| 摘要 |
A method for determination of functional state of human blood lymphocytes includes sampling venous blood with anticoagulant, separation of lymphocytes from it by means of density gradient phycol-verografin, setting reaction of rosette formation with ram erythrocytes conjugated with monoclonal antibodies for each separate CD-structure of diagnostic kit for lymphocyte phenotyping, centrifugation of the mixture of cells, thermal and cold incubation of those. Preparations are prepared, microscopic investigation and determination of cells that have bound 3 and more ram erythrocytes is performed. Characteristics obtained are compared to the age physiological norm. Functional state of lymphocytes is determined by phenotyping data. Culture mixture comprising: medium 199 and embryonic calf serum at volumetric ratio 4:1 with pH 7.2 is prepared. Additional centrifugation of erythrocytic diagnosticum “Anti-CD” is performed with following replacement of its liquid fraction with the same volume of cultural mix. Fixation of preparations is carried out with 0.6 % solution of glutaric aldehyde prepared on cultural mix, after that removal of supernatant is performed. Dyed preparations are differentiated, with determination among rosette-forming cells of activated lymphocytes that bind 8 and more ram erythrocytes. Activation index is calculated, and functional state of human blood lymphocytes is determined according to activation index and phenotyping data. |
