vang-like 2 (van gogh, Drosophila). -0.25. 568 Fam158a family with sequence similarity 158, member A. -0.59. 569 Lrrc8a leucine rich repeat containing 8A.
Immune signatures of protective spleen memory CD8 T cells
Lilia Brinza, Sophia Djebali, Martine Tomkowiak, Julien Mafille, Céline Loiseau, PierreEmmanuel Jouve, Simon de Bernard, Laurent Buffat, Bruno Lina, Michèle Ottmann, Manuel Rosa-Calatrava, Stéphane Schicklin, Nathalie Bonnefoy, Grégoire Lauvau, Morgan Grau, Mélanie Wencker, Christophe Arpin, Thierry Walzer, Yann Leverrier and Jacqueline Marvel
!
S100a6
Gzm k 15
re la tive m R N A le ve l
HP3
relative m R N A level
0.8 0.6 0.4 0.2 0.0
10
5
0
N a ive
T IM
F lu -T M V V -T M
N a ive
T IM
CCR9
Asns 0.15
relative m R N A level
relative m R N A level
0.06
HP2
0.04
0.02
0.10
0.05
0.00
0.00
N a ive
T IM
N a ive
F lu -T M V V -T M
T IM
CCR2 1.5
re la tive m R N A le ve l
re la tive m R N A le ve l
F lu -T M V V -T M
F g l2
3
HP1
F lu -T M V V -T M
2
1
0
1.0
0.5
0.0
N a ive
T IM
F lu -T M V V -T M
N a ive
T IM
F lu -T M V V -T M
Supplementary Fig. S1. Validation of transcriptomic signature by quantitative RT-PCR assay. mRNA of Naive, TIM or Flu-TM CD8 T cells were analyzed ex vivo (homeostatic condition - dashed bar). The expression of each selected gene was measured in triplicate with TLDA technology. Data show one out of 2 independent experiments.
Supplementary Fig. S2. Expression heatmap of the top 55 HP1 genes with a minimal FC of 4 when comparing Listeria MonocytogenesNP68 -induced memory cells (List WT) to naive cells. C57BL/6J female mice (8-12 weeks old) that had received 2 × 105 naive F5xLy5.1 CD8 T cells one day before were infected i.v. with 3x103 Listeria Monocytogenes strain 10403S expressing NP68 epitope. Specific memory CD8 T cells were purified at least 6 weeks after immunization and micro arrays data were generated as described in the methods
A BAL
BAL
maximum
maximum
ndLN
LUNG
BAL maximum
ndLN
LUNG
NAIVE minimum
dLN
ndLN
LUNG
Naive
minimum
minimum TIM
TIM
Flu
Flu-TM
Flu-TM
SPLEEN
dLN
Day 3
SPLEEN
Day 4
dLN
SPLEEN
Day 6
Number of virus-specific CD8+ T cells (day 6 spleen)
B *
1.0¥10 7
**
8.0¥10 6 6.0¥10 6 4.0¥10 6 2.0¥10 6 0
NAIVE
TIM
Naive
TIM
FLU-TM
Supplementary Fig. S3. Flu-TM localize more rapidly to the draining lymph node and the infected lung. A total of 5x104 naive, TIM or Flu-TM (CD45.1+) CD8 T cells were transferred i.v. into C57BL/6J mice that were infected i.n. 24 h later with 2x105 TCID50 Flu-NP68. (A) Mice were sacrificed at day 3, 4, 6 and the number of F5 CD8 T cells present in the BAL, lung, spleen, draining (mediastinal) and non-draining (inguinal) lymph nodes was determined. Data show one out of 6 independent experiments with similar results. (B) Mice were sacrificed at day 6 and the number of virus specific F5 CD8 T cells present in the spleen was determined. Data show mean of 4 independent experiments. *p
