title: Biodegradable PLGA85/15 nanoparticles as a delivery vehicle for Chlamydia trachomatis recombinant MOMP-187 peptide.
abstract: Development of a Chlamydia trachomatis vaccine has been a formidable task partly because of an ineffective delivery system. Our laboratory has generated a recombinant peptide of C. trachomatis major outer membrane protein (MOMP) (rMOMP-187) and demonstrated that it induced at 20 μg ml(-1) maximal interleukin (IL)-6 and IL-12p40 Th1 cytokines in mouse J774 macrophages. In a continuous pursuit of a C. trachomatis effective vaccine-delivery system, we encapsulated rMOMP-187 in poly(d,l-lactic-co-glycolic acid) (PLGA, 85:15 PLA/PGA ratio) to serve as a nanovaccine candidate. Physiochemical characterizations were assessed by Fourier transform-infrared spectroscopy, atomic force microscopy, Zetasizer, Zeta potential, transmission electron microcopy and differential scanning calorimetry. The encapsulated rMOMP-187 was small (∼200 nm) with an apparently smooth uniform oval structure, thermally stable (54 °C), negatively charged ( - 27.00 mV) and exhibited minimal toxicity at concentrations <250 μg ml (-1) to eukaryotic cells (>95% viable cells) over a 24-72 h period. We achieved a high encapsulation efficiency of rMOMP-187 (∼98%) in PLGA, a loading peptide capacity of 2.7% and a slow release of the encapsulated peptide. Stimulation of J774 macrophages with a concentration as low as 1 μg ml (-1) of encapsulated rMOMP-187 evoked high production levels of the Th1 cytokines IL-6 (874 pg ml(-1)) and IL-12p40 (674 pg ml(-1)) as well as nitric oxide (8 μM) at 24 h post-stimulation, and in a dose-response and time-kinetics manner. Our data indicate the successful encapsulation and characterization of rMOMP-187 in PLGA and, more importantly, that PLGA enhanced the capacity of the peptide to induce Th1 cytokines and NO in vitro. These findings make this nanovaccine an attractive candidate in pursuit of an efficacious vaccine against C. trachomatis.
authors: Center for NanoBiotechnology and Life Science Research (CNBR), Alabama State University, Montgomery, AL 36104, USA.
authors: Murtada A
authors: MA
authors: Taha
authors: Shree R
authors: SR
authors: Singh
authors: Vida A
authors: VA
authors: Dennis
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000276:immunology / Q000493:pharmacokinetics; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002692:Chlamydia trachomatis / Q000737:chemistry / Q000378:metabolism*; D004306:Dose-Response Relationship, Immunologic; D004337:Drug Carriers / Q000737:chemistry* / Q000494:pharmacology; D018664:Interleukin-12 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D019344:Lactic Acid / Q000737:chemistry* / Q000494:pharmacology; D051379:Mice; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry*; D009569:Nitric Oxide / Q000378:metabolism; D010316:Particle Size; D010455:Peptides / Q000737:chemistry* / Q000276:immunology / Q000493:pharmacokinetics; D011100:Polyglycolic Acid / Q000737:chemistry* / Q000494:pharmacology; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000493:pharmacokinetics; D014614:Vaccines, Synthetic
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004337:Drug Carriers; D015850:Interleukin-6; D010455:Peptides; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C078167:OMPA outer membrane proteins; D018664:Interleukin-12; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D009569:Nitric Oxide; D019344:Lactic Acid