Abstract
BackgroundInvestigating the neutralizing antibody (NAb) titer against HSV-1 is essential for monitoring the immune protection against HSV-1 in susceptible populations, which would facilitate the development of vaccines against herpes infection and improvement of HSV-1 based oncolytic virotherapy.ResultsIn this study, we have developed a neutralization test based on the enzyme-linked immunospot assay (ELISPOT-NT) to determine the neutralizing antibody titer against HSV-1 in human serum samples. This optimized assay employed a monoclonal antibody specifically recognizing glycoprotein D to detect the HSV-1 infected cells. With this test, the neutralizing antibody titer against HSV-1 could be determined within one day by automated interpretation of the counts of cell spots. We observed good correlation in the results obtained from ELISPOT-NT and plaque reduction neutralization test (PRNT) by testing 22 human serum samples representing different titers. Moreover, 269 human serum samples collected from a wide range of age groups were tested, the average neutralizing antibody titer (log2NT50) was 8.3 ± 2.8 and the prevalence of NAbs was 83.6 % in this cohort, it also revealed that the average neutralizing antibody titer in different groups increased with the age, and no significant difference in neutralizing antibody titers was observed between males and females.ConclusionsThese results prove that this novel assay would serve as an accurate and simple assay for the assessment of the neutralizing antibody titers against HSV-1 in large cohorts.
Highlights
Investigating the neutralizing antibody (NAb) titer against Herpes simplex virus type 1 (HSV-1) is essential for monitoring the immune protection against HSV-1 in susceptible populations, which would facilitate the development of vaccines against herpes infection and improvement of HSV-1 based oncolytic virotherapy
It was found that 2G5 harbored good reactivity with other HSV-1 strains, such as strain F and strain 17, and cross-reacted weakly with HSV-2 (Fig. 1c). 12 viral glycoproteins were employed to determine the target protein of 2G5 MAb, it revealed that 2G5 MAb recognized denature or naïve form of glycoprotein D without any background (Fig. 1d and e)
Feasibility of the ELISPOT-NT Based on the previous findings, we evaluated the feasibility of ELISPOT based neutralizing test (ELISPOT-NT) using 2G5 antibody on determining the a it was necessary to investigate whether the neutralizing antibody titers of human serum samples can be reliably measured by use of ELISPOT-NT under different experimental variations, such as infectious dose
Summary
Investigating the neutralizing antibody (NAb) titer against HSV-1 is essential for monitoring the immune protection against HSV-1 in susceptible populations, which would facilitate the development of vaccines against herpes infection and improvement of HSV-1 based oncolytic virotherapy. Surveillance of seroprevalence against HSV-1 in susceptible populations is essential for monitoring the seroconversion rates in countries where HSV-1 is endemic to evaluate the effectiveness of vaccination and anti-viral therapy against herpes infection, especially to HSV-2 infection. NAbs against HSV-1 are one of the barriers oncolytic viruses encounter during viral replication in vivo It is largely unknown whether the circulating NAbs in cancer patients will restrict the efficacy of HSV-1 based virotherapy, especially when the viruses are administrated through intravenous delivery [13]. It’s intriguing for us to know the average NAb titers in general population, the relationship between HSV-1 serostatus and NAb titers, and which amount of NAbs will affect the efficacy of HSV-2 vaccine, as well as HSV-1 based oncolytic virotherapy
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