scholarly journals A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

2019 ◽  
Author(s):  
Lihua Wang ◽  
Shijiang Mi ◽  
Rachel Madera ◽  
Llilianne Ganges ◽  
Manuel V. Borca ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Confidence Interval ◽  
Excellent Agreement ◽  
Neutralizing Antibodies ◽  
Vaccination Campaign ◽  
Classical Swine Fever ◽  
Enzyme Linked Immunosorbent Assay ◽  
Live Virus ◽  
Post Vaccination ◽  
Neutralizing Monoclonal Antibody

Abstract Background: Virus neutralization test (VNT) is widely used for serological survey of classical swine fever (CSF) and efficacy evaluation of CSF vaccines. However, VNT is a time consuming procedure that requires cell culture and live virus manipulation. C-strain CSF vaccine is the most frequently used vaccine for CSF control and prevention. In this study, we presented a neutralizing monoclonal antibody (mAb) based competitive enzyme-linked immunosorbent assay (cELISA) with the emphasis on the replacement of VNT for C-strain post–vaccination monitoring. Results: One monoclonal antibody (6B211) which has potent neutralizing activity against C-strain was generated. A novel cELISA was established and optimized based on the strategy that 6B211 can compete with C-strain induced neutralizing antibodies in pig serum to bind capture antigen C-strain E2. By testing C-strain VNT negative pig sera (n=445) and C-strain VNT positive pig sera (n=70), the 6B211 based cELSIA showed 100% sensitivity (95% confidence interval: 94.87 to 100%) and 100% specificity (95% confidence interval: 100 to 100%). The C-strain antibody can be detected in pigs as early as 7 days post vaccination with the cELISA. By testing pig sera (n=139) in parallel, the cELISA showed excellent agreement (Kappa=0.957) with VNT. The inhibition rate of serum samples in the cELISA is highly correlated with their titers in VNT (r 2 =0.903, p<0.001). In addition, intra- and inter-assays of the cELISA exhibited acceptable repeatability with low coefficient of variations (CVs). Conclusions: This novel cELISA demonstrated excellent agreement and high level correlation with VNT. It is a reliable tool for sero-monitoring of C-strain vaccination campaign because it is a rapid, simple, safe and cost effective assay that can be used to monitor vaccination-induced immune response at the population level.

scholarly journals A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

2020 ◽  
Author(s):  
Lihua Wang ◽  
Shijiang Mi ◽  
Rachel Madera ◽  
Llilianne Ganges ◽  
Manuel V. Borca ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Confidence Interval ◽  
Excellent Agreement ◽  
Neutralizing Antibodies ◽  
Vaccination Campaign ◽  
Classical Swine Fever ◽  
Enzyme Linked Immunosorbent Assay ◽  
Live Virus ◽  
Post Vaccination ◽  
Neutralizing Monoclonal Antibody

Abstract Background: Virus neutralization test (VNT) is widely used for serological survey of classical swine fever (CSF) and efficacy evaluation of CSF vaccines. However, VNT is a time consuming procedure that requires cell culture and live virus manipulation. C-strain CSF vaccine is the most frequently used vaccine for CSF control and prevention. In this study, we presented a neutralizing monoclonal antibody (mAb) based competitive enzyme-linked immunosorbent assay (cELISA) with the emphasis on the replacement of VNT for C-strain post–vaccination monitoring. Results: One monoclonal antibody (6B211) which has potent neutralizing activity against C-strain was generated. A novel cELISA was established and optimized based on the strategy that 6B211 can compete with C-strain induced neutralizing antibodies in pig serum to bind capture antigen C-strain E2. By testing C-strain VNT negative pig sera (n=445) and C-strain VNT positive pig sera (n=70), the 6B211 based cELSIA showed 100% sensitivity (95% confidence interval: 94.87 to 100%) and 100% specificity (95% confidence interval: 100 to 100%). The C-strain antibody can be detected in pigs as early as 7 days post vaccination with the cELISA. By testing pig sera (n=139) in parallel, the cELISA showed excellent agreement (Kappa=0.957) with VNT. The inhibition rate of serum samples in the cELISA is highly correlated with their titers in VNT (r 2 =0.903, p<0.001). In addition, intra- and inter-assays of the cELISA exhibited acceptable repeatability with low coefficient of variations (CVs). Conclusions: This novel cELISA demonstrated excellent agreement and high level correlation with VNT. It is a reliable tool for sero-monitoring of C-strain vaccination campaign because it is a rapid, simple, safe and cost effective assay that can be used to monitor vaccination-induced immune response at the population level.

scholarly journals A neutralizing monoclonal antibody-based competitive ELISA for classical swine fever C-strain post–vaccination monitoring

2019 ◽  
Author(s):  
Lihua Wang ◽  
Shijiang Mi ◽  
Rachel Madera ◽  
Llilianne Ganges ◽  
Manuel V. Borca ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Confidence Interval ◽  
Excellent Agreement ◽  
Neutralizing Antibodies ◽  
Classical Swine Fever ◽  
Competitive Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Live Virus ◽  
Post Vaccination ◽  
Neutralizing Monoclonal Antibody

Abstract Background: Virus neutralization test (VNT) is widely used for serological survey of classical swine fever (CSF) and efficacy evaluation of CSF vaccines. However, VNT is a time consuming procedure that requires cell culture and live virus manipulation. C-strain CSF vaccine is the most frequently used vaccine for CSF control and prevention. In this study, we presented a neutralizing monoclonal antibody (mAb) based competitive enzyme-linked immunosorbent assay (cELISA) with the emphasis on the replacement of VNT for C-strain post–vaccination monitoring. Results: One monoclonal antibody (6B211) which has potent neutralizing activity against C-strain was generated. A novel competitive ELISA (cELISA) was established and optimized based on the strategy that 6B211 can compete with C-strain induced neutralizing antibodies in pig serum to bind capture antigen C-strain E2. By testing C-strain VNT negative pig sera (n=445) and C-strain VNT positive pig sera (n=70), the 6B211 based cELSIA showed 100% sensitivity (95% confidence interval: 94.87 to 100%) and 100% specificity (95% confidence interval: 100 to 100%). By testing pig sera (n=139) in parallel, the cELISA showed excellent agreement (95.7%) with VNT. The inhibition rate of serum samples in the cELISA is highly correlated with their titers in VNT (r 2 =0.903, p<0.001). In addition, intra- and inter-assays of the cELISA exhibited acceptable repeatability with low coefficient of variations (CVs). Conclusions: This novel cELISA demonstrated excellent agreement and high level correlation with VNT. It is a reliable tool for sero-monitoring of C-strain vaccination campaign because it is a rapid, simple, safe and cost effective assay that can be used to monitor vaccination-induced immune response at the population level.

scholarly journals Development of specific immunity in laboratory animals after co-immunization against seasonal influenza and COVID-19

2022 ◽  
Vol 98 (6) ◽  
pp. 648-656
Author(s):  
G. M. Ignatyev ◽  
I. A. Leneva ◽  
A. V. Atrasheuskaya ◽  
L. I. Kozlovskaya ◽  
N. P. Kartashova ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Seasonal Influenza ◽  
Neutralization Test ◽  
Elisa Test ◽  
Control Measures ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Post Vaccination ◽  
Specific Immunity ◽  
Influenza Prevention

Introduction. In clinical practice, the differential diagnosis of COVID-19 can be challenging during the flu season, entailing serious consequences such as delays in appropriate control measures against the SARS-CoV-2 pandemic. Another problem is posed by co-infection of SARS-CoV-2 and influenza virus (IV), which significantly contributes to the severity of the COVID-19 disease. This study was aimed to explore the cross-impact of co-administration of Russian influenza and COVID-19 vaccines on development of specific immunity in laboratory animals.Materials and methods. The study was conducted on BALB/c mice. The animals were inoculated intramuscularly with the vaccine for COVID-19 prevention (CoviVac) and the vaccine for influenza prevention (Flu-M). The sera from the immunized animals were examined separately. Three IV strains were used in the hemagglutination inhibition assay. Antibodies (Abs) against SARS-CoV-2 were detected by an enzyme-linked immunosorbent assay (ELISA). The neutralization test was performed to detect virus neutralizing antibodies against SARS-CoV-2 and IV.Results. Relatively high titers of specific Abs were found in the groups of animals inoculated with one vaccine and with two vaccines concurrently. In the groups of animals inoculated with CoviVac and with two vaccines concurrently, both in the ELISA test and in the neutralization test, the average titers of specific Abs against SARSCoV- 2 did not demonstrate any statistical difference. The group of animals inoculated concurrently with two vaccines demonstrated statistically higher titers of Abs against IV after the second immunization compared to the group of animals inoculated with Flu-M.Discussion. The study has shown that post-vaccination immunity both to IV and to SARS-CoV-2 develops after co-vaccination with two vaccines. The observed enhanced post-vaccination immune response to IV in the coimmunized laboratory animals needs further research.Conclusion. The performed studies suggest the possibility of co-administration of two vaccines to prevent influenza and COVID-19.

