Method for detecting lymphocyte transformation by utilizing medicament cytochalasins B
A technology of lymphocyte transformation and cytochalasin, which is applied in the field of medical testing, can solve the problems of limited laboratory research, high laboratory requirements, and limited wide application, and achieve the effect of low equipment requirements, low experimental cost, and accurate results
Inactive Publication Date: 2012-06-20
FUDAN UNIV
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Problems solved by technology
Due to the use of radioactive materials, this method is demanding on the laboratory and may cause radioactive contamination of the environment and workplace
At the same time, this method reflects the cellular immune function through radioactive counting, and the number of cells in the experimental group and the control group must be absolutely the same, otherwise the experimental results will not be credible
The shortcomings of these two aspects limit the wide application of this method
[0006] Due to the defects of the above conventional methods themselves, a series of problems will be encountered in practical application; in addition, due to their complexity and precision, these conventional methods are absolutely not suitable for clinical use and can only be limited to laboratory research
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[0027] (1) Materials.
[0028] 1 Phytohemagglutinin (Sigma Company), cytochalasin B (Sigma Company).
[0029] 2 The culture medium is RPMI1640 (PAA), containing 10% calf serum (Sijiqing), and 1% double antibody (Geno).
[0030] 3 Heparin anticoagulation tubes.
[0031] 4 Normal saline, 087% NH 4 Cl solution, Giemsa staining solution.
[0032] (2) Operation method.
[0033] 1. Sterile blood collection, 0.5ml each, divided into 2Gy irradiation group (immunosuppressive group) and non-irradiation group, respectively injected into 4.5ml RPMI1640 culture medium containing PHA, the final concentration of PHA was 50μg / ml, mixed well, placed in Incubate in a 37°C incubator for 2 days.
[0034] 2. On the third day, cytochalasin B was added to the culture medium with a final concentration of 3 μg / ml, and the cells were cultured for 24 hours.
[0035] 3. Lyse red blood cells: Discard the supernatant on day 4, add 0.87% NH prewarmed at 37°C 4 Cl solution 3ml, incubate in a 37°C i...
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The invention relates to the technical field of medical detection, and particularly relates to a method for detecting lymphocyte transformation by utilizing a medicament cytochalasins B. According to the invention, the principle that peripheral blood lymphocytes can be transformed into lymphoblasts which are divisive after being stimulated by phytohemagglutinin in vitro and the principle that the cytochalasins B can retard cell division but can not retard cell nucleus division are utilized, 2 Gyg ray irradiation is carried out on the peripheral blood of a human body for further realizing immunodepression, and PHA (phytohemagglutinin) stimulation is carried out and the cytochalasins B is added for treatment, so that the difference between the dikaryon rate of radiated lymphocytes and the dikayron rate of non-irradiated lymphocytes is detected. The result shows that transformed lymphocytes can form dikaryocytes, the dikaryon rate of the radiated lymphocytes can be obviously reduced, the immunity of radiated T lymphocytes can be obviously inhibited, and the dikaryon rate can better reflect the immune function of the lymphocytes. The method is intuitive, simple in operation, rapid, economical, and accurate in result during detection of indexes.
Description
technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a method for detecting lymphocyte transformation. Background technique [0002] Lymphocyte transformation test is one of the in vitro test methods for detecting cellular immune function. It is a necessary experimental item for students majoring in medicine, especially students majoring in testing. It is mainly used clinically for tumors, delayed allergy, cellular immune deficiency, Diagnosis of organ transplantation and certain diseases that may involve cellular immunity. [0003] Activation or sensitization of lymphocytes is known to refer to the in vitro association of a process that normally occurs in vivo when antigens react with sensitized lymphocytes. Lymphocytic transformation is a term first used by Nowell in 1960 and later by Hirschorn to describe the morphological process by which small resting lymphocytes are transformed into lymphoblasts by expo...
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IPC IPC(8): C12Q1/06
Inventor 邵春林袁德晓潘燕叶爽张江虹白杨
Owner FUDAN UNIV
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