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Poultry Science Jun 2023An immunization experiment was conducted in specific pathogen-free chickens with the inactivated Newcastle disease virus (NDV) vaccine encapsulated in the...
An immunization experiment was conducted in specific pathogen-free chickens with the inactivated Newcastle disease virus (NDV) vaccine encapsulated in the poly-(lactic-co-glycolic) acid (PLGA) nanoparticles (NP) to evaluate its immunogenicity and protective efficacy. The NDV vaccine was prepared by inactivating one virulent Indian strain of NDV belonging to Genotype VII by using beta-propiolactone. PLGA nanoparticles encapsulating inactivated NDV were prepared by the solvent evaporation method. Scanning electron microscopy and zeta sizer analysis revealed that the (PLGA+NDV) NP were spherical, with an average size of 300 nm, having a zeta potential of -6 mV. The encapsulation efficiency and loading efficiency were 72% and 2.4%, respectively. On immunization trial in chicken, the (PLGA+NDV) NP induced significantly (P < 0.0001) higher levels of HI and IgY antibodies with the peak HI titer of 2 and higher expression of IL-4 mRNA. The consistency of higher antibody levels suggests slow and pulsatile release of the antigens from the (PLGA+NDV) NP. The nano-NDV vaccine also induced cell mediated immunity with higher expression of IFN-γ indicating strong Th1 mediated immune responses in contrast to the commercial oil adjuvanted inactivated NDV vaccine. Further, the (PLGA+NDV) NP afforded 100% protection against the virulent NDV challenge. Our results suggested that PLGA NP have adjuvant potential on induction of humoral as well as Th1 biased cell mediated immune responses and also enhanced protective efficacy of the inactivated NDV vaccine. This study provides an insight for development of PLGA NP based inactivated NDV vaccine using the same genotype circulating in the field as well as for other avian diseases at exigencies.
Topics: Animals; Newcastle disease virus; Newcastle Disease; Chickens; Vaccines, Inactivated; Glycols; Adjuvants, Immunologic; Immunity, Cellular; Nanoparticles; Viral Vaccines
PubMed: 37116285
DOI: 10.1016/j.psj.2023.102679 -
Fish & Shellfish Immunology Feb 2016Herpesviral haematopoietic necrosis (HVHN) of gibel carp (Carassius gibelio) is a newly emerged infectious disease caused by cyprinid herpesvirus 2 (CyHV-2) and has...
Herpesviral haematopoietic necrosis (HVHN) of gibel carp (Carassius gibelio) is a newly emerged infectious disease caused by cyprinid herpesvirus 2 (CyHV-2) and has caused huge economic losses in aquaculture operations. Currently, no effective methods are available for the control of the disease. In this study, β-propiolactone inactivated cyprinid herpesvirus 2 (CyHV-2) vaccine was prepared, and the immune response and protection in cultured gibel carp after vaccination was thoroughly investigated. This included blood cell counting and classification, phagocytic activity, lysozyme and superoxide dismutase activity, neutralizing antibody titration, immune gene expression analysis, and determination of the relative percent survival in vaccinated gibel carp. The results of blood cell counts indicated that the numbers of the red and white blood cells in the peripheral blood of immunized gibel carp increased significantly at day 4 and day 7 after vaccination (p < 0.01). The differential leukocyte count of neutrophils and monocytes were significantly different compared to the control group at day 4 and 7 and the percentage of lymphocytes reached a peak at day 21. The phagocytic percentage and phagocytic index peaked at day 4 post-vaccination. The lysozyme activity and superoxide dismutase activity were significantly increased compared to the control group (p < 0.01). The serum neutralizing antibody titer peaked (203.03 ± 13.44) at day 21. The qPCR analysis revealed that the expression of the immune genes interlukin 11 and complement component C3 were significantly up-regulated in the immunized group. The challenge test demonstrated that the immunized group had a relative survival rate of 71.4%. These results indicate that the inactivated CyHV-2 vaccine induced both non-specific and specific anti-viral immune responses that resulted in significant protection against HVHN disease and mortality in gibel carp.
