Journal: Cell reports

Article Title: Mass Cytometry Analysis Reveals that Specific Intratumoral CD4 + T Cell Subsets Correlate with Patient Survival in Follicular Lymphoma

doi: 10.1016/j.celrep.2019.01.085

Figure Lengend Snippet: (A) The tSNE maps of CD3 + , CD4 + , and CD8 + subsets from tonsil, reactive lymph node (rLN) and spleen (rSP) tissues, and FL biopsy specimens. The t-SNE analysis was performed on a concatenated file (31 FL, 6 tonsils, 3 rLNs, or 4 rSPs) using escalated parameter (surface makers). (B) Plots of principal-component analysis of CD3 + T cells from tonsil, rLN, rSP tissues, and FL biopsy specimens. (C) Plots from a representative sample of tonsil, rLN, rSP, or FL showing expression of CD45RO and CCR7. Graphs show the percentages of naive, central memory, effector memory, and terminally differentiated T cells from CD3 + T cells in tonsil, rLN, rSP, and FL. p value indicates a comparison between Tonsil and FL. See also .

Article Snippet: CD3 + or CD4 + T cells were isolated using positive selection with CD3 or CD4 microbeads kit (StemCell Technologies, Vancouver, Canada).

Techniques: Expressing, Comparison

Journal: Cell reports

Article Title: Mass Cytometry Analysis Reveals that Specific Intratumoral CD4 + T Cell Subsets Correlate with Patient Survival in Follicular Lymphoma

doi: 10.1016/j.celrep.2019.01.085

Figure Lengend Snippet: (A) Plots from a representative sample of FL showing expression of PD-1 on CD3 + , CD4 + , or CD8 + T cells. (B) The tSNE map of PD-1 + cells from CD3 + concatenated file (31 FL). (C) Graphs showing the percentages of PD-1 high or PD-1 low cells from CD4 + or CD8 + T cells from 31 FL specimens. (D) The tSNE maps of CD4 + T cells from a representative FL specimen showing the gating strategy to identify PD-1 + subsets . PD-1 expression on subsets S3–S6 in red circles was shown. (E) Graphs showing the percentages of surface markers from S3, S4, S5, or S6 in FL. (F) The tSNE maps of S3–S6 from the concatenated file of 31 FL specimens. (G) Kaplan-Meier curves for overall survival of FL patients (n = 31) by the number of total PD-1 + cells, S3, S4, S5, or S6 with a cutoff of 65.4%, 5.92%, 2.86%, 3.73%, or 14%, respectively. See also .

Article Snippet: CD3 + or CD4 + T cells were isolated using positive selection with CD3 or CD4 microbeads kit (StemCell Technologies, Vancouver, Canada).

Techniques: Expressing

Journal: Cell reports

Article Title: Mass Cytometry Analysis Reveals that Specific Intratumoral CD4 + T Cell Subsets Correlate with Patient Survival in Follicular Lymphoma

doi: 10.1016/j.celrep.2019.01.085

Figure Lengend Snippet: (A) The tSNE map of CD27 or CD28 + cells from CD3 + concatenated file (31 FL and 6 tonsils). CD4 + or CD8 + T cells were gated based on expression levels from CD4 or CD8 staining. (B) Graphs showing the percentages of CD27 − or CD28 − or CD27 − CD28 − cells from CD3 + T cells from 31 FL specimens and 6 tonsil tissues. (C) Graphs showing the percentages of CD27 − , CD28 − , or CD27 − CD28 − cells from CD25 − , CD25 + , PD-1 high , PD-1 low , PD-1 neg , T N , T CM , T EM , or T EMRA cells from 31 FL specimens. (D) Graphs showing the percentages of surface markers from CD27 − , CD27 + , CD28 − , or CD28 + cells from 31 FL specimens. (E and F) Kaplan-Meier curves for overall survival of FL patients (n = 31) by the number of CD3 + CD27 − , CD3 + CD28 − , CD3 + CD27 − CD28 − (E), CD4 + CD25 + , CD4 + CD25 + CD27 − , CD4 + PD-1 + , or CD4 + PD-1 + CD27 − (F) T cells. Bottom: p value indicates a comparison between T N and T EMRA . See also .

