Opposing effects of T cell receptor signal strength on CD4 T cells responding to acute versus chronic viral infection
- Immune Cell Biology Laboratory, Department of Biomedicine, University of Basel, University Hospital Basel, Switzerland
- Division of Experimental Virology, Department of Biomedicine – Haus Petersplatz, University of Basel, Switzerland
Figures
Figure 1 with 3 supplements
Generation and viral fitness of GP61 lymphocytic choriomeningitis virus (LCMV) variants.
(A) Scheme of GP61 wt and altered peptide ligand (APL) sequences with mutations highlighted in red ordered hierarchically according to T cell receptor (TCR) signal strength. (B) Peptide …
-
Figure 1—source data 1
Generation and viral fitness of GP61 lymphocytic choriomeningitis virus (LCMV) variants.
- https://cdn.elifesciences.org/articles/61869/elife-61869-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
GP61 altered peptide ligand (APL) screening.
Peptide dose–activation curves of overnight cultured SMARTA cells with peptide pulsed splenocytes using the percentage of CD69+ SMARTA cells as a readout for activation. Data represent one of n = 2 …
Figure 1—figure supplement 2
Interferon (IFN) type-I signaling exerts minor effects on the expression of activation markers in vitro.
Naïve SMARTA T cells were cultured in the presence of splenocytes and various concentrations of GP61 wt and altered peptide ligands (APLs). Proportion of CD69+ SMARTA cells 6 hr after stimulation …
Figure 1—figure supplement 3
Viral load in organs and blood of DBA/2 mice.
Viral load (focus forming units [FFU]) in indicated organs per gram tissue 21 days post infection and in blood over time in DBA/2 mice. Data points represent the mean ± SEM of 4–10 samples pooled …
Figure 2 with 7 supplements
T cell receptor (TCR) signal strength positively correlates with Th1 cell differentiation during lymphocytic choriomeningitis virus (LCMV) Armstrong variant infection.
(A) Number of SMARTA (left), NP309+ (middle), and CD44+ cells (right) 10 days (top) or >30 days (bottom) post infection (p.i.). (B) Histograms (top) and relative mean fluorescence intensity (MFI) …
-
Figure 2—source data 1
T cell receptor (TCR) signal strength positively correlates with T follicular helper (Tfh) cell differentiation during lymphocytic choriomeningitis virus (LCMV) Armstrong variant infection.
- https://cdn.elifesciences.org/articles/61869/elife-61869-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
SMARTA cell numbers 4 days post infection (p.i.).
Number of SMARTA (left) and CD44+ cells (right) 4 days p.i. Data are pooled from n = 2 independent experiments with seven to eight samples per group. Bars represent the mean and symbols represent …
Figure 2—figure supplement 2
T cell receptor (TCR) signal strength positively correlates with early Th1 cell differentiation during lymphocytic choriomeningitis virus (LCMV) Armstrong variant infection.
Histograms (left) and proportion of CD25+ SMARTA T cells (right) 4 days post infection (p.i.). Data are pooled from n = 2 independent experiments with seven to eight samples per group. Bars …
Figure 2—figure supplement 3
Germinal center B cell differentiation 4 days post infection (p.i.) is unaffected by lymphocytic choriomeningitis virus (LCMV) Armstrong variant infection.
Identification (left), proportion (middle), and quantification (right) of GL7+FAS+ B cells 4 days p.i. Data are pooled from n = 2 independent experiments with seven to eight samples per group. Bars …
Figure 2—figure supplement 4
The endogenous nucleoprotein (NP)-specific CD4 response is unaffected by lymphocytic choriomeningitis virus (LCMV) Armstrong variant infection.
Proportion of T follicular helper (Tfh) (left), Th1 cells (middle), and the Th1:Tfh ratio (right) of the NP309+ compartment 10 days post infection (p.i.). Data are pooled from n = 2 independent …
Figure 2—figure supplement 5
Alternative gating strategy to identify SMARTA T follicular helper (Tfh) cells.
