Journal: Advanced Science

Article Title: RBM10 Deficiency Promotes Anti‐PD‐1 Resistance in LUAD via STING Alternative Splicing‐Driven CCL7 Signaling and Macrophage Polarization

doi: 10.1002/advs.202522159

Figure Lengend Snippet: CCL7‐CCR2 axis inhibition potentiates anti‐PD‐1 therapy efficacy in RBM10‐low LUAD. (A) mIHC analysis of CCR2 expression on macrophages in tumor tissues from LUAD patients with high and low RBM10 expression. (B) FC analysis of CCR2 expression on macrophages (live + CD45 + CD11b + F4/80 + CD206 + ) in tumor tissues from mice with high and low RBM10 expression (n = 3). (C) Treatment schema for LLC shRBM10 tumor‐bearing mice treated with anti‐PD‐1 antibody, RS102895 or not (n = 5). [Created in BioRender. Gao, W. (2026) https://BioRender.com/awmu085 ] (D) Subcutaneous tumor formation in C57BL/6 injected with shRBM10 LLC cells treated with anti‐PD‐1 antibody, RS102895 or not (n = 5). (E) Tumor growth curves of indicated groups. (F) Survival curves of indicated groups. (G,H) IHC staining of CCL7, CD8, CD68, CD86, and CD206 in tumor tissues from each group. All data are presented as the mean ± SEM (n ≥ 3). The P values in panels (B) were calculated using two‐tailed unpaired Student's t ‐test. The P values in panels (E) were calculated using two‐way ANOVA. Survival curves (F) were calculated using log‐rank test. ns (not significant), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: In a separate cohort for CCR2 antagonist combination (Figure S11K), shRBM10 tumor‐bearing mice received daily intraperitoneal injections of RS102895 (5 mg/kg, MCE, HY‐18611) together with anti‐PD‐1 (twice weekly) or vehicle controls for 10 consecutive days.

Techniques: Inhibition, Expressing, Injection, Immunohistochemistry, Two Tailed Test

Journal: Hepatology Communications

Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

doi: 10.1097/HC9.0000000000000928

Figure Lengend Snippet: CD14 + CCR2 + (c0) MoMFs represent a functionally heterogeneous population driving fibrosis, inflammation, and lipid accumulation in MASLD. (A) Feature plots displaying the expression of F13a1 , Hp , Fn1 , Cd14 , Trem2 , and Ccr2 . (B) Flow cytometry analysis showing CD14 + MoMFs proportions among total MoMFs across NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice, with representative plots (upper) and quantitative data (lower). (C) Correlation between the proportion of CD14 + MoMFs and the NAFLD activity score in the combined cohort of NCD-fed and MCD-fed mice. (D) Feature plots displaying the expression of Ccr2 in NCD and MCD. (E) Feature plots displaying the expression of Mmp8 , Sell , S100a10 , Clec5a , C5ar1 , Tnf , Treml4 , Cd36 , Trem3 , H2-Eb1 , H2-Aa , and Cd74 . (F) Violin plots displaying the expression of representative marker genes of each subcluster. (G–L) Statistical analysis of the percentages of CD14 + MMP8 + (G), CD14 + DCFH-DA + (H), CD14 + TNFα + (I), CD14 + CD36 + (J), CD14 + BODIPY + (K), and CD14 + MHC-II + MoMFs (L) among the total MoMFs in NCD-fed, CDHFD-fed, MCD-fed, and HFD-fed mice. (M) Percentage of CD14 + cells among total intrahepatic MoMFs in NCD, MCD+vehicle, and MCD+RS102895 groups. (N, O) Plasma ALT and AST levels in each group. (P–R) Representative images (P) and quantification of H&E staining (Q) and Sirius red staining (R) in liver paraffin sections. Scale bars, 200 μm. Abbreviations: ALT, alanine transaminase; AST, aspartate aminotransferase; CDHFD, choline-deficient high-fat diet; DEGs, differentially expressed genes; GO, Gene Ontology; HFD, high-fat diet; H&E, Hematoxylin and Eosin; KEGG, Kyoto Encyclopedia of Genes and Genomes; MASLD, metabolic dysfunction–associated steatotic liver disease; MCD, methionine/choline-deficient diet; MoMFs, monocyte-derived macrophages; mRNA, messenger RNA; NAFLD, nonalcoholic fatty liver disease; NCD, normal control diet.

