Journal: Glia

Article Title: Activated microglia do not increase 18 kDa translocator protein ( TSPO ) expression in the multiple sclerosis brain

doi: 10.1002/glia.24052

Figure Lengend Snippet: Antibodies for immunohistochemistry

Article Snippet: CD68 , Rabbit , Atlas Antibody , HPA048982 , 1:4000.

Techniques:

Journal: Cellular and Molecular Life Sciences

Article Title: Neutrophils play a major role in the destruction of the olfactory epithelium during SARS-CoV-2 infection in hamsters

doi: 10.1007/s00018-022-04643-1

Figure Lengend Snippet: Iba1 + (microglia/monocyte lineage), CD68 + (macrophages) and MPO + (neutrophils) cells presence in the olfactory epithelium before and during SARS-CoV-2 infection. Immunostaining on successive slides of the olfactory epithelium from a non-infected ( A ) or 1 dpi hamster ( B ). Only Iba1 + cells are present in the uninfected olfactory epithelium (OE) and in the lamina propria (LP). In the infected epithelium, Iba1 + cells are massively present in the OE while CD68 + and MPO cells are mostly present in the desquamated cells (red asterisk) in the lumen of the nasal cavity (white asterisk)

Article Snippet: The sections were then incubated overnight with primary antibodies directed against SARS nucleocapsid protein (1/500; mouse monoclonal; clone 1C7C7; Sigma-Aldrich), ionized calcium-binding adapter molecule 1 (Iba1) (1/500; rabbit monoclonal; clone EPR16588; Abcam), myeloperoxidase protein (MPO) (1/500; rabbit monoclonal; clone EPR20257; Abcam), CD68 (1/200; rabbit polyclonal; PA1518; Boster), cleaved caspase 3 (C3C) (1/200; rabbit polyclonal; #9661; Cell signaling), G olf (1/300; rabbit polyclonal; C-18; Santa Cruz), Olfactory Marker Protein (OMP) (1/500; goat polyclonal; 544-10001; Wako) and CK18 (1:50; mouse polyclonal; MAB3234—RGE53, Sigma-Aldrich).

Techniques: Infection, Immunostaining

Journal: Cellular and Molecular Life Sciences

Article Title: Neutrophils play a major role in the destruction of the olfactory epithelium during SARS-CoV-2 infection in hamsters

doi: 10.1007/s00018-022-04643-1

Figure Lengend Snippet: CD68 + macrophage and MPO + neutrophil cells are associated with damage of the olfactory epithelium during SARS-CoV-2 infection. CD68 + ( A 1 ) and MPO + ( B 2 ) signal in the olfactory epithelium (OE, left) and lamina propria (LP, right) in either control animals (CTL) or at 1 or 2 days post-infection (dpi) (Mean normalized to control ± SEM, n = 4, * p < 0.05 (Mann–Whitney test)). Correlation between score damage and percentage of CD68 + ( A 1 ) and MPO + ( B 2 ) signal in the olfactory epithelium (left panel) and the lamina propria (right panel). Spearman test p value

Article Snippet: The sections were then incubated overnight with primary antibodies directed against SARS nucleocapsid protein (1/500; mouse monoclonal; clone 1C7C7; Sigma-Aldrich), ionized calcium-binding adapter molecule 1 (Iba1) (1/500; rabbit monoclonal; clone EPR16588; Abcam), myeloperoxidase protein (MPO) (1/500; rabbit monoclonal; clone EPR20257; Abcam), CD68 (1/200; rabbit polyclonal; PA1518; Boster), cleaved caspase 3 (C3C) (1/200; rabbit polyclonal; #9661; Cell signaling), G olf (1/300; rabbit polyclonal; C-18; Santa Cruz), Olfactory Marker Protein (OMP) (1/500; goat polyclonal; 544-10001; Wako) and CK18 (1:50; mouse polyclonal; MAB3234—RGE53, Sigma-Aldrich).

Techniques: Infection, Control, MANN-WHITNEY

Journal: Cellular and Molecular Life Sciences

Article Title: Neutrophils play a major role in the destruction of the olfactory epithelium during SARS-CoV-2 infection in hamsters

