Heterologous prime-boost immunization induces protection against dengue virus infection in cynomolgus macaques.

IMPORTANCE: Currently licensed dengue vaccines do not induce long-term protection in children without previous exposure to dengue viruses in nature. These vaccines are based on selected attenuated strains of the four dengue serotypes and employed in combination for two or three consecutive doses. In our search for a better dengue vaccine candidate, live attenuated strains were followed by non-infectious virus-like particles or the plasmids that generate these particles upon injection into the body. This heterologous prime-boost immunization induced elevated levels of virus-specific antibodies and helped to prevent dengue virus infection in a high proportion of vaccinated macaques. In macaques that remained susceptible to dengue virus, distinct mechanisms were found to account for the immunization failures, providing a better understanding of vaccine actions. Additional studies in humans in the future may help to establish whether this combination approach represents a more effective means of preventing dengue by vaccination.

Blockade-of-Binding Activities toward Envelope-Associated, Type-Specific Epitopes as a Correlative Marker for Dengue Virus-Neutralizing Antibody.

Humans infected with dengue virus (DENV) acquire long-term protection against the infecting serotype, whereas cross-protection against other serotypes is short-lived. Long-term protection induced by low levels of type-specific neutralizing antibodies can be assessed using the virus-neutralizing antibody test. However, this test is laborious and time-consuming. In this study, a blockade-of-binding enzyme-linked immunoassay was developed to assess antibody activity by using a set of neutralizing anti-E monoclonal antibodies and blood samples from dengue virus-infected or -immunized macaques. Diluted blood samples were incubated with plate-bound dengue virus particles before the addition of an enzyme-conjugated antibody specific to the epitope of interest. Based on blocking reference curves constructed using autologous purified antibodies, sample blocking activity was determined as the relative concentration of unconjugated antibody that resulted in the same percent signal reduction. In separate DENV-1-, -2-, -3-, and -4-related sets of samples, moderate to strong correlations of the blocking activity with neutralizing antibody titers were found with the four type-specific antibodies 1F4, 3H5, 8A1, and 5H2, respectively. Significant correlations were observed for single samples taken 1 month after infection as well as samples drawn before and at various time points after infection/immunization. Similar testing using a cross-reactive EDE-1 antibody revealed a moderate correlation between the blocking activity and the neutralizing antibody titer only for the DENV-2-related set. The potential usefulness of the blockade-of-binding activity as a correlative marker of neutralizing antibodies against dengue viruses needs to be validated in humans. IMPORTANCE This study describes a blockade-of-binding assay for the determination of antibodies that recognize a selected set of serotype-specific or group-reactive epitopes in the envelope of dengue virus. By employing blood samples collected from dengue virus-infected or -immunized macaques, moderate to strong correlations of the epitope-blocking activities with the virus-neutralizing antibody titers were observed with serotype-specific blocking activities for each of the four dengue serotypes. This simple, rapid, and less laborious method should be useful for the evaluation of antibody responses to dengue virus infection and may serve as, or be a component of, an in vitro correlate of protection against dengue in the future.

Clinical manifestations and pathological correlation of immunoglobulin A nephropathy in children.

BACKGROUND: IgA nephropathy in children has various clinical manifestations. Kidney biopsy is a gold standard for diagnosis by using Oxford classification 2016 with few studies about the correlation between clinical and pathology manifestations. This study aims to find these correlations at the time of diagnosis and during short-term follow-up. METHOD: In this retrospective cohort study, 47 pediatric patients who underwent renal biopsy from 2010 to 2021 in Thailand, were included. Oxford classification 2016 has been used to score patients' pathology. Univariate and multivariate associations have been used for correlation between clinical and pathologic parameters. RESULTS: The most common clinical manifestations were microscopic hematuria and proteinuria. There were 68% of children with mesangial hypercellularity (M1), 42% with segmental glomerulosclerosis (S1), 25% with moderate to severe crescent (C1/C2), 23% with endocapillary hypercellularity (E1), and 14% with moderate to a severe tubular atrophy/interstitial fibrosis (T1/T2). Microscopic hematuria was strongly associated with mesangial hypercellularity (M1) OR 7.14 (95%CI 1.83 - 27.88, p-value 0.005) and hypertension was strongly associated with segmental glomerulosclerosis (S1) adjusted OR 7.87 (95%CI 1.65 - 37.59, p-value 0.01). Intensive treatment was used more in the patients with tubular atrophy/interstitial fibrosis lesion on renal biopsy than other lesions from MEST-C scores OR 4.98 (95%CI 1.17-21.24, p-value 0.03). Furthermore, pulse methylprednisolone and cyclophosphamide were used in patients with crescentic lesions significantly than other lesions with OR 15.5 (95%CI 3.16- 75.93, p-value 0.001) and OR 5.75 (95%CI 1.31-25.29, p-value 0.021), respectively. CONCLUSION: Tubular atrophy/interstitial fibrosis and crescent lesions were correlated to intensive treatment in short-term outcomes.