Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: (A) Immunofluorescence of SKmel147 cells stably expressing AMIGO2-GFP (green), stained with AMIGO2 antibody (red) and Hoechst 33342 (blue). Scale bar, 20 μm. (B) Functional annotation of AMIGO2-interacting proteins detected by GFP pull-down followed by MS in SKmel147 cells stably expressing AMIGO2-GFP (see Table S4). (C) PTK7 and GFP immunoblots following GFP pull-down from 501MEL cells stably expressing AMIGO2-GFP. (D) Full-length PTK7 (FL-PTK7), C-terminal fragments CTF1- and CTF2-PTK7, and FOXM1 immunoblots of 501MEL cells 72 hr post-infection with shSCR or shPTK7 (shP7 #1 and #2). Actin was used as a loading control. (E) Relative growth curves of 501MEL (left) and SKmel147 (right) cells stably transduced with shSCR or shPTK7 (shP7 #1 and #2). Values are normalized to seeding control (n = 3). (F) Percent Annexin V-positive cells 6 days post-transduction for same cells as in (E). (G) FL-PTK7, CTF-PTK7, and FOXM1 immunoblots of 501MEL cells 48 hr post-transduction with shSCR or shAMIGO2 (shA2 #1 and #2). Actin was used as a loading control. (H) FL-PTK7, CTF-PTK7, FOXM1, and AMIGO2 immunoblots of 501MEL cells untreated or treated with JQ1 (JQ1[+]) for 72 hr. Tubulin was used as a loading control. (I) CTF2-PTK7 immunoblot of nuclear lysates from same cell as in (G) (left). Lamin B1 was used as loading control. Signal quantification (right), normalized to Lamin B1, relative to shSCR (n = 3). All values and error bars represent mean ± SD or ± SEM. See also Figures S3 and S4.

Article Snippet: LAMIN B1 , Santa Cruz , SC-6217.

Techniques: Immunofluorescence, Stable Transfection, Expressing, Staining, Functional Assay, Western Blot, Infection, Control, Transduction

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: LAMIN B1 , Santa Cruz , SC-6217.

Techniques: Microarray, Derivative Assay, Recombinant, Magnetic Beads, Flow Cytometry, Caspase Activity Assay, DNA Library Preparation, Blocking Assay, Extraction, TA Cloning, Sequencing, RNA Sequencing, Western Blot, Mass Spectrometry, Expressing, Software

Journal: Cell Reports Medicine

Article Title: Ligand-mediated PAI-1 inhibition in a mouse model of peritoneal carcinomatosis

doi: 10.1016/j.xcrm.2022.100526

Figure Lengend Snippet: Ascites in peritoneal carcinomatosis (PC) is biologically active (A) Kaplan-Meier curve showing survival of PC patients who underwent cytoreductive surgery (n = 121), stratified by the presence or absence of ascites. (B and C) Proliferation curves of (B) Colo-205 and (C) SNU-C1 cells treated with varying concentrations of ascites, measured by CellTitre-Glo assay. Data are represented as cell viability at day 5 relative to day 0. Dotted line represents cell viability of Colo-205 treated with 10% fetal bovine serum (FBS) at day 5 relative to day 0. Graph shows mean ± SD. (D and E) Migration of (D) Colo-205 and (E) SNU-C1 cells pre-treated with serum-free media (SFM), 10% FBS medium or 5% cell-free ascites (CFA) medium, assessed by transwell migration assay. Significant difference detected via unpaired two-sided t test between migration of cells pre-treated with SFM and 10% FBS or 5% CFA was denoted by ∗. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Significant difference detected via unpaired two-sided t test between migration of cells pre-treated with 10% FBS and SFM or 5% CFA was denoted by # . # p < 0.05, ## p < 0.01, ### p < 0.001. Graph shows mean ± SD. (F) Enriched signaling pathways in colorectal PC cell lines upon treatment with CFA, identified by comparing gene expressions of cells treated with 5% versus 0.1% CFA. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (G and H) Treatment of (G) Colo-205 and (H) SNU-C1 cells with 5% CFA activated STAT3 signaling pathway through phosphorylation at Tyr705.

Article Snippet: Total STAT3 ELISA , Cell Signalling Technology , 7305C.