Topics: Animals; Cyprinidae; DNA Virus Infections; DNA Viruses; Fish Diseases; Immunity, Humoral; Immunity, Innate; Propiolactone; Vaccines, Inactivated; Viral Vaccines
PubMed: 26772479
DOI: 10.1016/j.fsi.2016.01.003 -
JAMA Sep 2020A vaccine against coronavirus disease 2019 (COVID-19) is urgently needed. (Randomized Controlled Trial)
Randomized Controlled Trial
IMPORTANCE
A vaccine against coronavirus disease 2019 (COVID-19) is urgently needed.
OBJECTIVE
To evaluate the safety and immunogenicity of an investigational inactivated whole-virus COVID-19 vaccine in China.
INTERVENTIONS
In the phase 1 trial, 96 participants were assigned to 1 of the 3 dose groups (2.5, 5, and 10 μg/dose) and an aluminum hydroxide (alum) adjuvant-only group (n = 24 in each group), and received 3 intramuscular injections at days 0, 28, and 56. In the phase 2 trial, 224 adults were randomized to 5 μg/dose in 2 schedule groups (injections on days 0 and 14 [n = 84] vs alum only [n = 28], and days 0 and 21 [n = 84] vs alum only [n = 28]).
DESIGN, SETTING, AND PARTICIPANTS
Interim analysis of ongoing randomized, double-blind, placebo-controlled, phase 1 and 2 clinical trials to assess an inactivated COVID-19 vaccine. The trials were conducted in Henan Province, China, among 96 (phase 1) and 224 (phase 2) healthy adults aged between 18 and 59 years. Study enrollment began on April 12, 2020. The interim analysis was conducted on June 16, 2020, and updated on July 27, 2020.
MAIN OUTCOMES AND MEASURES
The primary safety outcome was the combined adverse reactions 7 days after each injection, and the primary immunogenicity outcome was neutralizing antibody response 14 days after the whole-course vaccination, which was measured by a 50% plaque reduction neutralization test against live severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).
RESULTS
Among 320 patients who were randomized (mean age, 42.8 years; 200 women [62.5%]), all completed the trial up to 28 days after the whole-course vaccination. The 7-day adverse reactions occurred in 3 (12.5%), 5 (20.8%), 4 (16.7%), and 6 (25.0%) patients in the alum only, low-dose, medium-dose, and high-dose groups, respectively, in the phase 1 trial; and in 5 (6.0%) and 4 (14.3%) patients who received injections on days 0 and 14 for vaccine and alum only, and 16 (19.0%) and 5 (17.9%) patients who received injections on days 0 and 21 for vaccine and alum only, respectively, in the phase 2 trial. The most common adverse reaction was injection site pain, followed by fever, which were mild and self-limiting; no serious adverse reactions were noted. The geometric mean titers of neutralizing antibodies in the low-, medium-, and high-dose groups at day 14 after 3 injections were 316 (95% CI, 218-457), 206 (95% CI, 123-343), and 297 (95% CI, 208-424), respectively, in the phase 1 trial, and were 121 (95% CI, 95-154) and 247 (95% CI, 176-345) at day 14 after 2 injections in participants receiving vaccine on days 0 and 14 and on days 0 and 21, respectively, in the phase 2 trial. There were no detectable antibody responses in all alum-only groups.
CONCLUSIONS AND RELEVANCE
In this interim report of the phase 1 and phase 2 trials of an inactivated COVID-19 vaccine, patients had a low rate of adverse reactions and demonstrated immunogenicity; the study is ongoing. Efficacy and longer-term adverse event assessment will require phase 3 trials.
TRIAL REGISTRATION
Chinese Clinical Trial Registry Identifier: ChiCTR2000031809.
Topics: Adjuvants, Immunologic; Adolescent; Adult; Aluminum Hydroxide; Antibodies, Neutralizing; Antibodies, Viral; Betacoronavirus; COVID-19; COVID-19 Vaccines; Coronavirus Infections; Dose-Response Relationship, Immunologic; Double-Blind Method; Female; Humans; Immunogenicity, Vaccine; Injections, Intramuscular; Male; Pandemics; Pneumonia, Viral; Propiolactone; SARS-CoV-2; Vaccines, Inactivated; Viral Vaccines; Young Adult
PubMed: 32789505
DOI: 10.1001/jama.2020.15543 -
Vaccine Nov 2023The H9N2 subtype avian influenza virus (AIV) is a low pathogenic AIV that infects avian species and lead to huge economical losses in the poultry industry. The unique...
