Journal: Journal of Virology

Article Title: Human Cytomegalovirus Decreases Major Histocompatibility Complex Class II by Regulating Class II Transactivator Transcript Levels in a Myeloid Cell Line

doi: 10.1128/JVI.01901-19

Figure Lengend Snippet: HCMV downregulates surface expression of MHC class II in a myeloid progenitor cell line. (A) Flow cytometry analysis of unstained Kasumi-3 cells or cells stained for HLA-DR, HLA-DQ, and HLA-DP. (B) Flow cytometry scatter plot of HCMV-infected (72 hpi) and uninfected cells showing the relationship between mCherry (a marker of HCMV infection) and HLA-DR. (C) Histograms of HCMV-infected (mCherry) and uninfected Kasumi-3 cells stained for surface HLA-DR at 72 and 120 h postinfection. (D) Bar graph of the geometric mean (GM) fluorescence values for the samples used in the assay whose results are presented in panel C, displayed as a percentage of the value for the uninfected sample. (E) Geometric mean fluorescence values from uninfected or HCMV-infected CD14+ human peripheral blood monocytes (HPBM) at 72 hpi. Values are averages from a minimum of three independent experiments. *, P < 0.05.

Article Snippet: Human peripheral blood monocytes (catalog number 6906K-50A; Cell Applications Inc.) were maintained in the manufacturer’s supplied human blood cell culture medium.

Techniques: Expressing, Flow Cytometry, Staining, Infection, Marker, Fluorescence

Journal: Cancer research

Article Title: Age-Associated Modulation of TREM1/2-Expressing Macrophages Promotes Melanoma Progression and Metastasis

doi: 10.1158/0008-5472.CAN-24-4317

Figure Lengend Snippet: Aging is conducive to the establishment of tolerogenic macrophages in melanoma. Csfr1 + cells [clusters C8–C11 from global Uniform Manifold Approximation and Projection (UMAP) in Fig. 1D ] from scRNA-seq analysis of CD45 + cells were reclustered to obtain higher resolution clustering of myeloid cells. A, Integrated UMAP plot of myeloid cells from tumors of 4- ( n = 5,473 cells), 12- ( n = 4,244 cells), and 20-month (M)-old mice ( n = 5,148 cells) showing nine subclusters. Mac, macrophages; Prolif., proliferative. B, Dot plots showing scaled expression of marker genes used to identify myeloid cell subclusters. C, Bar graph showing the percentages of various myeloid cell subclusters for each cluster from 4-, 12-, and 20-month-old mice. D, Dot plots showing scaled expression of indicated genes used to identify specific functions. ECM, extracellular matrix. E, Frequencies of CD11b + F480 low among CD11b + cells from CD45 + cells of tumors from 4- and 20-month-old. F, Frequencies of TREM1 + among CD11b + F480 low cells and representative flow cytometry plots showing TREM1 + expression by a gated subpopulation (CD11b + cells) of CD45.2 + cells from 4- and 20-month-old mice. G, Frequencies of F480 + among CD45 + cells in tumors from 4- and 20-month-old mice. H, Frequencies of TREM2 + among F480 + cells and representative flow cytometry plots showing TREM2 + expression by a gated subpopulation (F480 + cells) of CD45.2 + cells from 4- and 20-month-old mice. n = 7 for 4 month and n = 6 for 20 month for E – H . Data in D – H were analyzed by using the unpaired t test. C, control.

Article Snippet: For pharmacologic inhibition of TREM1, mice were randomly divided into two groups and 20 mg/kg of VJDT (MedChemExpress, catalog #HY-157122) or vehicle (DMSO) was administered (intraperitoneally) in them on day 4 after the melanoma cells injection and continued on alternate days until day 20, as previously reported ( 33 ).

Techniques: Expressing, Marker, Flow Cytometry, Control

Journal: Cancer research

Article Title: Age-Associated Modulation of TREM1/2-Expressing Macrophages Promotes Melanoma Progression and Metastasis

doi: 10.1158/0008-5472.CAN-24-4317

Figure Lengend Snippet: Function of TREM1 and TREM2 in age-associated melanoma progression and metastasis. A and B, YUMM1.7 melanoma cells (500,000) were injected subcutaneously into the flanks of 4- (young) and 20-month (M)-old mice (old). Four days later, mice were injected intraperitoneally with either TREM1 inhibitor VJDT or vehicle (control; A ), and injections were repeated every other day until day 20. B, A separate group of melanoma-bearing mice that were treated with anti-TREM2 antibody starting at day 5 after melanoma cell injection and repeated every 3 days for a total of four treatments. Isotype control served as the control ( B ). n = 4 for 4 month control-treated mice, n = 5 for 4 month TREM1-treated mice, and n = 4 for 20 month control-treated mice in A ; n = 8 for 4 month control-treated mice, n = 5 for 4 month TREM2-treated mice, n = 7 for 20 month control-treated mice, and n = 5 for 20 month TREM2-treated mice in B . C, Representative H&E staining of lung tissues from 20 month melanoma tumor–bearing mice treated either with isotype control (left) or anti-TREM2 antibody (right). Lung tissues were analyzed 22 days after melanoma cell injection. D, Quantification of lung metastases (mets) after treatment of mice with control or anti-TREM2 antibody as described in B . Tissues were subjected to IHC staining for S100 to detect metastatic melanoma cells (more than ten S100+ cells per lesion). n = 5 for each group. Scale bar, 300 μm. E, Frequencies of CD206 + and MHCI + among CD11b + and F480 + cells, and expression of CD11c + cells on F480 + cells of tumors from 4- and 20-month-old mice treated with isotype control or anti-TREM2 antibodies. F, Frequencies of CD4 + and CD8 + among CD45.2 + cells, and expression of CD44 + cells on CD4 + or CD8 + cells of tumors from 4- and 20-month-old mice treated with isotype control or anti-TREM2 antibodies. n = 8 for 4 month control-treated mice, n = 4 for 4 month TREM2-treated mice, n = 7 for 20 month control-treated mice, and n = 5 for 20 month TREM2-treated mice in E and F . Data in A , B , and D–F are presented as mean ± SEM. Data in A were analyzed by using two-way ANOVA. Data in D–F were analyzed by using the unpaired t test and compared with controls of the same age group. MFI, mean fluorescent intensity.

