Pathogen inactivation of Dengue virus in red blood cells using amustaline and glutathione

Abstract

BACKGROUND Dengue virus (DENV) is an arbovirus primarily transmitted through mosquito bite; however, DENV transfusion‐transmitted infections (TTIs) have been reported and asymptomatic DENV RNA–positive blood donors have been identified in endemic countries. DENV is considered a high‐risk pathogen for blood safety. One of the mitigation strategies to prevent arbovirus TTIs is pathogen inactivation. In this study we demonstrate that the amustaline and glutathione (S‐303/GSH) treatment previously found effective against Zika virus in red blood cells (RBCs) is also effective in inactivating DENV. STUDY DESIGN AND METHODS Red blood cells were spiked with high levels of DENV. Viral RNA loads and infectious titers were measured in the untreated control and before and after pathogen inactivation treatment of RBC samples. DENV infectivity was also assessed over five successive cell culture passages to detect any potential residual replicative virus. RESULTS The mean ± SD DENV titer in RBCs before inactivation was 6.61 ± 0.19 log 50% tissue culture infectious dose (TCID50)/mL and the mean viral RNA load was 8.42 log genome equivalents/mL. No replicative DENV was detected either immediately after completion of treatment using S‐303/GSH or after cell culture passages. CONCLUSION Treatment using S‐303/GSH inactivated high levels of DENV in RBCs to the limit of detection. In combination with previous studies showing the effective inactivation of DENV in plasma and platelets using the licensed amotosalen/UVA system, this study demonstrates that high levels of DENV can be inactivated in all blood components.

Amustaline (S‐303) treatment inactivates high levels of Chikungunya virus in red‐blood‐cell components

“…We used a protocol previously published with the same experimental design (Fig. ) as previously used to demonstrate the inactivation of DENV and ZIKV in RBCs using S‐303/GSH .…”

“…Inactivation of CHIKV has been demonstrated in plasma and platelets using different systems. Recently, we demonstrated that the INTERCEPT™ Blood System using amustaline (S‐303) and glutathione (GSH) was able to inactivate high levels of DENV and ZIKV in RBCs .…”

“…The study utilizes the same protocol as previously published to demonstrate the efficacy of inactivation of DENV and ZIKV in RBCs . We report the inactivation of high levels of CHIKV in RBCs using S‐303/GSH.…”

Inactivation of Plasmodium falciparum in whole blood using the amustaline and glutathione pathogen reduction technology

“…Glutathione (GSH) is used as a quencher to prevent non‐specific protein reactions and extracellular protein damage. Amustaline/GSH has been shown to be highly efficient at reducing high levels of a wide range of parasites, viruses, and bacteria in RBC . The current study investigated the efficacy of the amustaline/GSH PRT to inactivate P. falciparum in WB.…”

“…Amustaline/GSH has been shown to be highly efficient at reducing high levels of a wide range of parasites, viruses, and bacteria in RBC. [22][23][24] The current study investigated the efficacy of the amustaline/ GSH PRT to inactivate P. falciparum in WB.…”