scholarly journals Recombinant Filamentous Bacteriophages Encapsulated in Biodegradable Polymeric Microparticles for Stimulation of Innate and Adaptive Immune Responses

2020 ◽  
Vol 8 (5) ◽  
pp. 650 ◽  
Author(s):  
Rezvan Jamaledin ◽  
Rossella Sartorius ◽  
Concetta Di Natale ◽  
Raffaele Vecchione ◽  
Piergiuseppe De Berardinis ◽  
...  
Keyword(s):  
Morphological Properties ◽  
Antigen Delivery ◽  
Bacteriophage Display ◽  
Oil In Water ◽  
Filamentous Bacteriophages ◽  
Multiple Copies ◽  
Display Technology ◽  
Polymeric Microparticles ◽  
Covalently Linked ◽  
Immunogenic Properties

Escherichia coli filamentous bacteriophages (M13, f1, or fd) have attracted tremendous attention from vaccinologists as a promising immunogenic carrier and vaccine delivery vehicle with vast possible applications in the development of vaccines. The use of fd bacteriophage as an antigen delivery system is based on a modification of bacteriophage display technology. In particular, it is designed to express multiple copies of exogenous peptides (or polypeptides) covalently linked to viral capsid proteins. This study for the first time proposes the use of microparticles (MPs) made of poly (lactic-co-glycolic acid) (PLGA) to encapsulate fd bacteriophage. Bacteriophage–PLGA MPs were synthesized by a water in oil in water (w1/o/w2) emulsion technique, and their morphological properties were analyzed by confocal and scanning electron microscopy (SEM). Moreover, phage integrity, encapsulation efficiency, and release were investigated. Using recombinant bacteriophages expressing the ovalbumin (OVA) antigenic determinant, we demonstrated the immunogenicity of the encapsulated bacteriophage after being released by MPs. Our results reveal that encapsulated bacteriophages are stable and retain their immunogenic properties. Bacteriophage-encapsulated PLGA microparticles may thus represent an important tool for the development of different bacteriophage-based vaccine platforms.

scholarly journals Chimeric Fusion (F) and Attachment (G) Glycoprotein Antigen Delivery by mRNA as a Candidate Nipah Vaccine

2021 ◽  
Vol 12 ◽  
Author(s):  
Rebecca J. Loomis ◽  
Anthony T. DiPiazza ◽  
Samantha Falcone ◽  
Tracy J. Ruckwardt ◽  
Kaitlyn M. Morabito ◽  
...  
Keyword(s):  
Nipah Virus ◽  
Neutralizing Antibody ◽  
T Cell Response ◽  
Antigen Delivery ◽  
Design Elements ◽  
C Terminus ◽  
Glycoprotein Antigen ◽  
Pandemic Threat ◽  
Covalently Linked ◽  
Selection Of

Nipah virus (NiV) represents a significant pandemic threat with zoonotic transmission from bats-to-humans with almost annual regional outbreaks characterized by documented human-to-human transmission and high fatality rates. Currently, no vaccine against NiV has been approved. Structure-based design and protein engineering principles were applied to stabilize the fusion (F) protein in its prefusion trimeric conformation (pre-F) to improve expression and increase immunogenicity. We covalently linked the stabilized pre-F through trimerization domains at the C-terminus to three attachment protein (G) monomers, forming a chimeric design. These studies detailed here focus on mRNA delivery of NiV immunogens in mice, assessment of mRNA immunogen-specific design elements and their effects on humoral and cellular immunogenicity. The pre-F/G chimera elicited a strong neutralizing antibody response and a superior NiV-specific Tfh and other effector T cell response compared to G alone across both the mRNA and protein platforms. These findings enabled final candidate selection of pre-F/G Fd for clinical development.

