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Purification of Viral DNA for the Identification of Associated Viral and Cellular Proteins
Published on: August 31, 2017
Co-purification of soluble membrane cofactor protein (CD46) and human herpesvirus 6 variant A genome in serum from
A Fogdell-Hahn1, S S Soldan, S Shue
1Neuroimmunology Branch, National Institute of Neurological Disorders and Stroke (NINDS), National Institutes of Health (NIH), 9000 Rockville Pike, Building 10, Room 5B-16, Bethesda, MD 20892, USA.
Abstract:
The association of human herpesvirus 6 (HHV-6) and multiple sclerosis (MS) has been supported by several immunological and molecular studies. Recently, membrane cofactor protein (CD46) has been identified as the cellular receptor for the A and B variants of HHV-6. Elevated levels of soluble CD46 (sCD46) have been reported in the serum and CSF of MS patients. The aim of this study was to investigate a possible correlation between elevated levels of soluble CD46 and the presence of serum HHV-6 DNA in MS patients. An immunoaffinity column comprised of immobilized monoclonal antibodies to CD46 was developed to isolate sCD46 from cell free body fluids of MS patients and controls. After immunoaffinity purification, DNA was extracted from anti-CD46 column eluates and subjected to PCR amplification. Of the 42 MS samples tested, 4 serum samples were HHV-6 positive, 3 of which were typed as HHV-6A. The co-purification of sCD46 and HHV-6 DNA from MS sera indicates that HHV-6 is tightly connected to its receptor, CD46, in the serum of MS patients.
Insights
Human herpesvirus 6 (HHV-6) DNA was found in the serum of multiple sclerosis (MS) patients, specifically associated with soluble CD46 (sCD46). This suggests a direct link between HHV-6 and its receptor in MS patients.
Area of Science:
- Neuroimmunology
- Virology
Background:
- The association between human herpesvirus 6 (HHV-6) and multiple sclerosis (MS) is supported by immunological and molecular evidence.
- Membrane cofactor protein (CD46) is the cellular receptor for HHV-6 variants A and B.
- Elevated soluble CD46 (sCD46) levels are observed in the serum and cerebrospinal fluid (CSF) of MS patients.
Purpose of the Study:
- To investigate the correlation between elevated sCD46 levels and the presence of HHV-6 DNA in the serum of MS patients.
Main Methods:
- Development of an immunoaffinity column using monoclonal antibodies to CD46 for sCD46 isolation from patient body fluids.
- DNA extraction from anti-CD46 column eluates.
- Polymerase Chain Reaction (PCR) amplification to detect HHV-6 DNA.
Main Results:
- Of 42 MS samples, 4 tested positive for HHV-6 DNA.
- Three of the positive samples were identified as HHV-6A.
- Co-purification of sCD46 and HHV-6 DNA from MS sera was observed.
Conclusions:
- The findings indicate a tight connection between HHV-6 and its receptor, CD46, within the serum of multiple sclerosis patients.
- This study provides evidence for the presence of HHV-6 DNA associated with its receptor in MS serum.
