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Burkholderia pseudomallei  is the causative agent of melioidosis, a disease with high morbidity that is endemic in South East Asia and northern Australia. An unusual feature of the bacterium is its ability to induce multinucleated giant cell formation (MNGC), which appears to be related to bacterial pathogenicity. The mechanism of MNGC formation is not fully understood, but host cell factors as well as known bacterial virulence determinants are likely to contribute. Since members of the tetraspanin family of membrane proteins are involved in various types of cell:cell fusion, their role in MNGC formation induced by  Burkholderia thailandensis , a mildly pathogenic species closely related to  B. pseudomallei , was investigated. The effect of antibodies to tetraspanins CD9, CD81, and CD63 in MNGC formation induced by  B. thailandensis  in infected mouse J774.2 and RAW macrophage cell lines was assessed along with that of recombinant proteins corresponding to the large extracellular domain (EC2) of the tetraspanins.  B. thailandensis -induced fusion was also examined in macrophages derived from CD9 null and corresponding WT mice, and in J774.2 macrophages over-expressing CD9. Antibodies to CD9 and CD81 promoted MNGC formation induced by  B. thailandensis , whereas EC2 proteins of CD9, CD81, and CD63 inhibited MNGC formation. Enhanced MNGC formation was observed in CD9 null macrophages, whereas a decrease in MNGC formation was associated with overexpression of CD9. Overall our findings show that tetraspanins are involved in MNGC formation induced by  B. thailandensis  and by implication,  B. pseudomallei,  with CD9 and CD81 acting as negative regulators of this process.

A role for tetraspanin proteins in regulating fusion induced by Burkholderia thailandensis