scholarly journals Comparative kinetics of SARS-CoV-2 anti-spike protein RBD IgGs and neutralizing antibodies in convalescent and naïve recipients of the BNT162b2 mRNA vaccine versus COVID-19 patients

BMC Medicine ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Ioannis P. Trougakos ◽  
Evangelos Terpos ◽  
Christina Zirou ◽  
Aimilia D. Sklirou ◽  
Filia Apostolakou ◽  
...  
Keyword(s):  
Immune Response ◽  
Neutralizing Antibodies ◽  
Mononuclear Cells ◽  
Natural Infection ◽  
Severe Disease ◽  
Vaccination Campaign ◽  
Hospitalized Patients ◽  
Peripheral Blood Mononuclear ◽  
Post Vaccination ◽  
Antibody Titers

Abstract Background Coronavirus SARS-CoV-2, the causative agent of COVID-19, has caused a still evolving global pandemic. Given the worldwide vaccination campaign, the understanding of the vaccine-induced versus COVID-19-induced immunity will contribute to adjusting vaccine dosing strategies and speeding-up vaccination efforts. Methods Anti-spike-RBD IgGs and neutralizing antibodies (NAbs) titers were measured in BNT162b2 mRNA vaccinated participants (n = 250); we also investigated humoral and cellular immune responses in vaccinated individuals (n = 21) of this cohort 5 months post-vaccination and assayed NAbs levels in COVID-19 hospitalized patients (n = 60) with moderate or severe disease, as well as in COVID-19 recovered patients (n = 34). Results We found that one (boosting) dose of the BNT162b2 vaccine triggers robust immune (i.e., anti-spike-RBD IgGs and NAbs) responses in COVID-19 convalescent healthy recipients, while naïve recipients require both priming and boosting shots to acquire high antibody titers. Severe COVID-19 triggers an earlier and more intense (versus moderate disease) immune response in hospitalized patients; in all cases, however, antibody titers remain at high levels in COVID-19 recovered patients. Although virus infection promotes an earlier and more intense, versus priming vaccination, immune response, boosting vaccination induces antibody titers significantly higher and likely more durable versus COVID-19. In support, high anti-spike-RBD IgGs/NAbs titers along with spike (vaccine encoded antigen) specific T cell clones were found in the serum and peripheral blood mononuclear cells, respectively, of vaccinated individuals 5 months post-vaccination. Conclusions These findings support vaccination efficacy, also suggesting that vaccination likely offers more protection than natural infection. Graphical abstract

scholarly journals Identification of Neutralizing and Nonneutralizing Epitopes in the Porcine Reproductive and Respiratory Syndrome Virus GP5 Ectodomain

2002 ◽  
Vol 76 (9) ◽  
pp. 4241-4250 ◽  
Author(s):  
M. Ostrowski ◽  
J. A. Galeota ◽  
A. M. Jar ◽  
K. B. Platt ◽  
F. A. Osorio ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Phage Display ◽  
Neutralizing Antibodies ◽  
Phage Display Library ◽  
The Other ◽  
Respiratory Syndrome Virus ◽  
Rapid Rise ◽  
Neutralizing Activity ◽  
Neutralizing Monoclonal Antibody ◽  
Neutralizing Epitopes

