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Dengue viral RNA levels in peripheral blood mononuclear cells are associated with disease severity and preexisting dengue immune status.

Srikiatkhachorn A, Wichit S, Gibbons RV, Green S, Libraty DH, Endy TP, Ennis FA, Kalayanarooj S, Rothman AL - PLoS ONE (2012)

Bottom Line: Viral RNA levels in CD14+ cells and plasma were significantly higher in DHF compared to DF, and in cases with a secondary infection compared to those undergoing a primary infection.The distribution of viral RNA among cell subpopulations was similar in DF and DHF cases.Small amounts of negative strand RNA were found in a few cases only.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America. anon.srikiatkhachorn@umassmed.edu

ABSTRACT

Background: Infection with dengue viruses (DENV) causes a wide range of manifestations from asymptomatic infection to a febrile illness called dengue fever (DF), to dengue hemorrhagic fever (DHF). The in vivo targets of DENV and the relation between the viral burden in these cells and disease severity are not known.

Method: The levels of positive and negative strand viral RNA in peripheral blood monocytes, T/NK cells, and B cells and in plasma of DF and DHF cases were measured by quantitative RT-PCR.

Results: Positive strand viral RNA was detected in monocytes, T/NK cells and B cells with the highest amounts found in B cells. Viral RNA levels in CD14+ cells and plasma were significantly higher in DHF compared to DF, and in cases with a secondary infection compared to those undergoing a primary infection. The distribution of viral RNA among cell subpopulations was similar in DF and DHF cases. Small amounts of negative strand RNA were found in a few cases only. The severity of plasma leakage correlated with viral RNA levels in plasma and in CD14+ cells.

Conclusions: B cells were the principal cells containing DENV RNA in peripheral blood, but overall there was little active DENV RNA replication detectable in peripheral blood mononuclear cells (PBMC). Secondary infection and DHF were associated with higher viral burden in PBMC populations, especially CD14+ monocytes, suggesting that viral infection of these cells may be involved in disease pathogenesis.

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Related in: MedlinePlus

Positive and negative strand DENV RNA levels in unfractionated PBMC.Quantitative PCR for positive and negative strand viral RNA was performed with RNA extracted from unfractionated PBMC collected on fever day-2 from subjects infected with DENV1 (A), DENV2 (B), DENV3 (C), or DENV4 (D). The range of the positive strand DENV RNA was 0 to 6069 copies/million cells. Negative strand DENV RNA was detected in 6 of 18 samples with detectable positive strand RNA and the levels ranged from 5 to 35 copies/million cells.
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pone-0051335-g003: Positive and negative strand DENV RNA levels in unfractionated PBMC.Quantitative PCR for positive and negative strand viral RNA was performed with RNA extracted from unfractionated PBMC collected on fever day-2 from subjects infected with DENV1 (A), DENV2 (B), DENV3 (C), or DENV4 (D). The range of the positive strand DENV RNA was 0 to 6069 copies/million cells. Negative strand DENV RNA was detected in 6 of 18 samples with detectable positive strand RNA and the levels ranged from 5 to 35 copies/million cells.

Mentions: We performed quantitative RT-PCR for both positive and negative strand genomes using RNA samples extracted from PBMC preserved in RNA preserving solution at the time of collection (fever day −2). As shown in figure 3, we detected very low copy numbers of negative strand DENV RNA in a few of these PBMC samples, indicating that very little active viral replication is detectable in PBMC.


Dengue viral RNA levels in peripheral blood mononuclear cells are associated with disease severity and preexisting dengue immune status.

Srikiatkhachorn A, Wichit S, Gibbons RV, Green S, Libraty DH, Endy TP, Ennis FA, Kalayanarooj S, Rothman AL - PLoS ONE (2012)

Positive and negative strand DENV RNA levels in unfractionated PBMC.Quantitative PCR for positive and negative strand viral RNA was performed with RNA extracted from unfractionated PBMC collected on fever day-2 from subjects infected with DENV1 (A), DENV2 (B), DENV3 (C), or DENV4 (D). The range of the positive strand DENV RNA was 0 to 6069 copies/million cells. Negative strand DENV RNA was detected in 6 of 18 samples with detectable positive strand RNA and the levels ranged from 5 to 35 copies/million cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3526575&req=5

pone-0051335-g003: Positive and negative strand DENV RNA levels in unfractionated PBMC.Quantitative PCR for positive and negative strand viral RNA was performed with RNA extracted from unfractionated PBMC collected on fever day-2 from subjects infected with DENV1 (A), DENV2 (B), DENV3 (C), or DENV4 (D). The range of the positive strand DENV RNA was 0 to 6069 copies/million cells. Negative strand DENV RNA was detected in 6 of 18 samples with detectable positive strand RNA and the levels ranged from 5 to 35 copies/million cells.
Mentions: We performed quantitative RT-PCR for both positive and negative strand genomes using RNA samples extracted from PBMC preserved in RNA preserving solution at the time of collection (fever day −2). As shown in figure 3, we detected very low copy numbers of negative strand DENV RNA in a few of these PBMC samples, indicating that very little active viral replication is detectable in PBMC.

Bottom Line: Viral RNA levels in CD14+ cells and plasma were significantly higher in DHF compared to DF, and in cases with a secondary infection compared to those undergoing a primary infection.The distribution of viral RNA among cell subpopulations was similar in DF and DHF cases.Small amounts of negative strand RNA were found in a few cases only.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America. anon.srikiatkhachorn@umassmed.edu

ABSTRACT

Background: Infection with dengue viruses (DENV) causes a wide range of manifestations from asymptomatic infection to a febrile illness called dengue fever (DF), to dengue hemorrhagic fever (DHF). The in vivo targets of DENV and the relation between the viral burden in these cells and disease severity are not known.

Method: The levels of positive and negative strand viral RNA in peripheral blood monocytes, T/NK cells, and B cells and in plasma of DF and DHF cases were measured by quantitative RT-PCR.

Results: Positive strand viral RNA was detected in monocytes, T/NK cells and B cells with the highest amounts found in B cells. Viral RNA levels in CD14+ cells and plasma were significantly higher in DHF compared to DF, and in cases with a secondary infection compared to those undergoing a primary infection. The distribution of viral RNA among cell subpopulations was similar in DF and DHF cases. Small amounts of negative strand RNA were found in a few cases only. The severity of plasma leakage correlated with viral RNA levels in plasma and in CD14+ cells.

Conclusions: B cells were the principal cells containing DENV RNA in peripheral blood, but overall there was little active DENV RNA replication detectable in peripheral blood mononuclear cells (PBMC). Secondary infection and DHF were associated with higher viral burden in PBMC populations, especially CD14+ monocytes, suggesting that viral infection of these cells may be involved in disease pathogenesis.

Show MeSH
Related in: MedlinePlus