Activated K^dM2₈₂ T cells up-regulate expression of inhibitory receptors.

<p><b>(a)</b> Transcriptional expression of genes encoding inhibitory receptors with FC > ±1.3, p < 0.05 and FDR < 0.25 were listed. Data were pooled from 10 or 11 individual mice in each group. (<b>b</b>) Post-transcriptional expression of inhibitory receptors at 7 dpi was assessed by flow cytometry. The frequencies are shown as mean with independent data point and compared by Student’s <i>t</i>-test. Data represent 5 independent experiments (n = 5/group/experiment). Each symbol represents one mouse.</p

Activated K^dM2₈₂ T cells are apoptotic.

<p><b>(a)</b> Transcriptional expression of genes encoding pro- and anti-apoptosis molecules with FC > ±1.3, p < 0.05 and FDR < 0.25 were listed. Data were pooled from 10 or 11 individual mice in each group. (<b>b</b>) Apoptotic cells were identified by Annexin V staining, and post-transcriptional expression of Bcl-2 was identified by monoclonal antibody with flow cytometry at 7 dpi. The frequencies are shown as mean with independent data point and compared by Student’s <i>t</i>-test. Data represent 3 or 4 independent experiments (n = 5/group/experiment). Each symbol represents one mouse.</p

The D^bM₁₈₇ T cells efficiently control viral replication.

<p><b>(a)</b> Adoptive transfer of pMHC-specific donor cells increases precursor of CD8 T effector cells during early infection. Live D^bM₁₈₇, K^dM2₈₂ and bulk (with neither specificity) CD8 T cells from spleen lymphocytes of RSV-infected mice at 7dpi were sorted with FACS, and transferred into naive recipients respectively. The recipients were then challenged with RSV next day, and were evaluated for the donor D^bM₁₈₇ and K^dM2₈₂ T cell frequencies in the right lung at 4 dpi by flow cytometry. <b>(b)</b> Viral activity in RSV challenged recipients. Left lungs of the RSV-challenged recipients were assessed for virus replication. The virus titers are expressed as log₁₀ PFU/gram of lung tissue. Data are shown as mean with independent data point and compared by Student’s <i>t</i>-test. Data represent 3 independent experiments (n = 4 or 5/group/experiment). Each symbol represents one mouse.</p

The D^bM₁₈₇ T cells express high avidity TCR and signaling pathways promoting cytotoxic function.

<p><b>(a)</b> The TCR avidity was assessed by dissociation of D^bM₁₈₇ and K^dM2₈₂ from CD8 T cells. CD8 T cells were labeled with pMHCs and assessed cell-bound median fluorescence intensity (MFI) at indicated time point by flow cytometry. The MFI at 0 min was defined as the maximum measurement (100%). Data were analyzed with one-phase exponential decay using nonlinear regression, and shown at mean ± SEM of three independent experiments (n = 5/group/experiment). <b>(b)</b> Transcriptional expression of genes that are up-regulated after RSV infection and associated with conventional signaling pathways. The D^bM₁₈₇, K^dM2_82, and bulk CD8 T cells were sorted from spleen lymphocytes by FACS at 7 dpi. The mRNAs were isolated, amplified and labeled, then hybridized onto Illumina Mouse Chips. The quantitative gene expression were analyzed and normalized. Genes with Log₂ Fold Change (FC) > 1.3, p < 0.05 and FDR < 0.25 (listed on left side of the chat) and associated signaling pathways were shown (Pathways 1: Altered T Cell and B Cell Signaling in Rheumatoid Arthritis; 2: T Helper Cell Differentiation; 3: Dendritic Cell Maturation; 4: Type I Diabetes Mellitus Signaling; 5: Roe of NFAT in Regulation of the Immune Response; 6: Role of Pattern Recognition Receptors in Recognition of Bacteria and Viruses; 7: IL-10 Signaling; 8: Role of CHK Proteins in Cell Cycle Checkpoint Control; 9: TREM1 Signaling; 10: Communication between Innate and Adaptive Immune Cells; 11: CD40 Signaling; 12: Production of Nitric Oxide and Reactive Oxygen Species in Macrophages; 13: Cell Cycle Control of Chromosomal Replication; 14: Acute Phase Response Signaling; 15: CD28 Signaling in T Helper Cells; 16: PKC Signaling in T Lymphocytes; 17: IL-12 Signaling and Production in Macrophages.). Data were pooled from 10 or 11 individual mice in each group.</p