Article Snippet: During the final 2 weeks, they received daily intraperitoneal (i.p.) injections of the CCR2 antagonist RS102895 (5 mg/kg, HY-18611A, MedChemExpress) or vehicle control., RS102895 was prepared as a 25 mg/mL stock in DMSO and diluted to 1 mg/mL in a vehicle of 40% PEG300 (HY-Y0873, MedChemExpress), 5% Tween 80 (HY-Y1891, MedChemExpress), and 51% saline.

Techniques: Expressing, Flow Cytometry, Activity Assay, Marker, Clinical Proteomics, Staining, Derivative Assay, Control

Journal: Hepatology Communications

Article Title: Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease

doi: 10.1097/HC9.0000000000000928

Figure Lengend Snippet: Heterogeneity and dynamics of intrahepatic MoMFs during MASLD progression. This scRNA-seq analysis delineated 7 distinct MoMFs clusters (c0–c6) in the liver with dynamic changes during steatohepatitis progression. CD14 + (c0) MoMFs, the predominant CCR2 + population, exhibited multifaceted functions beyond inflammation, including roles in fibrosis, lipid uptake, and antigen presentation. The basal cluster (c1) is depleted during MASLD progression and acquires CD14 + and CCR7⁺ phenotypes under inflammatory stimulation in vitro. Lipid overload drives mitochondrial dysfunction and apoptosis in CD206 + (c2) MoMFs in MASLD. CCR3 + (c3) MoMFs drive MASLD progression through lipid accumulation, oxidative stress, and CCL5-mediated recruitment. CCL19/CCL21-recruited CCR7 + (c4) MoMFs exacerbate MASLD pathogenesis through enhanced antigen presentation. In addition, Ki67 + (c5) proliferative and CD3 + (c6) TCR-related MoMFs maintained stable proportions but exhibited functional alterations during MASLD. Abbreviations: MASLD, metabolic dysfunction–associated steatotic liver disease; MoMFs, monocyte-derived macrophages; scRNA-seq, single-cell RNA sequencing.

Article Snippet: During the final 2 weeks, they received daily intraperitoneal (i.p.) injections of the CCR2 antagonist RS102895 (5 mg/kg, HY-18611A, MedChemExpress) or vehicle control., RS102895 was prepared as a 25 mg/mL stock in DMSO and diluted to 1 mg/mL in a vehicle of 40% PEG300 (HY-Y0873, MedChemExpress), 5% Tween 80 (HY-Y1891, MedChemExpress), and 51% saline.

Techniques: Immunopeptidomics, In Vitro, Functional Assay, Derivative Assay, Single Cell, RNA Sequencing

Journal: Blood

Article Title: Heme-induced activation of the TLR3/TRIF–IFN-I–CCL2 pathway contributes to kidney injury in sickle cell disease

doi: 10.1182/blood.2025031060

Figure Lengend Snippet: IFN-I/CCL2 signaling contributes to kidney injury in SCD. (A) Absolute numbers of CMo in kidneys from control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 6). (B) Frequencies of MoMϕ among total kidney macrophages in control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 6-9). (C) Kidney Tnfa mRNA expression levels in control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 6). (D) uACR levels in control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 8-14). (E) uKIM1 levels in control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 7-11). (F-G) Plasma levels of BUN (F) and cystatin C (G) in control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 7-12). (H) MFI analysis of KIM-1 expression on renal PTECs in control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 6). Male mice aged 6 to 8 months were used. Data are presented as mean ± SEM. Statistical significance was determined using 1-way ANOVA. ∗ P < .05. BUN, blood urea nitrogen; MFI, mean fluorescence intensity; uKIM1, urinary KIM-1.

Article Snippet: For IFN-I/CCL2 signaling blocking studies, mice were injected with anti-IFNAR1 antibody (Leinco Technologies, St Louis, MO) or isotype control antibody for 3 consecutive days (day 0, 2 mg per 20 g body weight; days 1 and 2, half of the day 0 dose) or with CCR2 antagonist RS 504393 (MedChemExpress; 4 mg/kg body weight) for 3 consecutive days.

