Journal: Frontiers in Immunology

Article Title: A tale of two conditions: when people living with HIV meet three doses of inactivated COVID-19 vaccines

doi: 10.3389/fimmu.2023.1174379

Figure Lengend Snippet: (A) The frequency of IFN-γ + CD4 + T cells among PLWH and HC at multiple time points. (B) The frequency of IFN-γ + CD8 + T cells among PLWH and HC at multiple time points. (C) The frequency of TNF-α + CD4 + T cells among PLWH and HC at multiple time points. (D) The frequency of TNF-α + CD8 + T cells among PLWH and HC at multiple time points. D14A2: Day 14 after the second dose; D0B3: Day 0 before the booster dose; D14A3: Day 14 after the booster dose; D30A3: Day 30 after the booster dose; D180A3: Day 180 after the booster dose.

Article Snippet: Surface marker staining was performed with anti-CD3-PerCP-cy5.5 (clone UCHT1; Biolegend), anti-CD8-APC-Cy7 (clone SK1; Biolegend), and anti-CD4-PE-Cy7 (clone RPA-T4; Biolegend) for 30 min at 4°C in the dark.

Techniques:

Journal: Frontiers in Immunology

Article Title: A tale of two conditions: when people living with HIV meet three doses of inactivated COVID-19 vaccines

doi: 10.3389/fimmu.2023.1174379

Figure Lengend Snippet: (A) The frequencies of IFN-γ-secreting and TNF-α-secreting CD4 + and CD8 + T cells in PLWH at multiple time points. (B) The frequencies of IFN-γ-secreting and TNF-α-secreting CD4 + and CD8 + T cells in HC at multiple time points. D14A2: Day 14 after the second dose; D0B3: Day 0 before the booster dose; D14A3: Day 14 after the booster dose; D30A3: Day 30 after the booster dose; D180A3: Day 180 after the booster dose.

Article Snippet: Surface marker staining was performed with anti-CD3-PerCP-cy5.5 (clone UCHT1; Biolegend), anti-CD8-APC-Cy7 (clone SK1; Biolegend), and anti-CD4-PE-Cy7 (clone RPA-T4; Biolegend) for 30 min at 4°C in the dark.

Techniques:

Journal: Journal of Translational Medicine

Article Title: First-line chemoimmunotherapy in metastatic breast carcinoma: combination of paclitaxel and IMP321 (LAG-3Ig) enhances immune responses and antitumor activity

doi: 10.1186/1479-5876-8-71

Figure Lengend Snippet: IMP321 increases the numbers of monocytes, NK and activated CD8 T cells in blood (panel A) . Fresh blood samples were collected pre-dosing, at D1, D85 and D170 and directly stained with fluorochrome-conjugated antibodies in tubes containing a precise number of fluorescent control beads. The results show the mean ± sd of the absolute numbers of CD45 + CD14 + (monocytes), CD3 - CD56 + (NK cells) and CD38 + HLA-DR + CD8 + (activated CD8 + cells) cells. The paired non-parametric Wilcoxon signed rank test was used to compare increases observed between D85 or D170 and D1. When significant (< 0.05), p values are indicated. IMP321 increases the percentages of dendritic cells and cytotoxic CD45RA + Effector-Memory CD8 + T cells (EMRA) in PBMCs (panel B). PBMCs cells collected pre-dosing, at D1, D85 and D170 were isolated and stained with fluorochrome-conjugated antibodies and analyzed by flow cytometry. The results show the mean ± sd of the percentages of plasmacytoid dendritic cells (pDC, CD45 + CD14 - CD16 - HLA-DR + CD123 + CD11c - ) and myeloid dendritic cells (mDC, CD45 + CD14 - CD16 - HLA-DR + CD123 - CD11c + ) in CD45 + cells, as well as the percentages of CD45RA + CD45RO - CD62L - in the CD8 + T cell population (CD45RA + EM CD8 T cells or EMRA). The significant Wilcoxon p values are indicated.

Article Snippet: Blood samples were collected pre-dosing at D1, D85 and D170 and directly stained with BD Multitest CD8-FITC/CD38-PE/CD3-PerCP/HLA-DR-APC, with BD Multitest 6-color TBNK (CD3-FITC/CD16-PE+CD56-PE/CD45-PerCPCy5.5/CD4PE-Cy7/CD19-APC/CD8-APC-Cy7), BD Simultest LeucoGate (CD45-FITC/CD14-PE) in tubes containing a precise number of fluorescent control beads (BD Trucount™tubes, BD Biosciences).

