scholarly journals Immunogenicity and Protective Efficacy of a Recombinant Subunit West Nile Virus Vaccine in Rhesus Monkeys

2009 ◽  
Vol 16 (9) ◽  
pp. 1332-1337 ◽  
Author(s):  
Michael M. Lieberman ◽  
Vivek R. Nerurkar ◽  
Haiyan Luo ◽  
Bruce Cropp ◽  
Ricardo Carrion ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Cellular Immunity ◽  
Cytokine Production ◽  
Protective Efficacy ◽  
Expression System ◽  
High Yield ◽  
Separate Experiment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
West Nile

ABSTRACT The immunogenicity and protective efficacy of a recombinant subunit West Nile virus (WNV) vaccine was evaluated in rhesus macaques (Macaca mulatta). The vaccine consisted of a recombinant envelope (E) protein truncated at the C-terminal end, resulting in a polypeptide containing 80% of the N-terminal amino acids of the native WNV protein (WN-80E), mixed with an adjuvant (GPI-0100). WN-80E was produced in a Drosophila melanogaster expression system with high yield and purified by immunoaffinity chromatography using a monoclonal antibody specific for flavivirus E proteins. Groups of monkeys were vaccinated with formulations containing 1 or 25 μg of WN-80E antigen, and both humoral and cellular immunity were assessed after vaccination. The results demonstrated potent antibody responses to vaccination, as determined by both enzyme-linked immunosorbent assay and virus-neutralizing antibody assays. All vaccinated animals responded favorably, and there was little difference in response between animals immunized with 1 or 25 μg of WN-80E. Cellular immunity was determined by lymphocyte proliferation and cytokine production assays using peripheral blood mononuclear cells from vaccinated animals stimulated in vitro with WN-80E. Cell-mediated immune responses varied from animal to animal within each group. About half of the animals responded with lymphoproliferation, cytokine production, or both. Again, there was little difference in response between animals immunized with a 1- or 25-μg dose of WN-80E in the vaccine formulations. In a separate experiment, groups of monkeys were immunized with the WN-80E/GPI-0100 vaccine or an adjuvant-only control formulation. Animals were then challenged by inoculation of wild-type WNV, and the level of viremia in each animal was monitored daily for 10 days. The results showed that whereas all animals in the control group had detectable viremia for at least 3 days after challenge, all of the vaccinated animals were negative on all days after challenge. Thus, the WN-80E vaccine was 100% efficacious in protecting monkeys against infection with WNV.

scholarly journals West Nile Virus and Related Flavivirus in European Wild Boar (Sus scrofa), Latium Region, Italy: A Retrospective Study

Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 494
Author(s):  
Angela Petruccelli ◽  
Tiziana Zottola ◽  
Gianmarco Ferrara ◽  
Valentina Iovane ◽  
Cristina Di Russo ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Wild Boar ◽  
Specific Antibody ◽  
Central Italy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
West Nile ◽  
Wild Boars ◽  
European Wild Boar

Background: A retrospective sero-survey for evidence of West Nile virus (WNV) infection in European wild boar (Sus scorfa) was conducted in the Latium region, Italy, on stored serum samples of the period November 2011 to January 2012. Methods: Sera were collected from 168 European wild boars and screened for antibodies to WNV and other Flaviviruses by competitive enzyme linked immunosorbent assay (cELISA). All sera positive for Flavivirus antibodies by cELISA were further examined by virus neutralization test (VNT). To test the presence of Flavivirus RNA in samples, an RT-PCR was performed using a pan-Flavivirus primers pair. Results: Thirteen wild boars (7.73%) were seropositive for Flaviviruses. The hemolysis of serum samples limited the interpretation of the VNT for 7 samples, confirming the presence of specific antibody against WNV in a single European wild boar serum sample. The presence of ELISA positive/VNT negative samples suggests the occurrence of non-neutralizing antibodies against WNV or other antigen-related Flaviviruses. No samples resulted positive for Flavivirus by RT-PCR assay. Conclusion: Although a moderately high percentage of animals with specific antibody for WNV has been detected in wild boar in other surveillance studies in Europe, this has not been reported previously in Italy. Together, these data indicate that European wild boar are exposed to WNV and/or other related-Flavivirus in central Italy and confirm the usefulness of wild ungulates, as suitable Flavivirus sentinels.

