:VCARD BEGIN:VCARD VERSION:3.0 N:Saalmüller;Armin; FN:Armin Saalmüller X-ORCID:https://orcid.org/0000-0002-7703-3252 END:VCARD BEGIN:VCARD VERSION:3.0 N:Schmitt;Michael W.; FN:Michael W. Schmitt END:VCARD BEGIN:VCARD VERSION:3.0 N:Olgiati;Simone; FN:Simone Olgiati END:VCARD BEGIN:VCARD VERSION:3.0 N:Pernold;Clara; FN:Clara Pernold END:VCARD BEGIN:VCARD VERSION:3.0 N:Viano;Marta; FN:Marta Viano END:VCARD https://w3id.org/kim/hcrt/scheme https://w3id.org/kim/hcrt/text Textdokument Text LOMv1.0 yes https://creativecommons.org/licenses/by/4.0 application/pdf 6679833 https://phaidra.vetmeduni.ac.at/o:1806 https://phaidra.vetmeduni.ac.at/api/object/o:1806/thumbnail LOMv1.0 discipline