Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Gating strategy for isolating CD3+CD4+ primary human T cells. PBMCs were isolated from healthy donors and labelled with antibodies directed against CD3, CD4 and CD8. CD3 + CD4 + T cells were sorted by subsequent gating on (1) lymphocytes, (2) single cells, (3) CD3 + cells and (4) CD4 + cells. CD3 + CD8 + T cells were sorted by subsequent gating on (1) lymphocytes, (2) single cells, (3) CD3 + cells and (4) CD8 + cells (not depicted).

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Isolation

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Differential H 2 O 2 induction in CD3 + CD4 + and CD3 + CD8 + T cells. PBMCs were stimulated with 10 ng/ml PMA for 1 hour (+) in the presence or absence of catalase (C), and intracellular H 2 O 2 was analyzed in CD3 + T cells (CD3), CD3 + CD4 + T cells (CD4) and CD3 + CD8 + T cells (CD8) by measuring H 2 DCFDA using flow cytometry (MFI). Non-stimulated cells (-) were used are controls. Statistical analysis was performed using a two-sided, paired T-test, * p<0.05, ** p<0.01, n=3.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Flow Cytometry

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: PMA stimulation induced mtROS signaling without inducing calcium signaling. Isolated CD3 + CD4 + were activated with 10 ng/ml PMA for 1 hour (A-C) and analyzed for T cell activation as seen by CD69 surface-expression (A), and mtROS production as seen by H 2 DCFDA fluorescence (B). (C) Representative flow cytometry data showing CD69 surface expression (left panel) and mtROS production (right panel). (D) Intracellular calcium was measured using Fluo-4FF. Arrow indicates the application of PMA or Ionomycin. (A, B) Statistical analysis was performed using a two-sided, paired T-test, * p<0.05, ** p<0.01, n=3.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Isolation, Activation Assay, Expressing, Fluorescence, Flow Cytometry

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Stress response, H 2 O 2 induction and mtROS confirmation in CD4 + T cells. Isolated CD3 + CD4 + cells were activated with 10 ng/ml PMA for 1 hour (A-C) and analyzed for T cell stress response as seen by 7AAD fluorescence (A), and H 2 O 2 production as seen by H 2 DCFDA fluorescence in the presence and absence of catalase (B). (C) Fluorescence microscopy images showing co-localization of mitochondria (upper panel) and intracellular H 2 O 2 (lower panels). Statistical analysis was performed using a two-sided, paired T-test, * p<0.05, ** p<0.01, n=3.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Isolation, Fluorescence, Microscopy

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Multiplexed thiol-specific proteomic workflow that allows the quantification of residue specific cysteine oxidations. (A) CD3 + CD4 + T cells were isolated from human PBMC by FACS-based cell sorting (1) and were either activated with 10 ng/ml PMA for 1 hour (A) or left inactive (I). Cells were lysed in the presence of a thiol reactive (iodoTMT) tag, which labeled any free cysteine thiolate residues (2). After the removal of excess label, samples were reduced by TCEP resulting in the conversion of all reversibly oxidized cysteine residues (such as disulfide bonds or sulfenic acids) to free cysteine thiolate residues, which were subsequently labeled with another iodoTMT tag with a differing reporter mass (3). Lysates were mixed and digested before being subjected to offline high pH reverse phase chromatography to generate multiple fractions, which were then analyzed under conventional low pH reverse phase nanoLC-MS (4). For simplicity, (2 and 3) only shows the iodoTMT tag strategy for the activated (A) sample. (B) MS1 peptide mass fingerprints were taken for subsequent activation experiments utilizing Higher-energy C-trap dissociation (HCD) fragmentation to generate high quality MS 2 spectra. By the use of the reporter tags that were identified in the MS 2 , cysteine-containing peptides were identified and quantified using automated software, and quantitative information for 9598 cysteine-containing peptides could be generated from 3284 proteins. (C) Formulas used for the calculation of (i) the total % change in oxidation of each residue between stimulated versus unstimulated conditions or (ii) the fold change in oxidation between stimulated over unstimulated conditions.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Isolation, FACS, Labeling, Reversed-phase Chromatography, Activation Assay, Software, Generated