Techniques: Control, Expressing, Clinical Proteomics, Fluorescence

Journal: Blood

Article Title: Heme-induced activation of the TLR3/TRIF–IFN-I–CCL2 pathway contributes to kidney injury in sickle cell disease

doi: 10.1182/blood.2025031060

Figure Lengend Snippet: Improved renal histopathology in Ifnar1 −/− and Ccr2 −/− sickle mice. (A) Representative images of kidney sections stained by TUNEL assay (top) and H&E (bottom) from control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice. Scale bars, 50 μm (TUNEL) and 80 μm (H&E). The black arrows indicate TUNEL-positive cells in the top panels and glomeruli in the bottom panels. (B) Frequencies of TUNEL-positive cells in the kidney sections from control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 6-9). (C-D) Glomerular size (area) (C) and nuclei per glomerulus (D) in H&E-stained kidney sections from control, sickle, Ifnar1 −/− sickle, and Ccr2 −/− sickle mice (n = 5-9). Male mice aged 6 to 8 months were used. Data are presented as mean ± SEM. Statistical significance was determined using 1-way ANOVA. ∗ P < .05.

Article Snippet: For IFN-I/CCL2 signaling blocking studies, mice were injected with anti-IFNAR1 antibody (Leinco Technologies, St Louis, MO) or isotype control antibody for 3 consecutive days (day 0, 2 mg per 20 g body weight; days 1 and 2, half of the day 0 dose) or with CCR2 antagonist RS 504393 (MedChemExpress; 4 mg/kg body weight) for 3 consecutive days.

Techniques: Histopathology, Staining, TUNEL Assay, Control

Journal: bioRxiv

Article Title: MAIT cell responses to S. aureus and sensitivity to HlgAB are modulated by activation and tissue-dependent virulence effects

doi: 10.64898/2026.03.17.712383

Figure Lengend Snippet: (A) UMAP plots of total live PBMCs showing expression of the indicated markers. (B) UMAP plots of total PBMCs exposed to HlgAB for 1 h or not (n=6). (C) Relative percentages of total CD3 + cells, MAIT cells, CD56 + and CD56 - MAIT cells, and CD14 + cells after exposure to a range of HlgAB concentrations for 1 h. (D) CCR2, CXCR1 and CXCR2 expression across CD3 + cells, MAIT cells, CD56 + and CD56 - MAIT cells, and CD14 + cells. (E) Relative percentages of MAIT cells after exposure to a high dose (10 µg/ml) and low dose (0.01 µg/ml) of HlgAB during a time course from 30 min to 4 h (n=5). (F) Relative percentages of CD3 + cells, MAIT cells and CD14 + cells over a range of HlgAB concentrations after 1 h of exposure. The IC50 is indicated in red (n=5). Statistical comparison between conditions with the Wilcoxon matched-pairs signed-rank test (C-D) and p-values were adjusted using the Benjamini-Hochberg method (C). Statistical comparison between conditions with the Wilcoxon matched-pairs signed-rank test for (C and D). *p<0.05, **p<0.01 and ***p<0.001.

Article Snippet: To investigate possible CCR2 antagonist-mediated protection against HlgAB toxicity, cells were pre-treated with Cenicriviroc (MedChem Express) or BMS CCR2 22 (MedChem Express) for 1 h before HlgAB addition and maintained in the medium.

Techniques: Expressing, Comparison

Journal: bioRxiv

Article Title: MAIT cell responses to S. aureus and sensitivity to HlgAB are modulated by activation and tissue-dependent virulence effects

doi: 10.64898/2026.03.17.712383

Figure Lengend Snippet: Total PBMCs under unstimulated conditions or following TCR and innate cytokine stimulation, in the presence or absence of HlgAB (0.31 µg/ml) for 1 h. The corresponding UMAP plots of total live PBMCs showing expression of the indicated markers is shown in Supplementary Figure 3. (A) UMAP visualization of PBMCs showing alterations in MAIT cell and CD14⁺ monocyte compartments following HlgAB-induced cytotoxicity (n=6). (B) MAIT cells, CD56⁺ and CD56⁻ MAIT cells, and CD14⁺ monocytes after 1 h exposure to HlgAB, plotted as % of the respective untreated control. (C) Expression of CCR2, CXCR1, and CXCR2 across MAIT cells, CD56⁺ and CD56⁻ MAIT subsets, and CD14⁺ monocytes under stimulated and unstimulated conditions. Statistical comparison between conditions with the Wilcoxon matched-pairs signed-rank test (B and C). *p<0.05, **p<0.01, and ***p<0.001.

