Application of an in vitro method for detecting antibody-dependent enhancement of Chikungunya virus infection

INTRODUCTION. The mechanism of antibody-dependent enhancement (ADE) caused by the Fcγ receptor on immune cells is a key factor. When the virus binds to specific immunoglobulin G

(IgG), it enhances cell penetration. ADE can also occur in vaccinated individuals, making its study crucial for evaluating vaccine safety.

AIM. This study aimed to apply the method of detection of antibody-dependent enhacement in vitro using cells of the K562 and Vero lines, as well as the Chikungunya virus strain Nika21.

MATERIALS AND METHODS. Chikungunya virus (ChikV) Nika21 strain and different dilutions sera containing anti-ChikV IgG were used. To evaluate ADE, K562 and Vero cells were challenged with a ChikV with specific IgG mixture. On the fourth day after infection, the biological ChikV titer and viral RNA content were determined. ChikV titer was determined by standard in vitro virus titration and RNA amount was determined by real-time fluorescence reverse transcription PCR. ADE was considered confirmed if the values of the biological ChikV titer in the presence of the tested serum were >3 SD relative to the mean ChikV titer in the control, which represented ChikV with serum of a healthy volunteer.

RESULTS. In Vero cells, both methods showed increased ChikV replication with an increase in the dilution of immune sera, which, however, did not exceed the values of ChikV titers in the control. In K562 cells, serum with anti-ChikV IgG in one dilution demonstrated statistically significant enhancement of ChikV replication relative to the corresponding control. Thus, in our study, ADE was detected in the presence of intermediate dilutions of immune sera with a relatively medium or low titer of anti-ChikV AT.

CONCLUSION. ChikV replication in FcR-negative Vero cell line confirmed the high sensitivity to the virus and the possibility of using it exclusively for assessing the neutralizing activity of specific IgG. FcR-expressing K562 cell line allowed to analyze both virus neutralizing and activating infection functions of IgG. Our results confirmed the possibility of conducting ADE studies in vitro, which greatly simplifies the study of the ADE phenomenon when assessing the safety of vaccines under development.

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