Techniques: Staining, Isolation, Flow Cytometry

Journal: Journal of Translational Medicine

Article Title: First-line chemoimmunotherapy in metastatic breast carcinoma: combination of paclitaxel and IMP321 (LAG-3Ig) enhances immune responses and antitumor activity

doi: 10.1186/1479-5876-8-71

Figure Lengend Snippet: IMP321 increases the expression of activation markers on blood monocytes . Blood samples were collected pre-dosing, at D1, D85 and D170 and directly stained with fluorochrome-conjugated CD45, CD14, anti-HLA-DR and CD11a, CD11b, CD16, CD35, CD54, CD64, CD80 or CD86 antibodies in tubes containing a precise number of fluorescent control beads. The expression of activation markers on monocytes was directly proportional to the cell-bound fluorescence. The results shown in panel A are the mean ± sd after normalization of the cell-bound fluorescence against the fluorescence of control beads. Statistically significant increases between D85 or D170 and D1 are analyzed using Wilcoxon signed rank test and significant p values (< 0.05) are shown. In panel B, the percentage of patients showing increases in the expression of the indicated numbers of activation markers at D85 or D170 compared to the baseline at D1 was calculated. The number of markers (n) displaying an increase by at least 50% was calculated for each patient in the 1.25 mg (7 patients) and 6.25 mg (12 patients) groups. The pie charts represent the percentages of patients with increases in the indicated number of markers.

Article Snippet: Blood samples were collected pre-dosing at D1, D85 and D170 and directly stained with BD Multitest CD8-FITC/CD38-PE/CD3-PerCP/HLA-DR-APC, with BD Multitest 6-color TBNK (CD3-FITC/CD16-PE+CD56-PE/CD45-PerCPCy5.5/CD4PE-Cy7/CD19-APC/CD8-APC-Cy7), BD Simultest LeucoGate (CD45-FITC/CD14-PE) in tubes containing a precise number of fluorescent control beads (BD Trucount™tubes, BD Biosciences).

Techniques: Expressing, Activation Assay, Staining, Fluorescence

Journal: Biomedicines

Article Title: Esterification of p -Coumaric Acid Improves the Control over Melanoma Cell Growth

doi: 10.3390/biomedicines11010196

Figure Lengend Snippet: N -butyl p -coumarate stimulates the NK and CD8 T cell activation and controls the tumor growth. ( A , B ) splenocytes from C57BL/6 were incubated with p -coumaric acid ( p -CA, white), ethyl p -coumarate ( 1 , red) or n-butyl p -coumarate ( 2 , blue) at 0.1 mM for 6 h and the CD69 expression was analyzed by flow cytometry in the NK ( A ) and CD8 T ( B ) cells. The figure represents mean ± SD of one single experiment (n = 5/group). C-D: C57Bl/6 mice were injected i.p. with a saline solution containing p -CA or compound 2 (2 mg/animal) for 5 consecutive days. Control animals received the corresponding DMSO concentration. At day three of the treatment, 2 × 10 5 B16-F10 melanoma cells were inoculated i.v., and the number of tumor nodules was assessed fifteen days later. ( C ) representative figure from one animal/group. ( D ) The figure represents the mean ± SEM of the lung nodule counts from two individual experiments (n = 4–5/group). * p < 0.05. **** p < 0.0001.

Article Snippet: The total splenic cells were cultured with p -CA or compounds 1 or 2 (0.1 mM) for 6 h. Then, the cells were incubated with LIVE/DEAD Fixable Aqua Dead Stain Kit (ThermoFischer, Eugene, OR, USA) in the presence of Fc blocking (anti-CD16/32), washed, and labeled with anti-CD3 (APC-Cy7), anti-CD8 (PE-Cy7), anti-NK1.1 (FITC), and anti-CD69 (eFluor TM 450) (All antibodies are from eBioscience, San Diego, CA, USA).