doi: 10.1007/s00018-022-04643-1

Figure Lengend Snippet: Immunosuppression induced by cyclophosphamide reduces damage of the olfactory epithelium as well as OE infection area. ( A ) Expression of innate immune genes in the nasal turbinates with or without cyclophosphamide treatment at 1 and 2 days post-infection (dpi). Iba1, CD68 and Ncf2 are related to the presence of microglia/macrophages, monocytes/macrophages and neutrophils, respectively; TNFα and IL6 are two cytokines expressed during inflammation; SARS-CoV-2 N expression is related to the SARS-CoV-2 infection. Results represent the Mean ± SEM relative to vehicle-treated hamsters ( n = 4, * p < 0.05; Mann–Whitney test). Representative images of the infected olfactory epithelium immunostained for MPO (neutrophil marker) and SARS-CoV-2 N protein in ( B ) vehicle and ( C ) cyclophosphamide treated animal (olfactory epithelium (OE), lamina propria (LP)). In the vehicle condition, the lumen (white asterisk) is filled with desquamated cells (red asterisk) containing MPO signal. In the cyclophosphamide condition, MPO signal is absent and the lumen is mostly free of cellular debris . Quantification in the OE of ( D 1 ) MPO + neutrophil presence ( D 2 ) damage score ( D 3 ) SARS-CoV-2-infected area and in the lumen of the nasal cavity of ( D 4 ) desquamated cells area and ( D 5 ) percentage of SARS-CoV-2-infected area in the desquamated cells (Mean ± SEM, n = 8 areas of the nasal cavity from 4 different animals, * p < 0.05, ** p < 0.01, *** p < 0.001 (Mann–Whitney))

Article Snippet: The sections were then incubated overnight with primary antibodies directed against SARS nucleocapsid protein (1/500; mouse monoclonal; clone 1C7C7; Sigma-Aldrich), ionized calcium-binding adapter molecule 1 (Iba1) (1/500; rabbit monoclonal; clone EPR16588; Abcam), myeloperoxidase protein (MPO) (1/500; rabbit monoclonal; clone EPR20257; Abcam), CD68 (1/200; rabbit polyclonal; PA1518; Boster), cleaved caspase 3 (C3C) (1/200; rabbit polyclonal; #9661; Cell signaling), G olf (1/300; rabbit polyclonal; C-18; Santa Cruz), Olfactory Marker Protein (OMP) (1/500; goat polyclonal; 544-10001; Wako) and CK18 (1:50; mouse polyclonal; MAB3234—RGE53, Sigma-Aldrich).

Techniques: Infection, Expressing, MANN-WHITNEY, Marker

Journal: The Journal of Experimental Medicine

Article Title: Immune homeostasis and regulation of the interferon pathway require myeloid-derived Regnase-3

doi: 10.1084/jem.20181762

Figure Lengend Snippet: Regnase-3 –deficient mice develop hypertrophic lymph nodes. (A) Frequency of mice showing lymphadenopathy in a cohort of 24 Regnase-3 −/− mice and 24 Regnase-3 +/+ littermate controls at 3–6.5 mo of age. (B) Photography of skin-draining lymph nodes of four Regnase-3 −/− mice and their Regnase-3 +/+ littermate controls at 5 mo of age. (C) Representative photography of inguinal lymph nodes of a Regnase-3 −/− mouse at 5 mo of age. Arrow indicates hypertrophic lymph node. (D) H&E staining and immunohistochemical analysis of B cells (B220), T cells (CD3), and macrophages (F4/80) in skin-draining lymph nodes of Regnase-3 −/− mice with lymphadenopathy and Regnase-3 +/+ littermate controls (representative images from n = 3/3). Magnification of images is indicated in brackets. Bars, 1,000 µm. (E) Immunohistochemical analysis of macrophages (CD68) in skin-draining lymph nodes of Regnase-3 −/− mice with lymphadenopathy and Regnase-3 +/+ littermate controls (representative images from n = 6/6). Images of enlarged and small lymph nodes are taken from the identical Regnase-3 −/− mouse. Top right: Frequency of strong positive (pos.) pixels in CD68 immunohistochemical sections of the lymph nodes was determined by Definiens software ( n = 6/6). Bars, 500 µm. (F) Top: Frequencies of B cells (CD19 + ) and T cells (CD90 + ) in enlarged and normal-sized lymph nodes of the same Regnase-3 −/− mouse and its Regnase-3 +/+ littermate control at 6 mo of age, assessed by flow cytometry (representative blots of n = 6/6). Number of total cells in lymph nodes of Regnase-3 +/+ mice and Regnase-3 −/− littermates ( n = 6/6). Bottom: Frequencies of B cells (CD19 + ), T cells (CD90 + ), CD4 + and CD8 + T cells, and CD11b + cells in enlarged lymph nodes of Regnase-3 −/− mice and their Regnase-3 +/+ littermate controls at 6 mo of age, assessed by flow cytometry ( n = 6/6). Data are represented as mean ± SEM and were compared by Mann–Whitney U test (*, P ≤ 0.05; **, P ≤ 0.01; ns, not significant).

Article Snippet: Staining antibodies were anti-B220 (RA3-6B2, rat-IgG2a; BD), anti-CD3 (SP7, rabbit IgG; Zytomed), anti-F4/80 (BM8, rat-IgG2a; Linaris), anti-MHC-II (M5/114.15.2, rat IgG; Novus Biologicals), anti-CD68 (Ab125212, rabbit IgG; Abcam) anti-ki67 (SP6, rabbit IgG; Thermo Fisher Scientific), and anti-Tyr701-phospho-STAT1 (58D6, rabbit IgG; Cell Signaling).