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Absolute intensities of each label used to quantify the oxidation state of all cysteine-containing peptides. Distribution of absolute reporter ion intensities (in Log 2 ) of each label as used in our multiplex thiol-specific workflow revealed a general oxidation status of cysteine-containing peptides in non-induced (I) and PMA-induced (A) CD3 + T cells from 3 independent donors (D1-D3). Note that the absolute intensities of oxidized peptides (CysOx, red boxes) is lower than the absolute intensities of reduced peptides (CysFree, blue boxes).

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Multiplex Assay

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Distribution of average oxidation of cysteine residue-containing peptides in (A) unstimulated and (B) stimulated CD4+ T cells. PBMCs were isolated from healthy donors and stimulated with 10 ng/ml PMA for 1hour. Average oxidation of cysteine residues was calculated according the formula given in , which is ((CysOx l / CysOx l +CysFree l )*100) for unstimulated (inactive) cells (A) and ((CysOx A /CysOx A +CysFree A )*100 for stimulated (active) cells (B). Selected candidate proteins found with significantly differentially oxidized cysteine residues between stimulated and unstimulated conditions are marked with an asterisk. Statistical analysis was performed using a two-sided, paired T-test, * p <0.05, n=3.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Isolation

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Regulation of redox-modified CD4 + T cell proteins following PMA stimulation. Volcano plot showing proteins with regulated reduced (left) and oxidized (right) cysteine residues upon PMA stimulation of CD4 + T cells. PBMCs were isolated from healthy donors and stimulated with 10 ng/ml PMA for 1 hour, fold changes (Oxratio) in cysteine oxidation were analyzed as depicted in . In total, 4784 cysteine-containing peptides could be quantitatively analyzed of which 76 cysteine-containing peptides originating from 71 proteins were found either differentially oxidized or differentially reduced upon activation (highlighted in red) in at least 2 of 3 donors. X-axis shows the fold change (log 2 ) for each cysteine residue, y-axis shows the p-value (−log10) for each cysteine residue. The cut-off for fold change was set at log 2 0.5, and statistical significance was determined with a two-sided, paired t-test, and a p-value of −log10 1.3 (p< 0.05) was taken as significant, n=3.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Modification, Isolation, Activation Assay

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Cellular processes modified by oxidation or reduction in mtROS-producing CD4 + T cells. Enrichment analyses considered the 71 proteins with differentially redox-modified cysteine residues (from ) and revealed cellular processes mostly affected by oxidation (A) or reduction (B). The class “immune function” is presented for oxidized proteins only, whereas the majority of reduced proteins were related to immunity before. As such, the left most number indicates the actual value of proteins per category while the number of immune related proteins in (b) indicate the number of immune related proteins per category.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Modification

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Localization of free zinc ions (Zn 2+ ) and mitochondria in mtROS-producing T cells. PBMCs were stimulated with 10 ng/ml PMA for 1 hour in the presence of Zinpyr-1 and MitoTracker, prepared for fluorescence microscopy, and cross-sectional analyses of fluorescent signals were analyzed in CD3 + T cells (n=40). Representative images showing the local distribution of free Zn 2+ (top left panel, and green in merged image) and spatial clustering of mitochondria (top right panel, and red in merged image). Fluorescence intensity plots revealed local co-emerging of free zinc at the mitochondria (lower right panel). Arrow indicates the plane of the intensity plots, scale bar = 2μm.

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Fluorescence, Microscopy

Journal: bioRxiv

Article Title: Identification of mtROS-sensitive processes in activated CD4 + T cells

doi: 10.1101/2020.06.15.152116

Figure Lengend Snippet: Free zinc (Zn 2+ ) and mitochondria intensity profiles showing co-localization in 6 representative human CD3 + T cells. PBMCs were stimulated and imaged as described in . Representative fluorescence intensity plots of 6 single human CD3 + T cells revealing local co-emerging of free zinc (green line) at mitochondria (red line).

