Dengue virus neutralizing antibody levels associated with protection from infection in thai cluster studies

Abstract

Background: Long-term homologous and temporary heterologous protection from dengue virus (DENV) infection may be mediated by neutralizing antibodies. However, neutralizing antibody titers (NTs) have not been clearly associated with protection from infection.

Methodology/principal findings: Data from two geographic cluster studies conducted in Kamphaeng Phet, Thailand were used for this analysis. In the first study (2004-2007), cluster investigations of 100-meter radius were triggered by DENV-infected index cases from a concurrent prospective cohort. Subjects between 6 months and 15 years old were evaluated for DENV infection at days 0 and 15 by DENV PCR and IgM ELISA. In the second study (2009-2012), clusters of 200-meter radius were triggered by DENV-infected index cases admitted to the provincial hospital. Subjects of any age ≥6 months were evaluated for DENV infection at days 0 and 14. In both studies, subjects who were DENV PCR positive at day 14/15 were considered to have been "susceptible" on day 0. Comparison subjects from houses in which someone had documented DENV infection, but the subject remained DENV negative at days 0 and 14/15, were considered "non-susceptible." Day 0 samples were presumed to be from just before virus exposure, and underwent plaque reduction neutralization testing (PRNT). Seventeen "susceptible" (six DENV-1, five DENV-2, and six DENV-4), and 32 "non-susceptible" (13 exposed to DENV-1, 10 DENV-2, and 9 DENV-4) subjects were evaluated. Comparing subjects exposed to the same serotype, receiver operating characteristic (ROC) curves identified homotypic PRNT titers of 11, 323 and 16 for DENV-1, -2 and -4, respectively, to differentiate "susceptible" from "non-susceptible" subjects.

Conclusions/significance: PRNT titers were associated with protection from infection by DENV-1, -2 and -4. Protective NTs appeared to be serotype-dependent and may be higher for DENV-2 than other serotypes. These findings are relevant for both dengue epidemiology studies and vaccine development efforts.

Figure 1. Pre-exposure neutralizing antibody titers (NTs) in dengue virus serotype 1 (DENV-1) “susceptible” and “non-susceptible subjects.

Graphs show: (a) six DENV-1 PCR positive (“susceptible”) subjects; (b) 13 DENV-1 PCR negative (“non-susceptible”) subjects. NTs for each subject are against the reference strains for DENV-1, -2, -3, -4 and Japanese encephalitis virus (JEV) (red, blue, green, purple and orange, respectively).

Figure 2. Pre-exposure neutralizing antibody titers (NTs) in dengue virus serotype 4 (DENV-4) “susceptible” and “non-susceptible” subjects.

Graphs show: (a) six DENV-4 PCR positive (“susceptible”) subjects; (b) nine DENV-4 PCR negative (“non-susceptible”) subjects. NTs for each subject are against the reference strains for DENV-1, -2, -3, -4 and Japanese encephalitis virus (JEV) (red, blue, green, purple and orange, respectively).

Figure 3. Pre-exposure neutralizing antibody titers (NTs) in dengue virus serotype 2 (DENV-2) “susceptible” and “non-susceptible” subjects.

Each row shows one PCR positive (“susceptible”) subject and two matched PCR negative (“non-susceptible”) subjects. NTs for each subject are against (in order from left to right for each subject) the reference strains for DENV-1, -2, -3, -4 and Japanese encephalitis virus (JEV), and six different DENV-2 strains (Asian-American 1982 [#D82-165], Asian I 1974 [#D74-066], Asian I 1984 [#D84-501], Asian I 1994 [#D94-035], Asian I 2004 [#KDS00305]), and homologous DENV-2 Asian I strains).

Figure 4. Receiver operating characteristic (ROC) curves for homotypic neutralizing antibody titers (NTs) against reference strains.

Curves discriminate between dengue virus (DENV) PCR positive and negative subjects for: (a) all serotypes combined; (b) DENV-1; (c) DENV-4; (d) DENV-2.