Journal: Journal of experimental & clinical cancer research : CR

Article Title: Combinatorial macrophage induced innate immunotherapy against Ewing sarcoma: Turning "Two Keys" simultaneously.

doi: 10.1186/s13046-024-03093-w

Figure Lengend Snippet: Fig. 5 Characterization of markers for ES-phagocytic macrophages. A Gating strategy for evaluating cell surface protein expression on phagocytic versus non-phagocytic macrophages. In-vitro phagocytosis assay was carried out incubating A673 cells (Non- or DOX- or MAG- or D&M-treated) labeled with CMFDA (detected by FITC channel) and M-CSF Mφ. Cells were harvested and stained with APC-CD11b and PE-CD38 antibodies followed by flow cytometry analysis. The viable cells were plotted in FITC (CMFDA) x APC view to visualize cell populations in the in-vitro phagocytosis assay. A gate was drawn to include cells that were FITC positive and APC positive as the phagocytic macrophage population, and another gate was drawn to include cells that were FITC- APC + as the non-phagocytic macrophage population. The FITC + APC + and FITC- APC + cells were then plotted separately in a PE x APC view to visualize expression of CD38 in both cell populations. A quat gate was drawn to visualize APC + PE + as the CD38 + and APC + PE- as the CD38- macrophages. B Comparison of percent of CD14, CD38, CD163, CD206 or CD209 positive and negative cells in the phagocytic and non-phagocytic macrophages incubated with non- or DOX- or MAG- or D&M-treated A673 cells. Macrophages alone (no A673) served as a control. *p < 0.05, **p < 0.01, ***p < 0.001 (Fisher’s Exact test). Shown are results in a representative experiment. The same trend was seen in three independent biological repeats

Article Snippet: Antibodies used include α-CD47 (B6H12, ThermoFisher Scientific, 11–0479-42), α-Calreticulin (ThermoFisher Scientific, PA3-900), α-CD11b (Miltenyi Biotec, 130–110-554), α-CD14 (R&D Systems, FAB3832P-025), α-CD38 (R&D Systems, FAB2404P), α-CD68 (BioLegend, 333,819), α-CD80 (BioLegend, 305,220), α-CD163 (R&D Systems, FAB1607P-025), α-CD206 (R&D Systems, FAB25342P), α-CD209 (R&D Systems, FAB161P-025), Alexa Fluor 488 donkey α-rabbit IgG (Invitrogen, A21206).

Techniques: Expressing, In Vitro, Phagocytosis Assay, Labeling, Staining, Flow Cytometry, Comparison, Incubation, Control

Journal: Cells

Article Title: The Properties of Proinflammatory Ly6C hi Monocytes Are Differentially Shaped by Parasitic and Bacterial Liver Infections.

doi: 10.3390/cells11162539

Figure Lengend Snippet: Figure 4. Marked differences in expression of surface markers by Ly6Chi monocytes after infection with E. histolytica or L. monocytogenes (A) Heatmap depicting differential expression of mRNA encod- ing selected markers on the surface of liver Ly6Chi monocytes after infection with E. histolytica (E. his) or L. monocytogenes (L. m). (B) Normalized mRNA counts of selected surface marker genes (from transcriptome analysis). (C) Percentage of CD14+, CD38+, CD74+, and CD86+ Ly6Chi monocytes dur- ing the course of infection at the indicated time points post-infection (measured by flow cytometry). (D) Histogram and MFI of CD14+, CD38+, CD74+, and CD86+ Ly6Chi monocytes during the course of infection. Data in C were pooled from three independent experiments. Data in D are representative of one of these three experiments and all data are presented as the mean ± SEM (* p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001; Mann-Whitney U test).