Techniques: Activation Assay, Incubation, Expressing, Flow Cytometry, Injection, Saline, Control, Concentration Assay

Journal: Science Advances

Article Title: Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T cells

doi: 10.1126/sciadv.adc9346

Figure Lengend Snippet: ( A to G ) scRNA-seq data from B16F10 melanoma–infiltrating CD8 + TILs (GSE116390) were analyzed. Pdcd1 -expressing CD8 + TILs were filtered. (A) Clustering and uniform manifold approximation and projection (UMAP) visualization of Pdcd1 -expressing CD8 T cells from B16F10 melanoma tumor ( n = 2652 cells) on day 15. Colors denote transcriptional clusters, labeled with functional annotations. (B) UMAP visualization of transitory signature score within Pdcd1 -expressing CD8 + TILs. (C) Violin plot of transitory signature score within clusters. Cluster 5 denotes transitory effector cells. (D to G) Violin plot of the expression level of (D) Tox , (E) Tbx21 , (F) Tnf , and (G) Klf4 within clusters. Cluster 5 denotes transitory effector cells. ( H to K ) Five different CD8 T cell subpopulations were isolated from MC38 tumor–bearing C57BL/6 mice on day 18. (H) Schematic design of the experiment. (I) Histogram of Tox mRNA expression on five different CD8 T cell subpopulations ( n = 5 per group). (J) Histogram of ratio T-bet/Eomes (mRNA) on five different CD8 T cell subpopulations ( n = 4 per group). (K) Histogram of Klf4 mRNA expression on five different CD8 T cell subpopulations ( n = 5 per group). All data are means ± SEM. Statistical analysis was performed using Student’s t test. ns (nonsignificant), P > 0.05; * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: The following antibody conjugates were purchased from Invitrogen: CD44 (IM7)–APC, CD45 (HI30)–PerCP-Cy5.5, CD45.1 (A20)–APC eFluor780, CD45.2 (104)–PE-Cy7, CD8 (53-6.7)–fluorescein isothiocyanate (FITC), CD8 (53-6.7)–APC-Cy7, CD8 (53-6.7)–PE; CTLA-4 (UC10-4B9)–Biotin, Eomes (Dan11mag)–PE-Cy7, Ki-67 (SolA15)–Biotin, PD-1 (J43)–FITC, T-bet (4B10)–PE, CD69 (H1.2F3)–PerCP-Cy5.5, and TOX (TXRX10)–PE.

Techniques: Expressing, Labeling, Functional Assay, Isolation

Journal: Science Advances

Article Title: Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T cells

doi: 10.1126/sciadv.adc9346

Figure Lengend Snippet: ( A to N ) OT-I CD8 T cells were retrovirally transduced with MigRI/ Klf4 on day 1 during in vitro exhaustion process and were harvested on day 5. GFP + cells were gated for all analyses. (A) Representative flow cytometry plot of GzmB expression and the proportion of GzmB + cells in CD8 T cells transduced with MigRI/ Klf4 ( n = 6 per group). (B) Representative flow cytometry plot of IFN-γ expression and the proportion of IFN-γ + cells in CD8 T cells transduced with MigRI/ Klf4 ( n = 6 per group). (C to N) CD8 T cells transduced with MigRI/ Klf4 were isolated and RNA-seq was performed. (C) Volcano plot highlighting differential transcripts between CD8 T cells transduced with MigRI/ Klf4 ( n = 3 per group). (D) RNA expression heatmap of DEGs from CD8 T cells transduced with MigRI/ Klf4 ( n = 3 per group). (E) Gene Ontology (GO) (pathway and process) analysis of DEGs. (F) GO (transcriptional regulation network) analysis of DEGs. (G) RNA expression heatmap of AP-1 subunit genes from CD8 T cells transduced with MigRI/ Klf4 ( n = 3 per group). (H) Representative flow cytometry plot and histogram of relative MFI (mean fluorescence intensity) level of phospho–c-Jun in CD8 T cells transduced with MigRI/ Klf4 ( n = 4 per group). (I to M) GSEA between CD8 T cells transduced with MigRI/ Klf4 using gene sets of (I) Tcirc signature, (J) naïve T cell quiescence, (K) genes down-regulated in Tex int , (L) genes up-regulated in Tex int , and (M) genes up-regulated in Tex term . (N) Histogram of normalized enrichment score from GSEA using various gene sets. (A, B, and H) Data are means ± SEM. Statistical analysis was performed using Student’s t test. * P < 0.05; **** P < 0.0001. Log 2 FC, log 2 fold change; FDR, false discovery rate; NES, normalized enrichment score; FSC-H, forward scatter height.

