Multiplex cytokine profile from dengue patients: MIP-1beta and IFN-gamma as predictive factors for severity

Abstract

Background: Dengue virus pathogenesis is not yet fully understood and the identification of patients at high risk for developing severe disease forms is still a great challenge in dengue patient care. During the present study, we evaluated prospectively the potential of cytokines present in plasma from patients with dengue in stratifying disease severity.

Methods: Seventeen-cytokine multiplex fluorescent microbead immunoassay was used for the simultaneous detection in 59 dengue patients. GLM models using bimodal or Gaussian family were determined in order to associate cytokines with clinical manifestations and laboratory diagnosis.

Results: IL-1beta, IFN-gamma, IL-4, IL-6, IL-13, IL-7 and GM-CSF were significantly increased in patients with severe clinical manifestations (severe dengue) when compared to mild disease forms (mild dengue). In contrast, increased MIP-1beta levels were observed in patients with mild dengue. MIP-1beta was also associated with CD56+NK cell circulating rates. IL-1beta, IL-8, TNF-alpha and MCP-1 were associated with marked thrombocytopenia. Increased MCP-1 and GM-CSF levels correlated with hypotension. Moreover, MIP-1beta and IFN-gamma were independently associated with both dengue severity and disease outcome.

Conclusion: Our data demonstrated that the use of a multiple cytokine assay platform was suitable for identifying distinct cytokine profiles associated with the dengue clinical manifestations and severity. MIP-beta is indicated for the first time as a good prognostic marker in contrast to IFN-gamma that was associated with disease severity.

Figure 1

Cytokine levels in plasma from patients with mild and severe dengue. Box-and-whiskers graph. The box extends from the 25^th to the 75^th percentile and the line at the middle is the median. The error bars, or whiskers extend down to the lowest value and up to the highest. Factors were sorted according to their functional groups. Mann-Whitney U test was used to evaluate differences between cytokine concentration from severe and mild dengue patients. * P < 0.05, ** P < 0.01 and ** P < 0.001.

Figure 2

Cytokines detected in plasma as independent factors in predicting severity. Normal GLM model originated from logistic regression analysis using binomial family odds ratios (OD) and 95% confidence intervals (CI). Fifty-nine patients with dengue were grouped according with their clinical manifestations: Severe (N = 39), hypotension (N = 24) or thrombocytopenia (N = 18). Details on Table 2.

Figure 3

Hypothetic mechanism to explain cytokine models during dengue fever. MIP-1β is associated with a good prognostic and IFN-γ has a predictive value for severity. GM-CSF, MCP-1, IL-1β, IL-6, IL-8, IL-12, IL-13 are also playing important roles during dengue pathogenesis (see text for detailed description).