Identification and proportion of PD1hi FR4hi Tfh cells in the SMARTA compartment by flow cytometry. Data are pooled from n = 2 independent experiments with eight to nine samples per group. Bars …
Figure 2—figure supplement 6
T cell receptor (TCR) signal strength does not impact Bcl6 and Tbet expression in Ly6Clo Th1 SMARTA cells.
Relative mean fluorescence intensity (MFI) of Bcl6 and Tbet expression in the Ly6Clo Th1 SMARTA compartment. Data are pooled from n = 2 independent experiments with eight to nine samples per group. …
Figure 2—figure supplement 7
T cell receptor (TCR) signal strength does not impact the generation of Ly6Clo Th1 SMARTA cells.
Identification and proportion of Ly6Clo Th1 (Ly6CloPSGL1hi) in the SMARTA compartment by flow cytometry. Data are pooled from n = 2 independent experiments with eight to nine samples per group. Bars …
Figure 3 with 10 supplements
T cell receptor (TCR) signal strength positively correlates with T follicular helper (Tfh) cell differentiation during lymphocytic choriomeningitis virus (LCMV) Clone-13 variant infection. Spleens were harvested 7 days after infection with LCMV Clone-13 variants.
(A) Number of SMARTA (left), NP309+ (middle), and CD44+ cells (right). (B) Histograms (top) and relative mean fluorescence intensity (MFI) (bottom) of indicated phenotypic markers in the SMARTA …
-
Figure 3—source data 1
IT cell receptor (TCR) signal strength positively correlates with T follicular helper (Tfh) cell differentiation during lymphocytic choriomeningitis virus (LCMV) Clone-13 variant infection..
- https://cdn.elifesciences.org/articles/61869/elife-61869-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Lymphocytic choriomeningitis virus (LCMV) Armstrong and Clone-13 infection induce similar expansion of CD4 compartments.
Spleens were harvested 7 days after infection with LCMV Clone-13 variants. (A) Number of SMARTA (left), NP309+ (middle), and CD44+ cells (right). Data are pooled from n = 2 independent experiments …
Figure 3—figure supplement 2
Lymphocytic choriomeningitis virus (LCMV) Clone-13 infection results in a T follicular helper (Tfh)-based differentiation of SMARTA cells.
Spleens were harvested 7 days after infection with LCMV Clone-13 variants. (Left panel) Identification of Th1 (Ly6ChiPSGL1hi) and Tfh (Ly6CloPSGL1lo) subset in the SMARTA compartment by flow …
Figure 3—figure supplement 3
T cell receptor (TCR) signal strength negatively correlates with Ly6Clo Th1 differentiation during lymphocytic choriomeningitis virus (LCMV) Clone-13 variant infection.
Spleens were harvested 7 days after infection with LCMV Clone-13 variants. Identification and proportion of Ly6Clo Th1 (Ly6CloPSGL1hi) in the SMARTA compartment by flow cytometry. Data are pooled …
Figure 3—figure supplement 4
High viral load in kidneys 7 days post infection in all three viruses.
Viral load in kidneys (FFU/g) 7 days post infection. Data represent one of n = 2 independent experiments with four samples per group. Bars represent the mean and symbols represent individual mice.
Figure 3—figure supplement 5
The endogenous nucleoprotein (NP)-specific CD4 response is unaffected by lymphocytic choriomeningitis virus (LCMV) Clone-13 variant infection.
Spleens were harvested 7 days after infection with LCMV Clone-13 variants. Proportion of T follicular helper (Tfh) (left), Th1 cells (middle), and the Th1:Tfh ratio (right) of the NP309+ …
Figure 3—figure supplement 6
SMARTA cell numbers 4 and 14 days post infection (p.i.).
Number of SMARTA cells at day 4 (left) and 14 (right) p.i. Data are pooled from n = 2 independent experiments with six to nine samples per group. Bars represent the mean and symbols represent …
Figure 3—figure supplement 7
T cell receptor (TCR) signal strength negatively correlates with early Th1 cell differentiation during lymphocytic choriomeningitis virus (LCMV) Clone-13 variant infection.
Histograms and proportion of CD25+ SMARTA T cells (far left and left) or Ly6C+ cells (right and far right) 4 days post infection. Data are pooled from n = 2 independent experiments with seven to …
Figure 3—figure supplement 8
Th1 bias is maintained in Clone-13 variants 14 days post infection (p.i.).
