Tissue-Resident Memory T Cells in the Lungs Protect against Acute Respiratory Syncytial Virus Infection

Mitchell A. Luangrath; Megan E. Schmidt; Stacey M. Hartwig; Steven M. Varga

doi:10.4049/immunohorizons.2000067

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FIGURE 1.
RSV-specific CD4 and CD8 T cells localize to the lungs.
BALB/c mice were infected with RSV i.n., and lungs were harvested at days 8, 10, 15, 30, 79, and 149 p.i. (A) Frequency of CD4, CD49d⁻CD11a^lo, and CD49d⁺CD11a^hi CD4 T cells in the lung parenchyma after RSV infection. (B) Frequency and (C) number of virus-specific CD49d⁺CD11a^hi CD4 T cells in the lung parenchyma (IV⁻) and vasculature (IV⁺). (D) Frequency of CD8, CD11a^lo, CD11a^hi, and M2₈₂ tetramer–positive CD8 T cells in the lung parenchyma after RSV infection. (E) Frequency and (F) number of IV⁻ and IV⁺ CD11a^hi CD8 T cells in the lung. (G) Frequency and (H) number of IV⁻ and IV⁺ M2₈₂ tetramer⁺ CD8 T cells in the lung. Data are presented as mean ± SEM from two independent experiments that have been combined (n = 3–4 mice per experiment). *p < 0.05, **p < 0.01, ***p < 0.001, performed with Student t test. IV⁻, lung tissue intravascular stain.
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FIGURE 2.
CD4 and CD8 T cells numbers wane with time after RSV infection.
BALB/c mice were infected with RSV i.n., and lungs were harvested at days 8, 10, 15, 30, 79, and 149 p.i. Cells were analyzed by flow cytometry and gated on RSV-specific (A) TRM CD4 T cells and (B) TRM CD8 T cells in the lung parenchyma as shown. Representative staining panels are from day 30 p.i. (C) Frequency and (D) number of CD69⁺CD103⁻ CD49d⁺CD11a^hi CD4 T cells in the pulmonary parenchyma (IV⁻) and vasculature (IV⁺). (E) Number of IV⁻ CD69⁺CD103⁻ CD49d⁺CD11a^hi TRM and total CD49d⁺CD11a^hi CD4 T cells in the lung. (F) Frequency and (G) number of IV⁻ and IV⁺ CD69⁺CD103⁺ M2₈₂ tetramer⁺ CD8 T cells in the lung. (H) Number of CD69⁺CD103⁺ M2₈₂ tetramer⁺ TRM and total M2₈₂ tetramer⁺ CD8 T cells in the lung. Data are presented as mean ± SEM from two independent experiments that have been combined (n = 3–4 mice per experiment). *p < 0.05, **p < 0.01, ***p < 0.001, performed with Student t test. IV⁻, lung tissue intravascular stain.
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FIGURE 3.
Expression of phenotypic markers by TRM CD4 T cells.
(A) Expression of CD62L, CXCR3, and CD122 by CD69⁺CD103⁻ CD49d⁺CD11a^hi CD4 T cells was compared with circulating (IV⁺) CD49d⁺CD11a^hi CD4 T cells. Representative histograms from day 30 p.i. are shown. (B) Frequency and (C) number of CD62L⁺ cells in the lung. (D) Frequency and (E) number of CXCR3⁺ cells in the lung. (F) Frequency and (G) number of CD122⁺ cells in the lung. Data are represented as mean ± SEM from two independent experiments that have been combined (n = 3–4 mice per experiment). *p < 0.05, **p < 0.01, ***p < 0.001, performed with Student t test.
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FIGURE 4.
Expression of phenotypic markers by CD8 T cells.
(A) Expression of CD62L, CXCR3, and CD122 by CD69⁺CD103⁺ M2₈₂ tetramer⁺ CD8 T cells was compared with circulating (IV⁺) M2₈₂ tetramer⁺ CD8 T cells. Representative histograms from day 30 p.i. are shown. (B) Frequency and (C) number of CD62L⁺ cells in the lung. (D) Frequency and (E) number of CXCR3⁺ cells in the lung. (F) Frequency and (G) number of CD122⁺ cells in the lung. Data are presented as mean ± SEM from two independent experiments that have been combined (n = 3–4 mice per experiment). *p < 0.05, **p < 0.01, ***p < 0.001, performed with Student t test.
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FIGURE 5.
TRM CD4 and CD8 T cells protect against RSV infection.
(A) BALB/c mice were infected with RSV i.n. or left uninfected and then challenged with IAV-F₅₁ or IAV-M2₈₂ 1 mo later. Mice were treated daily with FTY720 i.p. for 7 d beginning 3 d prior to recombinant IAV challenge. Four days after IAV challenge, lungs were harvested, and plaque assays were performed. Viral titers of (B) IAV-PR8, (C) IAV-F₅₁, and (D) IAV-M2₈₂ in the lung are shown. The dotted line denotes the limit of detection (LOD) of the assay. Data are presented as mean ± SEM from two independent experiments that have been combined (n = 5 mice per experiment). *p < 0.05, **p < 0.01, ***p < 0.001, performed with Student t test.