Techniques: Glo Assay, Migration, Transwell Migration Assay

Journal: Cell Reports Medicine

Article Title: Ligand-mediated PAI-1 inhibition in a mouse model of peritoneal carcinomatosis

doi: 10.1016/j.xcrm.2022.100526

Figure Lengend Snippet: Identification and validation of prognostic putative activators of STAT3 signaling in ascites (A) Workflow to identify clinically significant putative secreted STAT3 activators. (B) Kaplan-Meier survival curve illustrating poorer survival in patients expressing three biomarkers as compared to patients expressing 0–2 biomarkers. (C) Characterization of cytokine profiles in colorectal PC ascites and benign ascites. Top 25% most highly abundant cytokines in colorectal PC ascites (by mean abundance) are shown. Heatmap displays Z score of normalized mean pixel density of duplicate cytokine spots on the array. (D) Downregulated signaling pathways upon PAI-1 inhibition in Colo-205 cells exposed to CFA with high PAI-1 levels (PC085), CFA with low PAI-1 levels (PC249) and no PAI-1 (FBS control), identified using RNA microarray. Only signaling pathways with differential downregulation are presented. IL6-JAK-STAT3 signaling pathway was significantly downregulated in high PAI-1 CFA-treated cells upon PAI-1 inhibition. Normalized enrichment scores <0 indicate pathway suppression and scores >0 indicate pathway activation. (D) is representative of two independent biological experiments. ∗p < 0.05.

Article Snippet: Total STAT3 ELISA , Cell Signalling Technology , 7305C.

Techniques: Expressing, Inhibition, Microarray, Activation Assay

Journal: Cell Reports Medicine

Article Title: Ligand-mediated PAI-1 inhibition in a mouse model of peritoneal carcinomatosis

doi: 10.1016/j.xcrm.2022.100526

Figure Lengend Snippet: Correlating PAI-1 level in ascites and intracellular STAT3 activation in cancer cells revealed distinct subgroups associated with differing susceptibility to PAI-1 inhibition (A) PAI-1 prevalence in colorectal PC ascites (n = 54). (B) Correlation between colorectal PC ascitic PAI-1 concentrations and ascites-treated Colo-205 cells p-STAT3(Y705) was determined by Pearson correlation coefficient test ( r = 0.4691, p = 0.0003). (C) PAI-1 prevalence in ascites of various histological PC subtypes (n = 156). † indicates benign ascites. (D) Correlation between various histological PC ascitic PAI-1 concentrations and ascites-treated Colo-205 cells p-STAT3(Y705) was determined by Pearson correlation coefficient test ( r = 0.6476, p < 0.0001). (A–D) PAI-1 concentrations are plotted on a log 2 scale to transform skewed data to normal distribution. p-STAT3(Y705) level was shown as optical density reading at 450 nm (OD450). (E) Untransformed values of PAI-1 and p-STAT3(Y705) levels from (D) were used for stratification strategy to identify patient subpopulations who might benefit from PAI-1 inhibition. Using 20 ng/mL PAI-1 level and 0.2 OD450 p-STAT3(Y705) level as cut-off values, three distinct subgroups of samples were observed: (i) high PAI-1 and high p-STAT3 levels, termed PAI-1 paracrine addicted (PPA) group (yellow region), (ii) low PAI-1 and high p-STAT3 levels, termed co-activators predominant (CAP) group (pink region) and (iii) low PAI-1 and low p-STAT3 levels, termed alternative pathways activation (APA) group (blue region). Each dot represents one patient ascites. Colors in each panel represent the various histological PC subtypes. All histological subtypes with less than five samples are grouped into other histological subtypes.

Article Snippet: Total STAT3 ELISA , Cell Signalling Technology , 7305C.

