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Encapsulation of Antigenic Secretory Proteins of Mycobacterium tuberculosis in Biopolymeric Nanoparticles for Possible Aerosol Delivery System

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Abstract:

Mycobacterium tuberculosis [M.tb.], a successful intracellular pathogen, persists inside the macrophages through modulation of host cell signalling that allows them to survive inside the host. Culture filtrate proteins (CFP) of M.tb. have been encapsulated in bio-polymeric nanoparticles as a novel delivery system against tuberculosis. The study proposed to evaluate whether antigenic secretory proteins in nanoparticles can stimulate an immune response similar to natural infections. Chitosan nanoparticles were prepared by ionotropic gelation method using tri polyphosphate (TPP) as cross-linker and characterized by DLS (∼250 nm–300 nm) and TEM. Positively charged particles were confirmed by the zeta potential (41 mV ±5.29). FTIR studies clearly show the characteristic peak of C–N (1250–1375 cm–1) in chitosan, but in chitosan nanoparticles the peak gets shifted to 1564.03 cm–1. Also the strong and sharp peak of phosphate at 1092 cm–1 in chitosan nanoparticles confirms the involvement of TPP while making the nanoparticles. The concentration of CFP10 and CFP21 entrapped in the nanoparticles was 257.936 μg/ml and 296.659 μg/ml respectively. Negligible cytotoxicity was observed by proteins encapsulated nanoparticles as compared to 60% cytotoxicity by CFP10 and CFP21 proteins per se.(5). Biodistribution studies of radiolabeled chitosan void nanoparticles showed maximum accumulation in kidney after an hour and 4 hours but encapsulated nanoparticles exhibited maximum accumulation in kidney after 1 hour, and was reduced by half after 4 hrs. The antigenic response of M.tb. CFPs were evaluated in-vivo. Higher levels of IFN-γ i.e., ∼300 pg/ml by CFP10 loaded nanoparticles and ∼400 pg/ml by CFP21 loaded nanoparticles were secreted by the stimulated splenocytes, while CFP10 and CFP21 per seshowed ∼100 pg/ml and ∼80 pg/ml respectively. Low levels of IL-4 were observed to be secreted by the splenocytes. This indicates a TH1 bias in T cell response that is an immune response analogous to natural infections.

Keywords: CFP-10 AND CFP-21; CHITOSAN NANOPARTICLES; CYTOKINES; DELIVERY SYSTEM; M.TB.

Document Type: Research Article

DOI: http://dx.doi.org/10.1166/jbns.2011.1042

Publication date: June 1, 2011

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  • Bionanoscience attempts to harness various functions of biological macromolecules and integrate them with engineering for technological applications. It is based on a bottom-up approach and encompasses structural biology, biomacromolecular engineering, material science, and engineering, extending the horizon of material science. The journal aims at publication of (i) Letters (ii) Reviews (3) Concepts (4) Rapid communications (5) Research papers (6) Book reviews (7) Conference announcements in the interface between chemistry, physics, biology, material science, and technology. The use of biological macromolecules as sensors, biomaterials, information storage devices, biomolecular arrays, molecular machines is significantly increasing. The traditional disciplines of chemistry, physics, and biology are overlapping and coalescing with nanoscale science and technology. Currently research in this area is scattered in different journals and this journal seeks to bring them under a single umbrella to ensure highest quality peer-reviewed research for rapid dissemination in areas that are in the forefront of science and technology which is witnessing phenomenal and accelerated growth.
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