Journal: bioRxiv

Article Title: Myeloid-Derived Suppressor Cells are relevant factors to predict the severity of multiple sclerosis

doi: 10.1101/2022.04.20.488896

Figure Lengend Snippet: A-C: CD14 + cells showed different levels of HLA-DR in demyelinating lesions. Based on the intensity of HLA-DR (á′, b′′, ć′) we identified two cell subpopulations: CD14 + HLA-DR -/low cells were considered as M-MDSC-like cells ( A , B ) and CD14 + HLA-DR hi cells were identified as putative inflammatory macrophages ( C ). The lack of CD15 expression in CD14 + cells (á′′, b′′′, ć′′) corroborated the exclusive presence of M-MDSC-like cells in MS lesions. D - G : Both CD14 + cell subpopulations were stained for CD11b, confirming their myeloid cell nature. Arrowheads point to myeloid cells with M-MDSC phenotype, i.e CD14 + CD11b + HLA-DR low , while arrows indicate CD11b + inflammatory macrophages. H - O : TMEM119 + cells with microglial morphology were detected in the NAWM ( H - K ). The lack of TMEM119 immunoreactivity observed in amoeboid CD14 + HLA-DR -/low M-MDSC-like cells (pointed by arrows on L-O) confirmed the peripheral origin of these cells. Yellow and white arrowheads in L-O point to inflammatory monocyte-derived macrophages (CD14 + HLA-DR hi ) and microglial-derived macrophages (CD14 - HLA-DR hi ), respectively. P : Quantification of the density of putative M-MDSC in all MS lesion types from SPMS and PPMS patients. The abundance of CD14 + CD15 - HLA-DR -/low cells was significantly higher in the AL and in the rAIL (n=46 AL, 27 AIL and 24 IL from 33 MS patients). Q: AL and the rAIL from SPMS and PPMS patients showing a greater presence of M-MDSC-like cells than in cAIL and IL (n = 24 AL, 14 AIL and 14 IL from 20 SPMS patients; 22AL, 12 AIL and 10 IL from 13 PPMS patients). Scale bar: A-C = 5 µm; insets in A-C = 10 µm; D-G = 40 µm; H-K = 20 µm; L-O = 37 µm. Comparative analyses were carried out with ANOVA and the mean + SEM data are shown: *, # p < 0.05. In Q, * represents the difference with respect to AL and # with respect to the rAIL. rAIL, rim of AIL; cAIL, center of AIL; High. Inf Area, high inflammatory area; Weak Inf Area, weak inflammatory area.

Article Snippet: Subsequently, immunohistochemistry (IHC) or immunofluorescence (IF) staining was performed by incubating sections overnight at 4°C with the following primary antibodies: rabbit anti-CD11b (1:100; Abcam, ab133357); rabbit anti-TMEM119 (1:50; Sigma-Aldrich, HPA0518870); biotinylated sheep anti-CD14 (1:25; R&D, BAF383), mouse anti-CD15 (1:25; Agilent, ISO62, carb-3 clone) and mouse anti-human leukocyte antigen (HLA)-DR (1:200 for IHC, 1:100 for IF; Agilent, M0746, TAL.1B5 clone).

Techniques: Expressing, Staining, Derivative Assay

Journal: bioRxiv

Article Title: Myeloid-Derived Suppressor Cells are relevant factors to predict the severity of multiple sclerosis

doi: 10.1101/2022.04.20.488896

Figure Lengend Snippet: A-L: The highest density of M-MDSC was always seen in high inflammatory areas, independent of the disease duration (long DD in A - C , short DD in G - I ). Both the cAIL and IL from SPMS with a long ( D - F ) and short disease duration ( J - L ) have a low density of M-MDSC. The arrows indicate M-MDSC (CD14 + CD15 - HLA-DR -/low ). M : Quantification of the M-MDSC distribution showing that the disease duration does not affect their density, which was always significantly higher in high inflammatory areas. N: Ratio between CD14 + HLA-DR hi cells/M-MDSCs (also referred as IIR in the main text) was significantly increased in high inflammatory areas of SPMS, independently on the disease duration. N = 11 SPMS with short DD and 9 SPMS with long DD. Scale bar: A-L = 20 µm. A comparative analysis was carried out with a Student’s t test, the means (± SEM) data are indicated: *p < 0.05, **p < 0.01 and ***p < 0.001. DD, disease duration. IIR: inflammatory-immunoregulatory ratio.

