Protein delivery to primary cells by protein transduction domain (PTD) serves as a novel measure for manipulation of the cells for biological study and for the treatment of various human conditions. Although the method has been employed to modulate cellular function in vitro, only limited reports are available on its application in the replacement of deficient signaling molecules into primary cells. We examined the potential of recombinant proteins to compensate for defective cytosolic components of the NADPH oxidase complex in chronic granulomatous disease (CGD) neutrophils in both p47 phox and p67 phox deficiency. The p47 phox or p67 phox protein linked to Hph-1 PTD was effectively expressed in soluble form and transduced into human neutrophils efficiently without eliciting unwanted signal transduction or apoptosis. The delivered protein was stable for more than 24h, expressed in the cytoplasm, translocated to the membrane fraction upon activation, and, most importantly able to restored reactive oxygen species (ROS) production. Although research on human primary neutrophils using the protein delivery system is still limited, our data show that the protein transduction approach for neutrophils may be applicable to the control of local infections in CGD patients by direct delivery of the protein product.
|Number of pages
|Biochemical and Biophysical Research Communications
|Published - 2012 Jan 6
Bibliographical noteFunding Information:
Authors thank Dr. Evan Rachlin for discussion and critical comment. This work was supported in part by grants from the Ministry of Health, Labour, and Welfare of Japan (TM) and from the Ministry of Education, Culture, Sports, Science and Technology of Japan (TM) .
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology