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Solubilization and characterization of Herpesvirus saimiri-induced membrane antigens
Abstract:
Treatment of Herpesvirus saimiri (HVS)-infected owl monkey cells by limited papain digestion removed the HVS-induced membrane antigen (MA) as determined by membrane immunofluorescence and antibody-dependent lymphocyte cytotoxicity (ADLC). Soluble antigenically active HVSMA was detected by inhibition of ADLC and by the decreased binding of 125I-labeled staphylococcus protein A to HVS-infected cells after absorption of an anti-MA-positive serum with papain extracts. Approximately 38% of the inhibitory activity of the papain extracts was sedimentable at 100,000 X g, indicating that the released MA was heterogeneous in size. Preliminary investigations by gel filtration chromatography identified a major peak of MA with a molecular weight between 20,000 and 50,000.
Insights
Papain digestion released Herpesvirus saimiri (HVS)-induced membrane antigen (MA) from infected cells. Soluble HVS-MA was detected, showing heterogeneous sizes, with a major peak between 20,000 and 50,000 molecular weight.
Area of Science:
- Virology
- Immunology
- Biochemistry
Background:
- Herpesvirus saimiri (HVS) is a primate rhadinovirus.
- HVS infection in owl monkeys can lead to lymphoproliferative disease.
- Understanding HVS-induced antigens is crucial for studying viral pathogenesis and immune responses.
Purpose of the Study:
- To characterize the Herpesvirus saimiri (HVS)-induced membrane antigen (MA).
- To determine if HVS-MA can be released in a soluble, antigenically active form.
- To investigate the physical properties, such as size, of the released HVS-MA.
Main Methods:
- Limited papain digestion of HVS-infected owl monkey cells.
- Membrane immunofluorescence assays to detect MA.
- Antibody-dependent lymphocyte cytotoxicity (ADLC) assays to assess MA activity.
- Ultracentrifugation to determine sedimentability of inhibitory activity.
- Gel filtration chromatography to estimate molecular weight.
Main Results:
- Papain digestion effectively removed HVS-MA from the cell surface.
- Soluble, antigenically active HVS-MA was detected in papain extracts, inhibiting ADLC and reducing 125I-Staphylococcus protein A binding.
- Approximately 38% of the inhibitory activity was sedimentable at 100,000 x g, indicating heterogeneity in size.
- Gel filtration identified a major peak of HVS-MA with a molecular weight between 20,000 and 50,000.
Conclusions:
- HVS-induced membrane antigen can be released from infected cells as a soluble, biologically active molecule.
- The released HVS-MA is heterogeneous in size, with a predominant component in the 20-50 kDa range.
- These findings contribute to the characterization of HVS antigens and their potential role in immune interactions.