monoclonal antibody against human cd8  (Bio X Cell)

Journal: JCI Insight

Article Title: Epstein-Barr virus infection induces tissue-resident memory T cells in mucosal lymphoid tissues

doi: 10.1172/jci.insight.173489

Figure Lengend Snippet: ( A and B ) Ratio of CD8/CD4 T cell frequencies and frequency of naive (CD45RA + CD62L + ) CD8 + T cells in blood over time. ( C ) Numbers of naive, central memory (TCM; CD45RA – CD62L + ), effector memory (TEM; CD45RA – CD62L – ), and TEMRA (CD45RA + CD62L – ) CD8 + T cells in blood over time ( n = 2–9 animals per group depicted as mean values with 95% CI). ( D ) Representative flow cytometry plots and gating for CD45RA and CD62L of CD8 + T cells and (top row) and (bottom row) for CD69 and CD103 of CD8 + TEM T cells in blood and NALT at sacrifice. ( E ) Heatmap of frequencies of CD8 + TEM (left) and TRM (right) (CD69 + TEM) T cells in indicated tissues at sacrifice ( n = 4–34 animals per group from a minimum of 2 independent experiments; quantification in , A and B). ( F ) Quantification of absolute cell numbers of CD69 + TEM (left) and CD69 + CD103 + TEM (right) in the NALT in PBS versus i.n. infected mice. ( G ) Representative histogram of geometric mean fluorescence intensity (MFI) of i.v. injected anti-CD45 in blood and NALT (left) and quantification (right) ( n = 14–28 animals per group from a minimum of 2 independent experiments). ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001, by Kruskal-Wallis test for 3 groups ( A and B ) or Mann Whitney U test for comparison of 2 groups ( F and G ), or using paired Wilcoxon signed-rank test comparing T cell subsets between weeks 4 and 5 ( C ).

Article Snippet: CD8 + T cells were depleted weekly, via i.p. injection beginning at week 3 after infection, with the monoclonal antibody against human CD8 (clone OKT-8; BioXCell).

Techniques: Flow Cytometry, Infection, Fluorescence, Injection, MANN-WHITNEY, Comparison

Journal: JCI Insight

Article Title: Epstein-Barr virus infection induces tissue-resident memory T cells in mucosal lymphoid tissues

doi: 10.1172/jci.insight.173489

Figure Lengend Snippet: ( A ) UMAP plot of single CD8 + TEM transcriptomes sorted from indicated tissues. ( B ) Heatmap showing log 2 (fold change) of the top 50 highest and lowest differentially expressed genes of NALT single-cell transcriptomes (rows). ( C ) Integrated UMAP plots of cells showing relative expression of indicated marker expression (first and third columns; yellow-green, high expression; violet, low expression) and (second and fourth columns) violin plot depicting single-cell expression of markers in indicated tissues as normalized residual values.

Article Snippet: CD8 + T cells were depleted weekly, via i.p. injection beginning at week 3 after infection, with the monoclonal antibody against human CD8 (clone OKT-8; BioXCell).

Techniques: Expressing, Marker

Journal: JCI Insight

Article Title: Epstein-Barr virus infection induces tissue-resident memory T cells in mucosal lymphoid tissues

doi: 10.1172/jci.insight.173489

Figure Lengend Snippet: ( A ) Integrated UMAP plot depicting unsupervised clustering of single CD8 + TEM transcriptomes sorted from blood, LN, NALT, and spleen. ( B ) Cellular trajectories in pseudo time (blue to yellow, earlier to later pseudo time point) of single CD8 + TEM transcriptomes sorted from blood, LN, NALT, and spleen. ( C ) Representative flow cytometry plots of example TCR-Vβ17 and TCR-Vβ2 staining in 2 EBV-infected animals. ( D ) Frequencies of indicated TCR-VB clones among splenic CD8 + TEMs following intranasal EBV infection. ( E ) Frequencies of the top 6 TCR-VB clones in NALT CD8 + TEMs in the same animals. ( F ) Representative flow cytometry plots of example TCR-Vβ17 and TCR-Vβ2 staining in 2 PBS control animals (left) frequencies of the top 6 TCR-VB clones in spleen CD8 + TEMs of PBS control animals (center) and frequencies of the top 6 TCR-VB clones in NALT CD8 + TEMs of PBS control animals (right).

Article Snippet: CD8 + T cells were depleted weekly, via i.p. injection beginning at week 3 after infection, with the monoclonal antibody against human CD8 (clone OKT-8; BioXCell).