Techniques: Activation Assay, Inhibition

Journal: Cell Reports Medicine

Article Title: Ligand-mediated PAI-1 inhibition in a mouse model of peritoneal carcinomatosis

doi: 10.1016/j.xcrm.2022.100526

Figure Lengend Snippet: Cells dependent on PAI-1 to activate STAT3 are most susceptible to PAI-1 inhibition (A) Effect of TM5441 (PAI-1 inhibitor) on CFA-treated Colo-205 cells. Representative inhibitor dose-response curves of PPA group (yellow), CAP group (pink), APA group (blue) and FBS (control, black) demonstrated a left shift, indicating responsiveness to PAI-1 inhibition. (B) Differential sensitivity to TM5441 corresponding to the three subgroups, with PPA (n = 18) being the most sensitive to PAI-1 inhibition, followed by CAP (n = 59) and APA (n = 17). Graph shows mean ± SD. (C–E) Effect of (C) Napabucasin (STAT3 inhibitor), (D) BEZ235 (dual PI3K/mTOR inhibitor) and (E) Mitomycin C (conventional chemotherapeutic agent – DNA crosslinker) on the three subgroups of ascites-treated Colo-205 cells. Representative inhibitor dose-response curves of PPA group (yellow; n = 3), CAP group (pink; n = 3), APA group (blue; n = 1) and FBS (control, black). Targeting PAI-1, a dominant paracrine factor in ascites, was more effective than targeting downstream signaling pathway activated by ascites, proliferation pathway or DNA synthesis. (F) Evaluation of STAT3 suppression in Colo-205 cells treated with PPA CFA (PC085 and PC383), CAP CFA (PC249), APA CFA (PC010) and various concentrations of TM5441 by ELISA. STAT3 activation was shown as p-STAT3(Y705) and total STAT3 ratio at the indicated concentration relative to DMSO vehicle. Cells exposed to PPA CFA relied on PAI-1 to activate STAT3 as they required lower concentrations of TM5441 to suppress STAT3 activation. Graph shows mean ± SEM. Data in (A-E) are representative of at least three independent biological experiments and (F) is representative of two independent biological experiments. ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Total STAT3 ELISA , Cell Signalling Technology , 7305C.

Techniques: Inhibition, DNA Synthesis, Enzyme-linked Immunosorbent Assay, Activation Assay, Concentration Assay

Journal: Cell Reports Medicine

Article Title: Ligand-mediated PAI-1 inhibition in a mouse model of peritoneal carcinomatosis

doi: 10.1016/j.xcrm.2022.100526

Figure Lengend Snippet:

Article Snippet: Total STAT3 ELISA , Cell Signalling Technology , 7305C.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Bradford Protein Assay, Sequencing, Microarray, Cell Culture, Software

Journal: Scientific Reports

Article Title: Clinicopathological features and prognostic value of SOX11 in childhood acute lymphoblastic leukemia

doi: 10.1038/s41598-020-58970-z

Figure Lengend Snippet: SOX11 expression in hematological malignancies. (a) Expression boxplots of SOX11 in healthy cells, leukemias, and mantle cell lymphoma. Data sources: Hemap microarray dataset , GSE47051 , and the PanALL study . See Supplementary Information for more details. (b) SOX11 expression in ALL cell lines and healthy B- and T-cells, as measured by RT-qPCR (N = 2, black lines indicate the median). ALL, acute lymphoblastic leukemia; AML, acute myeloid leukemia; BCL2/MYC, BCL2/MYC rearranged; BCP-ALL, B-cell precursor ALL; CRLF2, CRLF2 (non-Ph-like); DUX4, DUX4 rearranged; E/R, ETV6-RUNX1; HeH, high hyperdiploid; HLF, TCF3/TCF4-HLF; HSC, hematopoietic stem cell; iAMP21, intrachromosomal amplification of chromosome 21; IKZF1 N159Y, IKZF1 missense alteration encoding p.Asn159Tyr; KMT2A, KMT2A rearranged; MCL, mantle cell lymphoma; MEF2D, MEF2D rearranged; NUTM1, NUTM1 rearranged; PAX5alt, PAX5 alterations; PAX5 P80R, PAX5 p.Pro80Arg (P80R) alteration; Ph, Philadelphia chromosome; T/P, TCF3-PBX1; ZNF384, ZNF384 rearranged. Reproduced with permission .

Article Snippet: The cells were then stained with Alexa Fluor 647® conjugated rabbit anti-human SOX11 antibody [EPR8191 (2)] (ab198540, Abcam), while CD3-APC antibody (345767) (Becton Dickinson, Franklin Lakes, NJ, USA) served as a negative control.

Techniques: Expressing, Microarray, Quantitative RT-PCR, Amplification

Journal: Scientific Reports

Article Title: Clinicopathological features and prognostic value of SOX11 in childhood acute lymphoblastic leukemia

doi: 10.1038/s41598-020-58970-z

Figure Lengend Snippet: SOX11 protein expression and impact on the outcome in BCP-ALL. (a) Expression of SOX11 protein by immunohistochemistry. Mantle cell lymphoma (MCL), a strongly positive case for SOX11 (400×); ALL1, a negative B-ALL case (400×); ALL2, a positive B-ALL case (400×); ALL3, a strongly positive B-ALL case (400×). (b) Kaplan–Meier survival curves and Log-rank p-values for OS, EFS, and RFS in the SOX11-positive (high) and -negative (low) groups. Reproduced with permission .

Article Snippet: The cells were then stained with Alexa Fluor 647® conjugated rabbit anti-human SOX11 antibody [EPR8191 (2)] (ab198540, Abcam), while CD3-APC antibody (345767) (Becton Dickinson, Franklin Lakes, NJ, USA) served as a negative control.

Techniques: Expressing, Immunohistochemistry

Journal: Scientific Reports

Article Title: Clinicopathological features and prognostic value of SOX11 in childhood acute lymphoblastic leukemia

doi: 10.1038/s41598-020-58970-z

Figure Lengend Snippet: Summary of SOX11 protein expression by immunohistochemical staining in B-ALL.

Article Snippet: The cells were then stained with Alexa Fluor 647® conjugated rabbit anti-human SOX11 antibody [EPR8191 (2)] (ab198540, Abcam), while CD3-APC antibody (345767) (Becton Dickinson, Franklin Lakes, NJ, USA) served as a negative control.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: Clinicopathological features and prognostic value of SOX11 in childhood acute lymphoblastic leukemia

doi: 10.1038/s41598-020-58970-z

Figure Lengend Snippet: Multivariate and univariate analyses of the event-free survival based on expression level of SOX11 protein in pediatric B-ALL.

Article Snippet: The cells were then stained with Alexa Fluor 647® conjugated rabbit anti-human SOX11 antibody [EPR8191 (2)] (ab198540, Abcam), while CD3-APC antibody (345767) (Becton Dickinson, Franklin Lakes, NJ, USA) served as a negative control.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Clinicopathological features and prognostic value of SOX11 in childhood acute lymphoblastic leukemia

doi: 10.1038/s41598-020-58970-z

Figure Lengend Snippet: Survival analysis and methylation status of cases with either a high or low SOX11 mRNA expression. (a) Kaplan–Meier survival curves and p-values of Log-rank test for EFS in patients with a low or high expression of SOX11 and in patients with a low or high methylation of the SOX11 gene locus. (b) CpG sites at the SOX11 locus in chromosome 2 . (c) DNA methylation at the SOX11 gene locus among patients with either a low or high expression of SOX11 . (d) Effect of decitabine treatment on SOX11 mRNA expression in the KOPN-8 cell line. Reproduced with permission .

Article Snippet: The cells were then stained with Alexa Fluor 647® conjugated rabbit anti-human SOX11 antibody [EPR8191 (2)] (ab198540, Abcam), while CD3-APC antibody (345767) (Becton Dickinson, Franklin Lakes, NJ, USA) served as a negative control.

Techniques: Methylation, Expressing, DNA Methylation Assay

Journal: Scientific Reports

Article Title: Clinicopathological features and prognostic value of SOX11 in childhood acute lymphoblastic leukemia

doi: 10.1038/s41598-020-58970-z

Figure Lengend Snippet: Cell viability and transcriptomic changes after knockdown of SOX11 . (a) Expression level of SOX11 after knockdown, as analyzed by RT-qPCR (N = 3, black lines indicate the median). (b) Knockdown of SOX11 caused no evident changes in cell viability. Curves are drawn from the biological replicates using the median value at given time points. 697 and RCH-ACV cells represent the T/P subtype, and REH cells represent the E/R subtype. SOX11 knockdown was confirmed by Western blotting, and cell viability assessments were conducted with the AlamarBlue assay. Measured absorbance intensities are reported as x10 3 (697 N = 6; RCH-ACV and REH N = 4). Western blot gel figures are cropped per cell line from the original blot figures shown in Fig. S3. (c) SOX11 knockdown in 697 cells caused downregulation of genes that are known to be altered after MYC upregulation. (d) Venn diagram of differentially expressed genes in the REH, 697, and RCH-ACV cell lines after knockdown of SOX11 (adjusted p-value < 0.05). (e) Heatmap of 15 concordantly differentially expressed genes in all three cell lines after knockdown of SOX11 . Reproduced with permission .

Article Snippet: The cells were then stained with Alexa Fluor 647® conjugated rabbit anti-human SOX11 antibody [EPR8191 (2)] (ab198540, Abcam), while CD3-APC antibody (345767) (Becton Dickinson, Franklin Lakes, NJ, USA) served as a negative control.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Alamar Blue Assay

Journal: Scientific Reports

Article Title: Clinicopathological features and prognostic value of SOX11 in childhood acute lymphoblastic leukemia

doi: 10.1038/s41598-020-58970-z

Figure Lengend Snippet: Detection of SOX11-positive cells by flow cytometry in BCP-ALL. (a) Correlation of CD5 expression with SOX11 mRNA expression in the E/R and T/P subtypes of B-ALL and MCL . SOX11 expression was categorized into three groups, not detected (ND), low and not detected (ND). (b) Flow cytometry analysis of nuclear SOX11 expression in cell lines with either a low (NALM-6, KOPN-8) or high (REH, RCH-ACV) expression of SOX11. ND, not detected, MCL, mantle cell lymphoma. Reproduced with permission .

Article Snippet: The cells were then stained with Alexa Fluor 647® conjugated rabbit anti-human SOX11 antibody [EPR8191 (2)] (ab198540, Abcam), while CD3-APC antibody (345767) (Becton Dickinson, Franklin Lakes, NJ, USA) served as a negative control.

Techniques: Flow Cytometry, Expressing

Journal: Cancer cell

Article Title: Genomic Subtypes of Non-Invasive Bladder Cancer with Distinct Metabolic Profile, Clinical Outcome and Female Gender Bias in KDM6A Mutation Frequency

doi: 10.1016/j.ccell.2017.08.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rat monoclonal anti-alpha-tubulin , AbD Serotec , Cat# MCA77G, RRID: AB_325003.

Techniques: Formalin-fixed Paraffin-Embedded, Recombinant, Hybridization, Staining, DNA Extraction, Cloning, Microarray, Derivative Assay, Software, Expressing, Variant Assay

Journal: International journal of cancer

Article Title: CD4 T cells target colorectal cancer antigens upregulated by oxaliplatin.

doi: 10.1002/ijc.32620

Figure Lengend Snippet: Figure 1. Analysis of antigen expression in oxaliplatin resistant colorectal cancer cells. Three oxaliplatin resistant colorectal cell lines were established by treatment with increasing concentration of oxaliplatin. (a) A growth-inhibition MTT assay was performed to determine the percentage of viable cells. Absorbance was measured at 550 nm after the treatment of oxaR CRC cell lines or wild type (WT) counterpart with increasing dose of oxaliplatin. Student t-test, *p < 0.05, **p < 0.01. (b) Cancer testis antigen expression was compared between oxaR cell lines and WT counterpart by microarray analysis. FC, fold change; N/A, not available. (c) The expression of panel of genes expressed in colorectal cancer cell lines: TERT, mesothelin, COA-1, spermatogenesis-associated serine rich 2 like (SPATS2L), Mucin, Ephrin A1, Sperm protein 17 (P17), p53 and prostate stem cell antigen (PSCA) was assessed by RT-PCR analysis. (d) Comparative analysis of coa-1, mesothelin and TERT mRNA expression was performed by RT-PCR between chemotherapy-resistant cell lines and WT counterpart.

Article Snippet: Monocyte-derived DC and T-cell clones coculture experiment Monocyte-derived DC were generated as previously described.25 To use tumor cell lysate as a source of COA-1 or mesothelin protein, COA-1 and mesothelin expression were induced in M113 cell line by transfecting either COA-1 (Invitrogen, Cergy-Pontoise, France) or mesothelin DNA plasmid (Origene, Herford, Germany).

Techniques: Expressing, Concentration Assay, Inhibition, MTT Assay, Microarray, Reverse Transcription Polymerase Chain Reaction

Journal: International journal of cancer

Article Title: CD4 T cells target colorectal cancer antigens upregulated by oxaliplatin.

doi: 10.1002/ijc.32620

Figure Lengend Snippet: Figure 4. Characterization of HLA-DR restricted TERT, COA-1 and mesothelin-specific immune responses in mCRC patients at baseline. PBMC from mCRC patients of Epitopes-CRC02 cohort at baseline (n = 162) were cultured either with a mixture of three COA-1 peptides (COA-172-86, COA-1331-345, COA-1445-459) and two mesothelin peptides (Meso366-380 and Meso523-537), or eight TERT peptides (TERT44-58, TERT578-592, TERT921-935, TERT1055-1069, TERT541-555, TERT573-587, TERT613-627, TERT911-925) at 5 μg/ml. The T-cell reactivity against the individual peptide was detected by IFN-γ ELISPOT assay as described in material and methods. Stimulation with CEF peptide was used as control of T cell reactivity. (a) Results are shown as median of ΔIFN-γ spots number (magnitude) and frequency. (b) Frequency of negative and positive immune response (IR) against peptides derived from one antigen (1IR) or more than two antigens (≥2IR) among TERT, COA-1 and mesothelin. Results were expressed in percentage of mCRC patients at baseline with immunological data available for all antigens (n = 125). (c) Repartition of positive immune responses against TERT, COA-1 and/or mesothelin. Results were expressed in percentage of responding patients (n = 73).

Article Snippet: Monocyte-derived DC and T-cell clones coculture experiment Monocyte-derived DC were generated as previously described.25 To use tumor cell lysate as a source of COA-1 or mesothelin protein, COA-1 and mesothelin expression were induced in M113 cell line by transfecting either COA-1 (Invitrogen, Cergy-Pontoise, France) or mesothelin DNA plasmid (Origene, Herford, Germany).

Techniques: Cell Culture, Enzyme-linked Immunospot, Control, Derivative Assay

Journal: International journal of cancer

Article Title: CD4 T cells target colorectal cancer antigens upregulated by oxaliplatin.

doi: 10.1002/ijc.32620

Figure Lengend Snippet: Figure 5. Comparative analysis of spontaneous HLA-DR-restricted T-cell responses against TERT, COA-1 and mesothelin peptides before and after oxaliplatin-based chemotherapy exposure. PBMC from oxaliplatin-treated mCRC patients at baseline and after oxaliplatin-based chemotherapy treatment (Epitopes-CRC02 cohort) were cultured either with a mixture of three COA-1 peptides (COA-172-86, COA-1331-345, COA-1445-459) and two mesothelin peptides (Meso366-380, Meso523-537), or eight TERT peptides (TERT44-58, TERT578-592, TERT921-935, TERT1055-1069, TERT541-555, TERT573-587, TERT613-627, TERT911-925). The presence of antigen-specific T cells was detected using IFN-γ ELISPOT assay. (a) Magnitude and frequency of spontaneous T-cell responses against TERT, COA-1, mesothelin or CEF peptides before (T0) and 3 months (T3) after the end of oxaliplatin-based chemotherapy in mCRC patients (Oxa). Immune responses of mCRC patients who were not exposed to oxaliplatin during 3 months were also evaluated (n = 44; No Oxa). (b) Variation of immune responses intensity directed against TERT, COA-1, mesothelin or CEF peptides after 3 months of oxaliplatin exposure (T3). In order to pool patient’s immune responses, the specific immune responses before oxaliplatin treatment were normalized to zero. (c) Impact of oxaliplatin exposure on antitumor immune responses in mCRC patients. Frequency of negative and positive immune response (IR) against peptides derived from one antigen (1IR) or more than two antigens (≥2IR) among TERT, COA-1 and mesothelin in chemotherapy-naive (left) and 3 months-oxaliplatin exposed (right) mCRC patients. Results were expressed in percentage of mCRC patients with immunological data available at time point T0 and T3 (n = 62).

Article Snippet: Monocyte-derived DC and T-cell clones coculture experiment Monocyte-derived DC were generated as previously described.25 To use tumor cell lysate as a source of COA-1 or mesothelin protein, COA-1 and mesothelin expression were induced in M113 cell line by transfecting either COA-1 (Invitrogen, Cergy-Pontoise, France) or mesothelin DNA plasmid (Origene, Herford, Germany).

Techniques: Cell Culture, Enzyme-linked Immunospot, Derivative Assay