Introduction: The human cytomegalovirus (HCMV), a member of b-herpesviridae family, is a widespread pathogen that infect between 50% to 85% of human population. HCMV is able to establish a lifetime infection in the host with stages of latency followed by reactivation. Infection is usually benign in immunocompetent patients. However HCMV may cause a severe disease in organ transplant recipients, AIDS patients or hosts with defects of the immune systems. HCMV is a major cause of birth defects when infection occurs during pregnancy. After virusmembrane fusion events mediated by envelope proteins, both the DNA-containing capsid and the tegument proteins are released into the cell. Some tegument proteins play a critical roles in the disarming the immune system of the host (pp65, pp71) or they improve the viral genes expression (pp150, pp28). Nuclear sensing of viral DNA has emerged as an essential step in innate immune responses against herpesviruses, including HCMV. The DNA sensor protein IFI16 seems to have a key role in the cell defensive mechanism. We have been previously demonstrated that the viral protein kinase UL97 plays a crucial role in the regulation of IFI16 nucleo-cytoplasmic relocalization during HCMV infection. Materials and Methods: TR HCMV virions obtained by infection on human fibroblasts (HFF) were purified by ultracentrifugation on sucrose gradient. The purified virions were then lysated and checked on western blot for cellular residues. Immunoprecipitation was performed using anti-IFI16 policlonal antibodies as bait for virion proteins. The immunocomplexes (ICs) were then incubated and purified with commercial beads. ICs was analyzed by western blot. Results: Our group has newly obtained some experimental evidence that reveal a possible presence of IFI16 in the HCMV virion, probably associated with the viral protein UL83, but the significance of this data still remains unknown. Discussion and Conclusions: We have deeply investigated a possible interaction of virion proteins with IFI16. Our datas suggest a possible localization of IFI16 in HCMV virion through binding of viral pp65 and/or pp28, but the impact of this interaction on the innate immune evasion or other cell mechanisms still remains unclear.

HCMV TEGUMENT PROTEINS: NEW INSIGHTS ON INTERACTION BETWEEN THE DNA SENSOR PROTEIN IFI16 AND VIRION PARTICLES

GIORGIO, ALESSANDRO GIOVANNI;BIOLATTI, MATTEO;GATTI, DEBORAH;DELL'OSTE, Valentina;DE ANDREA, Marco;LANDOLFO, Santo Giuseppe
2014

Abstract

Introduction: The human cytomegalovirus (HCMV), a member of b-herpesviridae family, is a widespread pathogen that infect between 50% to 85% of human population. HCMV is able to establish a lifetime infection in the host with stages of latency followed by reactivation. Infection is usually benign in immunocompetent patients. However HCMV may cause a severe disease in organ transplant recipients, AIDS patients or hosts with defects of the immune systems. HCMV is a major cause of birth defects when infection occurs during pregnancy. After virusmembrane fusion events mediated by envelope proteins, both the DNA-containing capsid and the tegument proteins are released into the cell. Some tegument proteins play a critical roles in the disarming the immune system of the host (pp65, pp71) or they improve the viral genes expression (pp150, pp28). Nuclear sensing of viral DNA has emerged as an essential step in innate immune responses against herpesviruses, including HCMV. The DNA sensor protein IFI16 seems to have a key role in the cell defensive mechanism. We have been previously demonstrated that the viral protein kinase UL97 plays a crucial role in the regulation of IFI16 nucleo-cytoplasmic relocalization during HCMV infection. Materials and Methods: TR HCMV virions obtained by infection on human fibroblasts (HFF) were purified by ultracentrifugation on sucrose gradient. The purified virions were then lysated and checked on western blot for cellular residues. Immunoprecipitation was performed using anti-IFI16 policlonal antibodies as bait for virion proteins. The immunocomplexes (ICs) were then incubated and purified with commercial beads. ICs was analyzed by western blot. Results: Our group has newly obtained some experimental evidence that reveal a possible presence of IFI16 in the HCMV virion, probably associated with the viral protein UL83, but the significance of this data still remains unknown. Discussion and Conclusions: We have deeply investigated a possible interaction of virion proteins with IFI16. Our datas suggest a possible localization of IFI16 in HCMV virion through binding of viral pp65 and/or pp28, but the impact of this interaction on the innate immune evasion or other cell mechanisms still remains unclear.
42° Congresso della Società Italiana di Microbiologia
Torino
28-9/1-10/2014
42° Congresso della Società Italiana di Microbiologia
Edizioni Medico Scientifiche S.r.L.
142
142
Alessandro Giorgio; Matteo Biolatti; Deborah Gatti; Valentina Dell’Oste; Marco De Andrea; Santo Landolfo
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2318/150438
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