Journal: Translational Oncology

Article Title: Vaccine-induced ErbB (EGFR/HER2)-specific immunity in spontaneous canine cancer

doi: 10.1016/j.tranon.2021.101205

Figure Lengend Snippet: cEGFR p527 immunization mediates CD8 infiltration and antibody deposition into canine tumors. Postmortem bladder cancer tissue from a cEGFR p527 immune dog and tissue from healthy, non-cancerous bladder was examined for infiltration of CD8 T cells and endogenous antibody. A. CD8 isotype control. B. CD8 T cell staining (green). Arrows illustrate presence of CD8+ cells within the tumor tissue. C. & D. Antibody deposition in canine bladder tumor tissue from a cEGFR p527 immunized dog (two different fields). E. & F. Canine IgG deposition in normal canine bladder tissue (two different fields). Magnification is 40x (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.).

Article Snippet: CD8 T cells were stained in tissues using mouse anti-dog CD8 (MAB6709; R&D Systems, Minneapolis, MN), followed by goat anti-mouse IgG ( H + L ) Alexa Fluor 488 (A11017; Life Technologies, Eugene, OR) and counterstaining with DAPI (Molecular Probes; Eugene, OR).

Techniques: Control, Staining

Journal: Science China. Life sciences

Article Title: Total body irradiation of donors can alter the course of tolerance and induce acute rejection in a spontaneous tolerance rat liver transplantation model.

doi: 10.1007/s11427-012-4370-3

Figure Lengend Snippet: Figure 3 Detection of T cell subpopulations in the peripheral blood and CTL-mediated lytic activity. A, There was no difference in the percentage of CD3+ T and CD3+CD4+ T cells between the transplant groups. The number of CD3+CD8+ T cells in the acute rejection group increased significantly (P<0.05), while the number of CD4+/CD8+ cells decreased significantly compared to the other groups (P<0.05). B, The ratio of Foxp3+CD4+CD25+ Tregs and the expression of CD4+CD25+ T cells in the acute rejection group was significantly lower than those in the other groups (P<0.05). C, In the control group, GITR expression was not detectable on T cells. GITR expression on increased T cells from all groups after culture with phorbol ester and ionomycin. The GITR expression level of T cells in the acute rejection group was significantly higher than that in the other groups (P<0.05). D, The spleen CTL-mediated lytic activity of the recipients in the acute rejection group increased by 32.6%, and was significantly higher than those in the other three groups (P<0.05).

Article Snippet: A 50 μL sample of the whole blood obtained after stimulation was incubated with 10 μL of anti-CD3-PerCp and 5 μL of anti-CD8-APC-Cy7 mAbs in the dark at 4°C for 30 min. After haematolysis and washing with PBS, the resuspended cells were incubated in the dark at 4°C for 45 min with 10 μL anti-rat GITR-PE mAb (R&D Systems, Minneapolis, MN, USA) or 10 μL IgG1-PE (Beckman Coulter Inc) as an isotype-control.

Techniques: Activity Assay, Expressing, Control

Journal: Nature communications

Article Title: Monoacylglycerol lipase regulates cannabinoid receptor 2-dependent macrophage activation and cancer progression.