Article Snippet: The following antibodies were used: CD45.1 BUV395 (A20) (Becton Dickinson); CD11b BV510 (M1/70), CD14 BV421 (1Sa142), CD38 PeCy7 (90), CD45.2 PeCy5 (30-F11), CD74 AF647 (In1/CD74), CD86 AF700 (GL-1), Ly6C FITC/PE (HK1-4), Ly6G BV785 (1A8) (BioLegend); IRF2 AF488 (sc-374327) and mafb PerCP (OTI1E9) (Novus Biologicals).

Techniques: Expressing, Infection, Quantitative Proteomics, Marker, Cytometry, MANN-WHITNEY

Journal: Cells

Article Title: The Properties of Proinflammatory Ly6C hi Monocytes Are Differentially Shaped by Parasitic and Bacterial Liver Infections.

doi: 10.3390/cells11162539

Figure Lengend Snippet: Figure 5. New Ly6Chi monocyte subsets identified according to expression of selected surface markers and IRF2. (A) Gating strategy, histogram, and percentage of IRF2+ Ly6Chi monocytes from livers of naïve mice and from E. histolytica- and L. monocytogenes-infected mice on day 3 post infecion. (B) Gating strategy (exemplary for CD14+ gated cells) based on the FMO control to determine CD14+, CD38+, CD74+, and CD86+ monocytes within the IRF2+ Ly6Chi monocyte population from naïve mice and from both infection models on day 3 post infection. (C) UMAP plots of Ly6Chi

Article Snippet: The following antibodies were used: CD45.1 BUV395 (A20) (Becton Dickinson); CD11b BV510 (M1/70), CD14 BV421 (1Sa142), CD38 PeCy7 (90), CD45.2 PeCy5 (30-F11), CD74 AF647 (In1/CD74), CD86 AF700 (GL-1), Ly6C FITC/PE (HK1-4), Ly6G BV785 (1A8) (BioLegend); IRF2 AF488 (sc-374327) and mafb PerCP (OTI1E9) (Novus Biologicals).

Techniques: Expressing, Infection, Control

Journal: Cells

Article Title: The Properties of Proinflammatory Ly6C hi Monocytes Are Differentially Shaped by Parasitic and Bacterial Liver Infections.

doi: 10.3390/cells11162539

Figure Lengend Snippet: Figure 6. CD38+Ly6Chi monocytes represent the major monocytic source of ROS production and contribute to liver damage following E. histolytica infection. (A) Heat map of differentially regulated genes involved in ROS production and a graph showing fold changes in expression of Ncf1 mRNA in the liver during infection with E. histolytica or L. monocytogenes. (B) Gating scheme and histogram for ROS+ liver Ly6Chi monocytes. Percentage and mean fluorescent intensity (MFI) of ROS+Ly6Chi

Article Snippet: The following antibodies were used: CD45.1 BUV395 (A20) (Becton Dickinson); CD11b BV510 (M1/70), CD14 BV421 (1Sa142), CD38 PeCy7 (90), CD45.2 PeCy5 (30-F11), CD74 AF647 (In1/CD74), CD86 AF700 (GL-1), Ly6C FITC/PE (HK1-4), Ly6G BV785 (1A8) (BioLegend); IRF2 AF488 (sc-374327) and mafb PerCP (OTI1E9) (Novus Biologicals).

Techniques: Infection, Expressing

Journal: Molecular Neurobiology

Article Title: CD38 Coordinates with NF-κB to Promote Cochlear Inflammation in Noise-Induced Hearing Loss: the Protective Effect of Apigenin

doi: 10.1007/s12035-024-04675-7

Figure Lengend Snippet: Expression of p65 and CD38. A Nuclear transport of p65. In the control group (Ctrl), p65 was labeled around the nucleus of outer hair cell (white arrow). In the NE group (NE), p65 transferred into the nucleus (white arrow). B Immunofluorescence co-staining of CD38 and F4/80. Macrophages were labeled with Anti-F4/80 antibody (green). CD38 (red) was mainly expressed in basilar membrane macrophages (blue arrow). C CD38 protein expression in the control group (Ctrl) and the NE group (NE). Protein bands showed that CD38 protein expression in the NE group is higher than that in the control group. *Compared with control group, P < 0.05, n = 6 animals, 12 cochleae

Article Snippet: The cochlear basilar membrane was prepared with the surface preparation method, antibodies used in this experiment were rabbit anti-NF-κB p65 polyclonal antibody (Abcam, ab32536), rat anti-F4/80 polyclonal antibody (Abcam, ab6640), and mouse anti-CD38 monoclonal antibody (Proteintech, 60,006–1-lg).

