Journal: Frontiers in Immunology

Article Title: Multifocal Cerebral Microinfarcts Modulate Early Alzheimer’s Disease Pathology in a Sex-Dependent Manner

doi: 10.3389/fimmu.2021.813536

Figure Lengend Snippet: Dynamics of circulating monocytes are modulated upon multifocal microinfarction. (A) Gating strategy used to discriminate the circulating monocytes (CD11b + LyC6 + ) in leukocytes (CD45 + ) and the subsequent distribution of the inflammatory monocytes (Ly6C high ), patrolling monocytes (Ly6C low ) and neutrophils (Ly6G high ). (B) Flow cytometry analysis shows that the frequency of total monocytes in the blood circulation increases in MO-operated male WT mice while it remains unchanged in MO-operated male APP/PS1 mice compared to sham-operated male mice and decreases in MO-operated male APP/PS1 mice compared to MO-operated male WT mice. (C) Flow cytometry analysis shows that the frequency of Ly6C low monocytes decreases in the blood circulation of MO-operated male WT mice compared to sham-operated WT mice. (D) Flow cytometry analysis shows that the frequency of Ly6C int monocytes increases in the blood circulation of MO-operated male WT mice as well as MO-operated male APP/PS1 mice compared to sham-operated WT mice. (E) Flow cytometry analysis shows that the frequency of inflammatory Ly6C high monocytes decreases in the blood circulation of MO-operated male WT mice as well as MO-operated male APP/PS1 mice compared to sham-operated male mice. (F) Flow cytometry analysis shows that similarly to males, the frequency of total monocytes in the blood circulation increases in MO-operated female WT mice while it remains unchanged in MO-operated female APP/PS1 mice compared to sham-operated female mice and decreases in MO-operated female APP/PS1 mice compared to MO-operated female WT mice. (G) Flow cytometry analysis indicates that the frequency of Ly6C low monocytes increases in MO-operated female WT mice, while it decreases in the MO-operated female APP/PS1 mice compared to sham-operated female mice. (H) Flow cytometry analysis indicates that the frequency of Ly6C int monocytes increases in the blood circulation of MO-operated female WT mice and to a lesser extent MO-operated female APP/PS1 mice. (I) Flow cytometry analysis indicates that the frequency of Ly6C high monocytes is decreased in the blood circulation of MO-operated female WT mice as well as MO-operated female APP/PS1 mice compared to sham-operated female mice. (J) Flow cytometry analysis shows that the frequency of Ly6G neutrophils increases in MO-operated male WT mice compared to sham-operated male WT mice but to a lesser extent in MO-operated male APP/PS1 mice compared to sham-operated APP/PS1 male mice. (K) Flow cytometry analysis shows that the frequency of Ly6G neutrophils similarly increases in the blood circulation of MO-operated female WT mice compared to sham-operated female WT mice, whereas it remains unchanged in MO-operated female APP/PS1 mice compared to sham-operated female APP/PS1 mice. Data are mean ± SEM (n = 5-10 animals/group). *P < 0.05/**P < 0.01/****P < 0.01 compared with sham- and MO-operated male and female WT and APP/PS1 mice (two-tailed unpaired t-test).

Article Snippet: The following primary antibodies were used; anti-Aβ (6E10) (1:1000; mouse anti-human, Biolegend, 8030001), anti-ionized calcium binding adaptor molecule (IBA)-1 (1:1000, rabbit anti-mouse, WAKO, 019-19741), anti-cluster of differentiation (CD)-68 (1:1000, rat anti-mouse, Bio-Rad, Hercules, CA, USA, MCA1957) and anti-CD45 (1:500, rat anti-mouse, BD bioscience, 55376) and anti-nuclear receptor subfamily 4 group A member 1 (Nr4A1 or Nurr77) (1:250, rabbit, Abcam, ab13851).

Techniques: Flow Cytometry, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Multifocal Cerebral Microinfarcts Modulate Early Alzheimer’s Disease Pathology in a Sex-Dependent Manner

doi: 10.3389/fimmu.2021.813536

Figure Lengend Snippet: Infiltrated phagocytic monocytes are recruited to the lesion sites and Aβ plaques. (A) A tile representing a coronal brain section of MO-operated male APP/PS1/CX3CR1 GFP/+ mouse immunolabeled with CD45 (i.e. infiltrating cells, blue) and IBA1 (i.e. microglia; red) showing (A) the presence of CX3CR1 GFP/+ expressing Nurr77 + , highlighting their monocytic origin, at lesion site 1 week post-microinfarct induction. (B) A tile representing a coronal brain section of MO-operated male APP/PS1/CX3CR1 GFP/+ mouse immunolabeled with CD45 (blue) and 6E10 (Aβ plaques; red) showing (B’) the presence of CD45 + /CX3CR1 GFP/+ cells at the lesion site and (B’’) CD45 + /IBA1 + /CX3CR1 GFP/+ cells recruited to Aβ plaques. Scale bar = 20 µm (A) , 1000 µm (B) , 50 µm (B’) and 25 µm (B”) .

