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Transcriptome analysis of reactivated T H 1 cells reveal distinct differences between priming and reactivation processes
Author(s) -
Kiljan Martha,
Velázquez Camacho Oscar,
Ercanoglu Meryem,
Hesselmann Isabelle,
Ibruli Olta,
Sahbaz Yagmur,
Wagner Elena,
Cai Jiali,
Niu Lina,
Lackmann JanWilm,
Hillmer Axel,
Sorokin Lydia,
Engelbertsen Daniel,
Klein Florian,
Baues Christian,
MarnitzSchulze Simone,
HerterSprie Grit,
Herter Jan
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.02700
Subject(s) - nerve growth factor ib , priming (agriculture) , biology , t cell , effector , il 2 receptor , microbiology and biotechnology , cd28 , flow cytometry , t cell receptor , interleukin 21 , immune system , immunology , transcription factor , nuclear receptor , biochemistry , botany , germination , gene
As a key player of the adaptive immune system CD4+ T cells play a critical role in orchestrating an effective immune response during infections, autoimmunity or cancer. Few studies have addressed antigen‐mediated reactivation of T effector cells in peripheral tissues, as opposed to the well‐studied primary activation (priming) in lymph nodes. We hypothesize that T effector cell reactivation is distinct from naïve T cell priming beyond linage differentiation, focusing on T H 1 effector cells. We employed OT‐II/Nur77 GFP mice that allowed direct identification of T cell receptor (TCR) activated cells as Nur77 expression is up‐regulated by TCR stimulation and correlates with the strength of TCR stimulation. Isolated naïve CD4 T cells (naïve resting), were either stimulated (naïve stimulated) or differentiated into T H 1 (effector resting) and consecutively reactivated in an in vitro restimulation assay (effector stimulated). We correlated Nur77‐GFP expression to the expression of T H 1 lead cytokine IFNγ via qRT‐PCR and flow cytometry and found that the maximum of restimulation is reached after 6‐8 hours. While Nur77‐GFP expression declines slowly after 4 hours in effector cells, IFNγ expression increases for up to 8 hours. In naïve T cells we correlated Nur77‐GFP expression to IL2 expression and found that after 4 hours of stimulation IL2 expression reaches a saturation and nearly 90% of cells are Nur77‐GFP positive. Transcriptome analysis of sorted naïve (CD62L + , CD44 ‐ , CD69 ‐ , Nur77‐GFP ‐ ), active naïve (CD62L int , CD44 ‐ , CD69 + , Nur77‐GFP + ), T H 1 (CD62L + , CD44 + , Nur77‐GFP ‐ ) and reactivated T H 1 cells (CD62L + , CD44 + , Nur77‐GFP + ) revealed 11899 significantly differentially expressed genes of overall T cell differentiation and activation (naïve vs reactivated effector), 10840 of naïve T cell activation (naïve vs active naïve), 11583 of T H 1 differentiation (naïve vs effector) and 10736 of T H 1 reactivation (effector vs reactivated effector). Furthermore 3107 genes were predominantly specifically regulated in reactivated T H 1 cells. Of these, 447 are solely regulated in reactivated T H 1 cells and do not appear in the other comparisons. We here report a unique signature to T effector cell reactivation that goes beyond lineage alterations.