Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Overview of flow cytometry analyses. (A) Gating strategy for identification of monocytes and macrophages. The peripheral blood is shown. Designation of population shown within each dot-plot is indicated above the dot-plot. Leukocytes were identified as viable (a) non-doublet (b) cells with typical light scatter properties of leukocytes (c) . Then, macrophages were gated simply as CD203a hi leukocytes (d) and marked with blue color. Monocytes were gated as CD203a low/- SWC8 - (e) CD172a hi (f) leukocytes where the CD203a low/- region was defined as the complementary region to the CD203a hi region. Then, SLA-DR + monocytes were marked with red color and SLA-DR - monocytes were marked with green color (g) . SLA-DR - region was defined as the complementary region to the SLA-DR + region. Gating order is shown in the scheme (h) . (B) Representative CD163 vs. CD14 dot-plots of macrophages (blue) and monocyte subpopulations (green: SLA-DR – , red: SLA-DR + ) in various body compartments of control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Flow Cytometry, Infection

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Primers for chemokines, chemokine receptors, CD62L and reference gene used in quantitative real-time PCR.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques:

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Monocyte subpopulations in various body compartments from control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques:

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Representative pictures of immunohistochemical detection of CD163 + cells in tracheobronchial lymph node and spleen. CD163 + cells were detected in tracheobronchial lymph nodes from control (A) and APP-infected pigs (B) and in spleen from control (C) and APP-infected pigs (D) . Immunohistochemical visualization: horseradish peroxidase, brown substrate, hematoxylin counterstain; c, cortex; ca, central artery; e, ellipsoid; f, follicle; mz, marginal zone; pals, periarterial lymphatic sheath, rp, red pulp; s, subcapsular sinus.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Immunohistochemical staining, Infection

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Intensity of CD163, CD14 and SLA-DR expression by CD14 + /CD163 + monocytes in various body compartments. Data are shown as MFI (Median of fluorescence intensity) ± S.E.M. BM, bone marrow ( n = 5); PB, peripheral blood ( n = 5), UA, lungs-unaffected area ( n = 5), DZ, lungs-demarcation zone ( n = 4), NA, lungs-necrotic area ( n = 5), TBLN, tracheobronchial lymph node ( n = 5); MLN, mesenteric lymph node ( n = 4), spleen ( n = 4). The significant differences (Kruskal-Wallis test) amongst particular compartments are indicated.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing, Fluorescence

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Chemokine receptor, CD62L, CD163 and TBP1 expression by bone marrow and peripheral blood CD163 + monocytes from control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Chemokine receptor, CD62L and CD163 expression in lungs from control and APP-infected pigs.

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing

Journal: Veterinary Research

Article Title: Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

doi: 10.1186/1297-9716-44-98

Figure Lengend Snippet: Scheme of chemokines and corresponding chemokine receptor mRNA expression in various organs and CD163 + monocytes. Chemokines in bone marrow, necrotic area of the lungs and tracheobronchial lymph nodes and chemokine receptors in necrotic areas of the lungs and tracheobronchial lymph nodes which were up-regulated in the APP-infected pigs compared to control are highlighted in bold. Chemokine receptors in bone marrow and peripheral blood CD163 + monocytes which were expressed at relatively lower levels are highlighted in bold. Those expressed in relatively high levels are highlighted in bold and underlined. Arrangement of the scheme was created based on the review of Bonecchi et al. .

Article Snippet: BM and PB mononuclear cells, freshly isolated from BM and PB samples by Lympholyte-H gradient (Cedarlane, Burlington, Canada) centrifugation at 800 g , were labeled with anti-CD163 antibody for 15 min at 4 °C.

Techniques: Expressing, Infection