Article Snippet: To investigate possible CCR2 antagonist-mediated protection against HlgAB toxicity, cells were pre-treated with Cenicriviroc (MedChem Express) or BMS CCR2 22 (MedChem Express) for 1 h before HlgAB addition and maintained in the medium.

Techniques: Expressing, Control, Comparison

Journal: bioRxiv

Article Title: MAIT cell responses to S. aureus and sensitivity to HlgAB are modulated by activation and tissue-dependent virulence effects

doi: 10.64898/2026.03.17.712383

Figure Lengend Snippet: (A) Tissue samples were obtained from human organ donors and MAIT cells were subsequently identified as live CD45 + , CD3 + 5-OP-RU-MR1-tetramer + Vα7.2 + CD161 + lymphocytes. (B) MAIT cell frequency was calculated as the percentage of T cells for each donor and tissue and is shown on a logarithmic scale. Surface expression of (C) CD69 and (D) CD56 by MAIT cells. (E) Surface expression of leukocidin receptors CCR2, CXCR1, CXCR2 and CCR5 by total MAIT cells. The number of samples analyzed (donors=9): blood (n=9), spleen, (n=8), liver (n=4), ileum (n=5), cecum (n=5), colon (n=5), mLN (n=3), lung (n=6) and lungLN (n=7). In each tissue site, only samples with more than 20 gated MAIT cells were included in the phenotypic analysis. Differences in MAIT cell frequencies or marker expression between blood and each tissue were analyzed using Kruskal-Wallis test followed by Dunn’s post-hoc test for all pairwise comparisons. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001.

Article Snippet: To investigate possible CCR2 antagonist-mediated protection against HlgAB toxicity, cells were pre-treated with Cenicriviroc (MedChem Express) or BMS CCR2 22 (MedChem Express) for 1 h before HlgAB addition and maintained in the medium.

Techniques: Expressing, Marker

Journal: bioRxiv

Article Title: MAIT cell responses to S. aureus and sensitivity to HlgAB are modulated by activation and tissue-dependent virulence effects

doi: 10.64898/2026.03.17.712383

Figure Lengend Snippet: Peripheral blood and tissue samples were obtained from individuals undergoing tonsillectomy for recurrent infections or obstructive sleep apnea. (A) MAIT cells were identified as live CD45 + CD3 + 5-OP-RU-MR1-tetramer + Vα7.2 + CD161 + lymphocytes. (B) Frequencies of total T cells and their CD8⁺ and CD4⁺ subsets in peripheral blood versus tonsils, and (C) frequencies of MAIT cells, including CD56⁺ and CD56⁻ subsets, in peripheral blood versus tonsils (blood n=7 and tonsils n=25). (D) Expression of activation markers CD69, HLA-DR, CD25 and CD38 on tonsillar MAIT cells to assess their activation status (n=14). (E) Expression of CCR2, CXCR1, CXCR2 and CCR5 across tonsillar MAIT cells (n=14). (F) Relative percentages of tonsillar MAIT cells over a range of HlgAB concentrations after 1 h of exposure (n=4). Statistical comparisons between PBMCs and tonsillar cells were performed using the Wilcoxon rank-sum test (B, C). Comparisons between tonsillar MAIT cells under different treatment conditions were performed using the Wilcoxon matched-pairs signed-rank test (F). *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001.

Article Snippet: To investigate possible CCR2 antagonist-mediated protection against HlgAB toxicity, cells were pre-treated with Cenicriviroc (MedChem Express) or BMS CCR2 22 (MedChem Express) for 1 h before HlgAB addition and maintained in the medium.

Techniques: Expressing, Activation Assay