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CD4(+) T-cell inhibitory ligands: a tool for characterizing dysfunctional CD4(+) T cells during chronic infection - PubMed

CD4(+) T-cell inhibitory ligands: a tool for characterizing dysfunctional CD4(+) T cells during chronic infection

Courtney Dow et al. Immunology. 2013 Sep.

Erratum in

  • Immunology. 2013 Nov;140(3):390

Abstract

Activation of CD4(+) T cells helps to establish and maintain immune responses. During infection with lymphocytic choriomeningitis virus (LCMV) clone 13, the CD4(+) T-cell responses are lost. In this study, we were interested in the nature of the CD4(+) T-cell responses following infection with LCMV clone 13. To pursue this question, we infected C57BL/6 mice with LCMV clone 13. We used a GP66-80 MHC Class II tetramer to determine whether the CD4(+) T cells were present following infection with LCMV clone 13. We determined that the cells were present and antigen specific, but not functional. We attributed their dysfunction to the presence of CD4(+) T-cell inhibitory ligands. We further stained for the presence of CD4(+) T-cell inhibitory ligands. We found that the during chronic infection the number of CD4(+) T cells expressing programmed death-1 and CD160 were greater over the time-course study than the other CD4(+) T-cell inhibitory ligands. These data show that using CD4(+) T-cell inhibitory ligands as a reagent for characterization can help in understanding the complex immune responses associated with persistent infections.

Keywords: CD160; CD4+; lymphocytic choriomeningitis virus; programmed death-1; tetramer.

© 2013 The Authors. Immunology published by © 2013 John Wiley & Sons Ltd.

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Figures

Figure 1
Figure 1

Quantification of antigen-specific CD4+ T cells over a 30-day infection period. C57BL/6 mice were infected intraperitoneally with 2 × 105 plaque-forming units (PFU) of lymphocytic choriomeningitis virus (LCMV) Armstrong, retro-orbitally with 2 × 106 PFU of LCMV clone 13, or left uninfected. Eight days post-infection, CD4+ T cells from splenocyte samples were stained and tested against phycoerythrin-conjugated MHC II tetramers. Samples were then sorted using flow cytometry and data were analysed using F

low

J

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9.1. *P < 0·05.

Figure 2
Figure 2

Quantification of antigen-specific CD4+ GP66+ T cells was assessed over a 30-day time–course of infection. Splenocytes were collected and sorted using flow cytometry and data were analysed using F

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9.1.

Figure 3
Figure 3

Quantification of CD4+ T-cell expression of programmed death 1 (PD-1) and CD160 over a 30-day infection period. C57BL/6 mice were infected intraperitoneally with 2 × 105 plaque-forming units (PFU) of lymphocytic choriomeningitis virus (LCMV) Armstrong, retro-orbitally with 2 × 106 PFU of LCMV clone 13, or left uninfected. Eight, 15 and 30 days post-infection, CD4+ T cells from splenocyte samples were stained with (a) αPD-1 (b) αCD160 murine antibodies. Samples were then sorted using flow cytometry and data were analysed using F

low

J

o

9.1. *P < 0·05.

Figure 4
Figure 4

Quantification of CD4+ T-cell expression of cytotoxic T-lymphocyte antigen 4 (CTLA-4) and lymphocyte activation gene 3 (LAG-3) expression over a 30-day infection period. C57BL/6 mice were infected intraperitoneally with 2 × 105 plaque-forming units (PFU) of lymphocytic choriomeningitis virus (LCMV) Armstrong, retro-orbitally with 2 × 106 PFU of LCMV clone 13, or left uninfected. Eight, 15 and 30 days post-infection, CD4+ T cells from splenocyte samples were stained with (a) αCTLA-4 (b) αLAG-3 murine antibodies. Samples were then sorted using flow cytometry and data were analysed using

FlowJo

9.1.

Figure 5
Figure 5

Characterization of antigen-specific CD4+ T-cell expression of programmed death 1 (PD-1) over a 30-day infection period. C57BL/6 mice were infected intraperitoneally with 2 × 105 plaque-forming units (PFU) of lymphocytic choriomeningitis virus (LCMV) Armstrong, retro-orbitally with 2 × 106 PFU of LCMV clone 13, or left uninfected. Eight, 15 and 30 days post-infection, CD4+ T cells from splenocyte samples were stained with αPD-1 murine antibody. Samples were then sorted using flow cytometry and data were analysed using

FlowJo

9.1. *P < 0·05.

Figure 6
Figure 6

Characterization of antigen-specific CD4+ T-cell expression of (a) CD160, (b) cytotoxic T-lymphocyte antigen 4 (CTLA-4) and (c) lymphocyte activation gene 3 (LAG-3) over a 30-day infection period. C57BL/6 mice were infected intraperitoneally with 2 × 105 plaque-forming units (PFU) of lymphocytic choriomeningitis virus (LCMV) Armstrong, retro-orbitally with 2 × 106 PFU of LCMV clone 13 or left uninfected. Eight, 15 and 30 days post-infection, CD4+ T cells from splenocyte samples were stained with αCD160, αCTLA-4 and αLAG-3 murine antibodies. Samples were then sorted using flow cytometry and data were analysed using

FlowJo

9.1.

Figure 7
Figure 7

The concurrent expression of multiple inhibitory ligands [programmed death 1 (PD-1), cytotoxic T-lymphocyte antigen 4 (CTLA-4), lymphocyte activation gene 3 (LAG-3), CD160) on GP66+ CD4+ splenocytes from the three experimental groups at days 8, 15 and 30 post-infection. Individual populations were grouped based on total number of inhibitory ligands expressed using Boolean gating to analyse groups and SPICE for coexpression comparisons. The percentages of GP66+ CD4+ T-cells expressing combinations of inhibitory ligands are shown.

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