cd14 (epr3653) rabbit monoclonal antibody  (Cell Marque)

Journal: ACR Open Rheumatology

Article Title: CD14 + Dendritic‐Shaped Cells Functioning as Dendritic Cells in Rheumatoid Arthritis Synovial Tissues

doi: 10.1002/acr2.11670

Figure Lengend Snippet: CD14 high CD90 int cells in RA tissues. (A) and (B) show the sublining layer of RA synovial tissues. (C) and (D) show the cartilage/pannus junction where the synovial tissues invade the bone. (A) and (C) show HE staining. (B) and (D) show double immunofluorescence staining for CD14 (red) and CD90 (green). Scale bar, 100 μm. HE, hematoxylin and eosin; RA, rheumatoid arthritis.

Article Snippet: After heat‐induced antigen retrieval, fluorescent immunostaining was conducted using an anti‐CD14 rabbit monoclonal antibody (1:200; Abcam) and an anti‐CD90 mouse monoclonal antibody (1:1,000; ProteinTech), then visualized using goat anti‐rabbit immunoglobulin G (IgG) (1:200, Alexa Fluor 594; Abcam) and goat anti‐mouse IgG (1:200, Alexa Fluor 488; Abcam) as second antibodies.

Techniques: Staining, Double Immunofluorescence Staining

Journal: ACR Open Rheumatology

Article Title: CD14 + Dendritic‐Shaped Cells Functioning as Dendritic Cells in Rheumatoid Arthritis Synovial Tissues

doi: 10.1002/acr2.11670

Figure Lengend Snippet: Percentages of CD14 high CD90 int cells in peripheral blood samples and synovial tissues. (A) Percentages of CD14 high CD90 int cells in the peripheral blood samples are shown. (B) Percentages of CD14 high CD90 int cells in the synovial tissues are shown. Flow cytometry data shown in the bottom are representative data for each group. Each frame in data indicates CD14 high CD90 int cells. The regions of these frames were determined and fixed by comparison with OA controls. The bars show average values and SDs. The Mann–Whitney U test and Spearman's rank correlation coefficient were used to assess significant differences. * P < 0.05. OA, osteoarthritis (n = 5); RA in rem, RA in clinical remission (n = 10); RA, rheumatoid arthritis; untreated active RA, untreated and active RA (n = 10).

Article Snippet: After heat‐induced antigen retrieval, fluorescent immunostaining was conducted using an anti‐CD14 rabbit monoclonal antibody (1:200; Abcam) and an anti‐CD90 mouse monoclonal antibody (1:1,000; ProteinTech), then visualized using goat anti‐rabbit immunoglobulin G (IgG) (1:200, Alexa Fluor 594; Abcam) and goat anti‐mouse IgG (1:200, Alexa Fluor 488; Abcam) as second antibodies.

Techniques: Flow Cytometry, Comparison, MANN-WHITNEY

Journal: ACR Open Rheumatology

Article Title: CD14 + Dendritic‐Shaped Cells Functioning as Dendritic Cells in Rheumatoid Arthritis Synovial Tissues

doi: 10.1002/acr2.11670

Figure Lengend Snippet: (A) Dendritic cell–differentiation induction of RA synovial cells is shown. Subpanels show representative flow cytometry data from independent experiments (n = 7). Blue color in each subpanel indicates isotype control. (B) CD83 and HLA‐DR expression after dendritic cell–differentiation induction on day 7 is shown. The CD14 high CD90 int cell group showed significantly increased expression of CD83 and HLA‐DR compared with the non‐CD14 high CD90 int cell group. (C) Expression levels of IL‐6 and TNF‐α in the supernatant of both cell groups were significantly increased by dendritic cell–differentiation induction on day 4 and day 7 compared with day 1. These results suggest that CD14 high CD90 int cells have the potential to differentiate into CD83+ and HLA‐DR+ dendritic cells. The bars show average values and SDs. Scale bar, 100 μm. RA, rheumatoid arthritis; HLA‐DR, human leukocyte antigen‐DR; IL‐6, interleukin‐6; TNF‐α, tumor necrosis factor‐α.

