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-  2018 

Development of Poly-ε-caprolactone Based Nanoadjuvant For Effective Vaccination Against Tuberculosis - Development of Poly-ε-caprolactone Based Nanoadjuvant For Effective Vaccination Against Tuberculosis - Open Access Pub

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Abstract:

The aim of the study was to synthesize sub-100nm poly-ε-caprolactone nanoparticles (PCL NP), load them with the mycobacterial protein, ESAT 6 and study the resulting immune responses in CD4+ and CD8+ T cells when incubated with human peripheral blood monocyte derived macrophages that had internalized the PCL NP. The synthesized PCL NP were characterized for size, shape and charge. They were found to be about 60nm in size with spherical shape. MTT assay revealed that the particles were perfectly biocompatible when tested in vitro on THP1 human monocytic cell line. The particles had a slow protein release kinetics and did not degrade appreciably even after 30 days in buffer solution. ELISA was used to quantify the cytokine response of CD4+ and CD8+ T cells when incubated with the monocyte derived macrophages as antigen presenting cells. The result of antigen presentation assay revealed that the antigen loaded PCL NP enhanced Th1 and CD8+ T cell responses significantly compared to the pure antigen. Thus we conclude that PCL NP of 60nm size can be effectively tested as a vaccine adjuvant with resulting activation of Th1/Th2 immunity as well as cytotoxic T cell response. DOI10.14302/issn.2377-2549.jndc-13-329 Tuberculosis, caused by the causative agent M.tuberculosis, has become a global scourge claiming almost 2 million lives per year and infecting almost 9 million people annually. The widely used TB vaccine, M.bovisBacille Calmette-Geurin (BCG) used to immunize infants and young children to prevent disseminated TB fails to prevent pulmonary (reactivation) TB in adolescents and adults. 1 Protective immunity against M.tuberculosis depends on the generation of a strong Th1 and CD8+ T cell response. 2 At least 12 TB vaccine candidates are in clinical trials which include recombinant live vaccines and viral vectored vaccines but majority are recombinant protein subunit vaccines. Early secreted antigenic target, ESAT-6 has been identified as a highly immunogenic mycobacterial antigen that is conserved in a large number of mycobacterial strains. It is encoded by genes located in the RD1 region in the mycobacterial genome. 3 This region is known to be deleted in all strains of BCG. Hence this secreted mycobacterial protein is an ideal antigen for subunit vaccine development against TB with suitable adjuvants. Studies have indicated that this secreted protein gains entry into the cytosol from the bacterial phagosomes and gets presented on both MHC class-I and MHC class II molecules. Enhancing the cross-presentation of ESAT 6 antigen may help to develop a strong

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