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: bioRxiv

Article Title: Age-associated modulation of TREM1/2- expressing macrophages promotes melanoma progression and metastasis

doi: 10.1101/2024.11.20.624563

Figure Lengend Snippet: Csfr1 + cells (clusters C8-C11 from global UMAP in ) from scRNA-seq analysis of CD45 + cells were re-clustered to obtain higher resolution clustering of myeloid cells. ( A ) Integrated UMAP plot of myeloid cells from 4M (n= 5,473 cells), 12M (n= 4,244 cells) and 20M (n= 5,148 cells) tumors showing 9 sub-clusters. ( B ) Dot plots showing scaled expression of marker genes used to identify myeloid cell sub-clusters. ( C ) Bar graph showing the percentages of various myeloid cell sub-clusters of for each cluster from 4M, 12M and 20M old mice. ( D ) Dot plots showing scaled expression of indicated genes used to identify specific functions. ( E ) Frequencies of CD11b + F480 low among CD11b + cells from CD45 + cells of tumors from 4M and 20M old mice. ( 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 4M and 20M old mice. ( G ) Frequencies of F480 + among CD45 + cells in tumors from 4M and 20M 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 4M and 20M old mice. n = 7 for 4M and n = 6 for 20M for panels E-H. Data in panels D-H were analyzed by unpaired t -test.

Article Snippet: For pharmacological inhibition of TREM1, 20mg/kg of VJDT (MedChemexpress) or vehicle (DMSO), were administered (i.p.) in mice at day 4 after melanoma cells injection and continued alternate days util day 20 as previously reported .

Techniques: Expressing, Marker, Flow Cytometry

Journal: bioRxiv

Article Title: Age-associated modulation of TREM1/2- expressing macrophages promotes melanoma progression and metastasis

doi: 10.1101/2024.11.20.624563

Figure Lengend Snippet: Myeloid cell clusters (10 and 11 from global UMAP in ) from scRNA-seq analysis of CD45 + cells were re-clustered to obtain higher resolution clustering of myeloid cells. ( A-C ) Dot plots showing scaled expression of marker genes used to identify monocyte markers (A), genes involved in lipid metabolism (B) and chemokines (C) in each sub-clusters of myeloid cells. ( D-E ) Violin plots comparing expression of specific genes in 4M, 12M, 20M old macrophages subclusters. Statistical significance is reported for 20M relative to 4M performed using FindMarkers in Seurat. ( F-H ) Frequencies of CD45.2 + among total cells (F), frequencies of CD11b + among CD45.2 + cells (G) and frequencies of CD36 + among CD11b + cells (H) in tumors from 4M and 20M old mice. ( I ) Frequencies of CD11b + and F480 + among CD45.2 + cells, of TREM1 + among CD11b + cells and of MRC1 + among F480 + cells in tumors from 4M and 12M old mice. n = 7 for 4M and n = 6 for 20M in panels F-H. n = 4 for 4M and 12M in panel I. Data in panels F-I were analyzed by unpaired t -test.

Article Snippet: For pharmacological inhibition of TREM1, 20mg/kg of VJDT (MedChemexpress) or vehicle (DMSO), were administered (i.p.) in mice at day 4 after melanoma cells injection and continued alternate days util day 20 as previously reported .

Techniques: Expressing, Marker

Journal: bioRxiv

Article Title: Age-associated modulation of TREM1/2- expressing macrophages promotes melanoma progression and metastasis

doi: 10.1101/2024.11.20.624563

Figure Lengend Snippet: ( A-B ) YUMM1.7 melanoma cells (500,000) were injected s.c. into the flanks of 4M (young), and 20 months (old). Four days later mice were injected i.p. with either TREM1 inhibitor VJDT or vehicle (control) (A) and injections were repeated every other day until day 20. A separate group of melanoma-bearing mice were treated with anti-TREM2 antibody (B) starting at day 5 after melanoma cell injection and repeated every three days for a total of 4 treatments. Isotype control served as control (B). n = 5 for 4M control and TREM1 treated, n = 4 for 20M in panel A; n = 8 for 4M control, n = 5 for 4M TREM2 treated, n = 7 for 20M control, n = 5 for 20M TREM2 treated in panel B. ( C ) Representative H&E staining of lung tissues from 20M 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 following 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 10 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 4M and 20M 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 4M and 20M old mice treated with isotype control or anti-TREM2 antibodies. n = 8 for 4M control, n = 4 for 4M TREM2 treated, n = 7 for 20M control, n = 5 for 20M TREM2 treated in panels E and F. Data in panels A, B, D, E and F are presented as means + SEM. Data in panel A were analyzed by two-way ANOVA. Data in panel D, E and F were analyzed by unpaired t -test and compared to controls of the same age group.

