scholarly journals DEVELOPMENT OF AN IMPROVED EPSTEIN-BARR VIRUS (EBV) NEUTRALIZING ANTIBODY ASSAY TO FACILITATE DEVELOPMENT OF A PROPHYLACTIC GP350-SUBUNIT EBV VACCINE

2020 ◽  
Vol 12 (1) ◽  
pp. e2020016
Author(s):  
Hui Liu ◽  
Lorraine Gemmell ◽  
Rui Lin ◽  
Fengrong Zuo ◽  
Henry H. Balfour ◽  
...  
Keyword(s):  
High Throughput ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Epidemiologic Studies ◽  
Antibody Detection ◽  
Healthy Human ◽  
Barr Virus ◽  
Human Sera ◽  
Epstein Barr ◽  
Bioanalytical Assays

No licensed vaccine is available for prevention of EBV-associated diseases, and robust, sensitive, and high-throughput bioanalytical assays are needed to evaluate immunogenicity of gp350 subunit-based candidate EBV vaccines. Here we have developed and improved analytical tools for such a vaccine’s pre-clinical and clinical validation including a gp350-specific antibody detection assay and an EBV-GFP based neutralization assay for measuring EBV specific antibodies in human donors. The sensitivity of our previously published high-throughput EBV-GFP fluorescent focus (FFA)-based neutralization assay was further improved when guinea pig complement was supplemented using a panel of healthy human sera. Anti-gp350 antibody titers, which were evaluated using an anti-gp350 IgG ELISA assay optimized for capture and detection conditions, were moderately correlated to the FFA-based neutralization titers. Overall, these sensitive, and high-throughput bioanalytical assays are capable of characterizing the serologic response to natural EBV infection, with applications in evaluating EBV antibody status in epidemiologic studies and immunogenicity of candidate gp350-subunit EBV vaccines in clinical studies.

scholarly journals Purified Hexameric Epstein-Barr Virus-Encoded BARF1 Protein for Measuring Anti-BARF1 Antibody Responses in Nasopharyngeal Carcinoma Patients

2010 ◽  
Vol 18 (2) ◽  
pp. 298-304 ◽  
Author(s):  
E. K. Hoebe ◽  
S. H. Hutajulu ◽  
J. van Beek ◽  
S. J. Stevens ◽  
D. K. Paramita ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Epstein Barr Virus ◽  
Antibody Responses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Humoral Immune ◽  
Barr Virus ◽  
Humoral Immune Responses ◽  
Human Sera ◽  
Epstein Barr ◽  
Humoral Responses

ABSTRACTWHO type III nasopharyngeal carcinoma (NPC) is highly prevalent in Indonesia and 100% associated with Epstein-Barr virus (EBV). NPC tumor cells express viral proteins, including BARF1, which is secreted and is considered to have oncogenic and immune-modulating properties. Recently, we found conserved mutations in the BARF1 gene in NPC isolates. This study describes the expression and purification of NPC-derived BARF1 and analyzes humoral immune responses against prototype BARF1 (B95-8) and purified native hexameric BARF1 in sera of Indonesian NPC patients (n= 155) compared to healthy EBV-positive (n= 56) and EBV-negative (n= 16) individuals. BARF1 (B95-8) expressed inEscherichia coliand baculovirus, as well as BARF1-derived peptides, did not react with IgG or IgA antibodies in NPC. Purified native hexameric BARF1 protein isolated from culture medium was used in enzyme-linked immunosorbent assay (ELISA) and revealed relatively weak IgG and IgA responses in human sera, although it had strong antibody responses to other EBV proteins. Higher IgG reactivity was found in NPC patients (P= 0.015) than in regional Indonesian controls or EBV-negative individuals (P< 0.001). IgA responses to native BARF1 were marginal. NPC sera with the highest IgG responses to hexameric BARF1 in ELISA showed detectable reactivity with denatured BARF1 by immunoblotting. In conclusion, BARF1 has low immunogenicity for humoral responses and requires native conformation for antibody binding. The presence of antibodies against native BARF1 in the blood of NPC patients provides evidence that the protein is expressed and secreted as a hexameric protein in NPC patients.

