Journal: Journal of Virology

Article Title: Early Transcriptional Changes within Liver, Adrenal Gland, and Lymphoid Tissues Significantly Contribute to Ebola Virus Pathogenesis in Cynomolgus Macaques

doi: 10.1128/JVI.00250-20

Figure Lengend Snippet: Antibodies used for flow cytometric analysis of immune cell subsets a

Article Snippet: Gating strategies and antibody information were previously described ( 20 ) ( ). table ft1 table-wrap mode="anchored" t5 TABLE 3 caption a7 Flow panels Antibody Clone b Company b Catalog no. b Monocyte/DC basic Mouse anti-human CD3 FITC SP34-2 BD 556611 Mouse anti-human CD20 FITC 2H7 BD 556632 Mouse anti-human CD16 PE 3G8 BioLegend 302008 Mouse anti-human CD11c PE-Cy7 3.9 BioLegend 301608 Mouse anti-human CD14 APC M5E2 BioLegend 301808 Mouse anti-human CD123 PerCP-Cy5.5 6H6 BioLegend 306016 Mouse anti-human HLA-DR APC-Cy7 L243 BioLegend 307618 NK Mouse anti-human CD3 FITC – – – Mouse anti-human CD20 FITC – – – Mouse anti-human CD14 FITC M5E2 BioLegend 301804 Mouse anti-human CD56 PE-Cy7 B159 BD 557747 Mouse anti-human granzyme B Alex647* GB11 BioLegend 515405 Mouse anti-human CD159a PE Z199 Beckman Coulter IM3291U Mouse anti-human CD8a APC-Cy7 RPA-T8 BioLegend 301016 Mouse anti-human CD16 PerCP-Cy5.5 3G8 BioLegend 302028 DC Activation Mouse anti-human CD3 FITC – – – Mouse anti-human CD20 FITC – – – Mouse anti-human CD14 FITC M5E2 BioLegend 301804 Mouse anti-human HLA-DR APC-Cy7 – – – Mouse anti-human CD11c PE-Cy7 – – – Mouse anti-human CD123 PerCpCy5.5 – – – Mouse anti-human CD1c APC AD5-8E7 Miltenyi 130-090-903 Mouse anti-human CD86 PE IT2.2 BioLegend 305406 T cells Mouse anti-human CD4 PerCp-Cy5.5 OKT4 Tonbo 65-0048 Mouse anti-human CD8b PE 2ST8.5H7 BD 641057 Mouse anti-human CD28 PE-Cy7 CD28.2 Tonbo 60-0289 Mouse anti-human CD95 APC DX2 BD 558814 Mouse anti-human CCR7 APC-Cy7 G043H7 BioLegend 353212 Mouse anti-human Ki67 FITC* B56 BD 556026 B cells Mouse anti-human CD20 PE-Cy7 2H7 eBioscience 25-0209-73 Mouse anti-human CD27 APC O323 Tonbo 20-0279 Mouse anti-human IgD PE IA6-2 Biolegend 348204 Mouse anti-human CD40 APC-Cy7 5C3 BioLegend 334324 Mouse anti-human Ki67 FITC* – – – Mouse anti-human IgG PerCP-Cy5.5 HP6017 BioLegend 409312 Open in a separate window a *Denotes intracellular antibody. b –, Same as monocyte/DC basic panel.

Techniques: Activation Assay

Journal: Frontiers in Immunology

Article Title: Tracking of an Oral Salmonella -Based Vaccine for Type 1 Diabetes in Non-obese Diabetic Mice

doi: 10.3389/fimmu.2020.00712

Figure Lengend Snippet: In vivo detection of the red fluorescent protein in different organs of vaccinated NOD mice. NOD mice were given a single oral dose of Salmonella -containing plasmid (pSBbi-RP/SB100, n = 10) compared to Salmonella only ( n = 10). Four weeks post-vaccination, red fluorescent protein was detected in different organs. (A) Representative FACS plots gated the frequency of live red-labeled CD11c + and CD11c – cells in the spleens, pancreatic lymph nodes (PLN), mesenteric lymph nodes (MLN), and Peyer’s patches (PP) of mice from each group. (B) Quantification of the percentages of the CD11c + and CD11c – red-labeled cells in all organs. (C) Quantification of the percentages of red-labeled CD11c + MHCII + in total live cells isolated from the spleen, PLN, MLN, and PP. The data shown are as the average with standard deviation (SD) obtained from two independent experiments. Statistical analysis using two-way ANOVA shows significance between groups (**** p < 0.0001).

Article Snippet: Dendritic cells (DCs) were isolated from pooled splenocytes (5 mice) isolated from either vaccinated or non-vaccinated mice using PE-conjugated CD11c antibody (Miltenyi, 130-110-701) followed by anti-PE microbeads (Miltenyi Biotec, 130-091-041).

Techniques: In Vivo, Plasmid Preparation, Labeling, Isolation, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Tracking of an Oral Salmonella -Based Vaccine for Type 1 Diabetes in Non-obese Diabetic Mice

doi: 10.3389/fimmu.2020.00712

Figure Lengend Snippet: Vaccine-delivered TGFβ alters levels of DCs in secondary lymphoid tissues. NOD mice were given a single oral dose of Salmonella (pSBbi-RP-TGFβ/SB100, n = 10) or Salmonella (pSBbi-RP/SB100, n = 10). Four weeks post-vaccination, red fluorescent protein was detected in DCs of different organs. (A) Representative FACS plots gated the frequency of MHCII + CD11c + Td-Tomato + cells in the spleens, PLN, MLN, and PP of mice from each group. (B) Quantification of the percentages of MHCII + CD11c + Td-Tomato + cells in the spleen, PLN, MLN, and PP. The data shown are the average with SD from two independent experiments. Statistical analysis using two-way ANOVA shows significance between Salmonella (pSBbi-RP-TGFβ/SB100) and Salmonella (pSBbi-RP/SB100) groups (*** p < 0.001, **** p < 0 0001).

Article Snippet: Dendritic cells (DCs) were isolated from pooled splenocytes (5 mice) isolated from either vaccinated or non-vaccinated mice using PE-conjugated CD11c antibody (Miltenyi, 130-110-701) followed by anti-PE microbeads (Miltenyi Biotec, 130-091-041).

Techniques:

Journal: Frontiers in Immunology

Article Title: Tracking of an Oral Salmonella -Based Vaccine for Type 1 Diabetes in Non-obese Diabetic Mice

doi: 10.3389/fimmu.2020.00712

Figure Lengend Snippet: Vaccine-delivered TGFβ alters levels of gut DCs. NOD mice were given a single oral dose of Salmonella (pSBbi-RP-TGFβ/SB100, n = 10) or Salmonella (pSBbi-RP/SB100, n = 10). Four weeks post-vaccination, red fluorescent protein was detected in DCs of different organs. (A) Representative FACS plots gated the frequency of the live red-labeled MHCII + CD11c + CD103 + cells in the spleens, PLN, MLN, and PP of mice from each group. (B) Quantification of the numbers of MHCII + CD11c + CD103 + in red-labeled cells in the spleen, PLN, MLN, and PP. The data shown are the average with SD from two independent experiments. Statistical analysis using two-way ANOVA shows significance between Salmonella (pSBbi-RP-TGFβ/SB100) and Salmonella (pSBbi-RP/SB100) groups (**** p < 0.0001).

Article Snippet: Dendritic cells (DCs) were isolated from pooled splenocytes (5 mice) isolated from either vaccinated or non-vaccinated mice using PE-conjugated CD11c antibody (Miltenyi, 130-110-701) followed by anti-PE microbeads (Miltenyi Biotec, 130-091-041).

Techniques: Labeling

Journal: Frontiers in Immunology

Article Title: Tracking of an Oral Salmonella -Based Vaccine for Type 1 Diabetes in Non-obese Diabetic Mice

doi: 10.3389/fimmu.2020.00712

Figure Lengend Snippet: Endogenous TGFβ alters PDL-1 and CTLA4 expression in secondary lymphoid tissues. NOD mice were given a single oral dose of Salmonella (pSBbi-RP-TGFβ/SB100) or Salmonella (pSBbi-RP/SB100). Four weeks post-vaccination, the fold changes in gene expression of (A) CD274 (PDL-1) in sorted CD11c cells and (C) CTLA4 in sorted CD4 cells isolated from specified organs was determined. Percentage of CD11c + cells expressing PDL-1 (B) or CD4 + cells expressing CTLA4 (D) were quantified in the spleen, PLN, MLN, and PP. The data shown are the average with SD from two independent experiments. Statistical analysis using two-way ANOVA shows significance between Salmonella (pSBbi-RP-TGFβ/SB100) and Salmonella (pSBbi-RP/SB100) groups (** p < 0.01, *** p < 0.001, **** p < 0 0001).

Article Snippet: Dendritic cells (DCs) were isolated from pooled splenocytes (5 mice) isolated from either vaccinated or non-vaccinated mice using PE-conjugated CD11c antibody (Miltenyi, 130-110-701) followed by anti-PE microbeads (Miltenyi Biotec, 130-091-041).

Techniques: Expressing, Gene Expression, Isolation

Journal: Frontiers in Immunology

Article Title: Tracking of an Oral Salmonella -Based Vaccine for Type 1 Diabetes in Non-obese Diabetic Mice

doi: 10.3389/fimmu.2020.00712

Figure Lengend Snippet: A Salmonella -based vaccine increase DCs in lymphoid organs. NOD mice were injected with 2.5 μg/mouse of anti-CD3 mAb for 5 days followed by 2 doses of oral vaccine containing PPI + TGFβ + IL10 ( n = 5). Four weeks post-vaccination, DCs were analyzed in different organs by FACS analysis. (A) Quantification of the percentage of CD11c + in total live cells of the spleen, PLN, MLN, and PP. (B) MLN, (C) PP, (D) PLN, and (E) spleen expression of CD80, CD86, and MHCII was measured in DCs from each organ. The data shown are the average with SD from two independent experiments. Statistical analysis using two-way ANOVA shows significance between vaccine and vehicle treated groups (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: Dendritic cells (DCs) were isolated from pooled splenocytes (5 mice) isolated from either vaccinated or non-vaccinated mice using PE-conjugated CD11c antibody (Miltenyi, 130-110-701) followed by anti-PE microbeads (Miltenyi Biotec, 130-091-041).

Techniques: Injection, Expressing