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Characterization of gene expression patterns in the developing natural killer cell Birro, Natasha Diane

Abstract

Relatively little is known about development of the Natural Killer (NK) cell. Studies in the past have focused on characterizing the phenotype of the NK cell, and evaluating its development by manipulating hematopoietic stem cells in vitro. When I began my study, an in vitro culture system had been recently developed in my laboratory in which embryonic stem cells are induced with various cytokines and growth factors to differentiate into NK cells. This system provides a medium where NK cell development can be researched from an earlier stage than previously possible. The objective of my work was to use this culture system to investigate the expression patterns of five genes which are known to play a role in lymphoid cell development based on gene knock-out studies. Id-2 is an inhibitor of DNA binding protein, Ikaros-1 and Ets-1 are transcription factors, Notch-1 is a transmembrane receptor and Jagged-1 is a ligand for Notch. Using RT-PCR and phospho-image analysis, the expression patterns of these genes were assessed from various numbers of cells, and at five different points along the developmental pathway of the NK cell. Id-2 seemed to be expressed consistently throughout the system. Ikaros-1 and Ets-1 were interesting in that they showed an on/off expression pattern, Ets-1 with little expression on days 8 and 14, and Ikaros-1 with no expression on days 8 and 14. This implies that perhaps these transcription factors are not needed at the initial stage of NK cell differentiation from a common lymphoid precursor. Notch-1 and Jagged-1 were particularly interesting in that their expression patterns did not parallel each other. These results suggest that Jagged-1 may not be the ligand of choice for proper NK cell development as Notch levels were lower than expected. These studies have begun to give us a better understanding of gene expression in our system, and provide clues as to what role certain gene may play.

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