Lymphocyte toxicity test

When specific effector T lymphocytes are contacted with target cells in vitro, they may exhibit properties that destroy and lyse target cells, called cytotoxicity. The target cell can be a tumor cell or other tissue cell. The way lymphocytes kill target cells can destroy target cells by direct killing or production of lymphokines. This test method has two methods: morphological examination and isotope release. The former does not require special equipment, and it is convenient to use the microscope to count the survival number of the target cells. The latter uses 125I-deoxyuridine or 51Cr to label target cells, and the release of radioisotopes is used as an indicator of damage to target cells. This method requires special equipment such as a liquid scintillation meter, but it can be automatically measured and the result is reproducible. Basic Information Specialist classification: Oncology examination classification: blood examination Applicable gender: whether men and women apply fasting: fasting Analysis results: Below normal: Normal value: no Above normal: negative: normal. Positive: No relevant data at this time. Tips: Keep a normal state of mind. Normal value The morphological examination method was compared between the test group and the control group, P<0.05. 125I or 51Cr method P>0.05. Clinical significance This test can be used as an indicator for measuring the cellular immunity of tumor patients in vitro; it can also determine the prognosis of tumor patients by measuring the ability of immune cells to kill tumor cells; and can also identify functional subpopulations of lymphocytes. Positive results may be diseases: kidney transplantation, drug dermatitis considerations (1) In measuring the ability of the subject lymphocytes to attack tumor cells, tumor cells and lymphocytes of the subject were added to the experimental group, and tumor target cells and culture medium were added to the control group. If other samples such as tumor antigens or immunogens, therapeutic drugs, etc. are involved in cellular immunity, a third group should be added. In this test group, lymphocytes are firstly allowed to react with the sample to be tested, and then washed and then observed. The cytotoxic effect of sensitized lymphocytes on tumor target cells. The first two groups became the control group at this time. (2) The survival rate of target cells and lymphocytes should be in an optimal state, generally >90%. (3) The calf serum added to the culture solution must be tested for toxicity first. (4) The count of inoculated target cells and lymphocytes should be accurate. (5) The experimental group and the control group should be arranged on the same board to reduce the error. (6) The pH of the culture solution is most preferably 6.8 to 7.2. Inspection process (1) Inoculation of target cells: Select tumor cells that can grow adherently, wash them with Hank solution, digest with 2.5 g/L trypsin for 2 to 3 min, pour the trypsin solution (the cells are still attached to the bottle wall), and use Hank. The cells were lightly washed 3 times and the Hank solution was poured. The adherent cells were washed with RPMI1640 solution containing 10% to 20% calf serum, and the cell pellet was removed by filtration through a 100-mesh filter to prepare a single tumor cell suspension of 1×10 6 /ml, and the cell suspension was pipetted with a dropper. Drop into a 40-well culture plate, 1 drop per well (about 100-150 cells), and place the plate in a 5% CO2 incubator at 37 ° C for 20 h. The culture plate was taken out and the culture solution was removed. After the Wright's staining, the average number of adherent tumor cells per 10 wells was counted. If the difference between the two groups was > 1/3, the error was too large to be used, and the error was not large. The culture plate can be formally tested. (2) Separation of lymphocytes: Take the heparin anticoagulation 3 ml of the subject, separate the lymphocytes with the sucrose-diazepam leaching solution, then wash them with Hank solution 3 times, and then mix them with RPMI1640 solution (2~3) ) × 105 / ml cell suspension. (3) Cytotoxicity test: lymphocytes and target cells are mixed at a ratio of 200:1, and (2~3)×104 lymphocytes (in 0.1 ml volume) are added to each well, and about 20% calf serum is added per well. 0.1 ml of RPMI 1640 solution was placed in a 37 ° C 5% CO 2 incubator for 40 h. The culture plate was taken out and the culture solution was removed. After Wright's staining, the number of target cells remaining at the bottom of the plate was counted under a microscope. Not suitable for the crowd There are no special taboos. Adverse reactions and risks Nothing.