Identification and proportion of Th1 (Ly6ChiPSGL1hi) and T follicular helper (Tfh) (Ly6CloPSGL1lo) subset and Th1:Tfh ratio in the SMARTA compartment 14 days p.i. Data are pooled from n = 2 …
Figure 3—figure supplement 9
Germinal center B cell differentiation is unaffected by lymphocytic choriomeningitis virus (LCMV) Clone-13 variant infection.
Identification (left), proportion (middle), and quantification (right) of GL7+FAS+ B cells 4 days post infection (p.i.) (top row) or 14 days p.i. (bottom row). Cells are pregated for B220+dump-. …
Figure 3—figure supplement 10
Antigen load exerts different effects on strongly versus weakly activated SMARTA T cells.
(Left) Experimental setup. Number of SMARTA cells (middle) and proportion of Th1 (Ly6ChiPSGL1hi) and T follicular helper (Tfh) (Ly6CloPSGL1lo) subset and Th1:Tfh ratio in the SMARTA compartment …
Figure 4 with 5 supplements
Increased T cell receptor (TCR) signal strength induces expression of markers associated with chronic T cell stimulation. Spleens were harvested 14 days after infection with lymphocytic choriomeningitis virus (LCMV) Clone-13-based variants.
(A) Histograms (left) and relative mean fluorescence intensity (MFI) (right) of PD1 in the total SMARTA compartment (left) or SMARTA Th1 and T follicular helper (Tfh) subsets (right). (B) Histograms …
-
Figure 4—source data 1
Increased T cell receptor (TCR) signal strength induces expression of markers associated with chronic T cell stimulation.
- https://cdn.elifesciences.org/articles/61869/elife-61869-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
Viral load in kidneys 14 days post infection.
Viral load in kidneys (FFU/kidney) 14 days post infection. Data represents one of n = 2 independent experiments with four to five samples per group. Bars represent the mean and symbols represent …
Figure 4—figure supplement 2
Similar activation marker expression of CD4+ CD44+ T cells across all viruses.
Spleens were harvested 14 days after infection with lymphocytic choriomeningitis virus (LCMV) Clone-13-based variants. Relative mean fluorescence intensity (MFI) (right) of PD1 and Lag3 in the CD44+ …
Figure 4—figure supplement 3
T cell receptor (TCR) signal strength impacts TOX expression in SMARTA Th1 and T follicular helper (Tfh) compartments.
Spleens were harvested 14 days after infection with lymphocytic choriomeningitis virus (LCMV) Clone-13-based variants. Relative mean fluorescence intensity (MFI) of TOX in SMARTA Th1 and Tfh. Data …
Figure 4—figure supplement 4
TOX expression is not affected in CD44+ CD4+ T cells by lymphocytic choriomeningitis virus (LCMV) Clone-13 variant infection.
Spleens were harvested 14 days after infection with LCMV Clone-13-based variants. Relative mean fluorescence intensity (MFI) (right) of TOX in the CD44+ compartment. Data are pooled from n = 2 …
Figure 4—figure supplement 5
T cell receptor (TCR) signal strength does not impact TCF1 expression.
Spleens were harvested 14 days after infection with lymphocytic choriomeningitis virus (LCMV) Clone-13-based variants. Relative mean fluorescence intensity (MFI) of TCF1 in the SMARTA Th1 and T …
Author response image 1
Germinal center B cell differentiation 7 days post LCMV Clone-13 variant infection.
Proportion and numbers of GL7+FAS+ B cells 7 days p.i. Data are pooled from n = 2 independent experiments with 8 samples per group. Bars represent the mean and symbols represent individual mice. …
Additional files
-
Supplementary file 1
Altered peptide ligands (APLs) with altered potential to activate SMARTA and corresponding EC50 values.
- https://cdn.elifesciences.org/articles/61869/elife-61869-supp1-v1.docx
-
Transparent reporting form
- https://cdn.elifesciences.org/articles/61869/elife-61869-transrepform-v1.docx
