1.Production of anti-human c-kit monoclonal antibodies by direct intra-spleen injection of DNA vaccine
Lin SHI ; Dashui HE ; Chunling FENG ; Xiangfei YUAN ; Hao QU ; Lihua HUANG ; Liyan ZHANG ; Dongmei WANG ; Yi ZHANG ; Yuguang ZHANG
Chinese Journal of Immunology 2001;0(07):-
Objective:To prepare anti-human c-kit monoclonal antibody(McAb) by genetic immunization in spleen,and to determine practicability of these means to produce McAbs based on the biological activity of anti-human c-kit antibody.Methods:Recombinant plasmid pcDNA3.1/c-kit extracellular domain was constructed by molecular cloning techniques,and was used to immunize BALB/c mice in spleen directly to prepare mAb against human c-kit by routine hybridoma technique.FASC、fluorescence microscope and Western blot were utilized to identify the prepared antibody.Results:c-kit extracellular region was cloned and insert pcDNA3.1 plasmid successfully.Three hybridoma cell lines 6C4、2C5 and 5D5 that secrete anti-human c-kit McAbs were obtained after using intra-spleen immunization with a DNA vaccine.The isotypes of these three antibodies were all IgM,and the epitopes were different with each other.Conclusion:The method of genetic immunization into spleen can be used to prepare anti-human c-kit monoclonal antibodies.
2.Expression of cMPO in 502 cases of acute myeloid leukemia (AML) and its diagnosis significance in AML subtypes.
Lei SHANG ; Xuejing CHEN ; Yuanyuan LI ; Guiqing GUO ; Dashui HE ; Xiaojin CAI ; Bin ZHENG ; Yingchang MI ; Jianxiang WANG ; Kun RU ; Huijun WANG
Chinese Journal of Hematology 2015;36(11):906-911
OBJECTIVETo investigate the myeloperoxidase (cMPO) expression pattern by flow cytometry (FCM) in patients with acute myeloid leukemia (AML) and its role in classifying AML.
METHODSEight- color multiparametric FCM with CD45/SSC gating was used to determine the cMPO expression in 502 AML patients.
RESULTSThe positive rate of cMPO in all patients was 58.0%, in which the proportion of normal positivity, dim positivity and partial positivity was 21.5%, 34.1% and 2.4%, respectively. The remaining case (42.0%) were all negative. In AML with t (15;17）(q22;q12)/PMLRARα, the positive rate was the highest (100%) and the intensity was similar to that of the normal granular leukocytes, followed by AML with t (8;21（q22;q22/RUNX1-RUNX1T1, the positive rate was 91.4% and the intensity was mostly dim. AML with minimal differentiation and acute megakaryoblastic leukemia were all cMPO negative. The positive rates of cMPO in the remaining subtypes were between 22.7% and 76.2%.
CONCLUSIONThe positive rate and intensity of cMPO were significantly different among different subtypes of AML.
Cell Differentiation ; Flow Cytometry ; Granulocytes ; Humans ; Leukemia, Myeloid, Acute ; classification ; genetics ; Peroxidase ; genetics
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