Article Snippet: The following antibody conjugates were purchased from Invitrogen: CD44 (IM7)–APC, CD45 (HI30)–PerCP-Cy5.5, CD45.1 (A20)–APC eFluor780, CD45.2 (104)–PE-Cy7, CD8 (53-6.7)–fluorescein isothiocyanate (FITC), CD8 (53-6.7)–APC-Cy7, CD8 (53-6.7)–PE; CTLA-4 (UC10-4B9)–Biotin, Eomes (Dan11mag)–PE-Cy7, Ki-67 (SolA15)–Biotin, PD-1 (J43)–FITC, T-bet (4B10)–PE, CD69 (H1.2F3)–PerCP-Cy5.5, and TOX (TXRX10)–PE.

Techniques: Transduction, In Vitro, Flow Cytometry, Expressing, Isolation, RNA Sequencing Assay, RNA Expression, Fluorescence

Journal: Science Advances

Article Title: Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T cells

doi: 10.1126/sciadv.adc9346

Figure Lengend Snippet: ( A ) Representative flow cytometry plot of CEACAM1 expression and the proportion of CEACAM1 + cells in CD8 T cells transduced with MigRI/ Klf4 ( n = 5 per group) from in vitro exhaustion experiment. GFP + cells were gated for the analysis. ( B to J ) CD8 + TILs were analyzed from MC38 tumor–bearing C57BL/6 mice on day 18. (B) Representative flow cytometry plot of CEACAM1 and TIM3 expression, and the proportion of each subpopulation in PD1 + CD8 + TILs ( n = 9 per group). (C to F) Representative flow cytometry plot and the proportion of (C) GzmB + ( n = 8 per group), (D) IFN-γ + ( n = 7 per group), (E) TNF-α + ( n = 6 per group), and (F) CX3CR1 + ( n = 7 per group) cells in CEACAM1-TIM3–defined subsets. (G to J) Histogram of (G) relative TOX (MFI) ratio ( n = 4 per group), (H) relative T-bet/Eomes (MFI) ratio ( n = 5 per group), (I) relative phospho–c-Jun ( n = 3 per group), and (J) relative expression of Klf4 ( n = 3 per group) in CEACAM1-TIM3–defined subsets. Relative values were calculated on the basis of the level of CEACAM1 + TIM3 + subset. All data are means ± SEM. Statistical analysis was performed using Student’s t test. ns, P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The following antibody conjugates were purchased from Invitrogen: CD44 (IM7)–APC, CD45 (HI30)–PerCP-Cy5.5, CD45.1 (A20)–APC eFluor780, CD45.2 (104)–PE-Cy7, CD8 (53-6.7)–fluorescein isothiocyanate (FITC), CD8 (53-6.7)–APC-Cy7, CD8 (53-6.7)–PE; CTLA-4 (UC10-4B9)–Biotin, Eomes (Dan11mag)–PE-Cy7, Ki-67 (SolA15)–Biotin, PD-1 (J43)–FITC, T-bet (4B10)–PE, CD69 (H1.2F3)–PerCP-Cy5.5, and TOX (TXRX10)–PE.

Techniques: Flow Cytometry, Expressing, Transduction, In Vitro

Journal: Science Advances

Article Title: Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T cells

doi: 10.1126/sciadv.adc9346

Figure Lengend Snippet: ( A to F ) PmelI CD8 T cells transduced with MigRI/ Klf4 were adoptively transferred into MC38-gp100–inoculated Rag2 KO mice on day 1. The tumor-infiltrating CD8 T cells were analyzed on day 15. GFP + cells were gated for all analyses. (A) Tumor growth curve of mice received CD8 T cells transduced with MigRI ( n = 6)/ Klf4 ( n = 7). (B) The proportion of Ki-67 + cells in CD8 T cells transduced with MigRI/ Klf4 ( n = 4 per group). (C to E) Representative flow cytometry plot and the proportion of (C) GzmB + , (D) IFN-γ + , and (E) TNF-α + cells in CD8 T cells transduced with MigRI ( n = 4)/ Klf4 ( n = 3 to 4). (F) Representative flow cytometry plot and the proportion of CEACAM1 + TIM3 + cells in CD8 T cells transduced with MigRI ( n = 4)/ Klf4 ( n = 5). ( G to K ) PmelI CD8 T cells transduced with MigRI/ Klf4 were adoptively transferred into MC38-gp100–inoculated Rag2 KO mice on day 7. The tumor-infiltrating CD8 T cells transduced with MigRI (from four mice) and Klf4 (from four mice) were pooled (GFP + cells were sorted) on day 15, and SMART-seq was performed. (G) GO (GO biological process) analysis of DEGs. (H to J) GSEA between CD8 T cells transduced with MigRI/ Klf4 using gene sets of (H) genes up-regulated in Tex int , (I) genes up-regulated in Tex term , and (J) naïve T cell quiescence. (K) Histogram of fold change of genes between CD8 T cells transduced with MigRI/ Klf4 . (A to F) Data are means ± SEM. Statistical analysis was performed using Student’s t test. ns, P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: The following antibody conjugates were purchased from Invitrogen: CD44 (IM7)–APC, CD45 (HI30)–PerCP-Cy5.5, CD45.1 (A20)–APC eFluor780, CD45.2 (104)–PE-Cy7, CD8 (53-6.7)–fluorescein isothiocyanate (FITC), CD8 (53-6.7)–APC-Cy7, CD8 (53-6.7)–PE; CTLA-4 (UC10-4B9)–Biotin, Eomes (Dan11mag)–PE-Cy7, Ki-67 (SolA15)–Biotin, PD-1 (J43)–FITC, T-bet (4B10)–PE, CD69 (H1.2F3)–PerCP-Cy5.5, and TOX (TXRX10)–PE.

