Journal: Disease Markers

Article Title: Evaluation of Immune Infiltration Based on Image Plus Helps Predict the Prognosis of Stage III Gastric Cancer Patients with Significantly Different Outcomes in Northeastern China

doi: 10.1155/2022/2893336

Figure Lengend Snippet: Expression of CD4, CD8, CD20, CD56, CD68, CD117, and CD177 by immune cells infiltrated tumor tissues of GC patients. Representative images of immune markers staining in immune cells from GC samples are shown at ×200 (100 mm) original magnification. Positive expression of CD4 (a), CD8 (b), CD20 (c), CD56 (d), CD68 (e), CD117 (f), and CD177 (g) infiltrated in tumor tissues with a regular distribution. In order to show the distribution from immune cells more clearly, we used computerized imaging system Image-Pro Plus version 6.2 to highlight positive areas in different colors.

Article Snippet: After cleaned in distilled water, the paraffin sections were pretreated with citrate buffer, pH 6.0 (CD177) and EDTA Antigen Retrieval Solution, pH 8.0 (CD4, CD8, CD20, CD56, CD68, and CD117) for 3 min at 120°C in a pressure cooker, and endogenous peroxidase was inhibited with 3% H 2 O 2 in PBS for 10 min. Nonspecific actions in the sections were also blocked with goat serum (BOSTER, USA) for 1 h at room temperature.

Techniques: Expressing, Staining, Imaging

Journal: Disease Markers

Article Title: Evaluation of Immune Infiltration Based on Image Plus Helps Predict the Prognosis of Stage III Gastric Cancer Patients with Significantly Different Outcomes in Northeastern China

doi: 10.1155/2022/2893336

Figure Lengend Snippet: The schematic diagram of the distribution of immune cells. The blue points represent CD20 + B cells, the yellow points represent CD4 + T cells, the orange points represent CD117 + mast cells, the green points represent CD68 + macrophages, the pink points represent CD8 + T cells, the black points represent CD56 + NK cells, and the red points represent CD177 + neutrophils. Although not all immune cells are distributed according to this fixed law, this pattern can basically reflect the general characteristics of their distribution.

Article Snippet: After cleaned in distilled water, the paraffin sections were pretreated with citrate buffer, pH 6.0 (CD177) and EDTA Antigen Retrieval Solution, pH 8.0 (CD4, CD8, CD20, CD56, CD68, and CD117) for 3 min at 120°C in a pressure cooker, and endogenous peroxidase was inhibited with 3% H 2 O 2 in PBS for 10 min. Nonspecific actions in the sections were also blocked with goat serum (BOSTER, USA) for 1 h at room temperature.

Techniques:

Journal: Disease Markers

Article Title: Evaluation of Immune Infiltration Based on Image Plus Helps Predict the Prognosis of Stage III Gastric Cancer Patients with Significantly Different Outcomes in Northeastern China

doi: 10.1155/2022/2893336

Figure Lengend Snippet: Differences in immune marker positive area/total area between the two groups (survival time of patients in group A was less than 1 year and group B survival time was more than 5 years) by the rank sum test. (a) CD4 + T cells ( P < 0.001). (b) CD8 + T cells ( P < 0.001). (c) CD20 + B cells ( P < 0.001). (d) CD68 + macrophages ( P < 0.001). (e) CD117 + mast cells ( P < 0.001). (f) CD177 + neutrophils ( P < 0.001).

Article Snippet: After cleaned in distilled water, the paraffin sections were pretreated with citrate buffer, pH 6.0 (CD177) and EDTA Antigen Retrieval Solution, pH 8.0 (CD4, CD8, CD20, CD56, CD68, and CD117) for 3 min at 120°C in a pressure cooker, and endogenous peroxidase was inhibited with 3% H 2 O 2 in PBS for 10 min. Nonspecific actions in the sections were also blocked with goat serum (BOSTER, USA) for 1 h at room temperature.

Techniques: Marker

Journal:

Article Title: Cultured basophils but not cultured mast cells induce human IgE synthesis in B cells after immunologic stimulation

doi: 10.1046/j.1365-2249.1998.00474.x

Figure Lengend Snippet: Production of IgE, IgG4 and soluble CD23 by normal B cells mixed with cultured human mast cells or basophils. Mast cells or basophils were stimulated for 2h with antigen after sensitization with murine IgE, washed, added to highly purified B cells, and then cultured for 12 days. Stimulated mast cells were incubated with B cells in the absence or presence of recombinant IL-4. Stimulated basophils were incubated with B cells in the absence or presence of two neutralizing anti-IL-4 and anti-IL-13 MoAbs or a soluble form of CD40. Supernatants were collected and measured for concentrations of IgE, IgG4 and soluble CD23. The net synthesis of IgE and IgG4 was determined, as described in Materials and Methods. Results represent the mean ± s.e.m. of triplicate cultures. One of two comparable experiments is shown. IgE production by B cells was neither induced by unstimulated basophils, nor significantly affected in the cultures containing B cells plus stimulated basophils by the addition of control antibody or IgM (data not shown).

Article Snippet: Reagents, antigen, and antibodies Recombinant human stem cell factor (SCF) and IL-6 were generous gifts from Kirin Brewery Co. (Maebashi, Japan).

Techniques: Cell Culture, Purification, Incubation, Recombinant, Control

Journal:

Article Title: Cultured basophils but not cultured mast cells induce human IgE synthesis in B cells after immunologic stimulation

doi: 10.1046/j.1365-2249.1998.00474.x

Figure Lengend Snippet: Analysis of germ-line and mature Cε mRNA expression in normal B cells mixed with cultured human mast cells or basophils. The cell pellets obtained in Fig. 5 were used for the analysis of germ-line and mature Cε transcription. Highly purified B cells were incubated with medium alone (lane 1) or with stimulated mast cells in the absence (lane 2) or presence (lane 3) of recombinant IL-4. Stimulated basophils were incubated with B cells in the absence (lane 4) or presence of two neutralizing anti-IL-4 and anti-IL-3 MoAbs (lane 5) or a soluble form of CD40 (lane 6). After extraction of total cellular RNA, mRNA for germ-line Cε, mature Cε, and G3PDH was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR), subjected to electrophoresis, and then visualized by ethidium bromide. One of two comparable experiments is shown. Neither germ-line nor mature transcripts were detected in the cultures containing B cells plus unstimulated basophils (data not shown).

Article Snippet: Reagents, antigen, and antibodies Recombinant human stem cell factor (SCF) and IL-6 were generous gifts from Kirin Brewery Co. (Maebashi, Japan).

Techniques: Expressing, Cell Culture, Purification, Incubation, Recombinant, Extraction, Amplification, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Electrophoresis