Isolation, expression and bioactivity of feline granulocyte-macrophage colony-stimulating factor

被引：6

作者：

Dunham, SP ^{[1
]}

Bruce, J ^{[1
]}

机构：

[1] Univ Glasgow, Sch Vet, Dept Vet Pathol, Inst Comparat Med,Retrovirus Res Lab, Glasgow G61 1QH, Lanark, Scotland

来源：

GENE | 2004年 / 332卷

基金：

英国医学研究理事会;

关键词：

feline GM-CSF; dendritic cell; nucleotide sequence; cytokine; adjuvant;

D O I：

10.1016/j.gene.2004.02.023

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A cDNA encoding feline granulocyte-macrophage colony stimulating factor was cloned from alveolar macrophages using the reverse transcriptase-polymerase chain reaction (RT-PCR). The cDNA is 426 bp in length and encodes a predicted mature protein of 127 amino acids and the majority of the signal peptide. The recombinant protein (rfGM-CSF) was expressed in both Escherichia coli, as a calmodulin fusion protein, and mammalian cells. Biological activity of both recombinant proteins was demonstrated using the human erythroleukaemic cell line, TF-1. In a soft agar clonogenic assay, rfGM-CSF supported the development of granulocyte, macrophage and granulocyte-macrophage colonies. In combination with phytohaemagglutin (PHA) lymphocyte-conditioned medium, the number and size of such colonies were increased. Culture of feline bone marrow cells with rfGM-CSF was an efficient method for producing cells with morphology typical of dendritic cells (DC). The availability of the recombinant cytokine will permit further studies, in particular, the evaluation of the role of dendritic cells in feline immunopathology and its potential as a vaccine adjuvant. (C) 2004 Elsevier B.V. All rights reserved.

引用

页码：97 / 106

页数：10