Techniques: Staining, Immunohistochemical staining, Software, Flow Cytometry, MANN-WHITNEY

Journal: The Journal of Experimental Medicine

Article Title: Immune homeostasis and regulation of the interferon pathway require myeloid-derived Regnase-3

doi: 10.1084/jem.20181762

Figure Lengend Snippet: Systemic IFN signaling in Regnase-3 −/− mice. (A) CD19 + B cells were isolated from enlarged lymph nodes of Regnase-3 −/− mice and their Regnase-3 +/+ littermates ( n = 3/3), and RNA was isolated and subjected to RNA sequencing. Heatmap of RNA sequencing data for all significantly up-regulated (≥2 log2 fold) genes in Regnase-3 −/− B cells is shown. GO term association to “response to IFNβ” and “response to IFNγ” is indicated for each gene. (B) Serum cytokines in Regnase-3 −/− mice and Regnase-3 +/+ littermates at 6 mo of age ( n = 8/8), assessed by multiplex assay. (C) Stat1 mRNA expression in tissues from Regnase-3 +/+ and Regnase-3 −/− mice at 8 mo of age, assessed by quantitative RT-PCR, normalized to Hprt relative (rel.) to their expression in Regnase-3 +/+ mice ( n = 5/5). (D) Left: Immunohistochemical analysis of MHC-II in liver sections of Regnase-3 −/− mice with lymphadenopathy and Regnase-3 +/+ controls (representative images). Magnification of images is indicated in brackets. Bars, 250 µm. Right: Frequency of strong positive pixels in MHC-II immunohistochemical sections of lung, kidney, and liver were determined by Definiens software ( n = 6 Regnase-3 −/− mice and 6 Regnase-3 +/+ littermate controls). (E) Frequency of MHC-II–positive macrophages (CD68 + ) of all CD68 + macrophages in the lung of Regnase-3 −/− mice with lymphadenopathy and their Regnase-3 +/+ littermate controls, assessed in immunohistochemical, consecutive sections ( n = 5/6). (F) Left: Immunohistochemical analysis of pSTAT1 in skin-draining lymph nodes from Regnase-3 −/− mice with lymphadenopathy and Regnase-3 +/+ littermate controls (representative images). Right: Frequency of strong positive pixels in pSTAT1 immunohistochemical sections of lymph nodes, determined by Definiens software ( n = 6 Regnase-3 −/− mice and 6 Regnase-3 +/+ littermate controls). Bars, 500 µm. Data are represented as mean ± SEM and were compared by Mann–Whitney U test (*, P ≤ 0.05; **, P ≤ 0.01; ns, not significant).

Article Snippet: Staining antibodies were anti-B220 (RA3-6B2, rat-IgG2a; BD), anti-CD3 (SP7, rabbit IgG; Zytomed), anti-F4/80 (BM8, rat-IgG2a; Linaris), anti-MHC-II (M5/114.15.2, rat IgG; Novus Biologicals), anti-CD68 (Ab125212, rabbit IgG; Abcam) anti-ki67 (SP6, rabbit IgG; Thermo Fisher Scientific), and anti-Tyr701-phospho-STAT1 (58D6, rabbit IgG; Cell Signaling).

Techniques: Isolation, RNA Sequencing Assay, Multiplex Assay, Expressing, Quantitative RT-PCR, Immunohistochemical staining, Software, MANN-WHITNEY

Journal: PLOS One

Article Title: Photobiomodulation therapy increases neural stem cell pool in aged 3xTg-AD mice

doi: 10.1371/journal.pone.0321668

Figure Lengend Snippet: A. Representative confocal images of Aβ plaques (6E10), microglia (Iba1) and CD68 showing diffuse microglial phagocytosis away from plaques in sham compared to localized phagocytosis around plaques in PBM-treated animals. B. Quantitative analysis of total and activated microglia show no overall differences in the neuroinflammatory environment surrounding Aβ plaques. (Unpaired t-test). C. Analysis of the distribution of CD68+ microglia shows a trend of higher concentration of phagocytic cells in contact with plaques, and decreasing phagocytosis away from plaques. (Two-way ANOVA).

Article Snippet: Up to six sections representing different regions of the hippocampus were immunostained with the following primary antibodies: Sox2 (Abcam), doublecortin (DCX, Santa Cruz), calretinin (CR, SWANT), β-amyloid (6E10, Biolegend), phospho-tau (pTau, AT8, Invitrogen), βIII-tubulin (Abcam), Iba1 (Synaptic Systems), and CD68 (Rockland).

Techniques: Concentration Assay