doi: 10.1038/s41467-018-04999-8

Figure Lengend Snippet: Fig. 2 Macrophage MGLL inhibits tumor progression via CD8+T cells. a Six-week-old WT and TgMGLL mice were subcutaneously inoculated with MC-38 cells, and tumor volumes were measured dynamically. This test was performed four times. (n = 10) b The survival time of the MC-38 tumor-bearing mice described in a. The data represent the means ± s.e.ms. (n = 10, ***P < 0.005, Gehan-Breslow-Wilcoxon test). c The growth curves of the subcutaneous MC-38 tumors in Myeloid-mgll-KO mice and the control littermates (fl/fl). (n = 6). d The survival time of the MC-38 tumor-bearing mice described in c. The data represent the means ± s.e.ms. (n = 6, **P < 0.01, Gehan-Breslow-Wilcoxon test). e Incidence of metastasis in lungs and livers of the intravenous tumor model. The 6-week-old C57BL/6 WT or transgenic mice were intravenously injected with MC-38 colorectal cancer cells (5 × 106 cells per mouse) via the tail vein. The mice were sacrificed 2 weeks after tumor inoculation. The lungs and livers were dissected for pathological observation of tumor lesions. Data are means ± s.e.ms. and polled from four individual experiments. Each symbol represents an experimental replicate (n = 8~11). f–h The MC-38 tumor lesions in lungs of WT and TgMGLL mice, as described in e. Representative images of gross anatomy (f) and H&E staining (g) are displayed. The blue dotted lines indicate the tumor lesions. The relative tumor lesions were calculated (h). Scale bars, 200 μm. (n = 8~9). i Macrophage MGLL inhibited tumor growth in a Rag-1 dependent mannaer. MC-38 tumors were inoculated subcutaneously in WT, TgMGLL, Rag1KO or TgMGLL + Rag1KO mice and the tumor volumes were measured dynamically. (n = 5) j mRNA levels of IFNγ in CD8+ or CD4+ T cells that were isolated from the MC-38 tumors inoculated subcutaneously in WT or TgMGLL mice for 3 weeks. (n = 5). k Six-week-old WT or TgMGLL mice were subcutaneously inoculated with MC-38 cells and treated with anti-CD8 antibody or IgG as a control. The tumor volume was measured dynamically. (n = 10). l The survival time of the MC-38 tumor- bearing mice treated as described in k. The data represent the means ± s.e.ms. (n = 10, *P < 0.05, Gehan-Breslow-Wilcoxon test; ns, not significant). Data in a, c and e–k represent the means ± s.e.ms. (*P < 0.05, **P < 0.01, ***P < 0.005; student’s t-test; ns not significant)

Article Snippet: The MC-38 tumor-bearing mice were treated with monoclonal antibodies anti-CD8 (MAB116, Novus Biologicals) or IgG (MAB002, Novus Biologicals) as control.

Techniques: Control, Transgenic Assay, Injection, Staining, Isolation

Journal: Nature communications

Article Title: Monoacylglycerol lipase regulates cannabinoid receptor 2-dependent macrophage activation and cancer progression.

doi: 10.1038/s41467-018-04999-8

Figure Lengend Snippet: Fig. 6 MGLL-CB2 axis regulates tumor progression in mice. a Relative mRNA levels of TNFα, IL-6 and IFNγ in the CD8+ T cells in the MC-38 tumors inoculated subcutaneously in WT, TgMGLL, TgCB2, and TgMGLL+CB2 mice. The data represent the means ± s.e.ms. (n = 5, ***P < 0.005, Student’s t-test; ns not significant). b Six-week-old WT, TgMGLL, TgCB2, and TgMGLL+CB2 mice were subcutaneously inoculated with MC-38 cells, and the tumor volume was measured dynamically. This experiment was repeated three times. The data represent the means ± s.e.ms. (n = 10, **P < 0.01, ***P < 0.005, Student’s t-test). c The survival time of the MC-38 tumor-bearing mice. The mice were described in b. The data represent the means ± s.e.ms. (n = 10, **P < 0.01, ***P < 0.005, Gehan-Breslow-Wilcoxon test). d The incidence of tumor metastasis in MC-38 tumor model. Six-week-old WT, TgMGLL, TgCB2, and TgMGLL

Article Snippet: The MC-38 tumor-bearing mice were treated with monoclonal antibodies anti-CD8 (MAB116, Novus Biologicals) or IgG (MAB002, Novus Biologicals) as control.

Techniques:

Journal: Life (Basel, Switzerland)

Article Title: Diets Differently Regulate Pulmonary Pathogenesis and Immune Signaling in Mice during Acute and Chronic Mycobacterium tuberculosis Infection.

doi: 10.3390/life13010228

Figure Lengend Snippet: Figure 3. Representative immunohistochemistry (IHC) images of the lung sections of mice during acute Mtb infection (30 DPI) stained by CD4+, CD8+, F4/80+, and IFNγ antibodies. (RD_con— uninfected mice fed an RD; MFD_con—uninfected mice fed an MFD; RD_Inf—infected mice fed an RD; and MFD_Inf—infected mice fed an MFD). Magnification-4×; Scale bar–100 µ. IHC sections were graded and presented as a bar graph (see Supplementary Figure S3a).