Article Snippet: CD3 + CD4 + T cells sorted (Aria-II SORP, BD Biosciences, Franklin Lakes, NJ, USA) with the help of Dr. Lothar Gröbe (HZI, Braunschweig).

Techniques: Fluorescence

Journal: Virology

Article Title: Regulatory CD4 T cells inhibit HIV-1 expression of other CD4 T cell subsets via interactions with cell surface regulatory proteins

doi: 10.1016/j.virol.2017.12.036

Figure Lengend Snippet: Tregs, but not Tconvs, decrease HIV-1 expression in co-cultured responder CD4 T cells. See Section 4 for details of co-culture and HIV-1 infection. PHA and IL-2 activated CD4 T cells (see Section 4) were infected with HIV-1 and co-cultured with Tregs or Tconvs. (A) Flow cytometry dot plots reveal the p24/gag protein levels of responder CD4 T cells (CFSE-labeled), as well as co-cultured (CFSE-negative) Tregs (above) or control Tconvs (below). (B) A bar graph depicts the mean values ± SEM (n=4), and statistically significant differences for the cell populations.

Article Snippet: The CD4 T subpopulations of Tregs (CD4+, CD25high, CD127low), conventional effectors or Tconvs (CD4+, CD25+, CD127high), memory or Tmems (CD4+, CD25−, CD45RO+), and Tnaives (CD4+, CD25−, CD45RA+) were purified from patient PBMC directly by staining for CD4 T cell surface markers CD3 (PerCP), CD4 (FITC), CD25 (APC), CD127 (PE), CD45RA (V450), and CD45RO (Alexa Fluor 700), and sorting the stained cells on an ARIA cell sorter (BD Biosciences, San Jose, CA, USA) as shown in .

Techniques: Expressing, Cell Culture, Co-Culture Assay, Infection, Flow Cytometry, Labeling

Journal: Virology

Article Title: Regulatory CD4 T cells inhibit HIV-1 expression of other CD4 T cell subsets via interactions with cell surface regulatory proteins

doi: 10.1016/j.virol.2017.12.036

Figure Lengend Snippet: Single or combinational antibody blockade of Tregs cell surface proteins (CTLA-4, PD-1, GARP) increases HIV-1 DNA levels and mRNA levels in co-cultured responder CD4 T cells. Tregs or Tconvs (control) were co-cultured with responding CD4 T cells during HIV-1 infection in vitro in the presence of blocking antibodies to cell surface molecules (see Section 4). Bar graph depictions show HIV-1 total DNA copies (A) and integrated DNA copies (B) of responding CD4 T cells co-cultured with Tconvs (left bars in pairs) or Tregs (right bars in pairs) in the presence of anti-CTLA-4, anti-PD-1, anti-GARP, anti-CTLA-4 plus anti-PD-1, or anti-CTLA-4 plus anti-PD-1 plus anti-GARP antibodies (see Section 4 for copy number calculation). (C) Real-time PCR amplification curves in single and combinational antibody blockings for detection of HIV-1 gag mRNA (18 S RNA as an internal reference control) from cells shown in (A) and (B). (D) Bar graphs depict HIV-1 relative gag mRNA levels (ΔΔCt values) from the responding CD4 T cells in the presence of Tconvs (control) or Tregs, plus the different antibody conditions. All bar graphs represent means ± SEM from 3 independent experiments.

Article Snippet: The CD4 T subpopulations of Tregs (CD4+, CD25high, CD127low), conventional effectors or Tconvs (CD4+, CD25+, CD127high), memory or Tmems (CD4+, CD25−, CD45RO+), and Tnaives (CD4+, CD25−, CD45RA+) were purified from patient PBMC directly by staining for CD4 T cell surface markers CD3 (PerCP), CD4 (FITC), CD25 (APC), CD127 (PE), CD45RA (V450), and CD45RO (Alexa Fluor 700), and sorting the stained cells on an ARIA cell sorter (BD Biosciences, San Jose, CA, USA) as shown in .