Article Snippet: CD3 + or CD4 + T cells were isolated using positive selection with CD3 or CD4 microbeads kit (StemCell Technologies, Vancouver, Canada).

Techniques: Expressing, Staining, Comparison

Journal: Cell reports

Article Title: Mass Cytometry Analysis Reveals that Specific Intratumoral CD4 + T Cell Subsets Correlate with Patient Survival in Follicular Lymphoma

doi: 10.1016/j.celrep.2019.01.085

Figure Lengend Snippet: (A) Heatmap showing clustering results for differentially expressed markers on CD3 + T cells from 31 FL specimens. Clustering was performed by Cluster 3.0 software. Each vertical column represents a patient sample. (B) Plot showing clustering results from FL patients divided by 2 groups (alive versus dead). Circles in red represent clusters that differed between two groups. Number in circles indicates a cluster ID. Clustering was performed by CITRUS from Cytobank. (C) Histogram plots in corners showing expression of selected markers by cells from 4 parent clusters overlapped to background. Expression level of each selected marker was expressed by Cluster (red) over Background (light blue). Graphs on the right showed quantitative results of abundance from 4 parent clusters between groups of Alive and Dead. The histograms and graphs were generated by CITRUS from Cytobank. (D) Kaplan-Meier curves for overall survival of FL patients (n = 31) by the clusters 21536 and 21523 using a cutoff point of abundance. See also .

Article Snippet: CD3 + or CD4 + T cells were isolated using positive selection with CD3 or CD4 microbeads kit (StemCell Technologies, Vancouver, Canada).

Techniques: Software, Expressing, Marker, Generated

Journal: Cell reports

Article Title: Mass Cytometry Analysis Reveals that Specific Intratumoral CD4 + T Cell Subsets Correlate with Patient Survival in Follicular Lymphoma

doi: 10.1016/j.celrep.2019.01.085

Figure Lengend Snippet: (A) Histograms showing CD27 or CD28 expression of CD27 + CD28 + T cells cultured in anti-CD3-coated plate plus anti-CD28 Ab for 2 or 5 days (activated). Cells cultured in uncoated plate were used as control (resting). (B) CD27 or CD28 expression from CD4 + T cells cultured in anti-CD3-coated plate plus anti-CD28 Ab in the presence or absence of TGF-β for 3 days. (C) Immunohistochemistry showing CD70 expression from FL (n = 8) and tonsil tissue. (D) Plots showing CD70 expression on CD19 + cells from representative lymphoma (n = 11) and tonsil tissues. (E) Histograms showing (i) CD70 expression on DoHH2 cells, (ii) expression of CD27 and CD28 on T cells co-cultured with DoHH2 cells for 3 days, (iii) CD70 expression of DoHH2 cells treated with either anti-CD70 blocking Ab or mIgG for 2 h, and (iv) expression of CD27 on CD4 + T cells co-cultured with DoHH2 cells pretreated with either anti-CD70 blocking Ab or mIgG for 3 days. (F) Histograms showing CFSE staining of CD27 + CD28 + , CD27 + CD28 − , CD27 − CD28 + , or CD27 − CD28 − T cells cultured in anti-CD3-coated plate plus anti-CD28 Ab for 3 days. Graph shows the numbers of CFSE dim cells of these subsets; n = 4. (G) Plots showing CD45RO and CCR7 expression from CD4 + CD27 − or CD4 + CD27 + T cells. Cells in box represent terminally differentiated T cells (CD45RO − CCR7 − ; T EMRA ). See also .

Article Snippet: CD3 + or CD4 + T cells were isolated using positive selection with CD3 or CD4 microbeads kit (StemCell Technologies, Vancouver, Canada).

Techniques: Expressing, Cell Culture, Control, Immunohistochemistry, Blocking Assay, Staining

Journal: Cell reports

Article Title: Mass Cytometry Analysis Reveals that Specific Intratumoral CD4 + T Cell Subsets Correlate with Patient Survival in Follicular Lymphoma

doi: 10.1016/j.celrep.2019.01.085

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD3 + or CD4 + T cells were isolated using positive selection with CD3 or CD4 microbeads kit (StemCell Technologies, Vancouver, Canada).

Techniques: Selection, Recombinant, Blocking Assay, Software