anti rabbit txred  (Vector Laboratories)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Neutrophil vaccination dynamics and their capacity to mediate B-cell help in Rhesus Macaques

doi: 10.4049/jimmunol.1800677

Figure Lengend Snippet: PMNs were isolated from whole blood of protected (n=4), chronically infected (n=4), and naïve macaques (n=4) and co-cultured with autologous PBMCs enriched for B-cells. Enriched B-cells were also cultured alone or with a stimulation cocktail in parallel. B-cells were analyzed by flow cytometry prior to co-culture, and 18, 42, and 66 hrs post-co-culture. % AID+B-cells from (A) protected, (B) infected, and (C) naïve animals. CD27 MFI on B-cells from (D) protected, (E) infected, and (F) naïve animals. % IgD+B-cells from (G) protected, (H) infected, and (I) naïve animals. % IgM+B-cells from (J) protected, (K) infected, and (L) naïve animals. % Ki67+B-cells from (M) protected, (N) infected, and (O) naïve animals. Area under the curve values (AUCs) calculated from either the raw data or the log- or arcsine-transformed values were consistent with the assumptions of repeated measures ANOVA. * and # indicates a significant difference between B-cells + PMNs and stimulated B-cells or B-cells alone respectively. All p values were p<0.0001 except (D) # (p=0.016), (G) # (p=0.0003), (M-O) * (p=0.019), and (M,O) # (p<0.0005). The p values were corrected for multiple comparisons by the Hochberg method.

Article Snippet: Whole Blood Staining Anti-human fluorochrome-conjugated mAbs known to cross-react with Rhesus macaque antigens were used in this study, including PE anti-CD182 (6C6), PE-Cy5 anti-IgG (G18–145), PE-CF594 anti-CD32 (FL18.26), PE-CF594 anti-CD64 (10.1), AX700 anti-Ki67 (B56), BV605 anti-CD162 (KPL-1), BV605 anti-CD138 (MI15), v450 anti-IgM (G20–127), BV711 anti-CD197 (150503), BV786 anti-CD45 (D058–1283), BV786 anti-CD14 (M5E2), BUV395 anti-CD184 (12G5), BUV737 anti-CD64 (10.1), and BUV805 anti-CD14 (M5E2) (all from BD Biosciences, San Jose, CA); PerCP-eFluor710 anti-CD274 (MIH1), PerCP-eFluor710 anti-CD27 (O323), APC-Cy7 anti-CD11b (ICRF44), efluor450 anti-MPO (MPO455–8E6), and eVolve655 anti-CD20 (2H7) (all from eBioscience, San Diego, CA); FITC anti-CD2 (RPA-2.10), PE anti-CD257 (T7–241), PE-Cy7 anti-CD80 (2D10), APC anti-HLA-DR (L243), Ax647 anti-IL-21 (3A3-N2), Ax700 anti-TNFα (MAb11), and Ax750 anti-CD69 (FN50) (all from BioLegend, San Diego, CA); Ax488 anti-AID (polyclonal Rab) (Bioss); TxRed anti-IgD (Goat IgG) (Southern Biotech); PE-Cy7 anti-CD66abce (TET2) (Miltenyi); and PE anti-CD38 (OKT10) (NIH Nonhuman Primate Reagent Resource, Boston, MA).

Techniques: Isolation, Infection, Cell Culture, Flow Cytometry, Co-Culture Assay, Transformation Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Neutrophil vaccination dynamics and their capacity to mediate B-cell help in Rhesus Macaques

doi: 10.4049/jimmunol.1800677

Figure Lengend Snippet: PMNs were isolated from whole blood of protected (n=4), chronically infected (n=4), and naïve macaques (n=4) and co-cultured with autologous PBMCs enriched for B-cells with and without IL-10, or with enriched B-cells separated by a transwell system. Enriched B-cells were also cultured alone in parallel. B-cells were analyzed by flow cytometry prior to co-culture, and 18, 42, and 66 hrs post-co-culture. % CD69+B-cells from (A) protected, (B) infected, and (C) naïve animals. % CD80+B-from (D) protected, (E) infected, and (F) naïve animals. % IgD+B-cells from (G) protected, (H) infected, and (I) naïve animals. % IgG+B-cells from (J) protected, (K) infected, and (L) naïve animals. Area under the curve values (AUCs) calculated from either the raw data or the log- or arcsine-transformed values were consistent with the assumptions of repeated measures ANOVA. In (C) # shows B+PMN+IL-10 is significantly greater than both B-cell only and B+transPMN (p<0.0001), and $ shows B+PMN+IL-10 is significantly greater than B+PMN (p=0.0018). In (D), * indicates B+PMN is significantly greater than B-cell only (p=0.040). & shows a significant difference between B+PMN +/− IL-10 and B-cells only and B+transPMN in (G, I) p<0.015 and in (J, L) p<0.035. The p values were corrected for multiple comparisons by the Hochberg method.

Article Snippet: Whole Blood Staining Anti-human fluorochrome-conjugated mAbs known to cross-react with Rhesus macaque antigens were used in this study, including PE anti-CD182 (6C6), PE-Cy5 anti-IgG (G18–145), PE-CF594 anti-CD32 (FL18.26), PE-CF594 anti-CD64 (10.1), AX700 anti-Ki67 (B56), BV605 anti-CD162 (KPL-1), BV605 anti-CD138 (MI15), v450 anti-IgM (G20–127), BV711 anti-CD197 (150503), BV786 anti-CD45 (D058–1283), BV786 anti-CD14 (M5E2), BUV395 anti-CD184 (12G5), BUV737 anti-CD64 (10.1), and BUV805 anti-CD14 (M5E2) (all from BD Biosciences, San Jose, CA); PerCP-eFluor710 anti-CD274 (MIH1), PerCP-eFluor710 anti-CD27 (O323), APC-Cy7 anti-CD11b (ICRF44), efluor450 anti-MPO (MPO455–8E6), and eVolve655 anti-CD20 (2H7) (all from eBioscience, San Diego, CA); FITC anti-CD2 (RPA-2.10), PE anti-CD257 (T7–241), PE-Cy7 anti-CD80 (2D10), APC anti-HLA-DR (L243), Ax647 anti-IL-21 (3A3-N2), Ax700 anti-TNFα (MAb11), and Ax750 anti-CD69 (FN50) (all from BioLegend, San Diego, CA); Ax488 anti-AID (polyclonal Rab) (Bioss); TxRed anti-IgD (Goat IgG) (Southern Biotech); PE-Cy7 anti-CD66abce (TET2) (Miltenyi); and PE anti-CD38 (OKT10) (NIH Nonhuman Primate Reagent Resource, Boston, MA).

Techniques: Isolation, Infection, Cell Culture, Flow Cytometry, Co-Culture Assay, Transformation Assay