Techniques: Cell Culture, Infection, In Vitro, Blocking Assay, Real-time Polymerase Chain Reaction, Amplification

Journal: Virology

Article Title: Regulatory CD4 T cells inhibit HIV-1 expression of other CD4 T cell subsets via interactions with cell surface regulatory proteins

doi: 10.1016/j.virol.2017.12.036

Figure Lengend Snippet: HIV-1 infection of CD4 T cells in vitro demonstrates lower levels of HIV-1 DNA and gag mRNA in Tregs compared to Tconvs and Tmems. (A) FLOW contour plot shows the cell sorting strategy. The in vitro bulk infected ((B) and (C)) or freshly isolated ((D)) CD4 T cells were stained and sorted into Tregs (CD25high CD127low), Tconvs (CD25low CD127high), Tmems (CD25(−) CD45RO(+)), and Tnaives (CD25(−)CD45RA(+)) populations. (B) Semi-quantitative real-time PCR plots of HIV-1 viral DNA (top) and mRNA (bottom) levels, relative to control β-globin DNA and 18 S RNA, for Tmems (Tm), Tconvs (Tc), Tregs (Tr), and Tnaives (Tn) populations. (C) Bar graphs depict the mean results of viral DNA (top) and mRNA (bottom) levels from three independent experiments. Standard error bars are shown along with statistically significant comparisons between CD4 T cell populations. (D) Bar graph shows the viral DNA levels over two different time points as indicated post infection (n=3).

Article Snippet: The CD4 T subpopulations of Tregs (CD4+, CD25high, CD127low), conventional effectors or Tconvs (CD4+, CD25+, CD127high), memory or Tmems (CD4+, CD25−, CD45RO+), and Tnaives (CD4+, CD25−, CD45RA+) were purified from patient PBMC directly by staining for CD4 T cell surface markers CD3 (PerCP), CD4 (FITC), CD25 (APC), CD127 (PE), CD45RA (V450), and CD45RO (Alexa Fluor 700), and sorting the stained cells on an ARIA cell sorter (BD Biosciences, San Jose, CA, USA) as shown in .

Techniques: Infection, In Vitro, FACS, Isolation, Staining, Real-time Polymerase Chain Reaction

Journal: Virology

Article Title: Regulatory CD4 T cells inhibit HIV-1 expression of other CD4 T cell subsets via interactions with cell surface regulatory proteins

doi: 10.1016/j.virol.2017.12.036

Figure Lengend Snippet: CD4 Tregs from HIV-1/AIDS patients express lower levels of HIV-1 DNA and viral mRNA than Tconvs and Tmems ex vivo. (Left panels) CD4 T cell populations were sorted from PBMC of newly infected (patient A) and chronically HIV-1 infected (patient B) individuals. Bar graphs depict HIV-1 total DNA (top), gag mRNA (middle), and host FoxP3 mRNA (bottom) for sorted Tregs (Tr), Tconvs (Tc), Tmems (Tm), and Tnaives (Tn) populations. Levels were measured in triplicate and the mean values are shown.

Article Snippet: The CD4 T subpopulations of Tregs (CD4+, CD25high, CD127low), conventional effectors or Tconvs (CD4+, CD25+, CD127high), memory or Tmems (CD4+, CD25−, CD45RO+), and Tnaives (CD4+, CD25−, CD45RA+) were purified from patient PBMC directly by staining for CD4 T cell surface markers CD3 (PerCP), CD4 (FITC), CD25 (APC), CD127 (PE), CD45RA (V450), and CD45RO (Alexa Fluor 700), and sorting the stained cells on an ARIA cell sorter (BD Biosciences, San Jose, CA, USA) as shown in .