West Nile Virus

2016 ◽  
Author(s):  
Matthew Finn
Keyword(s):  
West Nile Virus ◽  
Rna Virus ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mosquito Vector ◽  
West Nile ◽  
Csf Analysis ◽  
Neuroinvasive Disease ◽  
Breeding Grounds ◽  
Arboviral Disease

West Nile virus (WNV) is a single-stranded RNA virus of the Flavivirus family that is transmitted via a mosquito vector, typically causing fever and capable of causing meningoencephalitis. Although mortality is low, it can lead to debilitating neuroinvasive disease in some patients. WNV is a leading cause of domestically-acquired arboviral disease and most commonly occurs in late August and early September. Consider WNV in otherwise unexplained cases of meningitis or encephalitis. Initial testing should consist of cerebrospinal fluid (CSF) analysis and West Nile immunoglobulin M enzyme-linked immunosorbent assay in serum and/or CSF. WNV is a nationally notifiable disease. Prevention remains the key to controlling this disease. Reducing the breeding grounds of the Culex mosquito and using insect repellant to prevent bites are two important strategies.

scholarly journals Persistence of Antibodies to West Nile Virus in Naturally Infected Rock Pigeons (Columba livia)

2005 ◽  
Vol 12 (5) ◽  
pp. 665-667 ◽  
Author(s):  
Samantha E. J. Gibbs ◽  
Douglas M. Hoffman ◽  
Lillian M. Stark ◽  
Nicole L. Marlenee ◽  
Bradley J. Blitvich ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Immunosorbent Assay ◽  
Neutralization Test ◽  
Columba Livia ◽  
Maternal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
West Nile ◽  
Entire Study ◽  
Plaque Reduction Neutralization Test ◽  
Antibody Persistence

ABSTRACT Wild caught rock pigeons (Columba livia) with antibodies to West Nile virus were monitored for 15 months to determine antibody persistence and compare results of three serologic techniques. Antibodies persisted for the entire study as detected by epitope-blocking enzyme-linked immunosorbent assay and plaque reduction neutralization test. Maternal antibodies in squabs derived from seropositive birds persisted for an average of 27 days.

scholarly journals West Nile Virus Seroprevalence among Qatari and Immigrant Populations within Qatar

2020 ◽  
Author(s):  
Hasna Kunhipurayil ◽  
Muna Ahmed ◽  
Gheyath Nasrallah
Keyword(s):  
West Nile Virus ◽  
Blood Donation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Large Sample Size ◽  
Serum Samples ◽  
West Nile ◽  
Viral Neutralization ◽  
Healthy Donors ◽  
Confirmatory Tests ◽  
Sub Saharan

Background: West Nile virus (WNV) is one of the most widely spread arboviruses worldwide and a highly significant pathogen in humans and animals. Despite frequent outbreaks and endemic transmission being reported in the Middle East and North Africa (MENA), seroprevalence studies of WNV in Qatar are highly lacking. Aim: This study aims to investigate the actual prevalence of WNV among local and expatriate communities in the Qatar using a large sample size of seemingly healthy donors. Method: A total of 1992 serum samples were collected from donors of age 18 or older and were tested for the presence of WNV antibodies. Serion enzyme-linked immunosorbent assay (ELISA) commercial microplate kits were used to detect the presence of the WNV IgM and IgG. The seropositivity was statistically analyzed using SPSS software with a confidence interval of 95%. Results: The seroprevalence of anti-WNV IgG and IgM in Qatar was 10.3% and 3.4%, respectively. The country-specific seroprevalence according to nationality for WNV IgG and IgM, respectively, were Sudan (37.0%, 10.0%), Egypt (31.6%, 4.4%), India (13.4%, 3.2%), Yemen(10.2%, 7.0%), Pakistan (8.6%, 2.7%), Iran (10.6%, 0.0%), Philippines (5.4%, 0.0%), Jordan(6.8%, 1.1%), Syria (2.6%, 9.6%), Palestine (2.6%, 0.6%), Qatar (1.6%, 1.7%), and Lebanon (0.9%, 0.0%). The prevalence of both IgM and IgG was significantly correlated with the nationality (p≤0.001). Conclusion: Among these tested nationalities, Qatar national has a relatively low burden of WNV disease. The highest prevalence of WNV was found in the Sub Saharan African nationalities like Sudan and Egypt. The seroprevalence of WNV is different from the previously reported arboviruses such as CHIKV and DENV, which was highest among Asian countries (India and Philippines). Further confirmatory tests such as viral neutralization assays are needed to confirm the IgM seropositivity in these samples since these samples could be a source of viral transmission through blood donation.

scholarly journals Evaluation of Cross Protective Efficacy of Commercial Vaccines against Mannheimia haemolytica in Mice

2019 ◽  
Vol 43 (1) ◽  
pp. 165-170
Author(s):  
Waffa A. Ahmed
Keyword(s):  
Immunosorbent Assay ◽  
Protective Efficacy ◽  
Challenge Test ◽  
Cross Protection ◽  
Mannheimia Haemolytica ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Significant Difference ◽  
Protection Efficacy

