A Hyper-Glycosylation of HBV Surface Antigen Correlates with HBsAg-Negativity at Immunosuppression-Driven HBV Reactivation in Vivo and Hinders HBsAg Recognition In Vitro
Romina Salpini,
Lorenzo Piermatteo,
Arianna Battisti,
Luna Colagrossi,
Marianna Aragri,
Katia Yu La Rosa,
Ada Bertoli,
Patrizia Saccomandi,
Miriam Lichtner,
Massimo Marignani,
Sarah Maylin,
Constance Delaugerre,
Filomena Morisco,
Nicola Coppola,
Aldo Marrone,
Nerio Iapadre,
Carlotta Cerva,
Stefano Aquaro,
Mario Angelico,
Loredana Sarmati,
Massimo Andreoni,
Jens Verheyen,
Francesca Ceccherini-Silberstein,
Massimo Levrero,
Carlo Federico Perno,
Laura Belloni,
Valentina Svicher
Affiliations
Romina Salpini
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Lorenzo Piermatteo
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Arianna Battisti
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Luna Colagrossi
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Marianna Aragri
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Katia Yu La Rosa
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Ada Bertoli
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Patrizia Saccomandi
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Miriam Lichtner
Public Health and Infectious Disease Department, Sapienza University, 00185 Rome, Italy
Massimo Marignani
Department of Gastroenterology, S.Andrea Hospital, 00189 Rome, Italy
Sarah Maylin
Laboratoire de Virologie, AP-HP Hopital Saint-Louis, 75010 Paris, France
Constance Delaugerre
Laboratoire de Virologie, AP-HP Hopital Saint-Louis, 75010 Paris, France
Filomena Morisco
Department of Clinical Medicine and Surgery, Section of Infectious Diseases, University of Naples Federico II, 80138 Naples, Italy
Nicola Coppola
Department of Mental Health and Public Medicine, Section of Infectious Diseases, Second University of Naples, 80138 Naples, Italy
Aldo Marrone
Internal Medicine and Hepatology Unit, Second University of Naples, 80138 Naples, Italy
Nerio Iapadre
Infectious Diseases Unit, San Salvatore Hospital, 67100 L’Aquila, Italy
Carlotta Cerva
Infectious Diseases Unit, Tor Vergata University Hospital, 00133 Rome, Italy
Stefano Aquaro
Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, 87036 Rende, Italy
Mario Angelico
Hepatology Unit, Tor Vergata University Hospital, 00133 Rome, Italy
Loredana Sarmati
Infectious Diseases Unit, Tor Vergata University Hospital, 00133 Rome, Italy
Massimo Andreoni
Infectious Diseases Unit, Tor Vergata University Hospital, 00133 Rome, Italy
Jens Verheyen
Institute of Virology, University-Hospital, University Duisburg-Essen, 47057 Essen, Germany
Francesca Ceccherini-Silberstein
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Massimo Levrero
Department of Internal Medicine-DMISM, Sapienza University, 00185 Rome, Italy
Carlo Federico Perno
Department of Oncology and Haemato-oncology, University of Milan, 20122 Milan, Italy
Laura Belloni
Department of Internal Medicine-DMISM, Sapienza University, 00185 Rome, Italy
Valentina Svicher
Department of Experimental Medicine, University of Rome Tor Vergata, 00133 Rome, Italy
Immune-suppression driven Hepatitis B Virus (HBV)-reactivation poses serious concerns since it occurs in several clinical settings and can result in severe forms of hepatitis. Previous studies showed that HBV strains, circulating in patients with HBV-reactivation, are characterized by an enrichment of immune-escape mutations in HBV surface antigen (HBsAg). Here, we focused on specific immune-escape mutations associated with the acquisition of N-linked glycosylation sites in HBsAg (NLGSs). In particular, we investigated profiles of NLGSs in 47 patients with immunosuppression-driven HBV-reactivation and we evaluated their impact on HBsAg-antigenicity and HBV-replication in vitro. At HBV-reactivation, despite a median serum HBV-DNA of 6.7 [5.3−8.0] logIU/mL, 23.4% of patients remained HBsAg-negative. HBsAg-negativity at HBV-reactivation correlated with the presence of >1 additional NLGSs (p < 0.001). These NLGSs are located in the major hydrophilic region of HBsAg (known to be the target of antibodies) and resulted from the single mutation T115N, T117N, T123N, N114ins, and from the triple mutant S113N+T131N+M133T. In vitro, NLGSs strongly alter HBsAg antigenic properties and recognition by antibodies used in assays for HBsAg-quantification without affecting HBsAg-secretion and other parameters of HBV-replication. In conclusion, additional NLGSs correlate with HBsAg-negativity despite HBV-reactivation, and hamper HBsAg-antigenicity in vitro, supporting the role of NGSs in immune-escape and the importance of HBV-DNA for a proper diagnosis of HBV-reactivation.