Techniques: Ex Vivo, Infection

Journal: Virology

Article Title: Regulatory CD4 T cells inhibit HIV-1 expression of other CD4 T cell subsets via interactions with cell surface regulatory proteins

doi: 10.1016/j.virol.2017.12.036

Figure Lengend Snippet: Ex vivo expanded Tregs retain cell surface phenotype and suppressive function. See Section 4 for expansion of Tregs. (A) and (B) Flow cytometry profiles show the relative levels of expression for cell surface markers of the ex vivo expanded Tregs and Tconvs. (C) Tregs (top panels) or control Tconvs (bottom panels) were co-cultured with responding PBMC at varying effector to responder ratios as detailed. Proliferation in response to anti-CD3/CD28 coated bead is denoted by CFSE dilution along the X-axis.

Article Snippet: The CD4 T subpopulations of Tregs (CD4+, CD25high, CD127low), conventional effectors or Tconvs (CD4+, CD25+, CD127high), memory or Tmems (CD4+, CD25−, CD45RO+), and Tnaives (CD4+, CD25−, CD45RA+) were purified from patient PBMC directly by staining for CD4 T cell surface markers CD3 (PerCP), CD4 (FITC), CD25 (APC), CD127 (PE), CD45RA (V450), and CD45RO (Alexa Fluor 700), and sorting the stained cells on an ARIA cell sorter (BD Biosciences, San Jose, CA, USA) as shown in .

Techniques: Ex Vivo, Flow Cytometry, Expressing, Cell Culture

Journal: Virology

Article Title: Regulatory CD4 T cells inhibit HIV-1 expression of other CD4 T cell subsets via interactions with cell surface regulatory proteins

doi: 10.1016/j.virol.2017.12.036

Figure Lengend Snippet: Tregs inhibit HIV LTR transcription within Tregs themselves or in neighboring responding CD4 T cells. (A) Flow cytometry plots show that LTR transcriptional activity in in vitro expanded Tregs is lower than in in vitro expanded T subsets Tconvs and Tmems, and (C) bar plots show the means ± SD values of relative percentages after normalization to Tregs from three independent experiments. (B) Flow cytometry dot plots show that Tregs, but not Tconvs, inhibit LTR transcription in their respective co-cultured responding CD4 T cells; and the presences of antibodies against Tregs cell surface proteins (anti-CTLA-4, PD-1, and -GARP1) partially blocked the inhibition; and (D) the means ± SD values of relative percentages normalized to Tregs from three independent experiments are summarized in the bar plots.

Article Snippet: The CD4 T subpopulations of Tregs (CD4+, CD25high, CD127low), conventional effectors or Tconvs (CD4+, CD25+, CD127high), memory or Tmems (CD4+, CD25−, CD45RO+), and Tnaives (CD4+, CD25−, CD45RA+) were purified from patient PBMC directly by staining for CD4 T cell surface markers CD3 (PerCP), CD4 (FITC), CD25 (APC), CD127 (PE), CD45RA (V450), and CD45RO (Alexa Fluor 700), and sorting the stained cells on an ARIA cell sorter (BD Biosciences, San Jose, CA, USA) as shown in .

Techniques: Flow Cytometry, Activity Assay, In Vitro, Cell Culture, Inhibition

Journal: Journal of Oncology

Article Title: SFRP4 Is a Potential Biomarker for the Prognosis and Immunotherapy for Gastric Cancer

doi: 10.1155/2022/8829649

Figure Lengend Snippet: Clinicopathological characteristics of the patient cohort ( n = 137).