Techniques: Transduction, Flow Cytometry

Journal: Science Advances

Article Title: Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T cells

doi: 10.1126/sciadv.adc9346

Figure Lengend Snippet: ( A to L ) MC38 cells were subcutaneously injected into the right flank of control/ Klf4 cKO mice. CD8 + TILs were analyzed on day 14. (A) Tumor growth curve of control ( n = 4)/ Klf4 cKO ( n = 5) mice. (B) Histogram of tumor mass of control ( n = 4)/ Klf4 cKO ( n = 5) mice. (C to E) Representative flow cytometry plot and the proportion of (C) GzmB + , (D) IFN-γ + , and (E) TNF-α + cells in PD1 + CD8 + TILs from control/ Klf4 cKO mice ( n = 3 per group). ( F ) Representative flow cytometry plot and the proportion of CEACAM1 + TIM3 + cells in PD1 + CD8 + TILs from control ( n = 4)/ Klf4 cKO ( n = 5) mice. ( G to L ) PD1 + CD8 + TILs from tumor tissues of four control mice and six Klf4 cKO mice were pooled on day 14, and SMART-seq was performed. (G) GO (GO biological process) analysis of DEGs. rRNA, ribosomal RNA. (H to K) GSEA between control and Klf4 cKO PD1 + CD8 + TILs using gene sets of (H) adenosine triphosphate (ATP) metabolic process, (I) oxidative phosphorylation, (J) genes down-regulated in Tex int , and (K) genes up-regulated in Tex term . (L) Histogram of fold change of genes between control and Klf4 cKO PD1 + CD8 + TILs. (A to F) Data are means ± SEM. Statistical analysis was performed using Student’s t test. * P < 0.05; ** P < 0.01.

Article Snippet: The following antibody conjugates were purchased from Invitrogen: CD44 (IM7)–APC, CD45 (HI30)–PerCP-Cy5.5, CD45.1 (A20)–APC eFluor780, CD45.2 (104)–PE-Cy7, CD8 (53-6.7)–fluorescein isothiocyanate (FITC), CD8 (53-6.7)–APC-Cy7, CD8 (53-6.7)–PE; CTLA-4 (UC10-4B9)–Biotin, Eomes (Dan11mag)–PE-Cy7, Ki-67 (SolA15)–Biotin, PD-1 (J43)–FITC, T-bet (4B10)–PE, CD69 (H1.2F3)–PerCP-Cy5.5, and TOX (TXRX10)–PE.