Preparation of a synergistically stabilized oil-in-water paraffin Pickering emulsion for potential application in wood treatment

Holzforschung ◽  
2018 ◽  
Vol 72 (6) ◽  
pp. 489-497 ◽  
Author(s):  
Jun Jiang ◽  
Jinzhen Cao ◽  
Wang Wang ◽  
Haiying Shen
Keyword(s):  
Mechanical Properties ◽  
Cell Wall ◽  
Size Distribution ◽  
Droplet Size ◽  
Surface Hardness ◽  
Pickering Emulsion ◽  
Silica Particles ◽  
Morphological Properties ◽  
Pickering Emulsions ◽  
Oil In Water

AbstractPickering emulsions (emulsions stabilized by solid-state additives) are attractive as they have strong similarities with traditional surfactant-based emulsions. In this study, an oil-in-water (O/W) paraffin Pickering emulsion system with satisfying stability and small droplet size distribution was developed by hydrophilic silica particles and traditional surfactants as mixed emulsifiers. The droplet morphology and size distribution were observed by optical microscopy and a laser particle analyzer. The emulsion stability was improved and the droplet size was reduced after addition of a suitable amount of silica particles. The silica concentration of 1% showed the optimal effect among all the levels observed (0.1, 0.5, 1 and 2%). Wood was impregnated with the prepared emulsion, and the chemical and morphological properties of the product were investigated by Fourier-transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) combined with energy-dispersed X-ray analysis (SEM-EDXA). Moreover, the hydrophobicity, thermal properties, surface hardness, axial compression strength (CS) and dynamic mechanical properties were tested. The silica was evenly distributed in the wood cell wall and thus there was a synergistic positive effect from the paraffin and silica in the cell wall leading to better hydrophobicity, improved surface hardness and mechanical properties including the thermal stability.

scholarly journals Arming Filamentous Bacteriophage, a Nature-Made Nanoparticle, for New Vaccine and Immunotherapeutic Strategies

Pharmaceutics ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 437 ◽  
Author(s):  
Rossella Sartorius ◽  
Luciana D’Apice ◽  
Antonella Prisco ◽  
Piergiuseppe De Berardinis
Keyword(s):  
Antigen Expression ◽  
Antigenic Determinants ◽  
Antigen Delivery ◽  
Adaptive Responses ◽  
Bacterial Strains ◽  
Phage Display Technology ◽  
Filamentous Bacteriophage ◽  
Filamentous Bacteriophages ◽  
Delivery Strategies ◽  
Therapeutic Tools

The pharmaceutical use of bacteriophages as safe and inexpensive therapeutic tools is collecting renewed interest. The use of lytic phages to fight antibiotic-resistant bacterial strains is pursued in academic and industrial projects and is the object of several clinical trials. On the other hand, filamentous bacteriophages used for the phage display technology can also have diagnostic and therapeutic applications. Filamentous bacteriophages are nature-made nanoparticles useful for their size, the capability to enter blood vessels, and the capacity of high-density antigen expression. In the last decades, our laboratory focused its efforts in the study of antigen delivery strategies based on the filamentous bacteriophage ‘fd’, able to trigger all arms of the immune response, with particular emphasis on the ability of the MHC class I restricted antigenic determinants displayed on phages to induce strong and protective cytotoxic responses. We showed that fd bacteriophages, engineered to target mouse dendritic cells (DCs), activate innate and adaptive responses without the need of exogenous adjuvants, and more recently, we described the display of immunologically active lipids. In this review, we will provide an overview of the reported applications of the bacteriophage carriers and describe the advantages of exploiting this technology for delivery strategies.

Localization of lipoyl-bearing domains in 2-ketoglutarate dehydrogenase complex by electron microscopy and single-particle averaging methods

1983 ◽  
Vol 41 ◽  
pp. 756-757
Author(s):  
Terence Wagenknecht ◽  
Joachim Frank
Keyword(s):  
Support Surface ◽  
X Ray Diffraction ◽  
X Ray ◽  
E Coli ◽  
Catalytic Function ◽  
Multiple Copies ◽  
Covalently Linked ◽  
Polypeptide Chains ◽  
Dehydrogenase Complex ◽  
Ketoglutarate Dehydrogenase

The 2-ketoglutarate dehydrogenase complex (KGDC) from E. coli (Mr=2.5 x 106) consists of multiple copies of three component enzymes. In addition to its catalytic function one of the components, dihydrolipoyl transsuccinylase (E2), serves as a structural core (Mr=l x 106) to which the other two components are noncovalently bonded. X-ray diffraction studies have shown the E2 complex to be built from 24 identical polypeptide chains and to have the overall shape of a cube having truncated corners. In the electron microscope the E2 complexes adhere preferentially to the support surface by one of their faces and thus appear square-shaped with four morphological units at the vertices and a stain filled center (Fig 1A).Each of the E2 chains contains a lipoic acid cofactor covalently linked to a lysine residue. A domain (Mr=ll,000) containing the lipoyl moiety is removed by trypsin without disrupting the basic architecture of the complex. Micrographs of the trypsin-treated E2 complexes (Fig 1B) appear similar to those of the native complexes (Fig 1A), although the trypsintreated complexes appear slightly smaller and have a more sharply defined 4-fold symmetry.