Article Snippet: Next, SFRP4, CD3 + T , CD4 + T , CD8 + T , and PD-L1 antibodies (SFRP4: Proteintech, 15328-1-AP, dilution ratio 1 : 300; CD3 + T : Abcam, ab16669, dilution ratio 1 : 200; CD4 + T : Abcam, ab133616, dilution ratio 1 : 200; CD8 + T : Abcam, ab17147, dilution ratio 1 : 200; PD-L1: DAKO/Agilent, SK006, dilution ratio 1 : 50) were added, incubated overnight at 4°C; then slides were washed for 5 minutes three times in PBS.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: SFRP4 Is a Potential Biomarker for the Prognosis and Immunotherapy for Gastric Cancer

doi: 10.1155/2022/8829649

Figure Lengend Snippet: Association of SFRP4 expression with PD-L1 and tumor-infiltrating lymphocytes in gastric cancer. (a) Association between PD-L1 expression and SFRP4 expression in gastric cancer. (b) Correlation between CD3 + T cells and SFRP4 expression in gastric cancer. (c) Association between CD4 + T cells and SFRP4 expression in gastric cancer. (d) Association between CD8 + T cells and SFRP4 expression in gastric cancer. (e) Correlation between SFRP4 expression and immune cells in gastric adenocarcinoma in the TIMER database (purity-corrected partial Spearman's rho value and statistical significance).

Article Snippet: Next, SFRP4, CD3 + T , CD4 + T , CD8 + T , and PD-L1 antibodies (SFRP4: Proteintech, 15328-1-AP, dilution ratio 1 : 300; CD3 + T : Abcam, ab16669, dilution ratio 1 : 200; CD4 + T : Abcam, ab133616, dilution ratio 1 : 200; CD8 + T : Abcam, ab17147, dilution ratio 1 : 200; PD-L1: DAKO/Agilent, SK006, dilution ratio 1 : 50) were added, incubated overnight at 4°C; then slides were washed for 5 minutes three times in PBS.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: SFRP4 Is a Potential Biomarker for the Prognosis and Immunotherapy for Gastric Cancer

doi: 10.1155/2022/8829649

Figure Lengend Snippet: In gastric cancer, SFRP4 is an independent prognostic factor, and SFRP4 paired with CD8 + T cells can better predict prognosis. (a) Kaplan-Meier survival curves for OS in tumor tissues based on SFRP4 expression. (b) Kaplan-Meier survival curves for OS based on tumor tissue PD-L1 expression. (c) Kaplan-Meier survival curves for OS in tumor tissues based on CD3+ T -cell expression. (d) Kaplan-Meier survival curves for OS in tumor tissues based on CD4+ T-cell expression. (e) Kaplan-Meier survival curves for OS in tumor tissues based on CD8+ T -cell expression. (f) Kaplan-Meier survival curves for OS based on SFRP4 expression in tumor tissues in combination with CD8 + T expression.

Article Snippet: Next, SFRP4, CD3 + T , CD4 + T , CD8 + T , and PD-L1 antibodies (SFRP4: Proteintech, 15328-1-AP, dilution ratio 1 : 300; CD3 + T : Abcam, ab16669, dilution ratio 1 : 200; CD4 + T : Abcam, ab133616, dilution ratio 1 : 200; CD8 + T : Abcam, ab17147, dilution ratio 1 : 200; PD-L1: DAKO/Agilent, SK006, dilution ratio 1 : 50) were added, incubated overnight at 4°C; then slides were washed for 5 minutes three times in PBS.

Techniques: Expressing

Journal: Journal of Oncology

Article Title: SFRP4 Is a Potential Biomarker for the Prognosis and Immunotherapy for Gastric Cancer

doi: 10.1155/2022/8829649

Figure Lengend Snippet: Univariate analysis of prognostic parameters for survival in gastric cancer patients.

Article Snippet: Next, SFRP4, CD3 + T , CD4 + T , CD8 + T , and PD-L1 antibodies (SFRP4: Proteintech, 15328-1-AP, dilution ratio 1 : 300; CD3 + T : Abcam, ab16669, dilution ratio 1 : 200; CD4 + T : Abcam, ab133616, dilution ratio 1 : 200; CD8 + T : Abcam, ab17147, dilution ratio 1 : 200; PD-L1: DAKO/Agilent, SK006, dilution ratio 1 : 50) were added, incubated overnight at 4°C; then slides were washed for 5 minutes three times in PBS.

Techniques: