Protective Effect of an Anti-HMGB-1 Neutralizing Antibody on Hemozoin-Induced Alveolar Epithelial Cell in a Model of Malaria Associated ALI/ARDS

Tachpon  Techarang; Pitchanee  Jariyapong; Parnpen  Viriyavejakul; Supattra  Glaharn; Charit  Srisook; Chuchard  Punsawad

doi:10.18502/ijpa.v16i3.7089

Tachpon Techarang Department of Medical Sciences, School of Medicine, Walailak University, Nakhon Si Thammarat, Thailand
Pitchanee Jariyapong Department of Medical Sciences, School of Medicine, Walailak University, Nakhon Si Thammarat, Thailand
Parnpen Viriyavejakul Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
Supattra Glaharn Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
Charit Srisook Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand
Chuchard Punsawad Department of Medical Sciences, School of Medicine, Walailak University, Nakhon Si Thammarat, Thailand

DOI: https://doi.org/10.18502/ijpa.v16i3.7089

Keywords: HMGB1 protein; Malaria; Acute lung injury

Abstract

Background: We aimed to determine whether neutralizing high mobility group box-1 (HMGB-1) prevents the release of HMGB-1 and proinflammatory cytokines on hemozoin (Hz)-induced alveolar epithelial cell in a model of malaria associated ALI/ARDS.

Methods: This study was conducted in the Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand in 2020. Human pulmonary alveolar epithelial cells (HPAEpiCs) were exposed to medium alone or 20 µM Hz for 24 h and incubated with different concentrations (1, 5, and 10 µg/ml) of anti-HMGB-1 monoclonal antibody (mAb) for various times (0, 4, 12, 24, and 48 h). The levels of HMGB-1, TNF-α and IFN-γ in the supernatants were measured by ELISA. The mRNA expression of RAGE, TLR-2 and TLR-4 were analyzed by real-time PCR.

Results: The HPAEpiCs treated with 10 µg/ml anti-HMGB-1 mAb showed a significant reduction in HMGB-1 release into the supernatant compared with those treated with 1 and 5 µg/ml anti-HMGB-1 mAb. The levels of TNF-α and IFN-γ were significantly decreased in the supernatant of HPAEpiCs treated with 1, 5, and 10 µg/ml anti-HMGB-1 mAb for 4, 12, 24, and 48 h compared with those stimulated with Hz alone. The mRNA expression levels of RAGE, TLR-2, and TLR-4 were significantly decreased after 24 h of anti-HMGB-1 antibody treatment at all concentrations.

Conclusions: An anti-HMGB-1 antibody could be an effective agent for inhibiting the release of HMGB-1, TNF-α and IFN-γ. Furthermore, a neutralizing anti-HMGB-1 antibody could be applicable for the treatment of malaria-associated ALI/ARDS.