betadex and Tuberculosis

Multi-drug-resistant tuberculosis (TB) is a major public health problem, concerning about half a million cases each year. Patients hardly adhere to the current strict treatment consisting of more than 10 000 tablets over a 2-year period. There is a clear need for efficient and better formulated medications. We have previously shown that nanoparticles made of cross-linked poly-β-cyclodextrins (pβCD) are efficient vehicles for pulmonary delivery of powerful combinations of anti-TB drugs. Here, we report that in addition to being efficient drug carriers, pβCD nanoparticles are endowed with intrinsic antibacterial properties. Empty pβCD nanoparticles are able to impair Mycobacterium tuberculosis (Mtb) establishment after pulmonary administration in mice. pβCD hamper colonization of macrophages by Mtb by interfering with lipid rafts, without inducing toxicity. Moreover, pβCD provoke macrophage apoptosis, leading to depletion of infected cells, thus creating a lung microenvironment detrimental to Mtb persistence. Taken together, our results suggest that pβCD nanoparticles loaded or not with antibiotics have an antibacterial action on their own and could be used as a carrier in drug regimen formulations effective against TB.

Topics: Animals; Antitubercular Agents; beta-Cyclodextrins; Drug Carriers; Drug Delivery Systems; Female; Humans; Macrophages, Alveolar; Male; Mice, Inbred BALB C; Mice, Inbred C57BL; Mycobacterium tuberculosis; Nanoparticles; Tuberculosis

Membrane lipid rafts are enriched in cholesterol and play an important role as signalling platforms. However, the roles of lipid rafts and associated signalling molecules in the innate immune responses to mycobacteria remain unknown. Here we show that stimulation with Mycobacterium tuberculosis 19 kDa lipoprotein, a TLR2/1 agonist, results in translocation of TLR2 to lipid rafts, coalescence of lipid rafts and production of reactive oxygen species (ROS) that drive pro-inflammatory responses. Disruption of lipid raft organization markedly reduced lipoprotein-induced ROS and inflammatory responses. Remarkably, the atypical protein kinase C (PKC) zeta was specifically recruited into detergent-resistant membrane fractions and associated with TLR2. PKCzeta activity was critical for lipoprotein-dependent ROS generation, raft coalescence and the pro-inflammatory responses by macrophages. Moreover, lipid raft organization was required for 19 kDa mediated PKCzeta activation. These results demonstrate that TLR2 trafficking and raft coalescence play an essential role for the initiation of lipoprotein-induced innate immune responses via TLR2 and ROS signalling. In addition, PKCzeta targets to lipid rafts and may act as a critical adaptor molecule to regulate lipid raft dynamics during TLR2 signalling.

Topics: Animals; Bacterial Proteins; beta-Cyclodextrins; Cells, Cultured; Cytokines; Immunity, Innate; Inflammation; Lipoproteins; Macrophage Activation; Macrophages; Membrane Microdomains; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Knockout; Mycobacterium tuberculosis; Protein Kinase C; Protein Transport; Reactive Oxygen Species; Signal Transduction; Toll-Like Receptor 2; Tuberculosis