Article Snippet: After heat‐induced antigen retrieval, fluorescent immunostaining was conducted using an anti‐CD14 rabbit monoclonal antibody (1:200; Abcam) and an anti‐CD90 mouse monoclonal antibody (1:1,000; ProteinTech), then visualized using goat anti‐rabbit immunoglobulin G (IgG) (1:200, Alexa Fluor 594; Abcam) and goat anti‐mouse IgG (1:200, Alexa Fluor 488; Abcam) as second antibodies.

Techniques: Cell Differentiation, Flow Cytometry, Control, Expressing

Journal: ACR Open Rheumatology

Article Title: CD14 + Dendritic‐Shaped Cells Functioning as Dendritic Cells in Rheumatoid Arthritis Synovial Tissues

doi: 10.1002/acr2.11670

Figure Lengend Snippet: (A) Overview of the co‐culture experiments of RA synovial cells and lymphocytes is shown. (B) shows pictures of phase contrast microscopy and HE‐stained histology after co‐culture of CD14 high CD90 int cells, non‐CD14 high CD90 int cells, CD14 high CD90 int cells after DC induction, and non‐CD14 high CD90 int cells after DC induction, with lymphocytes, respectively. In co‐culture of synovial cells after DC induction and lymphocytes (Bc and Bd), both synovial cells and lymphocytes proliferated remarkably. (C) IL‐6 and TNF‐α in the supernatant were also most highly expressed in the groups of synovial cells after DC induction. Experiments were performed three times for each condition. (D) shows the percentages of cell area occupied by synovial cells and lymphocytes relative to culture dish. Scale bar = 100 μm. The bars show average values and SDs. DC, dendritic cell; HE, hematoxylin and eosin; IL‐6, interleukin‐6; RA, rheumatoid arthritis; TNF‐α, tumor necrosis factor‐α.

Article Snippet: After heat‐induced antigen retrieval, fluorescent immunostaining was conducted using an anti‐CD14 rabbit monoclonal antibody (1:200; Abcam) and an anti‐CD90 mouse monoclonal antibody (1:1,000; ProteinTech), then visualized using goat anti‐rabbit immunoglobulin G (IgG) (1:200, Alexa Fluor 594; Abcam) and goat anti‐mouse IgG (1:200, Alexa Fluor 488; Abcam) as second antibodies.

Techniques: Co-Culture Assay, Microscopy, Staining

Journal: ACR Open Rheumatology

Article Title: CD14 + Dendritic‐Shaped Cells Functioning as Dendritic Cells in Rheumatoid Arthritis Synovial Tissues

doi: 10.1002/acr2.11670

Figure Lengend Snippet: Diagram showing our hypothesis of RA chronic inflammation induced by CD14+ dendritic‐shaped cells. We hypothesized that the CD14+ cells in the bone marrow flow into the synovial tissues via the circulating blood. Some of these cells contribute to RA inflammation after differentiating into HLA‐DR+ dendritic cells. The yellow‐colored cells in the immunohistochemistry images indicate the CD14 high CD90 int cells. Scale bar, 100 μm. HLA‐DR, human leukocyte antigen‐DR; RA, rheumatoid arthritis.

Article Snippet: After heat‐induced antigen retrieval, fluorescent immunostaining was conducted using an anti‐CD14 rabbit monoclonal antibody (1:200; Abcam) and an anti‐CD90 mouse monoclonal antibody (1:1,000; ProteinTech), then visualized using goat anti‐rabbit immunoglobulin G (IgG) (1:200, Alexa Fluor 594; Abcam) and goat anti‐mouse IgG (1:200, Alexa Fluor 488; Abcam) as second antibodies.

Techniques: Immunohistochemistry

Journal: Journal of Immunology Research

Article Title: Effect of LPS on Cytokine Secretion from Peripheral Blood Monocytes in Juvenile Idiopathic Arthritis-Associated Uveitis Patients with Positive Antinuclear Antibody

doi: 10.1155/2021/6691681

Figure Lengend Snippet: Immunofluorescence staining of TLR4 and CD14. (a) CD14 identifies monocytes. The cells were nearly round. (b) Some monocytes expressed TLR4 without LPS stimulation; TLR4 mainly expressed on the cell membrane. (c) Without LPS stimulation, some monocytes coexpress TLR4 and CD14.

Article Snippet: After fixation, the cells were washed with PBS and then permeabilized with HEPES Triton buffer (20 mm HEPES, 300 mM sucrose, 50 mM NaCl, 3 mM MgCl 2 , 0.5% Triton X-100, pH 7.4) for 1 h. After rinsing with PBS, the cells were blocked with 10% BSA (PBS dilution) at room temperature for 1 h and then incubated with CD14 (rabbit monoclonal antibody; Santa Cruz Biotechnology) and TLR4 (mouse monoclonal antibody; Santa Cruz Biotechnology) overnight at 4°C (all antibodies in 10% BSA/PBS were 1 : 50).

Techniques: Immunofluorescence, Staining

Journal: Journal of Immunology Research

Article Title: Effect of LPS on Cytokine Secretion from Peripheral Blood Monocytes in Juvenile Idiopathic Arthritis-Associated Uveitis Patients with Positive Antinuclear Antibody

doi: 10.1155/2021/6691681

Figure Lengend Snippet: Immunofluorescence of TLR4 and CD14 in the ANA+ group after LPS stimulation. (a) Monocytes were identified by CD14. Some cells changed from round to fusiform. (b) TLR4 was mainly expressed in the cell membrane. (c) With LPS stimulation, the mononuclear cells coexpressed TLR4 and CD14.

Article Snippet: After fixation, the cells were washed with PBS and then permeabilized with HEPES Triton buffer (20 mm HEPES, 300 mM sucrose, 50 mM NaCl, 3 mM MgCl 2 , 0.5% Triton X-100, pH 7.4) for 1 h. After rinsing with PBS, the cells were blocked with 10% BSA (PBS dilution) at room temperature for 1 h and then incubated with CD14 (rabbit monoclonal antibody; Santa Cruz Biotechnology) and TLR4 (mouse monoclonal antibody; Santa Cruz Biotechnology) overnight at 4°C (all antibodies in 10% BSA/PBS were 1 : 50).

Techniques: Immunofluorescence

Journal: Journal of Immunology Research

Article Title: Effect of LPS on Cytokine Secretion from Peripheral Blood Monocytes in Juvenile Idiopathic Arthritis-Associated Uveitis Patients with Positive Antinuclear Antibody

doi: 10.1155/2021/6691681

Figure Lengend Snippet: Immunofluorescence of TLR4 and CD14 in the control group after LPS stimulation. (a) Monocytes were identified by CD14. A few cells changed. (b) TLR4 was mainly expressed on the cell membrane. (c) TLR4 and CD14 were expressed in mononuclear cells without LPS stimulation.

Article Snippet: After fixation, the cells were washed with PBS and then permeabilized with HEPES Triton buffer (20 mm HEPES, 300 mM sucrose, 50 mM NaCl, 3 mM MgCl 2 , 0.5% Triton X-100, pH 7.4) for 1 h. After rinsing with PBS, the cells were blocked with 10% BSA (PBS dilution) at room temperature for 1 h and then incubated with CD14 (rabbit monoclonal antibody; Santa Cruz Biotechnology) and TLR4 (mouse monoclonal antibody; Santa Cruz Biotechnology) overnight at 4°C (all antibodies in 10% BSA/PBS were 1 : 50).

Techniques: Immunofluorescence