The H9N2 subtype avian influenza virus (AIV) is a low pathogenic AIV that infects avian species and lead to huge economical losses in the poultry industry. The unique immunomodulatory properties of Retinoic acid (RA), an active component of vitamin A, highlights its potential to enhance chicken's resistance to infectious diseases and perhaps vaccine-induced immunity. Therefore, the present study evaluated the effects of in ovo supplementation of RA on the immunogenicity and protective efficacy of an inactivated avian influenza virus vaccine. On embryonic day 18, eggs were inoculated with either 90 μmol RA/200 μL/egg or diluent into the amniotic sac. On days 7 and 21 post-hatch, birds were vaccinated with 15 μg of β-propiolactone (BPL) inactivated H9N2 virus via the intramuscular route. One group received BPL in combination with an adjuvant, while the other group received saline solution and served as a non-vaccinated control group. Serum samples were collected on days 7, 14, 21, 28, 35, and 42 post-primary vaccination (ppv) for antibody analysis. On day 24 ppv, spleens were collected, and splenocytes were isolated to analyze cytokine expression, interferon gamma (IFN-γ) production, and cell population. On day 28 ppv, birds in all groups were infected with H9N2 virus and oral and cloacal swabs were collected for TCID (50 % Tissue Culture Infectious Dose) assay up to day 7 post-infection. The results demonstrated that in ovo administration of RA did not significantly enhance the AIV vaccine-induced antibody response against H9N2 virus compared to the group that received the vaccine alone. However, RA supplementation enhanced the frequency of macrophages (KUL01), expression of inflammatory cytokines and production of IFN-γ by splenocytes. In addition, RA administration reduced oral shedding of AIV on day 5 post-infection. In conclusion, these findings suggest that RA can be supplemented in ovo to enhance AIV vaccine efficacy against LPAIV.
Topics: Animals; Influenza Vaccines; Influenza A Virus, H9N2 Subtype; Influenza in Birds; Tretinoin; Chickens; Immunity, Cellular; Vaccines, Inactivated; Antibodies, Viral
PubMed: 37923694
DOI: 10.1016/j.vaccine.2023.10.059 -
Journal of Virological Methods Nov 2023To facilitate the development of effective viral detection techniques, a positive control material is required for validating their quantitative performance. Inactivated...
To facilitate the development of effective viral detection techniques, a positive control material is required for validating their quantitative performance. Inactivated viruses serve as viable control materials, as they can be handled without the constraints of biohazard safety facilities. However, inactivation alters the structure of viral component molecules, necessitating the selection of inactivation methods that have minimal effects on the target molecules relevant to molecular detection techniques. Only a limited number of studies have investigated inactivation methods to produce viral control materials. Therefore, the aim of this study was to investigate various virus inactivation methods and evaluate their impact on molecular detection techniques, with a specific focus on viral proteins and RNA. We evaluated the effects of ultraviolet (UV) irradiation, heat, beta-propiolactone (BPL), hydrogen peroxide (HO), and perchloric acid (HClO) inactivation methods to identify the most effective technique and its optimal conditions. Enzyme-linked immunosorbent assay (ELISA) and reverse transcription-digital polymerase chain reaction (RT-dPCR) were employed as model assays to assess the effects of these treatments on protein and RNA measurements. Among the evaluated methods, UV and heat treatments demonstrated minimal interference with ELISA, while heat treatment had the least impact on RT-dPCR measurements. Consequently, our findings revealed that heat inactivation holds the potential for producing inactivated viruses that can be effectively used in molecular detection techniques targeting both viral protein and RNA.
Topics: Viral Proteins; Hydrogen Peroxide; Virus Inactivation; Biological Assay; RNA
PubMed: 37625621
DOI: 10.1016/j.jviromet.2023.114801 -
Journal of Virological Methods Sep 2016Beta-propiolactone (BPL) is used as an inactivating reagent for influenza virus in a number of countries. However, the treatment of viruses with BPL occasionally results...
