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Clinical and Vaccine Immunology, October 2009, p. 1420-1428, Vol. 16, No. 10
1071-412X/09/$08.00+0     doi:10.1128/CVI.00472-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Structural and Biological Diversity of Lipopolysaccharides from Burkholderia pseudomallei and Burkholderia thailandensis{triangledown}

Vidhya Novem,1 Guanghou Shui,2 Dongling Wang,1 Anne K. Bendt,2 Siew Hoon Sim,1 Yichun Liu,1 Tuck Weng Thong,1 Suppiah Paramalingam Sivalingam,1 Eng Eong Ooi,1,4 Markus R. Wenk,2,3 and Gladys Tan1*

Defense Medical and Environmental Research Institute, DSO National Laboratories, Singapore 117510, Singapore,1 Yong Loo Lin School of Medicine, National University of Singapore, Department of Biochemistry,2 Department of Biological Sciences, Singapore 117597, Singapore,3 Duke-NUS Graduate Medical School, Singapore 169857, Singapore4

Received 11 December 2008/ Returned for modification 4 February 2009/ Accepted 13 August 2009

Burkholderia pseudomallei, the etiological agent of melioidosis, is a facultative intracellular pathogen. As B. pseudomallei is a gram-negative bacterium, its outer membrane contains lipopolysaccharide (LPS) molecules, which have been shown to have low-level immunological activities in vitro. In this study, the biological activities of B. pseudomallei LPS were compared to those of Burkholderia thailandensis LPS, and it was found that both murine and human macrophages produced levels of tumor necrosis factor alpha, interleukin-6 (IL-6), and IL-10 in response to B. pseudomallei LPS that were lower than those in response to B. thailandensis LPS in vitro. In order to elucidate the molecular mechanisms underlying the low-level immunological activities of B. pseudomallei LPS, its lipid A moiety was characterized using mass spectrometry. The major lipid A species identified in B. pseudomallei consists of a biphosphorylated disaccharide backbone, which is modified with 4-amino-4-deoxy-arabinose (Ara4N) at both phosphates and penta-acylated with fatty acids (FA) C14:0(3-OH), C16:0(3-OH), and either C14:0 or C14:0(2-OH). In contrast, the major lipid A species identified in B. thailandensis was a mixture of tetra- and penta-acylated structures with differing amounts of Ara4N and FA C14:0(3-OH). Lipid A species acylated with FA C14:0(2-OH) were unique to B. pseudomallei and not found in B. thailandensis. Our data thus indicate that B. pseudomallei synthesizes lipid A species with long-chain FA C14:0(2-OH) and Ara4N-modified phosphate groups, allowing it to evade innate immune recognition.

* Corresponding author. Mailing address: Defense Medical and Environmental Research Institute, DSO National Laboratories, 27 Medical Drive, Singapore 117510, Singapore. Phone: 65-64857239. Fax: 65-64857262. E-mail: tgekyen{at}dso.org.sg

{triangledown} Published ahead of print on 19 August 2009.

Clinical and Vaccine Immunology, October 2009, p. 1420-1428, Vol. 16, No. 10
1071-412X/09/$08.00+0     doi:10.1128/CVI.00472-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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