Journal: Cancers
Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation
doi: 10.3390/cancers13081845
Figure Lengend Snippet: Murlentamab/pembrolizumab combination accentuates the anti-tumoral effect of murlentamab monotherapy through the enhancement of T cell activation. ( A – C ) SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors stimulated with M-CSF and IL-10 (TAMs). After 3 days of co-culture, activated T cells coming from the same healthy donor were added in the culture well for 4 more days. Pembrolizumab was added into co-culture wells everyday from day 3 to day 10. ( A ) Opsonized-SKOV3-R2 + cell number was determined by flow cytometry after one and two days of co-culture with TAMs. Data shown (mean ± SEM) are the results from three different experiments (performed with one healthy donors). ** p < 0.01 compared 3C23K-FcKO vs. Murlentamab. # p < 0.05; ## p < 0.01 compared 3C23K-FcKO + anti-PD-1 vs. Murlentamab + anti-PD-1 as determined using one-way ANOVA analysis followed by Dunnett’s multiple comparisons test. ( B , C ) The CD4 + Th1/Th2 polarization profile and the activation of T CD8 + cells were determined by flow cytometry after four days of co-culture. Data shown (mean ± SEM) are the results from three different experiments (performed with one healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test. ( D , E ) 10 × 10 6 COV434-R2 + ovarian tumor cells were transplanted subcutaneously into humanized GM-CSF/IL3/IL4 hu-NOG (NOD/Shi-scid/IL2Rγ null ) mice (Taconic). After 35 days, when tumors were big enough, mice were i.p treated or not with murlentamab (5 mg/kg) +/− pembrolizumab (25 mg/kg) twice a week for 4 weeks. ( D ) Quantification of circulating CD86 + and CD163 + cells by flow cytometry from blood of tumor-bearing mice before treatment and after 24 days of treatment with murlentamab (5 mg/kg) or pembrolizumab (25 mg/kg) as single agents or murlentamab/pembrolizumab combo-therapy. Data are represented as boxplots. *** p < 0.001, **** p < 0.0001 in comparison to baseline. ( E ) In vivo tumor growth. Data are represented as mean + SEM.
Article Snippet: After dewax and pretreatment, FFPE (Formalin-Fixed Paraffin-Embedded) slides were incubated with primary antibody CD16 (clone SP175, Ventana Medical Systems, Mannheim, Germany), CD8 (clone C8/144B, Agilent, DAKO, Glostrup, Danemark), Granzyme B (clone GRB7, Agilent, DAKO, Glostrup, Danemark), ICOS (clone D1K2T, Cell Signaling Technologies, Leiden, The Netherlands), CD86 (clone E2/G8P, Cell Signalling Technologies, Leiden, The Netherlands) CD163 (clone E2/G8P, Cell Signalling Technologies, Leiden, The Netherlands) and CD14 (clone EPR3653, Roche Diagnostic, Mannheim, Germany).
Techniques: Activation Assay, Labeling, Cell Culture, Derivative Assay, Co-Culture Assay, Flow Cytometry, In Vivo
![Comparison of <t>ICOS</t> protein levels in aortic tissues from healthy and atherosclerotic rats. (A) Protein extracts were prepared from the aortas of rats fed a normal [WT(0) and ApoE-KO(0)] or a high-fat diet for 16 weeks [WT(16) and ApoE-KO(16)]. ICOS protein level was quantified by western blot. (B) Fluorescence confocal microscopy images showing ICOS protein (green) in aortas of WT(0), ApoE-KO(0), WT(16), and ApoE-KO(16) rats; nuclei were counterstained with DAPI. ICOS was revealed with a primary rabbit anti-rat ICOS antibody and a <t>secondary</t> <t>Alexa488-conjugated</t> goat anti-rabbit antibody, and control slides (CN) were stained without the primary anti-ICOS antibody. Fluorescence confocal microscopy images at 63× magnifications.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1234/pmc07791234/pmc07791234__atm-08-23-1597-f6.jpg)
