Sublytic concentrations of Staphylococcus aureus Panton-Valentine leukocidin alter human PMN gene expression and enhance bactericidal capacity.

CA-MRSA infections are often caused by strains encoding PVL, which can cause lysis of PMNs and other myeloid cells in vitro, a function considered widely as the primary means by which PVL might contribute to disease. However, at sublytic concentrations, PVL can function as a PMN agonist. To better understand this phenomenon, we investigated the ability of PVL to alter human PMN function. PMNs exposed to PVL had enhanced capacity to produce O(2)(-) in response to fMLF, but unlike priming by LPS, this response did not require TLR signal transduction. On the other hand, there was subcellular redistribution of NADPH oxidase components in PMNs following exposure of these cells to PVL--a finding consistent with priming. Importantly, PMNs primed with PVL had an enhanced ability to bind/ingest and kill Staphylococcus aureus. Priming of PMNs with other agonists, such as IL-8 or GM-CSF, altered the ability of PVL to cause formation of pores in the plasma membranes of these cells. Microarray analysis revealed significant changes in the human PMN transcriptome following exposure to PVL, including up-regulation of molecules that regulate the inflammatory response. Consistent with the microarray data, mediators of the inflammatory response were released from PMNs after stimulation with PVL. We conclude that exposure of human PMNs to sublytic concentrations of PVL elicits a proinflammatory response that is regulated in part at the level of gene expression. We propose that PVL-mediated priming of PMNs enhances the host innate immune response.

Flexicate molecules as a potential new class of antibiotics.

Helicates are α-helical, nonpeptide complexes that bind to DNA and exhibit antimicrobial activity. In the past, enthusiasm for the use of helicates in biological applications was limited, at least in part, by the presence of a racemic mixture of enantiomers or the formation of complexes that are insoluble in aqueous solutions. Recently, Howson et al. overcame the barriers associated with helicate synthesis by generating helicate-like complexes that are soluble and stable in water, optically pure and synthetically flexible. The mechanism synthesizes nonpeptide mimetic α-helical 'flexicates' that bind to DNA and show broad-spectrum antimicrobial activity against representative Gram-positive and Gram-negative bacterial pathogens. Although the application of flexicates as an antimicrobial therapy remains to be determined, this study provides important insight into flexicate activity and the prospective use of flexicates as microbicidal agents.

Asbestos activates CH12.LX B-lymphocytes via macrophage signaling.

The impact of asbestos exposure on the development and progression of autoimmunity is becoming increasingly recognized as a public health issue. Epidemiological studies have shown an association between exposure to airborne silicates, such as asbestos, and autoimmunity, but the etiology remains unresolved. B1a B-lymphocytes have been implicated in autoimmune responses in mice, and splenic B1a cell numbers are altered following asbestos exposure. The purpose of this study was to explore the possible role of B1a B-lymphocytes in the production of pathogenic autoantibodies by testing the hypothesis that B1a B-lymphocytes directly react with asbestos and increase production of antibodies. The B1a-like B-lymphocyte model, CH12.LX, was exposed to asbestos in vitro via direct and indirect mechanisms. The effect was determined of these exposures on the rate of proliferation and on production of various immunoglobulin classes. Direct exposure elicited no measurable response by the CH12.LX cells. Culturing the CH12.LX cells in media from asbestos-exposed RAW 264.7 macrophages, however, decreased the proliferation rate and stimulated the cells to increase production of the immunoglobulin isotypes IgG1, IgG3, and IgA. It was discovered that asbestos stimulated the macrophages to increase production of the cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α. Recombinant murine IL-6 caused similar results seen with the macrophage media, indicating a role of IL-6 in stimulating a response by the B1a B-lymphocytes to asbestos. In correlation with the in vitro data, it was determined ex vivo that exposure of peritoneal cells (from C57Bl/6 mice) to asbestos caused an increase in the expression of IL-6 and TNFα, as well as of surface expression of IgA on the peritoneal B1a B-lymphocytes. These data demonstrate that asbestos leads to immunologic changes consistent with activation of B1a B-lymphocytes. This study also provides a model for analyzing the critical steps that may be involved in asbestos-induced autoimmune responses.