Article Snippet: The blots were probed with the following primary antibodies: PPARα-specific mouse monoclonal antibody (#MA1-822, ThermoFisher Scientific), Perilipin 1-specific rabbit monoclonal antibody (#9349, Cell Signaling Technology), Phospho-Perilipin 1 (Ser522)-specific mouse monoclonal antibody (#4856, Vala Sciences), F4/80-specific rat monoclonal (#NB600-404, Novus Biologicals), adiponectinspecific mouse monoclonal antibody (#ab22554, Abcam), IFNγ-specific rabbit monoclonal antibody (#ab133566, Abcam), TNFα-specific rabbit polyclonal antibody (#ab6671, Abcam), CD4 specific rabbit polyclonal antibody (#NB100-56457SS, Novus Biologicals), CD8 specific rabbit polyclonal antibody (#NBP2-29475, Novus Biologicals), BNIP3 specific rabbit polyclonal antibody (#44060, Cell signaling Technology), Caspase-3-specific rabbit polyclonal antibody (#9662, Cell signaling Technology), IL-6-specific mouse monoclonal antibody (#66146-1-Ig, Proteintech), and IL-10-specific mouse monoclonal antibody (#60269-1-Ig, Proteintech).

Techniques: Immunohistochemistry, Infection, Staining

Journal: Life (Basel, Switzerland)

Article Title: Diets Differently Regulate Pulmonary Pathogenesis and Immune Signaling in Mice during Acute and Chronic Mycobacterium tuberculosis Infection.

doi: 10.3390/life13010228

Figure Lengend Snippet: Figure 4. MFD alters the level of infiltrated immune cells and inflammatory markers in the lungs during chronic Mtb infection. Immunoblot analysis of (a) immune cells (CD4+, CD8+, and F4/80+) and (b) inflammatory markers (IFNγ, TNFα, and IL-6) in the lung lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during chronic (90 DPI) infection. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups.

Article Snippet: The blots were probed with the following primary antibodies: PPARα-specific mouse monoclonal antibody (#MA1-822, ThermoFisher Scientific), Perilipin 1-specific rabbit monoclonal antibody (#9349, Cell Signaling Technology), Phospho-Perilipin 1 (Ser522)-specific mouse monoclonal antibody (#4856, Vala Sciences), F4/80-specific rat monoclonal (#NB600-404, Novus Biologicals), adiponectinspecific mouse monoclonal antibody (#ab22554, Abcam), IFNγ-specific rabbit monoclonal antibody (#ab133566, Abcam), TNFα-specific rabbit polyclonal antibody (#ab6671, Abcam), CD4 specific rabbit polyclonal antibody (#NB100-56457SS, Novus Biologicals), CD8 specific rabbit polyclonal antibody (#NBP2-29475, Novus Biologicals), BNIP3 specific rabbit polyclonal antibody (#44060, Cell signaling Technology), Caspase-3-specific rabbit polyclonal antibody (#9662, Cell signaling Technology), IL-6-specific mouse monoclonal antibody (#66146-1-Ig, Proteintech), and IL-10-specific mouse monoclonal antibody (#60269-1-Ig, Proteintech).

Techniques: Infection, Western Blot

Journal: Life (Basel, Switzerland)

Article Title: Diets Differently Regulate Pulmonary Pathogenesis and Immune Signaling in Mice during Acute and Chronic Mycobacterium tuberculosis Infection.

doi: 10.3390/life13010228

Figure Lengend Snippet: Figure 6. MFD alters immune signaling in adipose tissue during (a) acute and (b) chronic Mtb infection. Immunoblot analysis of immune cells (CD4, CD8, and F4/80) and inflammatory markers (IFNγ, TNFα, IL-6, and IL-10) in the WAT lysates of RD-fed and MFD-fed uninfected and infected C57BL/6 mice during acute infection (30 DPI) and chronic (3 months) post-infection is shown. The error bars represent standard error of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001 between indicated groups.