Emergence of Twisted Magnetic Flux Related Sigmoidal Brightening

2000 ◽  
Vol 179 ◽  
pp. 263-264
Author(s):  
K. Sundara Raman ◽  
K. B. Ramesh ◽  
R. Selvendran ◽  
P. S. M. Aleem ◽  
K. M. Hiremath
Keyword(s):  
Magnetic Field ◽  
Magnetic Fields ◽  
Magnetic Flux ◽  
Ultra Violet ◽  
Active Regions ◽  
Morphological Properties ◽  
Energy Storage And Conversion ◽  
The Sun ◽  
X Rays ◽  
The Magnetic Field

Extended AbstractWe have examined the morphological properties of a sigmoid associated with an SXR (soft X-ray) flare. The sigmoid is cospatial with the EUV (extreme ultra violet) images and in the optical part lies along an S-shaped Hαfilament. The photoheliogram shows flux emergence within an existingδtype sunspot which has caused the rotation of the umbrae giving rise to the sigmoidal brightening.It is now widely accepted that flares derive their energy from the magnetic fields of the active regions and coronal levels are considered to be the flare sites. But still a satisfactory understanding of the flare processes has not been achieved because of the difficulties encountered to predict and estimate the probability of flare eruptions. The convection flows and vortices below the photosphere transport and concentrate magnetic field, which subsequently appear as active regions in the photosphere (Rust & Kumar 1994 and the references therein). Successive emergence of magnetic flux, twist the field, creating flare productive magnetic shear and has been studied by many authors (Sundara Ramanet al.1998 and the references therein). Hence, it is considered that the flare is powered by the energy stored in the twisted magnetic flux tubes (Kurokawa 1996 and the references therein). Rust & Kumar (1996) named the S-shaped bright coronal loops that appear in soft X-rays as ‘Sigmoids’ and concluded that this S-shaped distortion is due to the twist developed in the magnetic field lines. These transient sigmoidal features tell a great deal about unstable coronal magnetic fields, as these regions are more likely to be eruptive (Canfieldet al.1999). As the magnetic fields of the active regions are deep rooted in the Sun, the twist developed in the subphotospheric flux tube penetrates the photosphere and extends in to the corona. Thus, it is essentially favourable for the subphotospheric twist to unwind the twist and transmit it through the photosphere to the corona. Therefore, it becomes essential to make complete observational descriptions of a flare from the magnetic field changes that are taking place in different atmospheric levels of the Sun, to pin down the energy storage and conversion process that trigger the flare phenomena.

Characterization of Tamm-Horsfall Urinary Glycoprotein

1973 ◽  
Vol 31 ◽  
pp. 420-421
Author(s):  
John P. Robinson ◽  
J. David Puett
Keyword(s):  
Electron Microscopy ◽  
Circular Dichroism ◽  
Secondary Structure ◽  
Quaternary Structure ◽  
Normal Adult ◽  
Morphological Properties ◽  
Adult Males ◽  
Circular Dichroïsm ◽  
Urinary Glycoprotein

Much work has been reported on the chemical, physical and morphological properties of urinary Tamm-Horsfall glycoprotein (THG). Although it was once reported that cystic fibrotic (CF) individuals had a defective THG, more recent data indicate that THG and CF-THG are similar if not identical.No studies on the conformational aspects have been reported on this glycoprotein using circular dichroism (CD). We examined the secondary structure of THG and derivatives under various conditions and have correlated these results with quaternary structure using electron microscopy.THG was prepared from normal adult males and CF-THG from a 16-year old CF female by the method of Tamm and Horsfall. CF female by the method of Tamm and Horsfall.