Beta-propiolactone (BPL) is used as an inactivating reagent for influenza virus in a number of countries. However, the treatment of viruses with BPL occasionally results in a decrease in the hemagglutinin (HA) titer, which complicates vaccine development. In the present study, we examined the biological and biochemical characteristics of human H1N1 and H3N2 viruses treated with BPL, and developed an inactivation method for BPL-sensitive viruses. A significant decrease in HA titer was detected in the H3N2 viruses examined. The decrease in the pH of the virus fluid was not associated with the decreased HA titer, indicating that the decrease in HA titer for the H3N2 virus is the result of the direct effect of BPL. Excessive modification of M1 by BPL and loss of virion diameter were observed in 0.1% BPL-treated H3N2 virus. Taken together, these results suggest that the BPL sensitivity of H3N2 virus results from disruption of the virion. By contrast, the H3N2 virus was successfully inactivated by 0.02% BPL without a significant decrease in the HA titer or disruption of virion structure. Furthermore, we found that the 0.02% BPL in the virion preparation was hydrolyzed successfully by incubation at 37°C for 7h. Thus, mild treatment with a low concentration of BPL enabled us to inactivate the H3N2 virus.
Topics: Animals; Dogs; Humans; Hydrolysis; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H3N2 Subtype; Madin Darby Canine Kidney Cells; Propiolactone; Virion; Virus Inactivation
PubMed: 27142111
DOI: 10.1016/j.jviromet.2016.04.013 -
Journal of the American Chemical Society Sep 2018Treatment of (PDI)Fe(N) (PDI, 2,6-(2,6-iPrCHN═CMe)CHN) with CO and ethylene resulted in the formation of a homologous series of saturated and unsaturated iron...
Treatment of (PDI)Fe(N) (PDI, 2,6-(2,6-iPrCHN═CMe)CHN) with CO and ethylene resulted in the formation of a homologous series of saturated and unsaturated iron carboxylate products, (PDI)Fe(OCR), the distribution of which depends on the ratio of the reagents. The solid-state and electronic structures of a saturated product, (PDI)Fe(OCCH), were elucidated. Product distributions, deuterium labeling studies, and stoichiometric experiments support initial formation of a five-membered metallalactone intermediate, which undergoes subsequent ethylene insertions to generate macrocyclic metallalactones. Competitive β-hydride elimination, CO insertion, or reaction with H determines the fate of the metallalactone, the latter accounting for formation of iron complexes with saturated carboxylates. Similar reactivity was observed upon addition of propiolactone and ethylene to (PDI)Fe(N), supporting C-O oxidative addition and C-C bond formation through metallacycle intermediates.
PubMed: 30173506
DOI: 10.1021/jacs.8b07558 -
Vaccine Sep 2016Coxsackievirus A10 (CVA10) has become one of the major causative agents of hand, foot and mouth disease (HFMD). It is now recognized that CVA10 should be targeted for...
Coxsackievirus A10 (CVA10) has become one of the major causative agents of hand, foot and mouth disease (HFMD). It is now recognized that CVA10 should be targeted for vaccine development. We report here that β-propiolactone inactivated whole-virus based CVA10 vaccines can elicit protective immunity in mice. We prepared two inactivated CVA10 experimental vaccines derived from the prototype strain CVA10/Kowalik and from a clinical isolate CVA10/S0148b, respectively. Immunization with the experimental vaccines elicited CVA10-specific serum antibodies in mice. The antisera from vaccinated mice could potently neutralize in vitro infection with either homologous or heterologous CVA10 strains. Importantly, passive transfer of the anti-CVA10 sera protected recipient mice against CVA10/Kowalik or CVA10/S0148b infections. Moreover, active immunization with the inactivated vaccines also conferred protection against homologous and heterologous infections in mice. Collectively, our results demonstrate the proof-of-concept for inactivated whole-virus based CVA10 vaccines.
