To Study Immunophenotyping Of Leukemia By A Cell Microarray

ObjectiveLeukemia is one of major malignant tumors resulted in the death. Adult deaths from acute leukemia account for the first among 35 years old. According to statistics, the mortality rate of leukemia is the highest in Western Europe and North American for 3.2-7.4/10 million population; the mortality rate of leukemia is 2.8-4.5/10 million population in Asia and South American. Chinese annual incidence and mortality rates are 2-4/10 million population with a significant difference from Europe and the United States. During the diagnosis and treatment, the type of leukemia is the key. To different types, different clinic program and prognosis should be performed. Before the 1980's leukemia is diagnosis by the FAB classification criteria. Now MICM typing is benefiting to leukemia diagnosis, standard detection of residual disease and the prognosis. At present, immunohistochemical techniques or FCM is usually choosed. But both of them need large specimen volume,complic procedures,and other restrictions such as analysis only 1-4 surface antigens. Reseachers have begun to use biochip technological platform which is high-throughput, paralleled for the integration of leukemia immunophenotyping. On the basis of previous experiments, I conducted in-depth research in the application of the cell microarray in immunophenotyping of acute leukemia, in order to decide the best spotting conditions , the best incubation conditions, to comfirm the stability, I have used the microarrays to immunophenotyped 72 leukemias with peripheral blood samples. Get a quick, simple and complete immunization data for the clinical diagnosis and antibodies-targ treatment.Methods1. To determine the best spotting conditions2. to determine the best incubating conditions.3. to confirm the stability. 4. Immunophenotyping clinical 72 samples.Results1. In 50μg/ml, signal intensity is the saturation, in 25μg/ml, 12.5μg/ml, 6.25μg/ml signal intensity is gradually weakened.2. When at room temperature (20℃),with the cell suspension (5×10~6/mL) ,being incubated for 45 minutes ,the chips capture lattice cell with the highest SNR.3. Make certain to the stability(1) For the three samples ,at (25.0±0.5)°C and relative humidity 60%±5%, after 6 months, the Cell chips did not change the ability to capture cells.(2) Stored at -20℃for 12 months, chips did not change the ability to capture cells.(3) Stored at(4.0±0.5)℃,relative humidity 75%±5%, after 12 months, chips did not change the ability to capture cells.4. Immunophenotyping results of 72 cases of acute leukemia: B-ALL : 14 cases express HLA-DR,CD19,5 cases express CD20,CD22, 8 cases express CD79b, 9 cases express CD10, 1 case express CD34. T-ALL: 8 cases express CD2, 12 cases express CD7, 10 cases express TCRαβor TCRγδ. AML: 46 cases express CD33, 30cases express CD13, 22 cases express CD15, 14 cases express CD11b and CD14, 1 case express CD235a, 1 case express CD4, 23 cases express HLA-DR.ConclusionsIn this study, using 50μg/ml monoclonal antibodies,at room temperature conditions (20℃) and cell suspension (5×10~6/ mL) incubated for 45 minutes,chips get to the highest SNR. This cell chips are of good stability. The experimental results meet with the immunophenotyping of leukemia reported. The cell microarray can be easy to use, good repeatability and stability. For clinical leukemia immunophenotyping, it provides a fast, high-throughput, parallel, integrated technology platform.