Techniques: Expressing, Control, Labeling, Immunofluorescence, Staining, Membrane

Journal: Molecular Neurobiology

Article Title: CD38 Coordinates with NF-κB to Promote Cochlear Inflammation in Noise-Induced Hearing Loss: the Protective Effect of Apigenin

doi: 10.1007/s12035-024-04675-7

Figure Lengend Snippet: Western blot results of p65 ( A ) and CD38 ( B ) expression in the control group (Ctrl), the NE group (NE), the NE + Api group and the NE + Ana group. Relative protein content is shown in the histogram. #Compared with the control group, P < 0.05; ##compared with the control group, P < 0.01; *compared with NE group, P < 0.05

Article Snippet: The cochlear basilar membrane was prepared with the surface preparation method, antibodies used in this experiment were rabbit anti-NF-κB p65 polyclonal antibody (Abcam, ab32536), rat anti-F4/80 polyclonal antibody (Abcam, ab6640), and mouse anti-CD38 monoclonal antibody (Proteintech, 60,006–1-lg).

Techniques: Western Blot, Expressing, Control

Journal: Molecular Neurobiology

Article Title: CD38 Coordinates with NF-κB to Promote Cochlear Inflammation in Noise-Induced Hearing Loss: the Protective Effect of Apigenin

doi: 10.1007/s12035-024-04675-7

Figure Lengend Snippet: CD38 coordinates with NF-κB to promote cochlear inflammation in NIHL. As the closest immune cell to organ of Corti, activated macrophages can express CD38 and consume NAD in organ of Corti, causing NAD + metabolism dysfunction and ultimately leads to cochlear inflammation. In this process, inflammatory factors such as IL-1, IL-6, and TNF-α promoted the activation of NF-κB signaling pathway, forming a positive feedback cycle

Article Snippet: The cochlear basilar membrane was prepared with the surface preparation method, antibodies used in this experiment were rabbit anti-NF-κB p65 polyclonal antibody (Abcam, ab32536), rat anti-F4/80 polyclonal antibody (Abcam, ab6640), and mouse anti-CD38 monoclonal antibody (Proteintech, 60,006–1-lg).

Techniques: Activation Assay

Journal: iScience

Article Title: SFTSV infection is associated with transient overproliferation of monoclonal lambda-type plasma cells

doi: 10.1016/j.isci.2023.106799

Figure Lengend Snippet: Characterization of plasma cell subsets in the marrow from SFTS and LMM patients (A) FACS gating strategy for measurement of plasma cell subsets. Plasma cells (CD38 + CD138 + ) and monoclonal plasma cells with cytoplasmic lambda light chain (CD38 + cLambda + ) and (CD38 + cKappa − ). (B–D) The frequency of different plasma cell subsets from LMM and SFTS bone marrow lymphocytes. (E) The ratio of CD38 + cLambda + /CD38 + cKappa + in total cell in three groups. LMM: lambda light chain multiple myeloma; MCP: SFTS patients with monoclonal plasma cells; NP: SFTS patients with normal plasma cells. The median (IQR) is shown for each group, and the differences between two groups were tested by t test for normally distributed data or Mann-Whitney test for non-normally distributed variables. Owing to the frequency of CD38 within the normal range (<0.5%), plasma cell phenotypes were not further detected, and 9 from 22 SFTS cases with normal plasma cells were used for analysis in the NP group. Statistical significance was represented by asterisks, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Anti-human CD38 , Proteintech , Cat# clone 3C6G4.

