Detection of the Human Platelet-Activating Factor Receptor mRNA in Human B Lymphocytes by Polymerase Chain Reaction on Reverse Transcripts (RT-PCR)

Abstract

The platelet activating factor receptor (hPAFR) is a GTP binding protein linked receptor of the rhodopsin family. The hPAFR gene maps to chromosome 1 and is characterized by the absence of introns in its coding region. Because PAF demonstrates unique properties as a growth factor in human B lymphoblastoid cell lines, we developed an RT-PCR in order to detect the presence of hPAFR mRNA. We examined the presence of the hPAFR in a series of B lymphoblastoid cell lines, as well as on fresh human B cells and the T-leukemia cell line Jurkat. All cells of B lineage expressed hPAFR mRNA, including a B cell line not previously shown to express functional hPAFR. In contrast, the T-leukemia cell line Jurkat expressed very low levels of hPAFR mRNA. We discuss the methodology and its validation in developing an RT-PCR for intronless genes such as the hPAFR.