Techniques: Flow Cytometry, Staining, Infection, Clone Assay, Control

Journal: JCI Insight

Article Title: Epstein-Barr virus infection induces tissue-resident memory T cells in mucosal lymphoid tissues

doi: 10.1172/jci.insight.173489

Figure Lengend Snippet: ( A ) Representative flow cytometry gating for CD8 + TEM and CD69 + and CD69 + CD103 + CD8 + TEM in human tonsils. ( B ) Quantification of CD8 + TEM and CD69 + and CD69 + CD103 + CD8 + TEM in human PBMCs and tonsils. ( C ) Representative histogram of geometric mean fluorescence intensity (MFI) and quantification of CD11a expression by TEM from spleen (blue) and NALT (orange) of EBV-infected animals (left 2 panels) and (right 2 panels) in TEM from human PBMCs (blue) and tonsils from EBV + donors (red). ( D ) MFI and quantification of PD1 expression by TEM in spleen (blue) and NALT (orange) of EBV-infected animals (left 2 panels) and (right 2 panels) in TEM from human PBMCs (blue) and tonsils from EBV + donors (red). ( E ) MFI and quantification of CD27 expression by TEM in spleen (blue) and NALT (orange) of EBV-infected animals (left 2 panels) and (right 2 panels) in TEM from human PBMCs (blue) and tonsils from EBV-positive donors (red). For animal data, n = 14–30 animals per group from at least 6 independent experiments; for human data, n = 3–5 individuals per tissue from 2 independent experiments, except CD11a data, which is derived from a single experiment. * P ≤ 0.05, ** P ≤ 0.01, by Wilcoxon matched-pairs signed-rank test for animal data (second column, C , D , and E ) and Mann-Whitney U test for human data ( B and fourth column C , D , and E ).

Article Snippet: CD8 + T cells were depleted weekly, via i.p. injection beginning at week 3 after infection, with the monoclonal antibody against human CD8 (clone OKT-8; BioXCell).

Techniques: Flow Cytometry, Fluorescence, Expressing, Infection, Derivative Assay, MANN-WHITNEY

Journal: JCI Insight

Article Title: Epstein-Barr virus infection induces tissue-resident memory T cells in mucosal lymphoid tissues

doi: 10.1172/jci.insight.173489

Figure Lengend Snippet: ( A ) Representative flow cytometry plots showing IFN-γ and TNF-α expression by CD8 + TEMs derived from PBS control or EBV-infected NALT after PMA/ionomycin stimulation (left panels), as well as the distribution (center panels) and quantification (right panels) of each population. ( B ) As for A but depicting CD107a and granzyme B (GzmB) expression. ( C ) Representative flow cytometry plots showing IFN-γ and TNF-α expression byCD69 – or CD69 + CD8 + TEMs derived from tonsils of EBV carriers after PMA/ionomycin stimulation (left panels), as well as the distribution (center panels) and quantification (right panels) of each population. ( D ) As for C , but depicting CD107a and GzmB expression. Corresponding unstimulated data for all samples can be found in . For animal data, n = 12–14 animals per group from 2 independent experiments; for human data, n = 3–5 individuals per tissue from 2 independent experiments. * P ≤ 0.05, *** P ≤ 0.001, by Wilcoxon matched-pairs signed-rank test for animal data ( A and B ) and Mann-Whitney U test for human data ( C and D ).

Article Snippet: CD8 + T cells were depleted weekly, via i.p. injection beginning at week 3 after infection, with the monoclonal antibody against human CD8 (clone OKT-8; BioXCell).

Techniques: Flow Cytometry, Expressing, Derivative Assay, Control, Infection, MANN-WHITNEY

Journal: JCI Insight

Article Title: Epstein-Barr virus infection induces tissue-resident memory T cells in mucosal lymphoid tissues

doi: 10.1172/jci.insight.173489

Figure Lengend Snippet: ( A ) CD8 + T cell depletion efficiency in NALT, spleen, and blood of control (blue) and OKT8-treated (orange) animals given as percentage of CD4 – CD3 + T cells ( n = 8–11 animals per group from 2 independent experiments). ( B and C ) EBV viral loads in IU/mg NALT, IU/1 × 10 6 splenocytes, or IU/mL blood and normalized to the mean of the corresponding nondepleted group. ( D ) CD103 depletion efficiency in NALT, spleen, and blood of control (blue) and Ber-OCT3–treated (orange) animals given as percentage of CD8 + T cells ( n = 10–11 animals per group from 2 independent experiments). ( E and F ) EBV viral loads in IU/mg NALT, IU/1x × 10 6 splenocytes, or IU/mL blood and normalized to the mean of the corresponding nondepleted group. ( G ) Representative histogram of Ki-67 expression on CD8 + TEM in spleen and NALT (left) and quantification (right). ( H ) Representative histogram of Ki-67 expression on CD19 + cells in spleen and NALT (left) and quantification (right) ( n = 13 animals per group from 2 independent experiments). * P ≤ 0.05, *** P ≤ 0.001, **** P ≤ 0.0001. ( A – F ) Mann Whitney U test. ( G and H ) Wilcoxon matched-pairs signed rank test; dotted line indicates limit of detection.

Article Snippet: CD8 + T cells were depleted weekly, via i.p. injection beginning at week 3 after infection, with the monoclonal antibody against human CD8 (clone OKT-8; BioXCell).

Techniques: Control, Expressing, MANN-WHITNEY