Article Snippet: For pharmacologic inhibition of TREM1, mice were randomly divided into two groups and 20 mg/kg of VJDT (MedChemExpress, catalog #HY-157122) or vehicle (DMSO) was administered (intraperitoneally) in them on day 4 after the melanoma cells injection and continued on alternate days until day 20, as previously reported ( 33 ).

Techniques: Injection, Control, Staining, Immunohistochemistry, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: MMP28 recruits M2-type tumor-associated macrophages through MAPK/JNK signaling pathway-dependent cytokine secretion to promote the malignant progression of pancreatic cancer

doi: 10.1186/s13046-025-03321-x

Figure Lengend Snippet: Anti-IL-8 and anti-VEGFA antibodies partially reverse the ability of MMP28 to promote the migration of TAMs and the polarization of M2 TAMs. A CM from MMP28-overexpressing pancreatic cancer cells was treated with an anti-IL-8 neutralizing antibody (5 μg/mL) or an anti-VEGFA neutralizing antibody (1 μg/mL). The migration level of TAMs was determined by the Transwell migration experiment. Scale bar, 100 μm. B qRT-PCR determination of CD86 and CD163 mRNA expression levels in TAMs cocultured with Vector group of pancreatic cancer cells transfected with empty vector virus, TAMs cocultured with OE-MMP28 cell group, TAMs cocultured with the OE-MMP28 + IgG cell group, TAMs cocultured with OE-MMP28 + Anti-IL-8 cell group, and TAMs cocultured with OE-MMP28 + Anti-VEGFA cell group. C The expression levels of the TAM markers CD86 and CD163 in different coculture groups were determined by immunofluorescence staining. Scale bar, 50 μm. D The proportions of CD11b + CD86 + TAMs and CD11b + CD163 + TAMs in different coculture groups were analysed by flow cytometry. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001

Article Snippet: When the tumors reached a volume of at least 80 mm 3 one week post-injection, the treatment groups received the following treatments: PBS, Clodronate Liposomes (200 μL/mouse, YEASEN), the JNK inhibitor SP600125 (15 mg/kg/mouse, MCE), the interleukin-8 (IL-8)-neutralizing antibody MAB208-SP (10 μg/mouse, R&D Systems), or vascular endothelial growth factor A (VEGFA)-neutralizing antibody MAB293-SP (1 μg/mouse, R&D Systems).

Techniques: Migration, Quantitative RT-PCR, Expressing, Plasmid Preparation, Transfection, Virus, Immunofluorescence, Staining, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: MMP28 recruits M2-type tumor-associated macrophages through MAPK/JNK signaling pathway-dependent cytokine secretion to promote the malignant progression of pancreatic cancer

doi: 10.1186/s13046-025-03321-x

Figure Lengend Snippet: MMP28 enhances M2 TAM infiltration and promotes pancreatic cancer growth in vivo. A Effects of the administration of Clodronate Liposomes, the JNK inhibitor SP600125, the anti-IL-8 neutralizing antibody MAB208-SP, and the anti-VEGFA neutralizing antibody MAB293-SP on subcutaneous tumors in nude mice in the experimental group. B - C Representative images of subcutaneous tumors in nude mice detected with fluorescein potassium salt and the fluorescence quantitative analysis. After the tumor volume reached at least 80mm 3 , PBS, Clodronate Liposomes (200 μg), the JNK inhibitor SP600125 (15 mg/kg), the anti-IL-8 antibody MAB208-SP (10 μg), and the anti-VEGFA antibody MAB293-SP (1 μg) were injected intraperitoneally into the nude mice. D Representative images of subcutaneous xenograft tumors (5 mice per group). E The volume of the tumor was measured every four days. F Tumor weight in both groups. G Representative images of MMP28 expression levels in xenograft tumors from the Vector and OE-MMP28 groups as determined by immunohistochemical staining. Scale bar, 50 μm. H Immunohistochemical staining detection of expression of Ki67 and CD86 + and CD163 + TAM infiltration in xenograft tumors in the Vector group, OE-MMP28 group, OE-MMP28 + SP600125 group, OE-MMP28 + MAB208-SP group, OE-MMP28 + MAB293-SP group, and Vector + Clodronate Liposomes group

Article Snippet: When the tumors reached a volume of at least 80 mm 3 one week post-injection, the treatment groups received the following treatments: PBS, Clodronate Liposomes (200 μL/mouse, YEASEN), the JNK inhibitor SP600125 (15 mg/kg/mouse, MCE), the interleukin-8 (IL-8)-neutralizing antibody MAB208-SP (10 μg/mouse, R&D Systems), or vascular endothelial growth factor A (VEGFA)-neutralizing antibody MAB293-SP (1 μg/mouse, R&D Systems).

Techniques: In Vivo, Liposomes, Fluorescence, Injection, Expressing, Plasmid Preparation, Immunohistochemical staining, Staining