ABSTRACT After infection of swine with porcine reproductive and respiratory syndrome virus (PRRSV), there is a rapid rise of PRRSV-specific nonneutralizing antibodies (NNA), while neutralizing antibodies (NA) are detectable not sooner than 3 weeks later. To characterize neutralizing epitopes, we selected phages from a 12-mer phage display library using anti-PRRSV neutralizing monoclonal antibody (MAb) ISU25-C1. In addition, phages carrying peptides recognized by swine antibodies with high seroneutralizing titer were isolated after subtracting from the library those clones binding to swine anti-PRRSV serum with no neutralizing activity. Two epitopes located in the ectodomain of PRRSV GP5 were identified. One of these epitopes, which we named epitope B, was recognized both by neutralizing MAb ISU25-C1 and swine neutralizing serum (NS) but not by swine nonneutralizing serum (NNS), indicating that it is a neutralizing epitope. Epitope B is sequential, conserved among isolates, and not immunodominant. Antibodies directed against it are detected in serum late after infection. In contrast, the other epitope, which we named epitope A, is hypervariable and immunodominant. Antibodies against it appear early after infection with PRRSV. This epitope is recognized by swine NNA but is not recognized by either neutralizing MAb ISU25-C1 or swine NA, indicating that it is not involved in PRRSV neutralization. During infection with PRRSV, epitope A may act as a decoy, eliciting most of the antibodies directed to GP5 and delaying the induction of NA against epitope B for at least 3 weeks. These results are relevant to the design of vaccines against PRRSV.

scholarly journals Durability and Cross-Reactivity of Immune Responses Induced by an AS03 Adjuvanted Plant-Based Recombinant Virus-Like Particle Vaccine for COVID-19

2021 ◽  
Author(s):  
Philipe A.M. Gobeil ◽  
Stéphane Pillet ◽  
Iohann Boulay ◽  
Nathalie Charland ◽  
Aurélien Lorin ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Phase 1 ◽  
Cross Reactivity ◽  
Interleukin 4 ◽  
Phase 3 ◽  
Live Virus ◽  
Post Vaccination ◽  
Virus Like Particle ◽  
Ifn Γ ◽  
Alpha Variant

As the SARS-COV-2 pandemic evolves, what is expected of vaccines extends beyond efficacy and includes evaluations of both durability and cross-reactivity to emerging variants. To complement an on-going Phase 3 efficacy study, this report expands on previously reported immunogenicity results from a Phase 1 trial of an AS03-adjuvanted, plant-based virus-like particle (VLP) displaying the spike glycoprotein of the Wuhan strain of SARS-CoV-2 virus (NCT04450004). Durability of the humoral and cellular responses against the ancestral strain was evaluated 6 months post-second dose (Day 201) at which time ~94% of vaccinated individuals remained seropositive. Interferon gamma (IFN-γ) and interleukin 4 (IL-4) responses remained detectable in ~94% and ~92% of vaccinated individuals respectively. Cross-reactivity of neutralizing antibodies to Alpha (B.1.17), Beta (B.1.351), and Gamma (P.1) variants of concern (VOC) were also measured. Twenty-one days after the second vaccination, detectable neutralizing antibodies were observed to the Alpha variant by both pseudovirion and wild-type assays for all vaccinated individuals, while 94.7% of individuals had detectable antibodies to the Beta variant in both assays. Neutralizing antibodies to the Gamma variant were detected in 100% and 94.7% of individuals using the pseudovirion and live virus neutralization assays, respectively. In all cases, the vaccine-induced neutralizing GMTs to the VOC 3 weeks post-vaccination were greater than the Wuhan-specific neutralization titers seen in individuals recovered from COVID-19.

scholarly journals Clinical and Virological Response to a Neutralizing Monoclonal Antibody for Hospitalized Patients with COVID-19

2021 ◽  
Author(s):  
Jens D. Lundgren ◽  
Keyword(s):  
Monoclonal Antibody ◽  
Neutralizing Antibodies ◽  
A Priori ◽  
Neutralizing Antibody ◽  
Viral Rna ◽  
Study Entry ◽  
Neutralizing Monoclonal Antibody ◽  
Rate Ratios ◽  
Sustained Recovery ◽  
Rna Levels