Novel nuclear antigens recognized by human sera in lymphocytes latently infected by Epstein-Barr virus

Virology ◽  
1987 ◽  
Vol 156 (1) ◽  
pp. 153-162 ◽  
Author(s):  
David T. Rowe ◽  
Paul J. Farrell ◽  
George Miller
Keyword(s):  
Epstein Barr Virus ◽  
Barr Virus ◽  
Nuclear Antigens ◽  
Latently Infected ◽  
Human Sera ◽  
Epstein Barr

scholarly journals Rabies neutralizing antibody detection by indirect immunperoxidase serum neutralization assay performed on chicken embryo related cell line

2004 ◽  
Vol 99 (5) ◽  
pp. 531-534 ◽  
Author(s):  
Tereza Cristina Cardoso ◽  
Luzia Helena Queiroz da Silva ◽  
Avelino Albas ◽  
Helena Lage Ferreira ◽  
Silvia Helena Venturoli Perri
Keyword(s):  
Cell Line ◽  
Chicken Embryo ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Antibody Detection ◽  
Serum Neutralization ◽  
Related Cell

scholarly journals Interpreting the Epstein-Barr Virus (EBV) Epigenome Using High-Throughput Data

Viruses ◽  
2013 ◽  
Vol 5 (4) ◽  
pp. 1042-1054 ◽  
Author(s):  
Aaron Arvey ◽  
Italo Tempera ◽  
Paul Lieberman
Keyword(s):  
High Throughput ◽  
Epstein Barr Virus ◽  
High Throughput Data ◽  
Barr Virus ◽  
Epstein Barr

scholarly journals Evaluation of Rapid Neutralizing Antibody Detection Test against Rabies Virus in Human Sera

2015 ◽  
Vol 43 (2) ◽  
pp. 111-116 ◽  
Author(s):  
Kieu Anh Thi Nguyen ◽  
Thu Tuyet Nguyen ◽  
Dong Vinh Nguyen ◽  
Giang Chau Ngo ◽  
Cam Nhat Nguyen ◽  
...  
Keyword(s):  
Rabies Virus ◽  
Neutralizing Antibody ◽  
Antibody Detection ◽  
Human Sera

scholarly journals Statistical Approach To Estimate Vaccinia-Specific Neutralizing Antibody Titers Using a High-Throughput Assay

2009 ◽  
Vol 16 (8) ◽  
pp. 1105-1112 ◽  
Author(s):  
Richard Kennedy ◽  
V. Shane Pankratz ◽  
Eric Swanson ◽  
David Watson ◽  
Hana Golding ◽  
...  
Keyword(s):  
Immune Responses ◽  
Vaccinia Virus ◽  
High Throughput ◽  
Large Scale ◽  
Statistical Approach ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Virus Neutralization Assay ◽  
Antibody Titers ◽  
High Throughput Assay

ABSTRACT Because of the bioterrorism threat posed by agents such as variola virus, considerable time, resources, and effort have been devoted to biodefense preparation. One avenue of this research has been the development of rapid, sensitive, high-throughput assays to validate immune responses to poxviruses. Here we describe the adaptation of a β-galactosidase reporter-based vaccinia virus neutralization assay to large-scale use in a study that included over 1,000 subjects. We also describe the statistical methods involved in analyzing the large quantity of data generated. The assay and its associated methods should prove useful tools in monitoring immune responses to next-generation smallpox vaccines, studying poxvirus immunity, and evaluating therapeutic agents such as vaccinia virus immune globulin.

scholarly journals Interleukin 13 Is Secreted by and Stimulates the Growth of Hodgkin and Reed-Sternberg Cells

1999 ◽  
Vol 189 (12) ◽  
pp. 1939-1946 ◽  
Author(s):  
Ursula Kapp ◽  
Wen-Chen Yeh ◽  
Bruce Patterson ◽  
Andrew J. Elia ◽  
David Kägi ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Expression Patterns ◽  
Neutralizing Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Barr Virus ◽  
Non Hodgkin Lymphoma ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Epstein Barr

Gene expression patterns can provide vital clues to the pathogenesis of neoplastic diseases. We investigated the expression of 950 genes in Hodgkin's disease (HD) by analyzing differential mRNA expression using microarrays. In two independent microarray experiments, the HD-derived cell lines L428 and KMH2 were compared with an Epstein-Barr virus (EBV)-immortalized lymphoblastoid B cell line, LCL-GK. Interleukin (IL)-13 and IL-5 were found to be highly expressed in the HD-derived cell lines. Examination of IL-13 and IL-5 expression by Northern blot analysis and enzyme-linked immunosorbent assay confirmed these results and revealed the expression of IL-13 in a third HD-derived cell line, HDLM2. Control LCL and EBV-negative non-Hodgkin lymphoma–derived cell lines did not express IL-13. In situ hybridization of lymph node tissue from HD patients showed that elevated levels of IL-13 were specifically expressed by Hodgkin/Reed-Sternberg (H/RS) tumor cells. Treatment of a HD-derived cell line with a neutralizing antibody to IL-13 resulted in a dose-dependent inhibition of H/RS cell proliferation. These data suggest that H/RS cells produce IL-13 and that IL-13 plays an important role in the stimulation of H/RS cell growth, possibly by an autocrine mechanism. Modulation of the IL-13 signaling pathway may be a logical objective for future therapeutic strategies.

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