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      CD44 targeted delivery of oncolytic Newcastle disease virus encapsulated in thiolated chitosan for sustained release in cervical cancer: a targeted immunotherapy approach

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          Abstract

          Introduction

          Cervical cancer accounts for one of most common cancers among women of reproductive age. Oncolytic virotherapy has emerged as a promising immunotherapy modality but it comes with several drawbacks that include rapid clearance of virus from body due to immune-neutralization of virus in host. To overcome this, we encapsulated oncolytic Newcastle disease virus (NDV) in polymeric thiolated chitosan nanoparticles. For active targeting of virus loaded nanoformulation against CD44 (cluster of differentiation 44) receptors which are overly expressed on cancer cells, these nanoparticles were surface functionalized with hyaluronic acid (HA).

          Methods

          Using half dose of NDV (TCID 50 (50% tissue culture infective dose) single dose 3 × 10 5), virus loaded nanoparticles were prepared by green synthesis approach through ionotropic gelation method. Zeta analysis was performed to analyse size and charge on nanoparticles. Nanoparticles (NPs) shape and size were analysed by SEM (scanning electron microscope) and TEM (transmission electron microscope) while functional group identification was done by FTIR (fourier transform infrared) and XRD (X-ray diffraction). Viral quantification was done by TCID 50 and Multiplicity of infection (MOI) determination while oncolytic potential of NPs encapsulated virus was analysed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and cell morphology analysis.

          Results

          Zeta analysis showed that average size of NDV loaded thiolated chitosan nanoparticles surface functionalized with HA (HA-ThCs-NDV) was 290.4nm with zeta potential of 22.3 mV and 0.265 PDI (polydispersity index). SEM and TEM analysis showed smooth surface and spherical features of nanoparticles. FTIR and XRD confirmed the presence of characteristic functional groups and successful encapsulation of the virus. In vitro release showed continuous but sustained release of NDV for up to 48 hours. TCID 50 for HA-ThCs-NDV nanoparticles was 2.63x 10 6/mL titter and the nanoformulation exhibited high oncolytic potential in cell morphology analysis and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay as compared to naked virus, in dose dependent manner.

          Discussion

          These findings suggest that virus encapsulation in thiolated chitosan nanoparticles and surface functionalization with HA is not only helpful in achieving active targeting while masking virus from immune system but, it also gives sustained release of virus in tumor microenvironment for longer period of time that increases bioavailability of virus.

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          Most cited references37

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          ClusPro is a web server that performs rigid-body docking of two proteins by sampling billions of conformations. Low-energy docked structures are clustered, and centers of the largest clusters are used as likely models of the complex.
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            Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria

            Summary Macrophages mediate crucial innate immune responses via caspase-1-dependent processing and secretion of IL-1β and IL-18. While wild type Salmonella typhimurium infection is lethal to mice, a strain that persistently expresses flagellin was cleared by the cytosolic flagellin detection pathway via NLRC4 activation of caspase-1; however, this clearance was independent of IL-1β and IL-18. Instead, caspase-1 induced pyroptotic cell death, released bacteria from macrophages and exposed them to uptake and killing by reactive oxygen species in neutrophils. Similarly, caspase-1 cleared unmanipulated Legionella and Burkholderia by cytokine-independent mechanisms. This demonstrates for the first time that caspase-1 clears intracellular bacteria in vivo independent of IL-1β and IL-18, and establishes pyroptosis as an efficient mechanism of bacterial clearance by the innate immune system.
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              Determination of 50% endpoint titer using a simple formula.

              Two commonly used methods for calculating 50% endpoint using serial dilutions are Spearman-Karber method and Reed and Muench method. To understand/apply the above formulas, moderate statistical/mathematical skills are necessary. In this paper, a simple formula/method for calculating 50% endpoints has been proposed. The formula yields essentially similar results as those of the Spearman-Karber method. The formula has been rigorously evaluated with several samples.
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                Author and article information

                Contributors
                Journal
                Front Immunol
                Front Immunol
                Front. Immunol.
                Frontiers in Immunology
                Frontiers Media S.A.
                1664-3224
                22 May 2023
                2023
                : 14
                : 1175535
                Affiliations
                [1] 1 Industrial Biotechnology, Atta-Ur-Rahman School of Applied Biosciences, National University of Sciences and Technology , Islamabad, Pakistan
                [2] 2 Shifa College of Pharmaceutical Sciences, Shifa Tameer e Millat University , Islamabad, Pakistan
                [3] 3 Immune Responses in Different Diseases Research Group, Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University , Jeddah, Saudi Arabia
                [4] 4 Center of Excellence in Genomic Medicine Research, King Abdulaziz University , Jeddah, Saudi Arabia
                [5] 5 Department of Biology, University of Hail , Hail, Saudi Arabia
                Author notes

                Edited by: Rohimah Mohamud, University of Science Malaysia (USM), Malaysia

                Reviewed by: Samuel Álvarez-Almazán, National Autonomous University of Mexico, Mexico; Surjendu Maity, Terasaki Institute for Biomedical Innovation, United States; Kamala Priya, Translational Health Science and Technology Institute (THSTI), India

                *Correspondence: Kousain Kousar, kousain777@ 123456outlook.com ; Tahir Ahmad, tahir@ 123456asab.nust.edu.pk
                Article
                10.3389/fimmu.2023.1175535
                10239954
                37283735
                b46171c0-ece0-489d-b5a7-8362c8105c80
                Copyright © 2023 Kousar, Naseer, Abduh, Anjum and Ahmad

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 27 February 2023
                : 09 May 2023
                Page count
                Figures: 16, Tables: 5, Equations: 0, References: 38, Pages: 18, Words: 7378
                Funding
                This research work was funded by Institutional Fund Projects under grant no. (IFPIP:360-290-1443). The authors gratefully acknowledge technical and financial support provided by the Ministry of Education and King Abdulaziz University, DSR, Jeddah, Saudi Arabia.
                Categories
                Immunology
                Original Research
                Custom metadata
                Cancer Immunity and Immunotherapy

                Immunology
                oncolytic newcastle disease virus,cervical cancer,green synthesis,sustained release,cd44,polymeric nanoparticles

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