BACKGROUND Bamlanivimab, a neutralizing monoclonal antibody given in combination with remdesivir, did not improve outcomes among hospitalized COVID-19 patients based on an early futility assessment. In this final study report, we evaluate an a priori hypothesis that greater benefit of bamlanivimab would be identified in those without detectable endogenous neutralizing antibody levels at study entry, especially if viral levels were high. METHODS Hospitalized COVID-19 patients were randomized to receive bamlanivimab (7000mg) or placebo and followed for 90 days for sustained recovery (home for 14 consecutive days); recovery rate ratios (RRRs) are cited. RESULTS Among 314 participants (163 on bamlanivimab and 151 on placebo), the median time to sustained recovery was 19 days and RRR=0.99 (95% CI: 0.79-1.22; p=0.89). At entry, 50% evidenced production of anti-spike neutralizing antibodies; 50% had SARS-CoV-2 nucleocapsid plasma antigen levels ≥ 1,000 ng/L. Among those without and with antibodies at study entry, the RRRs were 1.24 (95% CI: 0.90-1.70) and 0.74 (95% CI: 0.54-1.00) (p=0.02 for interaction). The RRRs were elevated for those with plasma antigen or nasal viral RNA levels above versus below median at entry, and was greatest for those without antibodies and with elevated antigen or viral RNA levels: 1.48 (95% CI: 0.99-2.23), 1.94 (1.25-3.00), respectively (p<0.05 for all interactions). Hazard ratios for safety outcomes also differed by serostatus at entry. CONCLUSIONS Sustained recovery after administration of bamlanivimab versus placebo differed by presence of neutralizing antibodies at study entry, especially if participants had markers of elevated viral replication. ClinicalTrials.gov number, NCT04501978.

scholarly journals Field safety and efficacy of a unique live virus vaccine for controlling avian encephalomyelitis and fowlpox in poultry

2019 ◽  
Vol 12 (8) ◽  
pp. 1291-1298
Author(s):  
Girish Sarma ◽  
Barry A. Kersting ◽  
Gary Spina
Keyword(s):  
Virus Vaccine ◽  
Adverse Reactions ◽  
Field Conditions ◽  
Enzyme Linked Immunosorbent Assay ◽  
Live Virus ◽  
Post Vaccination ◽  
Safety And Efficacy ◽  
Live Virus Vaccine ◽  
Field Virus ◽  
The Usa

Background and Aim: Infection of commercial poultry with avian encephalomyelitis (AE) and fowlpox (FP) virus causes heavy economic loss in endemic areas. Although vaccines are routinely used to control these two diseases, the problem still persists almost all over the world. This study aimed to evaluate safety and efficacy of a unique AE + FP + pigeon pox (PP) live virus vaccine in layer-type chickens under both laboratory and field conditions. Materials and Methods: The study was conducted using 289 specific-pathogen-free (SPF) chickens under the laboratory conditions and 185,648 commercial layer-type chickens under field conditions. In two consecutive laboratory trials, 8-week-old SPF chickens were vaccinated with the AE + FP + PP live virus vaccine through wing web route and challenged against virulent strains of FP and AE viruses at 3-week post-vaccination (WPV). Challenged chickens were observed for disease protection for 10-21 days. For field safety trials, commercial layer-type chickens in three different geographical areas in the USA were vaccinated with the AE + FP + PP vaccine and observed daily up to 21 days for vaccine "take". adverse reactions, and mortality. Results: The vaccine was found safe and efficacious under both laboratory and field conditions. Vaccine "take" and protection against FP challenge were 100%. Average protection against AE challenge was 97%. Mean AE enzyme-linked immunosorbent assay (ELISA) antibody titer in the field vaccinated chickens was >1200 at 10 WPV. Average daily post-vaccination mortality in the field vaccinated chickens was 0.04%. So far, more than 400 million chickens in the USA have been vaccinated with this vaccine. No vaccine-associated adverse reactions, other safety issues, or immunity breakdown cases in the vaccinated flocks due to field virus infection have been reported. Conclusion: This unique vaccine containing AE, FP, and PP viruses in a single preparation was found to be safe and efficacious in controlling the diseases caused by the virulent field strains of AE and FP. Besides being safe and efficacious, this vaccine also offered distinct advantages over the traditional vaccination practices in controlling these two diseases in poultry. Keywords: avian encephalomyelitis, efficacy, field safety, fowlpox, live virus vaccine, pigeon pox, protection.

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