Article Snippet: Subsequently, immunohistochemistry (IHC) or immunofluorescence (IF) staining was performed by incubating sections overnight at 4°C with the following primary antibodies: rabbit anti-CD11b (1:100; Abcam, ab133357); rabbit anti-TMEM119 (1:50; Sigma-Aldrich, HPA0518870); biotinylated sheep anti-CD14 (1:25; R&D, BAF383), mouse anti-CD15 (1:25; Agilent, ISO62, carb-3 clone) and mouse anti-human leukocyte antigen (HLA)-DR (1:200 for IHC, 1:100 for IF; Agilent, M0746, TAL.1B5 clone).

Techniques:

Journal: bioRxiv

Article Title: Myeloid-Derived Suppressor Cells are relevant factors to predict the severity of multiple sclerosis

doi: 10.1101/2022.04.20.488896

Figure Lengend Snippet: A: Representative flow cytometry plots for M-MDSCs in human PBMC samples. In both control and MS patients, live MNCs were gated after removing cellular aggregates and death cells based in Zombi-NIR expression. Next, immature myeloid cells were gated from monocyte subpopulation as CD33 + HLA-DR −/low populations. CD33 + HLA-DR −/low cells were assessed for CD14 and CD15 expression to identify the M-MDSC subset defined as CD14 + CD15 - (in this panel, the percentage of M-MDSCs refers to MNCs). B : The abundance of M-MDSCs measured in MS patients at an early stage of their disease course seemed to be higher than controls, although no significant differences were seen. C: The M-MDSC load in human peripheral blood was inversely correlated with the time elapsed from the first relapse to sampling. D : M-MDSCs were more abundant in MS patients at the time of their first referred relapse (MS-R, <30 days after the relapse) than in controls. E-F: In the sub-cohort of MS patients in relapse who were followed up for one year, the abundance of M-MDSCs at baseline was inversely correlated with the EDSS at baseline ( E ) and with the EDSS one year later ( F ). G : MS patients at relapse with full recovery (with an EDSS of 0 at 12 months) had a higher proportion of M-MDSCs than those with partial recovery. For correlation analysis a Spearman test was carried out (Control in B n= 26; MS in B and C n= 39; MS ≤30 days in D-G n = 23; MS >30 days in D n = 16; Total recovery n = 14 and partial recovery n = 9 MS patients in G). Comparative analysis in D was carried out with ANOVA and Student’s t test was used to compare groups of patients in B and G. The mean (± SEM) are represented, at *p < 0.05.

Article Snippet: Subsequently, immunohistochemistry (IHC) or immunofluorescence (IF) staining was performed by incubating sections overnight at 4°C with the following primary antibodies: rabbit anti-CD11b (1:100; Abcam, ab133357); rabbit anti-TMEM119 (1:50; Sigma-Aldrich, HPA0518870); biotinylated sheep anti-CD14 (1:25; R&D, BAF383), mouse anti-CD15 (1:25; Agilent, ISO62, carb-3 clone) and mouse anti-human leukocyte antigen (HLA)-DR (1:200 for IHC, 1:100 for IF; Agilent, M0746, TAL.1B5 clone).

Techniques: Flow Cytometry, Expressing, Sampling

Journal: Frontiers in Veterinary Science

Article Title: An isocitrate lyase gene-deleted strain of Nocardia seriolae in live attenuated vaccine development against fish nocardiosis

doi: 10.3389/fvets.2025.1664034

Figure Lengend Snippet: Immune response of hybrid snakehead after immunization with the attenuated NS-ΔICL vaccine. (A) Alkaline phosphatase (AKP), (B) acid phosphatase (ACP), (C) peroxidase (POD), (D) lysozyme (LZM) activities, and (E) specific IgM antibody levels in serum at 1, 4, 7, 14, 21, 28, and 35 days post-vaccination (d.p.v.). Data are presented as the mean ± SD ( N = 3 biological replicates; serum samples from three individual fish per group per time point, with each sample measured in technical triplicate). Asterisks indicate significant differences between the NS-ΔICL group and the PBS control group (* p < 0.05).