Mannheimia haemolytica together with Pasteurella multocida represents as a major bacterial causative agent of cattle, sheep and goats respiratory diseases and its one of the most important causes for economic losses to these animals .Commercially available vaccines were used to prevent infections caused by P. multocida and M. haemolytica. Thus, the aim of the present study was to evaluate the cross protection efficacy of two vaccines to protect mice against M.haemolytica, studying humeral immunity, using Enzyme-Linked Immunosorbent Assay. Forty five mice were divided into three equal groups, group one and two were inoculated subcutaneously  4μl\JOVAPAST® and 1μl of Al-kindy vaccines respectively, while the third group was with 0.5 ml sub cutaneous PBS. LD50for M.haemolytica was estimated as 2× 106 cfu \ml and challenge test was conducted by dropping 0.05 ml 2× 106 cfu \ml intranasally after three weeks of immunization for the three groups. The results of Enzyme-Linked Immunosorbent Assay, showed significant increase of antibody titters at (P<0.01) in (group 1 and 2) after first and second weeks post immunization, in comparison with control group. Also, the re-isolation of M.haemolytica from lungs tissue of all groups after challenged were positive with significant difference between control and immunized group, control group was 4× 108 cfu ∕ml which was higher than immunized group one and group two,which were 2.5×104 cfu∕ml and 3,5×105 cfu∕ml respectively after 24 hour of vaccine. In conclusion, the two commercial vaccines showed good cross protection efficacy against M. haemolytica, but JOVAPAST® vaccine showed higher efficacy than Alkindy vaccine, as that it contain  two  heterologous  killed strains and providing the basis for production a vaccine from the two  pathogen of local strains. 

scholarly journals Soluble Expression of a Neo2/15-Conjugated Single Chain Fv against PD-L1 in Escherichia coli

2022 ◽  
Vol 44 (1) ◽  
pp. 301-308
Author(s):  
Sun-Hee Kim ◽  
Hee-Jin Jeong
Keyword(s):  
Fusion Protein ◽  
Expression System ◽  
Antibody Fragment ◽  
Therapeutic Index ◽  
High Yield ◽  
Soluble Form ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain ◽  
Single Chain Fv ◽  
Programmed Death

Immunocytokines, antibody-cytokine fusion proteins, have the potential to improve the therapeutic index of cytokines by delivering the cytokine to the site of localized tumor cells using antibodies. In this study, we produced a recombinant anti-programmed death-ligand 1 (PD-L1) scFv, an antibody fragment against PD-L1 combined with a Neo2/15, which is an engineered interleukin with superior function using an E. coli expression system. We expressed the fusion protein in a soluble form and purified it, resulting in high yield and purity. The high PD-L1-binding efficiency of the fusion protein was confirmed via enzyme-linked immunosorbent assay, suggesting the application of this immunocytokine as a cancer-related therapeutic agent.

scholarly journals Protective Efficacy of a Chimeric Insect-Specific Flavivirus Vaccine against West Nile Virus

Vaccines ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 258 ◽  
Author(s):  
Laura J. Vet ◽  
Yin Xiang Setoh ◽  
Alberto A. Amarilla ◽  
Gervais Habarugira ◽  
Willy W. Suen ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Protective Efficacy ◽  
Neutralizing Antibody ◽  
Human Infection ◽  
Chimeric Virus ◽  
E Proteins ◽  
West Nile ◽  
Lethal Challenge ◽  
Virulent Strains ◽  
Flavivirus Vaccine

Virulent strains of West Nile virus (WNV) are highly neuro-invasive and human infection is potentially lethal. However, no vaccine is currently available for human use. Here, we report the immunogenicity and protective efficacy of a vaccine derived from a chimeric virus, which was constructed using the structural proteins (prM and E) of the Kunjin strain of WNV (WNVKUN) and the genome backbone of the insect-specific flavivirus Binjari virus (BinJV). This chimeric virus (BinJ/WNVKUN-prME) exhibits an insect-specific phenotype and does not replicate in vertebrate cells. Importantly, it authentically presents the prM-E proteins of WNVKUN, which is antigenically very similar to other WNV strains and lineages. Therefore BinJ/WNVKUN-prME represents an excellent candidate to assess as a vaccine against virulent WNV strains, including the highly pathogenic WNVNY99. When CD1 mice were immunized with purified BinJ/WNVKUN-prME, they developed robust neutralizing antibody responses after a single unadjuvanted dose of 1 to 5 μg. We further demonstrated complete protection against viremia and mortality after lethal challenge with WNVNY99, with no clinical or subclinical pathology observed in vaccinated animals. These data suggest that BinJ/WNVKUN-prME represents a safe and effective WNV vaccine candidate that warrants further investigation for use in humans or in veterinary applications.

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