Techniques: Clinical Proteomics, MANN-WHITNEY

Journal: iScience

Article Title: SFTSV infection is associated with transient overproliferation of monoclonal lambda-type plasma cells

doi: 10.1016/j.isci.2023.106799

Figure Lengend Snippet: The viral loads and antibody responses to the SFTSV nucleocapsid protein in both plasma and marrow liquid (A and B) The NP-specific IgM and IgG antibody titers in plasma among MCP and NP groups were diluted in 1:10 and 1:100, and OD 450 measurements were determined by a commercial ELISA kit. (C) The neutralization titers for different time points for the representative case 4 were recorded by real-time quantitative RT-PCR. The red, brown, and green curves represented collection date 2020.5.20 diluted 1: 20, collection date 2020.7.9 diluted 1: 20, and collection date 2020.7.9 diluted 1: 40, respectively. (D) Neutralization antibodies in acute or recovery phase plasma in paired samples were tested by 2-fold dilutions in the range 1:20-640, and paired t test was used for statistical analysis. The red and blue lines represent the MCP and NP groups, respectively. (E) Plasma viral loads of each acute phase patient were determined by real-time quantitative RT-PCR, and cycle threshold ( Ct ) values between the MCP and NP groups were compared. (F) Spearman’s correlation coefficients were calculated between plasma viral load ( Ct value), and the ratio of CD38 + cLambda + versus CD38 + cKappa + using viremic samples. X axis denotes the Ct value in plasma. r s and P indicate the correlation coefficient and significance, respectively. (G) Viral loads in the bone marrow lymphocytes of each acute phase patient were determined by real-time quantitative RT-PCR, and cycle threshold ( Ct ) values between the MCP and NP groups were compared. (H and I) Correlation of NP-specific IgM and IgG antibody levels between plasma and bone liquid. The differences between two groups were tested by Mann-Whitney test or t test, as appropriate. r s indicates the Spearman correlation coefficient. Statistical significance was represented by an asterisk, ∗p < 0.05; ∗∗p < 0.01; ns, no significance.

Article Snippet: Anti-human CD38 , Proteintech , Cat# clone 3C6G4.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Neutralization, Quantitative RT-PCR, MANN-WHITNEY

Journal: iScience

Article Title: SFTSV infection is associated with transient overproliferation of monoclonal lambda-type plasma cells

doi: 10.1016/j.isci.2023.106799

Figure Lengend Snippet: SFTSV infection induces transient overproliferation of MCP cells (A–C) The changes of different plasma subsets (CD38 + CD138 + , CD38 + cLambda + , and CD38 + cKappa + ) in both the acute and recovery phases in SFTS cases. The changes of different plasma subsets in both the acute and recovery phases for the representative MCP case 6 are shown, and paired t tests were employed to assess the significance. (D) The ratio of CD38 + cLambda + /CD38 + cKappa in the acute and recovery phases was compared, and paired t test was used to assess the significance. The green shade represents the normal region. Statistical significance was represented by asterisks, ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Anti-human CD38 , Proteintech , Cat# clone 3C6G4.

Techniques: Infection, Clinical Proteomics

Journal: iScience

Article Title: SFTSV infection is associated with transient overproliferation of monoclonal lambda-type plasma cells

doi: 10.1016/j.isci.2023.106799

Figure Lengend Snippet: Monoclonal plasma cells overexpressing lambda light chain in different severe interferon receptor-deficient mouse bone marrow (A) FACS gating strategy for measurements of plasma cell subsets in mice experiments. (B) The frequency of CD38 + CD138 + in different infection groups. (C and D) The frequency of CD38 + cLambda + and CD38 + cKappa + plasma cells in bone marrow by intracellular staining, respectively. (E) The ratio of CD38 + cLambda + /CD38 + cKappa in CD45 + lymphocytes in different infection groups. (F–I) The changes in different plasma cell subsets from 3 dpi to the 5 dpi in 10 PFU infection dose. Brown-Forsythe ANOVA tests with Holm-Sidak’s multiple comparisons test were used for the analyses, and the differences between two groups were tested by t test. Sample data collected for phosphate-buffered saline (PBS), 0.08, 0.4, 2, and 10 PFU in 5 dpi. The median (IQR) is shown for each group, and statistical significance is represented by asterisks, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; no significance was not shown for clear picture.

