Protocols: Naïve HA-specific CD8+ T cells were purified from the spleen and lymph nodes of CL4-TCR F1 mice. Purified HA-specific CD8+ T cells were then cultivated with irradiated syngeneic splenocytes in the presence of HA512-520-peptide, IL-2, and IL-12. After five days of culture, living cells were collected. 3×106 activated CD8+ T cells were adoptively transferred via i.v. in female adult (5 - 7 week old) Orex-HA mice. At day 30 post-transfer, the mice were sacrificed and the HA-specific CD8+ T cells (CD11b- NK1.1- CD45.2+ CD8+ TCRb+ CD45.1+) were isolated by FACS sorting (BD FACSAria Fusion) from the spleen and hypothalami of the 13 Orex-HA mice. Immediately after cell sorting, cells were subjected to scRNA-seq processing (Chromium Single Cell 3ʹ Reagent Kits v3, CG000183 Rev C) according to the manufacturer's instructions. Libraries were constructed using the 10X Genomics, Chromium Single Cell 3' v3 kit including : - Chromium Single Cell 3ʹ GEM, Library & Gel Bead Kit v3, 16 rxns PN-1000075; Chromium Single Cell B Chip Kit, 48 rxns PN-1000073 and Chromium i7 Multiplex Kit, 96 rxns PN-120262; according to manufacturer's instructions.
ENA first public
2022-04-14
ENA last update
2022-04-14
External Id
SAMEA12921926
INSDC center alias
Team EPIIC 'Eukaryotic Pathogens: Inflammation, T cell Immunity and Chemoresistance' Toulouse Institute for Infectious and Inflammatory Diseases (Infinity)-INSERM UMR1291 - CNRS UMR5051 - University Toulouse III
INSDC center name
Team EPIIC 'Eukaryotic Pathogens: Inflammation, T cell Immunity and Chemoresistance' Toulouse Institute for Infectious and Inflammatory Diseases (Infinity)-INSERM UMR1291 - CNRS UMR5051 - University Toulouse III
INSDC first public
2022-04-14T00:24:37Z
INSDC last update
2022-04-14T00:24:37Z
INSDC status
public
Submitter Id
E-MTAB-11416:SPLEEN
broker name
ArrayExpress
cell type
HA-specific CD8-positive T cell
common name
house mouse
developmental stage
adult
disease
normal
genotype
wild type genotype
growth condition
adoptively transferred in Orex-HA adult female mice
immunophenotype
CCD11b-; NK1.1-; CD45.2+; CD8+; TCRb+; CD45.1+
individual
pool of 13 mice
inferred cell type
activated CD8-positive, alpha-beta T cell
organism part
spleen; lymph node
sample name
E-MTAB-11416:SPLEEN
sampling site
spleen
sex
female
strain
CL4-TCR
time
30