There are many kinds of antigens on the surface of T lymphocytes, and there are many classifications of cluster differentiation (CD) antigens. In addition to the types and stages of differentiation of labeled cells, these antigens also have certain functions, or as receptors, adhesion molecules, signal transduction components, etc. Subpopulations can be identified by these surface characteristic antigens. At present, more monoclonal antibodies against anti-differentiation antigens, such as anti-CD4 monoclonal antibodies, are used to detect peripheral blood mononuclear cells, and the distribution of subpopulations is determined according to the positive rate. T lymphocyte subsets are important methods for observing the cellular immune level of the body, and have important value for the diagnosis, treatment and prognosis of malignant tumors, autoimmune diseases, immune deficiency diseases and blood system diseases. There are mutual constraints and mutual aids between T cell subsets, and the increase or decrease of either side affects the formation of another subgroup.

Basic Information

Specialist classification: growth and development check classification: immunological examination

Applicable gender: whether men and women apply fasting: not fasting

Tips: This can also be done with an immunofluorescence microscope. Normal value

55% to 65%.

Clinical significance

(1) Reduce hypogammaglobulinemia, bacterial infection, and certain viral infections.

(2) elevated acute graft-versus-host disease (GVH), sarcoma, scleroderma, skin mucosal lymph node syndrome.

Precautions

Same as B cell detection. In addition, due to the non-specific binding of sheep or rabbit anti-mouse fluorescent antibodies used in indirect immunofluorescence technology to the surface of some leukocytes, and the non-specific cellular reaction of Fc receptors and the amplification effect of polyclonal fluorescent antibodies, non-specific positive fluorescence is caused. Increased cells. For the FITC direct monoclonal antibody, the antibody composition, properties and structure are completely uniform, as long as the protein-FITC complex is not charged with excessive charge, no non-specific adsorption will occur. At present, foreign countries basically use direct immunofluorescence and flow cytometry to analyze T cell subsets. Due to the high price of the instrument, it has not been widely used in China. Therefore, for general clinical laboratories, this can also be done with an immunofluorescence microscope.

Inspection process

(1) Direct immunofluorescence:

1 Take heparin anticoagulated peripheral blood, obtain mononuclear cells with lymphocyte stratification solution, and Hank liquid is formulated into (1 ~ 2) × 106 cells / ml.

2 Take 1ml of cell suspension into a 5ml plastic tube, centrifuge 2000r / min, 2min, discard the supernatant.

3 plus FITC-CD420μl, control tube plus mIgG-FITC, reaction at 4 ° C for 30min.

4Hank was washed twice, 2000r/min, 2min.

5 flow cytometry analysis.

(2) Indirect immunofluorescence:

1 Isolation of PBMC, adjusted to (1 ~ 2) × 106 cells / ml.

2 Take 1 ml of the cell suspension in a 5 ml plastic tube, centrifuge at 2000 r/min for 2 min, and discard the supernatant.

3 Add anti-CD4 monoclonal antibody 100 μl, control tube plus antibody dilution or normal mouse IgG, and react at 4 ° C for 30 min.

4Hank was washed twice, centrifuged at 2000 r/min for 2 min, and the supernatant was discarded.

5 plus 50 μl of FITC-IgG, reacted at 4 ° C for 30 min.

6 Hank was washed twice, 2000 r/min, 2 min.

7 drops, fluorescence microscopy or flow cytometry analysis.

Not suitable for the crowd

There are no taboos.

Adverse reactions and risks

There are no related complications and hazards.