In vitro transfection of bone marrow-derived dendritic cells with TATp-liposomes

Juan Sebastián Pappalardo,1–3 Cecilia A Langellotti,2 Sebastián Di Giacomo,1 Valeria Olivera,1 Valeria Quattrocchi,2 Patricia I Zamorano,1,2 William C Hartner,3 Tatyana S Levchenko,3 Vladimir P Torchilin3 1Virology Institute, Center for Research in Veterinary and Agronomic Sciences, National Institute for Agricultural Technology (INTA), Hurlingham, BA, Argentina; 2National Council for Scientific and Technical Research (CONICET), Autonomous City of Buenos Aires, Argentina; 3Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, Boston, MA, USA Abstract: Dendritic cells (DC) are antigen-presenting cells uniquely capable of priming naïve T cells and cross-presenting antigens, and they determine the type of immune response elicited against an antigen. TAT peptide (TATp), is an amphipathic, arginine-rich, cationic peptide that promotes penetration and translocation of various molecules and nanoparticles into cells. TATp-liposomes (TATp-L) used for DC transfection were prepared using TATp derivatized with a lipid-terminated polymer capable of anchoring in the liposomal membrane. Here, we show that the addition of TATp to DNA-loaded liposomes increased the uptake of DNA in DC. DNA-loaded TATp-L increased the in vitro transfection efficiency in DC cultures as evidenced by a higher expression of the enhanced green fluorescent protein and bovine herpes virus type 1 glycoprotein D (gD). The de novo synthesized gD protein was immunologically stimulating when transfections were performed with TATp-L, as indicated by the secretion of interleukin 6. Keywords: dendritic cell transfection, green fluorescent protein, bovine herpes virus 1 glycoprotein D, liposomes, TAT peptide, interleukin

Dendrimeric peptides can confer protection against foot-and-mouth disease virus in cattle

Foot-and-mouth disease virus (FMDV) causes a highly contagious disease in cloven-hoofed animals. A synthetic vaccine candidate consisting of dendrimeric peptides harbouring two copies of a B-epitope [VP1(136–154)] linked to a T-cell epitope [3A(21–35)] of FMDV confers protection to type O FMDV challenge in pigs. Herein we show in cattle that novel dendrimeric peptides bearing a T-cell epitope [VP1(21–40] and two or four copies of a B-cell epitope [VP1(135–160)] from type O1 Campos FMDV (termed B2T and B4T, respectively) elicited FMDV specific immune responses to similar levels to a commercial vaccine. Animals were challenged with FMDV and 100% of vaccinated cattle with B2T or B4T were protected to podal generalization. Moreover, bovines immunized with B4T were completely protected (with no clinical signs) against FMDV challenge after three vaccine doses, which was associated with titers of viral neutralizing antibodies in serum higher than those of B2T group (p< 0.05) and levels of opsonic antibodies similar to those of animals immunized with one dose of FMDV commercial vaccine. Bovines vaccinated with both dendrimeric peptides presented high levels of IgG1 anti FMDV in sera and in mucosa. When IgA in nasal secretions was measured, 20% or 40% of the animals in B2T or B4T groups respectively, showed anti-FMDV IgA titers. In addition, B2T and B4T peptides evoked similar consistent T cell responses, being recognized in vitro by lymphocytes from most of the immunized cattle in the proliferation assay, and from all animals in the IFN-γ production assay. Taken together, these results support the potential of dendrimers B2T or B4T in cattle as a highly valuable, cost-effective FMDV candidate vaccine with DIVA potential.Work at INTA was supported by the National Institute of Agricultural Technology (grant number PNSA 1115052) and cooperation agreement INTA-INIA (Argentina-Spain SA 20938). Evaluation of vaccine immunogenicity and protection in cattle. Work at Universitat Pompeu Fabra and CBMSO was supported by MINECO, Spain (grant number AGL2014-52395-C2)

Anaerobic membrane bioreactors for industrial wastewater treatment

Human activities, particularly from industrial zones, produce a large volume of wastewater that contains high levels of toxic chemicals and causes serious environmental problems and detrimental impacts on human health. Therefore, industrial wastewater discharge must meet the stringent water quality permissible limits to prevent the problem associated with the effluent released to the environment. Recently, Anaerobic Membrane Bioreactor (AnMBR) has been considered as an attractive approach for the treatment of industrial wastewaters due to its ability to offset the disadvantages of conventional anaerobic treatment and aerobic membrane bioreactor (MBR). Besides, biogas production by AnMBR as a means of obtaining renewable energy can provide many benefits to fulfill partially the world’s energy needs. This book chapter discusses the fundamentals of AnMBR, application of AnMBR for various stream of industrial wastewater treatment, biogas production, membrane fouling, and future prospective for further research