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EXPRESSION OF P1A1 AND Yukb ANTIGENS ON MEGAKARYOCYTIC LEUKEMIA CELL LINES AND ITS GENOMIC ANALYSIS Ryukichi Ryo*, Mutsumi Yasunaga, Masashi Adachi, Wataru Sugano, Akinori Yoshida, Katsuyasu Saigo and Nobuo Yamaguchi *Blood Transfusion Service, Kobe University Hospital Department of Laboratory Medicine, Kobe University School of Medicine Yukb and P1A1 antigen expressions on megakaryocytic leukemia cell lines K562 and CMK were examined by Western blotting with alloantibodies against these antigens. Both K562 and CMK cells had a 110 kDaand 96 kDa-mol wt bands that reacted with alloantibodies against Yukb and P1A1, which were seen on P1A1 and Yukb positive platelets. We found no differences between PP1 and Yukb antigens on CMK and K562 by immunoblotting analysis. However, it is worth noting that since CMK and K562 can synthesize glycoprotein (GP)IIIa carrying Yukb and P1A1 antigens, these cells are useful for studying the different pathways of synthesizing alloantigens such as Yukb and P1A1. Furthermore, GPIIIa cDNA in CMK and K562 were analyzed using Southern hybridization with a 1.6 kb EcoRI fragment, representing the 3' portion of a cDNA clone containing a 4.0 kb GPIlIa isolated from a HEL cell library as a probe. Genomic DNA prepared from peripheral mononuclear cells of 7PP1 and Yukb positive normal indivduals and cells of K562 or CMK were digested with restriction enzymes HindSIII, BamHl, EcoRl, Pstl, and examined by Southern blotting. Identical restriction fragment patterns were obtained with the DNAs from normal individuals, CMK and K562. These studies indicate that the P1A1 and Yukb positive platelet phenotype can be regulated by GPIIIa gene obtained from our present studies.

Expression of P1A1 and Yukb antigens on megakaryocytic leukemia cell lines and its genomic analysis.