Limits...
Delayed antibody dependent enhancement of low passage dengue virus 4 isolates.

Wikan N, Libsittikul S, Yoksan S, Auewarakul P, Smith DR - BMC Res Notes (2015)

Bottom Line: The concept of antibody dependent enhancement (ADE) of dengue virus (DENV) infection is a cornerstone of our current understanding of dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage virus isolates.This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE.These results show that low passage DENV 4 viruses undergo ADE mediated infection, but that the process is significantly temporally delayed as compared to laboratory adapted DENV 4.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya Campus, 25/25 Phuttamonthol Sai 4, Salaya, Nakorn Pathom, 73170, Thailand. nitwara@hotmail.com.

ABSTRACT

Background: The concept of antibody dependent enhancement (ADE) of dengue virus (DENV) infection is a cornerstone of our current understanding of dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE.

Results: Little or no ADE of infection was observed on day 2 post infection with low passage isolates, while high enhancement of infection was seen with the laboratory adapted virus. However, both of the low passage isolates showed high levels of infection (60-100%) by day 5 post infection.

Conclusions: These results show that low passage DENV 4 viruses undergo ADE mediated infection, but that the process is significantly temporally delayed as compared to laboratory adapted DENV 4.

No MeSH data available.


Related in: MedlinePlus

ADE of infection of U937 cells with high and low passage DENVs. Antibody–virus complexes were generated with a DENV 4 LAB, b DENV 4 DF and c DENV 4 DHF and different dilutions of HB 114 antibodies before incubation with U937 cells and subsequent culture for 1–5 days post infection. The degree of infection was determined by flow cytometry. Experiment was undertaken independently in triplicate and error bars show SEM
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4556410&req=5

Fig1: ADE of infection of U937 cells with high and low passage DENVs. Antibody–virus complexes were generated with a DENV 4 LAB, b DENV 4 DF and c DENV 4 DHF and different dilutions of HB 114 antibodies before incubation with U937 cells and subsequent culture for 1–5 days post infection. The degree of infection was determined by flow cytometry. Experiment was undertaken independently in triplicate and error bars show SEM

Mentions: Undifferentiated U937 cells are highly susceptible to ADE mediated DENV infection, but are poorly susceptible to direct DENV infection [12, 13]. Previous studies have suggested that low passage DENVs or DENVs isolated directly from patients do not undergo ADE, or undergo ADE only poorly in comparison to laboratory adapted viruses [10, 11]. To investigate this further, three DENV 4 isolates were selected, one of which was a high passage laboratory (LAB) adapted virus (DENV 4 LAB), one a DENV 4 isolated from a dengue fever (DF) patient and passaged three times in C6/36 (DENV 4 DF) and one a DENV 4 isolated from a dengue hemorrhagic fever (DHF) patient and passaged three times in C6/36 (DENV 4 DHF). Under standard conditions, these three viruses were incubated with either no antibody or increasing dilutions of an antibody produced by hybridoma HB 114, which we have previously shown to be able to mediate ADE with DENV 4 LAB [12]. Cells were incubated under standard conditions and collected on days 1, 2, 3 and 5 post infection before analysis by flow cytometry to determine the degree of infection. Results (Fig. 1) show high levels infection enhancement with DENV 4 LAB from day 2 post infection, with maximum enhancement observed with antibody dilutions between 1:200 and 1:2000, consistent with our previous observations [12]. Significantly, both DENV 4 DF and DENV 4 DHF showed little or no ADE on day 2 post infection, but by day 3 (DENV 4 DF) or day 5 (DENV 4 DHF) post infection, cells infected with low passage isolates showed high degrees of infection, with maximum enhancement being observed with antibody dilutions of 1:200. Notably, cells infected with DENV 4 DF showed nearly 100 % infection by day 5 post infection, while cells infected with DENV 4 DHF showed approximately 60 % infection.Fig. 1


Delayed antibody dependent enhancement of low passage dengue virus 4 isolates.

Wikan N, Libsittikul S, Yoksan S, Auewarakul P, Smith DR - BMC Res Notes (2015)

ADE of infection of U937 cells with high and low passage DENVs. Antibody–virus complexes were generated with a DENV 4 LAB, b DENV 4 DF and c DENV 4 DHF and different dilutions of HB 114 antibodies before incubation with U937 cells and subsequent culture for 1–5 days post infection. The degree of infection was determined by flow cytometry. Experiment was undertaken independently in triplicate and error bars show SEM
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4556410&req=5

Fig1: ADE of infection of U937 cells with high and low passage DENVs. Antibody–virus complexes were generated with a DENV 4 LAB, b DENV 4 DF and c DENV 4 DHF and different dilutions of HB 114 antibodies before incubation with U937 cells and subsequent culture for 1–5 days post infection. The degree of infection was determined by flow cytometry. Experiment was undertaken independently in triplicate and error bars show SEM
Mentions: Undifferentiated U937 cells are highly susceptible to ADE mediated DENV infection, but are poorly susceptible to direct DENV infection [12, 13]. Previous studies have suggested that low passage DENVs or DENVs isolated directly from patients do not undergo ADE, or undergo ADE only poorly in comparison to laboratory adapted viruses [10, 11]. To investigate this further, three DENV 4 isolates were selected, one of which was a high passage laboratory (LAB) adapted virus (DENV 4 LAB), one a DENV 4 isolated from a dengue fever (DF) patient and passaged three times in C6/36 (DENV 4 DF) and one a DENV 4 isolated from a dengue hemorrhagic fever (DHF) patient and passaged three times in C6/36 (DENV 4 DHF). Under standard conditions, these three viruses were incubated with either no antibody or increasing dilutions of an antibody produced by hybridoma HB 114, which we have previously shown to be able to mediate ADE with DENV 4 LAB [12]. Cells were incubated under standard conditions and collected on days 1, 2, 3 and 5 post infection before analysis by flow cytometry to determine the degree of infection. Results (Fig. 1) show high levels infection enhancement with DENV 4 LAB from day 2 post infection, with maximum enhancement observed with antibody dilutions between 1:200 and 1:2000, consistent with our previous observations [12]. Significantly, both DENV 4 DF and DENV 4 DHF showed little or no ADE on day 2 post infection, but by day 3 (DENV 4 DF) or day 5 (DENV 4 DHF) post infection, cells infected with low passage isolates showed high degrees of infection, with maximum enhancement being observed with antibody dilutions of 1:200. Notably, cells infected with DENV 4 DF showed nearly 100 % infection by day 5 post infection, while cells infected with DENV 4 DHF showed approximately 60 % infection.Fig. 1

Bottom Line: The concept of antibody dependent enhancement (ADE) of dengue virus (DENV) infection is a cornerstone of our current understanding of dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage virus isolates.This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE.These results show that low passage DENV 4 viruses undergo ADE mediated infection, but that the process is significantly temporally delayed as compared to laboratory adapted DENV 4.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya Campus, 25/25 Phuttamonthol Sai 4, Salaya, Nakorn Pathom, 73170, Thailand. nitwara@hotmail.com.

ABSTRACT

Background: The concept of antibody dependent enhancement (ADE) of dengue virus (DENV) infection is a cornerstone of our current understanding of dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE.

Results: Little or no ADE of infection was observed on day 2 post infection with low passage isolates, while high enhancement of infection was seen with the laboratory adapted virus. However, both of the low passage isolates showed high levels of infection (60-100%) by day 5 post infection.

Conclusions: These results show that low passage DENV 4 viruses undergo ADE mediated infection, but that the process is significantly temporally delayed as compared to laboratory adapted DENV 4.

No MeSH data available.


Related in: MedlinePlus