Article Snippet: Antibody combined with the antigen which was detected by using the rabbit anti-hybrid snakehead IgM antibody previously prepared by our lab. Microplates were incubated with goat anti-rabbit IgG HRP conjugate (BOSTER Biological Technology, China).

Techniques: Control

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Odontocete Ear Canal-Associated Lymphoid Tissue (ECALT) and Lymph Nodes: Morphological and Pathological Description with Immuno-Phenotypic Characterisation

doi: 10.3390/ani12172235

Figure Lengend Snippet: The primary antibodies.

Article Snippet: The following primary antibodies were used: monoclonal mouse anti-human CD3 (Dako M7254), monoclonal rabbit anti-human CD20 (Thermo Scientific #RB-9013-P, Runcorn, UK), polyclonal rabbit anti-human Iba-1 (Wako #019-19741), monoclonal mouse anti-human HLA-Dr (Dako M0746, Glostrup, Denmark), monoclonal mouse anti-porcine Vimentin (clone V9, Dako M0725, Glostrup, Denmark), monoclonal mouse anti-human Cytokeratin (Dako M3515, Glostrup, Denmark), and monoclonal rabbit anti-human von Willebrand factor (factor VIII) (Dako A0082, Glostrup, Denmark) (for details, see ).

Techniques: Staining, Blocking Assay, Positive Control

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Odontocete Ear Canal-Associated Lymphoid Tissue (ECALT) and Lymph Nodes: Morphological and Pathological Description with Immuno-Phenotypic Characterisation

doi: 10.3390/ani12172235

Figure Lengend Snippet: The histological images: ( A ) HE staining; ( B ) CD3; ( C ) CD20; ( D ) HLA-DR; ( E ) Iba-1; ( F ) PanCK; ( G ) Vimentin of a mononuclear infiltrate (asterisks) between the left ear canal and cartilage in a bottlenose dolphin (444_L18). Scale bars = 50 µm.

Article Snippet: The following primary antibodies were used: monoclonal mouse anti-human CD3 (Dako M7254), monoclonal rabbit anti-human CD20 (Thermo Scientific #RB-9013-P, Runcorn, UK), polyclonal rabbit anti-human Iba-1 (Wako #019-19741), monoclonal mouse anti-human HLA-Dr (Dako M0746, Glostrup, Denmark), monoclonal mouse anti-porcine Vimentin (clone V9, Dako M0725, Glostrup, Denmark), monoclonal mouse anti-human Cytokeratin (Dako M3515, Glostrup, Denmark), and monoclonal rabbit anti-human von Willebrand factor (factor VIII) (Dako A0082, Glostrup, Denmark) (for details, see ).

Techniques: Staining

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Odontocete Ear Canal-Associated Lymphoid Tissue (ECALT) and Lymph Nodes: Morphological and Pathological Description with Immuno-Phenotypic Characterisation

doi: 10.3390/ani12172235

Figure Lengend Snippet: A histological image (274/18_8R) of the immunohistochemical analysis of a lymphoid follicle stained with ( A ) anti-CD3, ( B ) anti-CD20, ( C ) anti-HLA-DR, ( D ) anti-Iba1, ( E ) anti-vimentin ( E ). Scale = 100 µm.

Article Snippet: The following primary antibodies were used: monoclonal mouse anti-human CD3 (Dako M7254), monoclonal rabbit anti-human CD20 (Thermo Scientific #RB-9013-P, Runcorn, UK), polyclonal rabbit anti-human Iba-1 (Wako #019-19741), monoclonal mouse anti-human HLA-Dr (Dako M0746, Glostrup, Denmark), monoclonal mouse anti-porcine Vimentin (clone V9, Dako M0725, Glostrup, Denmark), monoclonal mouse anti-human Cytokeratin (Dako M3515, Glostrup, Denmark), and monoclonal rabbit anti-human von Willebrand factor (factor VIII) (Dako A0082, Glostrup, Denmark) (for details, see ).

Techniques: Immunohistochemical staining, Staining