Article Snippet: The following primary antibodies were used; anti-Aβ (6E10) (1:1000; mouse anti-human, Biolegend, 8030001), anti-ionized calcium binding adaptor molecule (IBA)-1 (1:1000, rabbit anti-mouse, WAKO, 019-19741), anti-cluster of differentiation (CD)-68 (1:1000, rat anti-mouse, Bio-Rad, Hercules, CA, USA, MCA1957) and anti-CD45 (1:500, rat anti-mouse, BD bioscience, 55376) and anti-nuclear receptor subfamily 4 group A member 1 (Nr4A1 or Nurr77) (1:250, rabbit, Abcam, ab13851).

Techniques: Immunolabeling, Expressing

Journal: Immunity

Article Title: Microbiota Sensing by Mincle-Syk Axis in Dendritic Cells Regulates Interleukin-17 and -22 Production and Promotes Intestinal Barrier Integrity

doi: 10.1016/j.immuni.2018.12.020

Figure Lengend Snippet: The Mincle-Syk Axis Contributes to Intestinal Barrier Function (A and B) Reg3g transcripts were measured by qPCR and normalized to Gapdh in the jejunum of the indicated genotypes during the steady state (A) or after L. plantarum gavage, as described in the scheme (B). (C) Quantification of total IgA measured by ELISA in the intestinal lumen of the indicated genotypes. (D) Frequency of IgA + bacteria by flow cytometry in the indicated genotypes. (E and F) Representative plots and summary graphs of CD45 + B220 − IgA + (E) or IgG + (F) plasmatic cells from the small intestine LP of Mincle-deficient ( Clec4e −/− ) mice and WT littermate controls. (G) Representative plots and summary graph of CD4 + PD-1 high T cells from PPs of the indicated genotypes. Data were pooled from at least two independent experiments. Individual mice and the arithmetic mean of each group are indicated. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 (A and B: one-way ANOVA and Bonferroni post hoc test; C–H: unpaired two-tailed Student’s t test). See also Figure S6 .

Article Snippet: anti-mouse CD45.1, eFluor 450, clone A20 , eBiosciences , Cat# 48-0453-82; RRID: AB_1272189.

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Two Tailed Test

Journal: Immunity

Article Title: Microbiota Sensing by Mincle-Syk Axis in Dendritic Cells Regulates Interleukin-17 and -22 Production and Promotes Intestinal Barrier Integrity

doi: 10.1016/j.immuni.2018.12.020

Figure Lengend Snippet: Mincle-Syk Pathway Promotes Commensal Bacteria Containment (A) Bacterial translocation into the liver of Mincle-deficient ( Clec4e −/− ) mice and WT littermates. On the left is the bacterial load as colony-forming units (CFUs) per organ, indicating the limit of detection (LOD). On the right, the frequencies of the mice of each genotype show more than 40 CFUs per organ (2× LOD). (B) 16S rRNA sequencing analysis of LB-grown commensals from the liver (left) or bacterial DNA directly extracted from the liver (right) in the indicated genotypes. The graph shows the percentage of total reads corresponding to each phylum. Each bar represents four pooled LB plates per mouse and four mice per sample of each genotype (LB-grown commensals) or six pooled mice per genotype (commensals from the liver). (C) ELISA of serum IgG against intestinal bacteria in the indicated genotypes. (D) Total numbers of CD45 + CD11b high cells infiltrated in the liver of the indicated genotypes in the steady state or after administration of an antibiotic cocktail (ABX). (E) Total (left) or direct (right) bilirubin in the indicated genotypes. (F) Acc , Fas , Scd1 , Srebpc , G6pase , Pepck , Cpt1a , and Ppara transcripts from the liver of 15 mice of the indicated genotypes were analyzed by qPCR and normalized to Gapdh in three independent experiments; the graph shows fold induction compared with the WT. (G and H) Individual lipid species of diacylglycerides (DAGs) (G) and the margaric (C17:0) and linoleic (C18:2) acid classes of free fatty acids (H) measured by a liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) metabolomics-based profiling approach. Data are presented as the metabolites’ abundance. Data were pooled from at least two independent experiments. Individual mice and the arithmetic mean of each group are shown (A, C–E, G, and H). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 (A–C and E–H [margaric acid]: unpaired two-tailed Student’s t test; H [linoleic acid]: Mann-Whitney U test; D: one-way ANOVA and Bonferroni post hoc test). See also Figure S7 .

Article Snippet: anti-mouse CD45.1, eFluor 450, clone A20 , eBiosciences , Cat# 48-0453-82; RRID: AB_1272189.

Techniques: Translocation Assay, Sequencing, Enzyme-linked Immunosorbent Assay, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Gas Chromatography, Gas Chromatography-Mass Spectrometry, Two Tailed Test, MANN-WHITNEY

Journal: Immunity

Article Title: Microbiota Sensing by Mincle-Syk Axis in Dendritic Cells Regulates Interleukin-17 and -22 Production and Promotes Intestinal Barrier Integrity

doi: 10.1016/j.immuni.2018.12.020

Figure Lengend Snippet:

Article Snippet: anti-mouse CD45.1, eFluor 450, clone A20 , eBiosciences , Cat# 48-0453-82; RRID: AB_1272189.

Techniques: Blocking Assay, Negative Control, Recombinant, Staining, Amplification, Enzyme-linked Immunosorbent Assay, PCR Cloning, Software