| Objective:To detect the cytotoxicity and quantity of NK cells in different stages of chemotherapy in children with acute leukemia by flow cytometry in order to explore its significance in leukemia immunotherapy and disease surveillance.Methods:NK cell was detected by flow cytometry with CD107a labeling antibody in peripheral blood from16cases of new diagnosis acute leukemia(AL) children and46cases children with complete remission.15cases of healthy children were used as normal control group.16cases were preliminary diagnosis patients including11cases of acute lymphoblastic leukemia (ALL) and5cases of acute non-lymphoblastic leukemia (ANLL) patients.There were46cases of complete remission in AL children (ALL39example, ANLL7cases), and divided into three groups according to duration of treatment, including16cases of children with complete remission from six to nine months (CR-A group),14cases of children with complete remission from1year to1.5years (CR-B group),16cases of complete remission over2years (CR-C group). NK activity of peripheral blood were collected and the number of changes in cells.The peripheral blood from them were collected to detect NK cell cytotoxicity and quantity.At the same time,we used the traditional dehydrogenase (LDH) release assay to understand the correlation between the two detection methods.Results:The quantity and activity of NK cells in new diagnosis AL children were respectively (3.86±2.14)%and (5.07±2.79)%, and the quantity and activity of NK cells in children with normal control group was respectively (10.80±2.01)%and (19.27±1.64)%, the quantity and activity of NK cells in new diagnosis AL children were significantly lower than the control group (P <0.05).The quantity and activity of NK cells in complete remission children were respectively (7.01±2.76)%,(7.11±3.14)%, and were significantly higher than preliminary diagnosis patients(P<0.05). In complete remission group: The quantity of NK cells in CR-A group was (5.97±2.19)%, the number of NK cells in CR-B group was (6.59±3.11)%, and in CR-C group was (8.36±2.42)%, CR-A, CR-B, CR-C significantly higher than the new diagnosis AL childreng(P<0.05),but there was not statistically significant difference(P>0.05) between CR-A group and CR-B group. The cytotoxicity of NK cells in CR-A group was (4.78±1.92)%.The cytotoxicity of NK cells in CR-B group was (7.11±2.27)%.The cytotoxicity of NK cells in CR-C group was (9.43±3.13)%.The difference between CR-A group and the new diagnosis AL children was not statistically significant (P>0.05), and it has also no different between CR-B groupand CR-C group. The new diagnosis AL children significantly lower than CR-B group,CR-C group(P<0.05), CR-B group significantly higher than CR-A group(P<0.05), CR-C group was significantly higher than the new diagnosis AL group, CR-A group(P<0.05) and the number and activity of NK cells in normal control group were higher than CR-C group (P<0.05).The CD107a labeling antibody by flow cytometry and LDH release assay were positively correlated, and the correlation coefficient was0.754.Conclusion:1.CD107a labeling antibody by flow cytometry correlates with the LDH release assay positively,and is a good specificity, simple,reproducibility detection method.2.New diagnosis leukemia activity and number of NK cells are lower than normal controls, with time gradually increased, and they increase at the time of complete remission6-9months and from one year after complete remission desperateditily, but activity and number of NK cells in over two years complete remission are still lower than the normal.3.The NK cell number of New diagnosis ALL is higher than children with ANLL, but the activity of ANLL is higher than that of ALL... |
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