Advanced

Purification of infectious human herpesvirus 6A virions and association of host cell proteins

Hammarstedt, Maria LU ; Ahlqvist, Jenny; Jacobson, Steven; Garoff, Henrik and Fogdell-Hahn, Anna (2007) In Virology Journal 4.
Abstract
BACKGROUND: Viruses that are incorporating host cell proteins might trigger autoimmune diseases. It is therefore of interest to identify possible host proteins associated with viruses, especially for enveloped viruses that have been suggested to play a role in autoimmune diseases, like human herpesvirus 6A (HHV-6A) in multiple sclerosis (MS). RESULTS: We have established a method for rapid and morphology preserving purification of HHV-6A virions, which in combination with parallel analyses with background control material released from mock-infected cells facilitates qualitative and quantitative investigations of the protein content of HHV-6A virions. In our iodixanol gradient purified preparation, we detected high levels of viral DNA by... (More)
BACKGROUND: Viruses that are incorporating host cell proteins might trigger autoimmune diseases. It is therefore of interest to identify possible host proteins associated with viruses, especially for enveloped viruses that have been suggested to play a role in autoimmune diseases, like human herpesvirus 6A (HHV-6A) in multiple sclerosis (MS). RESULTS: We have established a method for rapid and morphology preserving purification of HHV-6A virions, which in combination with parallel analyses with background control material released from mock-infected cells facilitates qualitative and quantitative investigations of the protein content of HHV-6A virions. In our iodixanol gradient purified preparation, we detected high levels of viral DNA by real-time PCR and viral proteins by metabolic labelling, silver staining and western blots. In contrast, the background level of cellular contamination was low in the purified samples as demonstrated by the silver staining and metabolic labelling analyses. Western blot analyses showed that the cellular complement protein CD46, the receptor for HHV-6A, is associated with the purified and infectious virions. Also, the cellular proteins clathrin, ezrin and Tsg101 are associated with intact HHV-6A virions. CONCLUSION: Cellular proteins are associated with HHV-6A virions. The relevance of the association in disease and especially in autoimmunity will be further investigated. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Virology Journal
volume
4
publisher
BioMed Central
external identifiers
  • Scopus:37549055125
ISSN
1743-422X
DOI
10.1186/1743-422X-4-101
language
English
LU publication?
yes
id
c65d760a-c874-498e-9f1a-f498d71720cc (old id 1298987)
date added to LUP
2009-07-09 14:30:44
date last changed
2016-11-15 13:14:30
@misc{c65d760a-c874-498e-9f1a-f498d71720cc,
  abstract     = {BACKGROUND: Viruses that are incorporating host cell proteins might trigger autoimmune diseases. It is therefore of interest to identify possible host proteins associated with viruses, especially for enveloped viruses that have been suggested to play a role in autoimmune diseases, like human herpesvirus 6A (HHV-6A) in multiple sclerosis (MS). RESULTS: We have established a method for rapid and morphology preserving purification of HHV-6A virions, which in combination with parallel analyses with background control material released from mock-infected cells facilitates qualitative and quantitative investigations of the protein content of HHV-6A virions. In our iodixanol gradient purified preparation, we detected high levels of viral DNA by real-time PCR and viral proteins by metabolic labelling, silver staining and western blots. In contrast, the background level of cellular contamination was low in the purified samples as demonstrated by the silver staining and metabolic labelling analyses. Western blot analyses showed that the cellular complement protein CD46, the receptor for HHV-6A, is associated with the purified and infectious virions. Also, the cellular proteins clathrin, ezrin and Tsg101 are associated with intact HHV-6A virions. CONCLUSION: Cellular proteins are associated with HHV-6A virions. The relevance of the association in disease and especially in autoimmunity will be further investigated.},
  author       = {Hammarstedt, Maria and Ahlqvist, Jenny and Jacobson, Steven and Garoff, Henrik and Fogdell-Hahn, Anna},
  issn         = {1743-422X},
  language     = {eng},
  publisher    = {ARRAY(0x7f5a400)},
  series       = {Virology Journal},
  title        = {Purification of infectious human herpesvirus 6A virions and association of host cell proteins},
  url          = {http://dx.doi.org/10.1186/1743-422X-4-101},
  volume       = {4},
  year         = {2007},
}