Article Snippet: Anti-human CD38 , Proteintech , Cat# clone 3C6G4.

Techniques: Clinical Proteomics, Infection, Staining, Saline

Journal: iScience

Article Title: SFTSV infection is associated with transient overproliferation of monoclonal lambda-type plasma cells

doi: 10.1016/j.isci.2023.106799

Figure Lengend Snippet:

Article Snippet: Anti-human CD38 , Proteintech , Cat# clone 3C6G4.

Techniques: Virus, Purification, Enzyme-linked Immunosorbent Assay, Modification, Lysis, Sequencing, Software

Journal: BMC immunology

Article Title: Cycling and activated CD8 + T lymphocytes and their association with disease severity in influenza patients.

doi: 10.1186/s12865-022-00516-1

Figure Lengend Snippet: Fig. 1 The percentage of cycling memory CD8+ and CD4+ T cells increased with disease severity of influenza infection. A, B Percentages of cycling CD8+ T cells, cycling CD8+ memory and effector T cells in healthy donors (HD, n = 19), moderate patients (M, n = 34) and severe patients (S, n = 27). C, D Percentages of cycling CD4+ T cells, cycling CD4+ memory and effector T cells in healthy donors and influenza patients. Data are displayed as mean ± SEM. Statistical significance is determined by unpaired t test. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The PBMCs were thawed and analyzed using a panel of antibodies as follows: PE-eFluor 610 anti-human Ki67 (Thermo/eBio); FITC anti-human CD8α (clone Hit8α), PE anti-human CD4 (clone RPA-T4), PE-Cyanine7 antihuman CD38 (clone HB7), violetFluorTM450 anti-human HLA-DR (clone LN3), APC-Cyanine7 anti-human CD27 (clone O323), APC anti-human CD279 (PD-1) (clone J110), PerCP-Cyanine5.5 anti-human CD45RO (clone UCHL1), redFluorTM 710 anti-human CD45RA (clone HI100) (TONBO biosciences).

Techniques: Infection

Journal: BMC immunology

Article Title: Cycling and activated CD8 + T lymphocytes and their association with disease severity in influenza patients.

doi: 10.1186/s12865-022-00516-1

Figure Lengend Snippet: Fig. 3 The kinetic changes of cycling and activated T cells during influenza infection were longitudinally characterized. Kinetics of cycling and activated T cells are shown as frequencies, A %Ki67+CD8+ T cells of total CD8+ T cells, B %Ki67+CD4+ T cells of total CD4+ T cells. C %CD38+HLA-DR+CD8+ T cells of total CD8+ T cells, D %CD38+HLA-DR+CD4+ T cells of total CD4+ T cells during the disease course. Statistical significance is determined by unpaired t test. *P < 0.05, **P < 0.01, ns, not significant

Article Snippet: The PBMCs were thawed and analyzed using a panel of antibodies as follows: PE-eFluor 610 anti-human Ki67 (Thermo/eBio); FITC anti-human CD8α (clone Hit8α), PE anti-human CD4 (clone RPA-T4), PE-Cyanine7 antihuman CD38 (clone HB7), violetFluorTM450 anti-human HLA-DR (clone LN3), APC-Cyanine7 anti-human CD27 (clone O323), APC anti-human CD279 (PD-1) (clone J110), PerCP-Cyanine5.5 anti-human CD45RO (clone UCHL1), redFluorTM 710 anti-human CD45RA (clone HI100) (TONBO biosciences).

Techniques: Infection