scholarly journals Antibody Responses Determined for Japanese Dengue Fever Patients by Neutralization and Hemagglutination Inhibition Assays Demonstrate Cross-Reactivity between Dengue and Japanese Encephalitis Viruses

2003 ◽  
Vol 10 (4) ◽  
pp. 725-728 ◽  
Author(s):  
Ken-Ichiro Yamada ◽  
Tomohiko Takasaki ◽  
Masaru Nawa ◽  
Sadao Yabe ◽  
Ichiro Kurane
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Japanese Encephalitis ◽  
Hemagglutination Inhibition ◽  
Neutralization Test ◽  
Neutralization Assay ◽  
Cross Reactivity ◽  
Antibody Responses ◽  
Dengue Virus Serotypes ◽  
Virus Antigens

ABSTRACT Titers of antibodies to infecting dengue virus serotypes determined by serum neutralization assay were higher than those of antibody to Japanese encephalitis (JE) virus in Japanese dengue patients after disease day 8. Titers of antibody to dengue virus antigens determined by hemagglutination inhibition (HI) assay were higher in only 1 of 23 serum specimens after disease day 11. Thus, the neutralization test is more reliable than the HI test for serological diagnosis of dengue in countries where JE vaccination is widely used or JE is endemic.

scholarly journals Optimization of Flow-Cytometry Based Assay for Measuring Neutralizing Antibody Responses against Each of the Four Dengue Virus Serotypes

Vaccines ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1339
Author(s):  
Pragati Sharma ◽  
Kaustuv Nayak ◽  
Elluri Seetharami Reddy ◽  
Humaira Farooqi ◽  
Kaja Murali-Krishna ◽  
...  
Keyword(s):  
Public Health ◽  
Flow Cytometry ◽  
Dengue Virus ◽  
Vaccine Development ◽  
Neutralizing Antibody ◽  
Neutralization Assay ◽  
Antibody Responses ◽  
Health Concern ◽  
Dengue Virus Serotypes ◽  
Focus Reduction

Dengue is an important public health problem worldwide, with India contributing nearly a third of global dengue disease burden. The measurement of neutralizing antibody responses is critical for understanding dengue pathophysiology, vaccine development and evaluation. Historically, dengue virus neutralization titers were measured using plaque reduction neutralization tests (PRNTs), which were later adapted to focus reduction neutralization tests (FRNTs). Given the slow and laborious nature of both these assays, there has been interest in adapting a high-throughput flow cytometry based neutralization assay. However, flow cytometry based assays typically underestimate neutralization titers, and in situations where the titers are low they can even fail to detect neutralization activity. In this study, by evaluating graded numbers of input Vero cell numbers and viral inoculum, we optimized the flow cytometry based neutralization assay in such a way that it is sensitive and scores titers that are in concordance with focus reduction neutralization tests for each of the four dengue virus serotypes (p < 0.0001). Given that dengue is a global public health concern, and several research groups are making efforts to understand its pathophysiology and accelerate vaccine development and evaluation both in India and worldwide, our findings have timely significance for facilitating these efforts.

scholarly journals Comparison of Complement Fixation and Hemagglutination Inhibition Assays for Detecting Antibody Responses following Influenza Virus Vaccination

2003 ◽  
Vol 10 (3) ◽  
pp. 481-482 ◽  
Author(s):  
Harry E. Prince ◽  
Amy L. Leber
Keyword(s):  
Influenza Virus ◽  
Antibody Response ◽  
Hemagglutination Inhibition ◽  
Complement Fixation ◽  
Antibody Responses ◽  
Virus Vaccination ◽  
Virus Antigens ◽  
Low Sensitivity

ABSTRACT Complement fixation (CF) was compared to hemagglutination inhibition (HI) as a method for identifying antibody responses to influenza virus vaccination. CF assays were performed at two different laboratories using paired (pre- and postvaccination) sera from 38 vaccinated laboratory employees; HI assays were performed at a third laboratory. As expected, most vaccinees (31/38 = 82%) responded to at least one of three influenza virus antigens as measured by HI. In contrast, only 21% (8/38) of vaccinees showed a response by CF at laboratory 1, and only 29% (11/38) showed a response by CF at laboratory 2. These findings indicate that due to low sensitivity, CF assays should not be used to assess the antibody response to influenza virus vaccination.

scholarly journals Antibodies to Envelope Glycoprotein of Dengue Virus during the Natural Course of Infection Are Predominantly Cross-Reactive and Recognize Epitopes Containing Highly Conserved Residues at the Fusion Loop of Domain II

2008 ◽  
Vol 82 (13) ◽  
pp. 6631-6643 ◽  
Author(s):  
Chih-Yun Lai ◽  
Wen-Yang Tsai ◽  
Su-Ru Lin ◽  
Chuan-Liang Kao ◽  
Hsien-Ping Hu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Primary Infection ◽  
Nonstructural Protein ◽  
Secondary Infection ◽  
Critical Role ◽  
Neutralization Assay ◽  
Antibody Responses ◽  
Conserved Residues ◽  
Nonstructural Protein 1 ◽  
Fusion Loop