Techniques: Injection, Flow Cytometry

Journal: Science Advances

Article Title: Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T cells

doi: 10.1126/sciadv.adc9346

Figure Lengend Snippet: ( A to H ) OT-I CD8 T cells were transduced with pRetroX-GFP/pRetroX- Klf4 on day 1 during in vitro exhaustion process. Exhausted CD8 T cells were harvested on day 5 and cultured with doxycycline for additional 2 days. GFP + cells were gated for all analyses. (A) Representative flow cytometry plot and the proportion of IFN-γ + cells in pRetro-GFP/Dox + and pRetro- Klf4 /Dox + cells ( n = 4 per group). (B) Representative flow cytometry plot of CEACAM1 expression and the proportion of CEACAM1 + cells in pRetro-GFP/Dox + and pRetro- Klf4 /Dox + cells ( n = 4 per group). (C to H) Histogram of relative mRNA expression of (C) Klf4 ( n = 4 per group), (D) Tox ( n = 4 per group), (E) Tcf7 ( n = 4 per group), (F) Tbx21 ( n = 3 per group), (G) Gzmb ( n = 4 per group), and (H) Ifng ( n = 4 per group) in pRetro-GFP/Dox + and pRetro- Klf4 /Dox + cells. ( I to Q ) ATAC-seq was performed on naïve, effector, pRetro-GFP/Dox + , and pRetro- Klf4 /Dox + cells. Effector cells were in vitro generated by peptide stimulation for 2 days. (I) Principal components analysis plot of ATAC-seq data from effector CD8 T cells ( n = 3), pRetro-GFP/Dox + ( n = 3), and pRetro- Klf4 /Dox + ( n = 2) cells. (J to L) GSEA on chromatin accessibility of genes adjacent to peak regions between pRetro-GFP/Dox + and pRetro- Klf4 /Dox + cells using gene sets of (J) genes up-regulated in Tex int , (K) genes down-regulated in Tex int , and (L) top 100 genes increased DEGs by Klf4 overexpression (RNA-seq from in vitro exhaustion model). (M to Q) Representative ATAC-seq genome track of (M) Tox , (N) Ifng , (O) Jun , (P) Klf3 , (Q) Cd160 in naïve, effector, pRetro-GFP/Dox + , and pRetro- Klf4 /Dox + cells. (A to H) Data are means ± SEM. Statistical analysis was performed using Student’s t test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: The following antibody conjugates were purchased from Invitrogen: CD44 (IM7)–APC, CD45 (HI30)–PerCP-Cy5.5, CD45.1 (A20)–APC eFluor780, CD45.2 (104)–PE-Cy7, CD8 (53-6.7)–fluorescein isothiocyanate (FITC), CD8 (53-6.7)–APC-Cy7, CD8 (53-6.7)–PE; CTLA-4 (UC10-4B9)–Biotin, Eomes (Dan11mag)–PE-Cy7, Ki-67 (SolA15)–Biotin, PD-1 (J43)–FITC, T-bet (4B10)–PE, CD69 (H1.2F3)–PerCP-Cy5.5, and TOX (TXRX10)–PE.

Techniques: Transduction, In Vitro, Cell Culture, Flow Cytometry, Expressing, Generated, Over Expression, RNA Sequencing Assay

Journal: Science Advances

Article Title: Krüppel-like factor 4 regulates the cytolytic effector function of exhausted CD8 T cells

doi: 10.1126/sciadv.adc9346

Figure Lengend Snippet: ( A and B ) Kaplan-Meier curve of patients with COAD and KIRC based on their (A) KLF4 and (B) CEACAM1 expression (upper, 25%; and lower, 25%). Analyzed by OncoLnc. ( C ) Pearson correlation between KLF4 and CEACAM1 expression on COAD ( n = 275) and KIRC ( n = 523) tumor tissue. Analyzed by GEPIA2. ( D ) Pearson correlation between the expression of GAPDH/KLF4/CEACAM1 and average expression of effector signature genes (top 100 genes induced by Klf4 overexpression from RNA-seq data) on COAD ( n = 275) and KIRC ( n = 523) tumor tissue. Analyzed by GEPIA2. ( E ) Correlation of KLF4 expression and CD8 T cell infiltration score on COAD ( n = 458) and KIRC ( n = 533) tumor tissue. Analyzed by TIMER2.0. ( F ) KLF4 fold change after anti-PD1 therapy in CR (complete response)/PR (partial response; n = 5), SD (stable disease; n = 4), and PD (progressive disease; n = 9) group from patients with melanoma. Data are means ± SEM. Statistical analysis was performed using Student’s t test. ns, P > 0.05; * P < 0.05. ( G ) Kaplan-Meier curve of melanoma patient groups distinguished by KLF4 fold change. P values less than 0.05 were considered to be significant for the log-rank test and Pearson correlation test.

Article Snippet: The following antibody conjugates were purchased from Invitrogen: CD44 (IM7)–APC, CD45 (HI30)–PerCP-Cy5.5, CD45.1 (A20)–APC eFluor780, CD45.2 (104)–PE-Cy7, CD8 (53-6.7)–fluorescein isothiocyanate (FITC), CD8 (53-6.7)–APC-Cy7, CD8 (53-6.7)–PE; CTLA-4 (UC10-4B9)–Biotin, Eomes (Dan11mag)–PE-Cy7, Ki-67 (SolA15)–Biotin, PD-1 (J43)–FITC, T-bet (4B10)–PE, CD69 (H1.2F3)–PerCP-Cy5.5, and TOX (TXRX10)–PE.

Techniques: Expressing, Over Expression, RNA Sequencing Assay

Journal: Cancer Management and Research

Article Title: Treatment-Related Lymphopenia is Possibly a Marker of Good Prognosis in Nasopharyngeal Carcinoma: a Propensity-Score Matching Analysis

doi: 10.2147/CMAR.S456717

Figure Lengend Snippet: Comparison Between High and Low Lymphocyte Count Groups (Wilcoxon Test)

Article Snippet: The blood samples underwent treatment with six-color fluorescent monoclonal antibodies ( Supplementary Table 1 ), namely CD3-FITC, CD16+56-PE, CD45-PerCP-Cy5.5, CD19-APC, CD8-APC-cy7, and CD4-PE-cy7 (Beijing Tongsheng Shidai Biotechnology, Beijing, China).

Techniques: Comparison

Journal: Cancer Management and Research

Article Title: Treatment-Related Lymphopenia is Possibly a Marker of Good Prognosis in Nasopharyngeal Carcinoma: a Propensity-Score Matching Analysis

doi: 10.2147/CMAR.S456717

Figure Lengend Snippet: Correlations Between Baseline and Δ Values (N = 746)

Article Snippet: The blood samples underwent treatment with six-color fluorescent monoclonal antibodies ( Supplementary Table 1 ), namely CD3-FITC, CD16+56-PE, CD45-PerCP-Cy5.5, CD19-APC, CD8-APC-cy7, and CD4-PE-cy7 (Beijing Tongsheng Shidai Biotechnology, Beijing, China).

Techniques:

Journal: Cancer Management and Research

Article Title: Treatment-Related Lymphopenia is Possibly a Marker of Good Prognosis in Nasopharyngeal Carcinoma: a Propensity-Score Matching Analysis

doi: 10.2147/CMAR.S456717

Figure Lengend Snippet: Kaplan-Meier survival curves based on high and low baseline counts of CD8 + ( A and B ), total ( C and D ), and CD4 + ( E and F ) T cells.

Article Snippet: The blood samples underwent treatment with six-color fluorescent monoclonal antibodies ( Supplementary Table 1 ), namely CD3-FITC, CD16+56-PE, CD45-PerCP-Cy5.5, CD19-APC, CD8-APC-cy7, and CD4-PE-cy7 (Beijing Tongsheng Shidai Biotechnology, Beijing, China).

Techniques:

Journal: Cancer Management and Research

Article Title: Treatment-Related Lymphopenia is Possibly a Marker of Good Prognosis in Nasopharyngeal Carcinoma: a Propensity-Score Matching Analysis

doi: 10.2147/CMAR.S456717

Figure Lengend Snippet: Kaplan-Meier survival curves for patients in the high and low CD8 + T cell count ( A and B ) and CD4 + /CD8 + T cell ratio ( C and D ) groups after treatment.

Article Snippet: The blood samples underwent treatment with six-color fluorescent monoclonal antibodies ( Supplementary Table 1 ), namely CD3-FITC, CD16+56-PE, CD45-PerCP-Cy5.5, CD19-APC, CD8-APC-cy7, and CD4-PE-cy7 (Beijing Tongsheng Shidai Biotechnology, Beijing, China).

Techniques: Cell Counting

Journal: Cancer Management and Research

Article Title: Treatment-Related Lymphopenia is Possibly a Marker of Good Prognosis in Nasopharyngeal Carcinoma: a Propensity-Score Matching Analysis

doi: 10.2147/CMAR.S456717

Figure Lengend Snippet: Kaplan-Meier survival curves for patients in the high and low CD4 + ( A and B ), total ( C and D ), and CD8 + ( E and F ) T cell groups, based on Δ values.

Article Snippet: The blood samples underwent treatment with six-color fluorescent monoclonal antibodies ( Supplementary Table 1 ), namely CD3-FITC, CD16+56-PE, CD45-PerCP-Cy5.5, CD19-APC, CD8-APC-cy7, and CD4-PE-cy7 (Beijing Tongsheng Shidai Biotechnology, Beijing, China).

Techniques: