Journal: International Journal of Molecular Sciences

Article Title: Intraclonal Enrichment of IL-23 Receptor Complex Expression in the Proliferative Fraction of Chronic Lymphocytic Leukemia

doi: 10.3390/ijms27031202

Figure Lengend Snippet: IL-23R and IL-12R expression in ex vivo CLL cells assessed by flow cytometry. FSC is proportional to cell size and SSC reflects intracellular complexity/granularity (each dot represents one cell). Axes in marker plots show fluorescence intensity for the indicated markers. Percentages within gates indicate the fraction of cells relative to the parent population. Lymphocytes were identified by FSC/SSC properties, followed by doublet exclusion using pulse height vs. width plots. Within the single-cell gate, CD19 + /CD5 + CLL cells were selected. This population was further sub-divided into RF and PF based on the differential expression of CD184 and CD5. ( A ) Gating strategy used to identify lymphocytes, single cells, CD19 + CLL cells, and to discriminate proliferative fraction (PF) and resting fraction (RF); ( B ) surface expression of IL-23R, IL-12Rβ1, and IL-12Rβ2 receptor subunits in ex vivo CLL cells, shown for CD19 + PF and RF populations; ( C ) representative flow cytometry plots showing sorting of PF and RF populations from six CLL samples according to the gates indicated; ( D ) IL-12Rβ1 mRNA expression in sorted RF and PF cells, measured by RT-qPCR. Data are shown as mean ± SEM. Statistical significance of the difference is evaluated using the two-sided Wilcoxon signed-rank test. * p < 0.05; ** p < 0.01, ns: not significant. FSC: forward scatter; SSC: side scatter; PE Phycoerythrin; PE-Cy7: Phycoerythrin–Cyanine 7; APC: Allophycocyanin; AF 488: Alexa Fluor 488; PE VIO 770: near-infrared fluorochrome emitting in the ~770–780 nm range.

Article Snippet: The following monoclonal antibodies were used: mouse anti human IL-23R-PE (Cat. #FAB14001P, R&D Systems, Minneapolis, MN, USA), mouse anti human IL-12Rβ1-BB515 (CD212, BD Hori-zon, Cat. #565043, BD Biosciences, San Jose, CA, USA, and mouse anti human IL-12Rβ2-PerCP (Cat. #FAB1959C, R&D Systems).

Techniques: Expressing, Ex Vivo, Flow Cytometry, Marker, Fluorescence, Single Cell, Quantitative Proteomics, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Intraclonal Enrichment of IL-23 Receptor Complex Expression in the Proliferative Fraction of Chronic Lymphocytic Leukemia

doi: 10.3390/ijms27031202

Figure Lengend Snippet: IL-23R and IL-12R expression in CLL cells after incubation with CpG and CpG + IL-15 for 72 h assessed by flow cytometry analyses. ( A ) Representative flow cytometry gating strategy used to identify CLL cells and to assess IL-23R and IL-12R complex expression under the indicated culture conditions (medium alone, CpG, or CpG + IL-15) after 72 h; ( B ) percentage of CLL cells expressing the IL-23R receptor subunit, IL-12Rβ1 receptor subunit, and IL-12Rβ2 receptor subunit following 72 h incubation under the indicated conditions; ( C ) percentage of CLL cells expressing the IL-23R complex and the IL-12R complex following 72 h incubation under the indicated conditions. Statistical significance of the difference is evaluated using the two-sided Wilcoxon signed-rank test. * p < 0.05; **** p < 0.0001. FSC: forward scatter; SSC: side scatter; PE Phycoerythrin; PE-Cy7: Phycoerythrin–Cyanine 7; APC: Allophycocyanin; AF 488: Alexa Fluor 488.

Article Snippet: The following monoclonal antibodies were used: mouse anti human IL-23R-PE (Cat. #FAB14001P, R&D Systems, Minneapolis, MN, USA), mouse anti human IL-12Rβ1-BB515 (CD212, BD Hori-zon, Cat. #565043, BD Biosciences, San Jose, CA, USA, and mouse anti human IL-12Rβ2-PerCP (Cat. #FAB1959C, R&D Systems).

Techniques: Expressing, Incubation, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Intraclonal Enrichment of IL-23 Receptor Complex Expression in the Proliferative Fraction of Chronic Lymphocytic Leukemia

doi: 10.3390/ijms27031202

Figure Lengend Snippet: Differential expression of IL-23R and IL-12R in CLL RF and PF subsets after stimulation with CpG and CpG + IL-15 assessed by flow cytometry analyses. ( A ) Representative flow cytometry gating strategy used to identify CLL cells and to distinguish RF and PF subsets, followed by assessment of IL-23R and IL-12R complex expression after 72 h incubation with CpG or CpG + IL-15; ( B ) percentage of RF and PF CLL cells expressing the IL-23R receptor subunit, IL-12Rβ1 receptor subunit, and IL-12Rβ2 receptor subunit after 72 h incubation under the indicated conditions; ( C ) percentage of RF and PF CLL cells expressing the IL-23R complex and the IL-12R complex after 72 h incubation under the indicated conditions. Each symbol represents an individual CLL sample; horizontal bars indicate mean ± SEM. Statistical significance of the difference is evaluated using the two-sided Wilcoxon signed-rank test. ** p < 0.01; **** p < 0.0001, ns: not significant. FSC: forward scatter; SSC: side scatter; PE Phycoerythrin; PE-Cy7: Phycoerythrin–Cyanine 7; APC: Allophycocyanin; AF 488: Alexa Fluor 488; BV421 = Brilliant Violet 421.

Article Snippet: The following monoclonal antibodies were used: mouse anti human IL-23R-PE (Cat. #FAB14001P, R&D Systems, Minneapolis, MN, USA), mouse anti human IL-12Rβ1-BB515 (CD212, BD Hori-zon, Cat. #565043, BD Biosciences, San Jose, CA, USA, and mouse anti human IL-12Rβ2-PerCP (Cat. #FAB1959C, R&D Systems).

Techniques: Quantitative Proteomics, Flow Cytometry, Expressing, Incubation

Journal: Cell Reports Medicine

Article Title: Genetic and pharmaceutical targeting of HIF1α allows combo-immunotherapy to boost graft vs. leukemia without exacerbation graft vs. host disease

doi: 10.1016/j.xcrm.2023.101236

Figure Lengend Snippet:

Article Snippet: PerCP conjugated anti-human HIF1α , RD Systems , IC1935C.

Techniques: Recombinant, Software

Journal: Immunity

Article Title: Differentiation and protective capacity of virus-specific CD8 + T cells suggest murine norovirus persistence in an immune-privileged enteric niche

doi: 10.1016/j.immuni.2017.09.017

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Dead cells were removed by gating on a LIVE/DEAD Aqua kit (Invitrogen, Carlsbad, CA) versus forward scatter (FSC-H). table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti CD8β-PerCP-Cy5.5 (clone YTS156.7.7) Biolegend Cat# 126609 Anti PD-1-PE-Cy7 (clone RMP1-30) Biolegend Cat# 109109 Anti CD103-PacificBlue (clone 2E7) Biolegend Cat# 121417 Anti CD44-BV785 (clone IM7) Biolegend Cat# 103041 Anti CD45.2-BV785 (clone 104) Biolegend Cat# 109839 Anti CXCR4-BV421(clone L276F12) Biolegend Cat# 146511 Anti Ly6c-BV785 (clone HK1.4) Biolegend Cat# 128041 Anti KLRG-1-BV605 (clone 2F1) Biolegend Cat# 138419 Anti CD62L-BV605 (clone MEI-14) Biolegend Cat# 104438 Anti TNFa-PacificBlue (clone MP6-XT22) Biolegend Cat# 506318 Anti CD107a-AF488 (clone 1D4B) Biolegend Cat# 121607 Anti CD28-PE-Cy7 (clone 37.51) Biolegend Cat# 102126 Anti CX3CR1-PE (clone SA011F11) Biolegend Cat# 149006 Anti CD11a-PE (clone 2D7) Biolegend Cat# 101008 Anti FOXP3-PE (clone MF-14) Biolegend Cat# 126404 Anti CD90.1 (Thy-1.1)-BV605 (clone OX-7) Biolegend Cat# 202537 Anti CD25-PE-Cy5 (clone PC61) Biolegend Cat# 102010 Anti CD8a-PE-eF610 (clone 53.6) eBioscience Cat# 61-0081-80 Anti CD69-PE-Cy7 (clone H1.2F3) eBioscience Cat# 25-0691-82 Anti 2B4-FITC (clone eBio244F4) eBioscience Cat# 11-2441-82 Anti CD49b-PE-Cy5 (clone DX5) eBioscience Cat# 15-5971-82 Anti CD49d-FITC (clone R1-2) eBioscience Cat# 11-0492-81 Anti CD90.2 (Thy1.2)-FITC (clone 30-H12) eBioscience Cat# 11-0903-82 Anti CD27-APC-eFluor 780 (clone LG.7F9) eBioscience Cat# 47-0271-82 Anti CD45.1-APC-R700 (clone A20) BD Biosciences Cat# 565814 Anti IFN -AlexaFluor700 (clone XMG1.2) BD Biosciences Cat# 557998 Anti GITR-FITC (clone DTA-1) BD Biosciences Cat# 558139 Anti Granzyme-B-PE (clone GB11) Invitrogen Cat# GRB04 Anti MIP-1a-APC (clone 39624) R&D Systems Cat# 1C450A Anti IL-12Rβ2-PRCP (clone 305719) R&D Systems Cat# FAB1959C Anti-CD8β (Lyt 3.2) (clone 53-5.8) Bio X Cell Cat # BE0223 Anti-Thy1.1 (clone 19E12) Bio X Cell Cat # BE0214 Bacterial and Virus Strains MNV-CW3 Tomov et al., 2013 GenBank EF014462 MNV-CR6 Tomov et al., 2013 GenBank EU004676 MNV-CW3 Y→F This study N/A MNV-CR6 F→Y This study N/A rVV 519Y This study N/A Biological Samples N/A N/A N/A Chemicals, Peptides, and Recombinant Proteins Deoxyribonuclease I from Bovine Pancreas Sigma-Aldrich Cat # D5025 Collagenase/Dispase Roche Cat # 11097113001 DTT (dithiothreitol) Thermo Fisher Cat # R0861 RPMI 1640 Corning Cat # 15-040-CM GemCell U.S.

Techniques: Virus, Recombinant, Saline, Reverse Transcription, Gene Expression, SYBR Green Assay, Isolation, Staining, Microarray, Plasmid Preparation, Software

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Myeloma-specific multiple peptides able to generate cytotoxic T lymphocytes: A potential therapeutic application in multiple myeloma and other plasma cell disorders

doi: 10.1158/1078-0432.CCR-11-2776

Figure Lengend Snippet: MP-CTL generated with a cocktail of XBP1 US, XBP1 SP, CD138, and CS1 peptides were evaluated for their respective peptide-specific responses. Specific activities of MP-CTL were measured in response to the individual peptide pulsed APC by detecting the increase of CD107a+ or IFN-γ+ cells.

Article Snippet: Reagents Mouse anti-human CD3, CD4, CD8, CCR7, CD45RO, CD69, CD107a, IFN-γ, and HLA-A2 mAbs conjugated with FITC, PE, PerCP, PerCP-Cy5.5, APC, Pacific Blue, APC-H7 or PE-Cy7 were purchased from Becton Dickinson (BD)/Pharmingen or BD/Biosciences (San Diego, CA).

Techniques: Generated