ABSTRACT The antibody response to the envelope (E) glycoprotein of dengue virus (DENV) is known to play a critical role in both protection from and enhancement of disease, especially after primary infection. However, the relative amounts of homologous and heterologous anti-E antibodies and their epitopes remain unclear. In this study, we examined the antibody responses to E protein as well as to precursor membrane (PrM), capsid, and nonstructural protein 1 (NS1) of four serotypes of DENV by Western blot analysis of DENV serotype 2-infected patients with different disease severity and immune status during an outbreak in southern Taiwan in 2002. Based on the early-convalescent-phase sera tested, the rates of antibody responses to PrM and NS1 proteins were significantly higher in patients with secondary infection than in those with primary infection. A blocking experiment and neutralization assay showed that more than 90% of anti-E antibodies after primary infection were cross-reactive and nonneutralizing against heterologous serotypes and that only a minor proportion were type specific, which may account for the type-specific neutralization activity. Moreover, the E-binding activity in sera of 10 patients with primary infection was greatly reduced by amino acid replacements of three fusion loop residues, tryptophan at position 101, leucine at position 107, and phenylalanine at position 108, but not by replacements of those outside the fusion loop of domain II, suggesting that the predominantly cross-reactive anti-E antibodies recognized epitopes involving the highly conserved residues at the fusion loop of domain II. These findings have implications for our understanding of the pathogenesis of dengue and for the future design of subunit vaccine against DENV as well.

scholarly journals Mapping the Human Memory B Cell and Serum Neutralizing Antibody Responses to Dengue Virus Serotype 4 Infection and Vaccination

2016 ◽  
Vol 91 (5) ◽  
Author(s):  
Usha K. Nivarthi ◽  
Nurgun Kose ◽  
Gopal Sapparapu ◽  
Douglas Widman ◽  
Emily Gallichotte ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Neutralizing Antibodies ◽  
Natural Infection ◽  
Neutralizing Antibody ◽  
Hemorrhagic Fever ◽  
Antibody Responses ◽  
Human Antibodies ◽  
E Protein ◽  
Virus Serotype

ABSTRACT The four dengue virus (DENV) serotypes are mosquito-borne flaviviruses responsible for dengue fever and dengue hemorrhagic fever. People exposed to DENV develop antibodies (Abs) that strongly neutralize the serotype responsible for infection. Historically, infection with DENV serotype 4 (DENV4) has been less common and less studied than infections with the other three serotypes. However, DENV4 has been responsible for recent large and sustained epidemics in Asia and Latin America. The neutralizing antibody responses and the epitopes targeted against DENV4 have not been characterized in human infection. In this study, we mapped and characterized epitopes on DENV4 recognized by neutralizing antibodies in people previously exposed to DENV4 infections or to a live attenuated DENV4 vaccine. To study the fine specificity of DENV4 neutralizing human antibodies, B cells from two people exposed to DENV4 were immortalized and screened to identify DENV-specific clones. Two human monoclonal antibodies (MAbs) that neutralized DENV4 were isolated, and their epitopes were finely mapped using recombinant viruses and alanine scan mutation array techniques. Both antibodies bound to quaternary structure epitopes near the hinge region between envelope protein domain I (EDI) and EDII. In parallel, to characterize the serum neutralizing antibody responses, convalescence-phase serum samples from people previously exposed to primary DENV4 natural infections or a monovalent DENV4 vaccine were analyzed. Natural infection and vaccination also induced serum-neutralizing antibodies that targeted similar epitope domains at the EDI/II hinge region. These studies defined a target of neutralizing antigenic site on DENV4 targeted by human antibodies following natural infection or vaccination. IMPORTANCE The four serotypes of dengue virus are the causative agents of dengue fever and dengue hemorrhagic fever. People exposed to primary DENV infections develop long-term neutralizing antibody responses, but these principally recognize only the infecting serotype. An effective vaccine against dengue should elicit long-lasting protective antibody responses to all four serotypes simultaneously. We and others have defined antigenic sites on the envelope (E) protein of viruses of dengue virus serotypes 1, 2, and 3 targeted by human neutralizing antibodies. The epitopes on DENV4 E protein targeted by the human neutralizing antibodies and the mechanisms of serotype 4 neutralization are poorly understood. Here, we report the properties of human antibodies that neutralize dengue virus serotype 4. People exposed to serotype 4 infections or a live attenuated serotype 4 vaccine developed neutralizing antibodies that bound to similar sites on the viral E protein. These studies have provided a foundation for developing and evaluating DENV4 vaccines.

scholarly journals Seroprevalence of Flavivirus Neutralizing Antibodies in Thailand by High-Throughput Neutralization Assay: Endemic Circulation of Zika Virus before 2012

mSphere ◽  
2021 ◽  
Author(s):  
Atsushi Yamanaka ◽  
Mami Matsuda ◽  
Tamaki Okabayashi ◽  
Pannamthip Pitaksajjakul ◽  
Pongrama Ramasoota ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Japanese Encephalitis Virus ◽  
Japanese Encephalitis ◽  
High Throughput ◽  
Zika Virus ◽  
Neutralizing Antibodies ◽  
Neutralization Assay ◽  
Encephalitis Virus ◽  
Antibody Detection ◽  
Luciferase Gene

Neutralization tests are the most reliable assay for flavivirus antibody detection; however, these assays are not suitable for high-throughput processing due to their time-consuming and labor-intensive nature. In this study, we developed single-round infectious particles (SRIPs) with a luciferase gene for dengue virus types 1 to 4, Japanese encephalitis virus, and Zika virus for use in a safe, high-throughput neutralization assay.

Serotype-specific and cross-reactive neutralizing antibody responses in cynomolgus monkeys after infection with multiple dengue virus serotypes

2011 ◽  
Vol 156 (6) ◽  
pp. 1073-1077 ◽  
Author(s):  
Mikako Ito ◽  
Yuko Katakai ◽  
Fumiko Ono ◽  
Hirofumi Akari ◽  
Ryo-zabro Mukai ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Neutralizing Antibody ◽  
Antibody Responses ◽  
Cynomolgus Monkeys ◽  
Dengue Virus Serotypes
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