Article Snippet: The blots were probed with the following primary antibodies: PPARα-specific mouse monoclonal antibody (#MA1-822, ThermoFisher Scientific), Perilipin 1-specific rabbit monoclonal antibody (#9349, Cell Signaling Technology), Phospho-Perilipin 1 (Ser522)-specific mouse monoclonal antibody (#4856, Vala Sciences), F4/80-specific rat monoclonal (#NB600-404, Novus Biologicals), adiponectinspecific mouse monoclonal antibody (#ab22554, Abcam), IFNγ-specific rabbit monoclonal antibody (#ab133566, Abcam), TNFα-specific rabbit polyclonal antibody (#ab6671, Abcam), CD4 specific rabbit polyclonal antibody (#NB100-56457SS, Novus Biologicals), CD8 specific rabbit polyclonal antibody (#NBP2-29475, Novus Biologicals), BNIP3 specific rabbit polyclonal antibody (#44060, Cell signaling Technology), Caspase-3-specific rabbit polyclonal antibody (#9662, Cell signaling Technology), IL-6-specific mouse monoclonal antibody (#66146-1-Ig, Proteintech), and IL-10-specific mouse monoclonal antibody (#60269-1-Ig, Proteintech).

Techniques: Infection, Western Blot

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A IOD of ACAT2 expression in CC tissues and adjacent tissues was examined using immunohistochemical staining ( n = 47 biologically independent samples). IOD of DHCR7 B and MSMO1 C expression in CC patients with high ( n = 27 biologically independent samples) or low ( n = 20 biologically independent samples) expression of ACAT2 was examined using immunohistochemical staining. The number of activated CD8 T cells (CD8A + GZMB + ) D or activated NK cells (CD56 + GZMB + ) E infiltrated in the tumor tissues of patients with high ( n = 27 biologically independent samples) and low ACAT2 ( n = 20 biologically independent samples) expression was detected. Data represent mean ± SEM. Statistical analysis was performed using the paired A or unpaired ( B – E ) t-test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: ACAT2 expression in HCeEpiC and CC cell lines was examined using RT-qPCR A and Western blot analysis B ( n = 5 independent experiments). C ACAT2, DHCR7, and MSMO1 expression in CC cells after infection with Scramble-sh, ACAT2-sh #1, and ACAT2-sh #2 was examined using Western blot analysis ( n = 5 independent experiments). D Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in CC cells ( n = 5 independent experiments). The proliferation of CC cells was examined using CCK8 ( E ) and colony formation assays F (n = 5 independent experiments). G CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 T cells for 6 h, respectively, and the death of CC cells was detected ( n = 5 independent experiments). H IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , B ) or two-way ( C - H ) ANOVA, followed by Tukey’s multiple comparisons test ( A – H ).

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Infection, Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A ACAT2 knockdown efficiency in U14 cells was examined using western blot analysis ( n = 10 independent experiments). B Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells (n = 10 animals). C The images and weight of the tumors harvested on day 21 ( n = 10 animals). D Protein expression of ACAT2, MSMO1, DHCR7, and PCNA in transplanted tumors was examined using western blot analysis ( n = 10 animals). E Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in transplanted tumors ( n = 10 animals). The gating strategy for GZMB + NK cells and CD8 + T cells F and quantification G were analyzed using flow cytometry ( n = 10 animals). H Survival of mice over 60 days after subcutaneous inoculation of U14 cells was analyzed using the log-rank test ( n = 20 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , C , E , G ) or two-way ( B , D ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Knockdown, Western Blot, Expressing, Flow Cytometry

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: The proliferation of CC cells was examined using CCK8 A and colony formation assays B ( n = 5 independent experiments). C CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 + T cells, and the death of CC cells was detected ( n = 5 independent experiments). D IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). E TGF-β1 released by CC cells was examined using ELISA ( n = 5 independent experiments). F PD-L1 expression levels in CC cells were observed using immunofluorescence staining ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the two-way ( A – F ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells ( n = 5 animals). B The images and weight of the tumors harvested on day 21 ( n = 5 animals). The gating strategy for GZMB + NK cells and CD8 + T cells C and quantification D were analyzed using flow cytometry ( n = 5 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( B , D ) or two-way A ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Flow Cytometry