Longitudinal serum profiling in Dengue fever patients with different severities by using multiplex flavivirus antigen microarrays

Dengue fever is one of the most common arbovirus-infected diseases and typically progresses through the febrile, critical, and convalescent phases. While most Dengue Fever patients have mild symptoms, a few develop severe symptoms due to hyperimmune responses. However, the role of the humoral responses during the Dengue fever progression, especially in severe cases, is still unclear. Here, we developed multiplex Flavivirus Antigen Microarrays (FAMs) for profiling antibody specificities in different infection phases and Dengue fever severities. The FAMs comprised 17 nonstructural and envelope proteins from Flaviviruses that frequently infect humans, e.g., 4 serotypes of Dengue viruses, Zika virus, Japanese encephalitis virus, West Nile virus, Yellow fever virus, and Tick-borne encephalitis virus. We collected serum samples from Dengue fever patients with different degrees of severity, including mild (n = 17), hospitalized non-severe (n = 21), and severe (n = 19), during febrile (Day < 7) and convalescent (Day > 7) phases. We simultaneously profiled the serum IgG and IgM against multiple antigens on the FAMs to demonstrate the immunological signatures in Dengue fever patients with different severities and two infection stages. Compared to febrile phase, the convalescent phase of mild but not hospitalized cases exhibiting evoked Dengue-specific IgM. In IgG profiling, the convalescent phase showed generally higher immune responses compared to febrile phase. Moreover, in the febrile phase, the hospitalized patients displayed higher IgG levels against 4 serotypes of Dengue viruses, Zika virus, and West Nile virus compared to mild patients, suggesting early and strong humoral responses. This study provides a valuable tool for monitoring antibody specificities and advances our understanding of humoral responses along with Dengue fever severities.