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Originally published online as doi:10.1189/jlb.1104684 on March 17, 2005

Published online before print March 17, 2005
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(Journal of Leukocyte Biology. 2005;77:999-1007.)
© 2005 by Society for Leukocyte Biology

Phospholipase D (PLD) gene expression in human neutrophils and HL-60 differentiation

Mauricio Di Fulvio and Julian Gomez-Cambronero1

Department of Physiology and Biophysics, Wright State University, School of Medicine, Dayton, Ohio

1 Correspondence: Department of Physiology and Biophysics, Wright State University, School of Medicine, 3640 Colonel Glenn Highway, Dayton, OH 45435. E-mail: julian.cambronero{at}wright.edu

Human neutrophils exhibit a regulated phospholipase D (PLD) activity that can be measured biochemically in vitro. However, the precise expression pattern of PLD isoforms and their specific biological role(s) are not well understood. Neutrophil mRNA is intrinsically difficult to isolate as a result of the extremely high content of lytic enzymes in the cell’s lysosomal granules. Reverse transcription coupled to polymerase chain reaction indicated that pure populations of human neutrophils had the CD16b+/CD115–/CD20–/CD3{zeta}–/interleukin-5 receptor {alpha}– phenotype. These cells expressed the following splice variants of the PLD1 isoform: PLD1a, PLD1b, PLD1a2, and PLD1b2. As for the PLD2 isoform, neutrophils expressed the PLD2a but not the PLD2b mRNA variant. The relative amount of PLD1/PLD2 transcripts exists in an approximate 4:1 ratio. The expression of PLD isoforms varies during granulocytic differentiation, as demonstrated in the promyelocytic leukemia HL-60 cell line. Further, the pattern of mRNA expression is dependent on the differentiation-inducing agent, 1.25% dimethyl sulfoxide causes a dramatic increase in PLD2a and PLD1b transcripts, and 300 nM all-trans-retinoic acid induced PLD1a expression. These results demonstrate for the first time that human neutrophils express five PLD transcripts and that the PLD genes undergo qualitative changes in transcription regulation during granulocytic differentiation.

Key Words: RT-PCR • ATRA • promyelocytic leukemia • CD markers • monocytes • COS-7 cells




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J. Gomez-Cambronero, M. Di Fulvio, and K. Knapek
Understanding phospholipase D (PLD) using leukocytes: PLD involvement in cell adhesion and chemotaxis
J. Leukoc. Biol., August 1, 2007; 82(2): 272 - 281.
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