Topics: Animals; Antibodies, Neutralizing; Antibodies, Viral; Enterovirus; Female; Hand, Foot and Mouth Disease; Immunization, Passive; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; Neutralization Tests; Vaccines, Inactivated; Viral Vaccines
PubMed: 27562093
DOI: 10.1016/j.vaccine.2016.08.033 -
Journal of Colloid and Interface Science Jan 2021The micellization behavior of nonionic surfactants is significantly influenced by substituting ethylene oxide (EO) units with CO in the head group of nonionic CEO...
HYPOTHESIS
The micellization behavior of nonionic surfactants is significantly influenced by substituting ethylene oxide (EO) units with CO in the head group of nonionic CEO surfactants. Incorporating hydrophobic units has a major effect on the driving forces of the micellization process by a reduced hydration affinity. Hence, the incorporation of CO moieties is favoring micellization and thereby adding a further tuning parameter.
EXPERIMENTS
The micellization of the surfactants was characterized in terms of thermodynamics and the assembly properties on the water/air interface by isothermal titration calorimetry (ITC) and surface tension measurements. The incorporation of CO moieties was compared to the effect of propylene oxide (PO) and propiolactone (PL) over the temperature range of 25-50 °C. From ITC measurements and the van't Hoff relation, we determined the thermodynamic parameters of micellization: enthalpy (ΔH), entropy (ΔS), and Gibbs free energy (ΔG).
FINDINGS
The incorporation of CO moieties reduces the critical micellization concentration (cmc) and a transfer energy of -0.36 kT/CO unit quantifies favored micellization for CO surfactants. The presence of PO or CO in the head group has a similar effect on the cmc, but for CO ΔH is substantially decreased, resulting in a largely reduced temperature sensitivity of the micellization process and indicating a reduced hydration affinity. This thermodynamic analysis reveals that CO and PO behave very differently concerning their effect on the micellization process.
PubMed: 32818681
DOI: 10.1016/j.jcis.2020.07.141 -
Voprosy Virusologii Mar 2021Hemorrhagic fever with renal syndrome (HFRS) holds a leading place among natural focal human diseases in Russian Federation. There is no etiotropic therapy for the...
INTRODUCTION
Hemorrhagic fever with renal syndrome (HFRS) holds a leading place among natural focal human diseases in Russian Federation. There is no etiotropic therapy for the disease now. The vaccine prophylaxis is the most effective method to control this infection. The main criteria for inactivated vaccines evaluation are its immunogenicity and specific activity.The study purposes were to develop a sensitive and specific real-time PCR method for viral RNA quantification in the inactivated vaccine and to study the correlation between the viral RNA amount and vaccine immunogenicity.
MATERIAL AND METHODS
L-segment fragments of the Puumala, Hantaan, and Sochi vaccine strains were selected as diagnostic targets for oligonucleotides and fluorescent probes. The immunogenicity of experimental vaccines was determined by the induction of neutralizing antibodies in BALB/c mice.
RESULTS
A highly specific, sensitive and reproducible real-time PCR method has been developed. The analytical sensitivity was 1.24 ± 1.5 x 102 copies/ml for Puumala virus; 1.16 ± 1.4 * 102 copies/ml for Hantaan; 1.32 ± 1.8 * 102 copies/ ml for Sochi, with a virus content of 1.5 ± 0.5 lg FFU/ml; 1.8 ± 0.5 lg FFU/ml and 2.2 ± 0.5 lg FFU/ml, respectively. The viral RNA amount in experimental vaccine preparations inactivated with β-propiolactone was proportional to the neutralizing antibodies titer observed in mice following the immunization.
DISCUSSION
It was found that different virus inactivators differently affects the detected viral RNA amount, but not the vaccine immunogenicity, which indicates the same degree of the immunogenic proteins damage. The direct relationship between the viral RNA copy number and vaccine immunogenicity makes it possible to use this criterion for vaccine dosage preparation.
CONCLUSION
The developed method for viral RNA quantification is a promising tool for the specific activity control of the HFRS vaccine.
Topics: Animals; Antibodies, Neutralizing; Antibodies, Viral; Hemorrhagic Fever with Renal Syndrome; Mice; Mice, Inbred BALB C; RNA, Viral; Real-Time Polymerase Chain Reaction; Vaccines, Inactivated; Viral Vaccines
PubMed: 33683067
DOI: 10.36233/0507-4088-30