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Clinical and Vaccine Immunology, March 2006, p. 309-313, Vol. 13, No. 3
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.3.309-313.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Endotoxin Contamination in Commercially Available Pokeweed Mitogen Contributes to the Activation of Murine Macrophages and Human Dendritic Cell Maturation

Jae Seung Yang,1,2,{dagger} Hye Jin Kim,1,{dagger} Young Hee Ryu,1 Cheol-Heui Yun,1 Dae Kyun Chung,3,4 and Seung Hyun Han1*

International Vaccine Institute, Seoul 151-600, Republic of Korea,1 Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejon 305-701, Republic of Korea,2 School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Suwon 449-701, Republic of Korea,3 RNA Inc., Suwon 449-701, Republic of Korea4

Received 8 September 2005/ Returned for modification 23 November 2005/ Accepted 28 December 2005

Commercially available pokeweed mitogen (PWM) has been reported to activate macrophages, leading to production of proinflammatory cytokines and nitric oxide (NO). However, we found that polymyxin B (PMB), a specific inhibitor of endotoxin activity, inhibited the PWM-induced expression of proinflammatory cytokines and NO and the activation of Toll-like receptor 4 (TLR4). A kinetic-turbidimetric Limulus amebocyte lysate assay demonstrated that commercial PWM contained substantial endotoxin, over 104 endotoxin units/mg of the PWM. A PWM repurified by PMB-coupled beads no longer induced the expression of proinflammatory cytokines, TLR4 activation, or dendritic cell maturation. However, the repurified PWM remained able to induce proliferation of human lymphocytes, which is a representative characteristic of PWM. These results suggest that commercial PWM might be contaminated with a large amount of endotoxin, resulting in the attribution of misleading immunological properties to PWM.

* Corresponding author. Mailing address: Humoral Immunology Section, International Vaccine Institute, SNU Research Park, San 4-8 Bongcheon-7 dong, Kwanak-gu, Seoul 151-818, Republic of Korea. Phone: 82 2 881 1189. Fax: 82 2 872 2803. E-mail address: shhan{at}ivi.int.

{dagger} J.S.Y. and H.J.K. contributed equally to this study.

Clinical and Vaccine Immunology, March 2006, p. 309-313, Vol. 13, No. 3
1071-412X/06/$08.00+0     doi:10.1128/CVI.13.3.309-313.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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