Studies on Leukemogenic and Sarcomagenic Viruses

1970 ◽  
Vol 28 ◽  
pp. 156-157
Author(s):  
Leon Dmochowski
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Morphological Properties ◽  
Mode Of Development ◽  
Relationship Of ◽  
The Relationship

Electron microscopy has proved to be an invaluable discipline in studies on the relationship of viruses to the origin of leukemia, sarcoma, and other types of tumors in animals and man. The successful cell-free transmission of leukemia and sarcoma in mice, rats, hamsters, and cats, interpreted as due to a virus or viruses, was proved to be due to a virus on the basis of electron microscope studies. These studies demonstrated that all the types of neoplasia in animals of the species examined are produced by a virus of certain characteristic morphological properties similar, if not identical, in the mode of development in all types of neoplasia in animals, as shown in Fig. 1.

Alkaline Dissociation Products of Lumbricus Terrestris Hemoglobin Viewed with the STEM

1981 ◽  
Vol 39 ◽  
pp. 434-435
Author(s):  
O. H. Kapp ◽  
M. Ohtsuki ◽  
N. Robin ◽  
S. N. Vinogradov ◽  
A. V. Crewe
Keyword(s):  
Carbon Film ◽  
Lumbricus Terrestris ◽  
Uranyl Acetate ◽  
Acetate Solution ◽  
Oxygen Carriers ◽  
Complex Molecules ◽  
Thin Carbon ◽  
Thin Carbon Film ◽  
Multiple Copies ◽  
Hill Coefficients

Annelid extracellular hemoglobins are among the largest known proteins (M.W = 3.9 x 106), and together with the hemocyanins are the largest known oxygen carriers. They display oxygen affinities generally higher than those o vertebrate hemoglobins with Hill coefficients ranging from slightly higher than unity to values as high as 5-6. These complex molecules are composed of multiple copies of as many as six different polypeptides and posse: approximately 150 hemes per molecule.The samples were diluted to 100-200 μg/ml with distilled water just before application to a thin carbon film (∽15 Å thick). One percent (w/v) uranyl acetate solution was used for negative staining for 2 minutes and dried in air. The specimens were examined with the high resolution STEM. Their general appearance is that of a hexagonal bilayer (Fig. 1), each layer consisting of six spheroidal subunits. The corner to corner hexagonal dimensic is approximately 300 Å and the bilayer thickness approximately 200 Å.

Large-cluster and combined fluorescent and gold immunoprobes

1996 ◽  
Vol 54 ◽  
pp. 892-893
Author(s):  
Richard D. Powell ◽  
James F. Hainfeld ◽  
Carol M. R. Halsey ◽  
David L. Spector ◽  
Shelley Kaurin ◽  
...  
Keyword(s):  
Metal Cluster ◽  
Sulfonyl Chloride ◽  
Gel Filtration ◽  
Cross Linking ◽  
Site Specific ◽  
Fab Fragments ◽  
Cluster Label ◽  
Covalently Linked ◽  
Texas Red ◽  
Fold Excess

Two new types of covalently linked, site-specific immunoprobes have been prepared using metal cluster labels, and used to stain components of cells. Combined fluorescein and 1.4 nm “Nanogold” labels were prepared by using the fluorescein-conjugated tris (aryl) phosphine ligand and the amino-substituted ligand in the synthesis of the Nanogold cluster. This cluster label was activated by reaction with a 60-fold excess of (sulfo-Succinimidyl-4-N-maleiniido-cyclohexane-l-carboxylate (sulfo-SMCC) at pH 7.5, separated from excess cross-linking reagent by gel filtration, and mixed in ten-fold excess with Goat Fab’ fragments against mouse IgG (obtained by reduction of F(ab’)2 fragments with 50 mM mercaptoethylamine hydrochloride). Labeled Fab’ fragments were isolated by gel filtration HPLC (Superose-12, Pharmacia). A combined Nanogold and Texas Red label was also prepared, using a Nanogold cluster derivatized with both and its protected analog: the cluster was reacted with an eight-fold excess of Texas Red sulfonyl chloride at pH 9.0, separated from excess Texas Red by gel filtration, then deprotected with HC1 in methanol to yield the amino-substituted label.

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