Article Snippet: For pharmacological inhibition of TREM1, 20mg/kg of VJDT (MedChemexpress) or vehicle (DMSO), were administered (i.p.) in mice at day 4 after melanoma cells injection and continued alternate days util day 20 as previously reported .

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

Journal: Aging Cell

Article Title: The activation of cGAS‐STING pathway causes abnormal uterine receptivity in aged mice

doi: 10.1111/acel.14303

Figure Lengend Snippet: CD14 protein levels and its effects on uterine receptivity in aged mice. (a) Proteomic analysis of young (D4) and aged mouse (AGED D4) uteri on day 4 of pregnancy. (b) Western blot analysis of CD14 protein levels in young and aged mouse uteri on day 4 of pregnancy. (c) CD68 and CD14 immunofluorescence in young and aged mouse uteri on day 4 of pregnancy. Scale bar, 50 μm. (d) Uterine CD14 concentration after ovariectomized mice were injected subcutaneously with 0, 3, 10, or 25 ng E 2 for 7 days. (e) CD14 concentration in the cultured medium after Progerin gene was overexpressed in mouse epithelial cells. Con, empty vector control; Progerin , Progerin overexpression. (f) CD14 concentration in the cultured medium after mouse epithelial cells were transfected with SDNA for 48 h. (g) CD14 concentration in the cultured medium after mouse epithelial cells were treated with ADU S100 for 48 h. (h) Immunofluorescence of the receptivity‐related proteins after mouse epithelial organoids were treated with CD14 for 48 h. Scale bar, 100 μm. (i) Western blot analysis of the receptivity‐related proteins after mouse epithelial organoids were treated with CD14 for 48 h. (j) A representative photograph showing the number of implantation sites on day 5 after recombinant CD14 was injected into the uterine lumen on day 4 of pregnancy. (k) Statistical analysis on the number of implantation sites on day 5 after recombinant CD14 was injected into the uterine lumen on day 4 of pregnancy. * p < 0.05; **, p < 0.01; *** p < 0.001.

Article Snippet: Luminal epithelial cells were treated with ADU‐S100 (2 and 20 μM, HY‐12885A, MedChemExpress), Salmon SDNA (0.2 and 2 μg/mL) or CD14 (0.1 and 1 μg/mL, HY‐P75444, MedChemExpress) in DMEM/F12 with 2% charcoal‐treated FBS (cFBS, Biological Industries, Cromwell, CT).

Techniques: Western Blot, Immunofluorescence, Concentration Assay, Injection, Cell Culture, Plasmid Preparation, Control, Over Expression, Transfection, Recombinant

Journal: Aging Cell

Article Title: The activation of cGAS‐STING pathway causes abnormal uterine receptivity in aged mice

doi: 10.1111/acel.14303

Figure Lengend Snippet: Effects of STING inhibitor (C176) on uterine receptivity and cGAS‐STING pathway. (a) Uterine CD14 protein levels young mice (D4), aged mice (AGED D4), and C176‐treated mice on day 4 of pregnancy. (b)CD14 immunofluorescence in young mice, aged mice, and C176‐treated mice on day 4 of pregnancy. Scale bar, 50 μm.(c) CD14 concentration in uteri of aged mice and C176‐treated mice. (d) CD14 concentration in serum of aged mice and C176‐treated mice. (e)Immunofluorescence of uterine AREG, MUC1 and MSX1 in young mice, aged mice, and C176‐treated mice. Scale bar, 50 μm. (f) Cytoplasmic dsDNA concentration in the uterus of young mice, aged mice, and C176‐treated mice. (g) Western blot analysis of the cGAS‐STING related proteins and receptivity‐related proteins in young mice, aged mice, and C176‐treated mice. (h) The protein levels CGAS‐STING pathway and uterine receptivity marker in mice on the D3 of pregnancy were injected with E2 (25 ng) and TLR4 inhibitor Resatorvid (10 mg/mL) subcutaneously for 24 h. (i) The protein levels CGAS‐STING pathway and uterine receptivity marker after epithelial organoids were overexpression of PROGERIN for 48 hr in the absence or presence of CD14 antibody. con, empty vector control; Progerin , Progerin overexpression. *, p < 0.05; **, p < 0.01; ****, p < 0.0001.

Article Snippet: Luminal epithelial cells were treated with ADU‐S100 (2 and 20 μM, HY‐12885A, MedChemExpress), Salmon SDNA (0.2 and 2 μg/mL) or CD14 (0.1 and 1 μg/mL, HY‐P75444, MedChemExpress) in DMEM/F12 with 2% charcoal‐treated FBS (cFBS, Biological Industries, Cromwell, CT).

Techniques: Immunofluorescence, Concentration Assay, Western Blot, Marker, Injection, Over Expression, Plasmid Preparation, Control

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: (A) Time-dependent changes in cytokine and chemokine production in LPS-stimulated THP-1 cells. After LPS (10 μg/ml) treatment for 15, 30, 60, or 120 min without any agents and with minocycline (50 μg/ml), tigecycline (50 μg/ml), or doxycycline (50 μg/ml), cytokines and chemokines were measured using Multi Plex according to the manufacturer’s protocols. * p <0.05 compared with LPS only at 60 min. ** p <0.05 compared with LPS only at 120 min. Mino: minocycline, Tige: tigecycline, Doxy: doxycycline. (B) The rate of cytokine and chemokine production in the THP-1 cell line compared to the production of cytokines and chemokines by LPS stimulation without tetracyclines. After LPS treatment (10 μg/ml) for 30, 60, 120 or 240 min without any agents and with minocycline (50 μg/ml), tigecycline (50 μg/ml), or doxycycline (50 mg/ml), cytokines and chemokines were measured with Multi Plex.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques:

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: Effects of minocycline, doxycycline, and tigecycline on the modulation of NF-κB, phospho-NF-κB, IKKα/β, phospho-IKKα/β, IκBα, and phospho-IκBα in LPS-stimulated THP-1 cells. THP-1 cells were incubated without or with 10 μg/ml LPS, or with LPS plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 30, 60, or 120 min. NF-κB, phospho-NF-κB, IKKα/β, phospho-IKKα/β, IκBα, and phospho-IκBα were assessed with Western blotting.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Incubation, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: Effects of tetracyclines (minocycline, doxycycline, and tigecycline) on the activation of phospho-ERK1/2 and phospho-p38 in LPS-stimulated THP-1 cells. THP-1 cells were incubated without or with 10 μg/ml LPS, or with LPS plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 30 or 60 min. Phospho-p38 and phospho-ERK1/2 were assessed with Western blotting.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Activation Assay, Incubation, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways

doi: 10.1016/j.bbrep.2015.11.003

Figure Lengend Snippet: SB203580, U0126 and BAY11-7082 suppressed TNF-α and IL-8 production in LPS-stimulated THP-1 cells on treatment with or without tetracyclines. THP-1 cells were pre-incubated by SB203580 (10 μM), U0126 (5 μM) and BAY11-7082 (5 μM) for 30 min, followed treatment without or with LPS (10 μg/ml), or with LPS (10 μg/ml) plus minocycline (50 μg/ml), doxycycline (50 μg/ml), or tigecycline (50 μg/ml) for 60 min. TNF-α were measured with ELISA. * p <0.05, ** p <0.01, *** p <0.001 compared to the measurement without the inhibitor in the same group. Abbreviation; SB: SB203580, U: U0126, BAY: BAY11-7082.

Article Snippet: The THP-1 human monocytic leukemia cell line was purchased from RIKEN Cell Bank (Wako, Japan).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay