Trypanosoma b. rhodesiense (WRATat Serodeme): Purification and Characterization of Surface Antigens for the Vaccine Development Program.
Abstract
Variant-specific surface antigens (VSSA) from Trypanosoma b. rhodesiense WRATat clones have been purified using Con A-lectins affinity chromotography. The molecular weights of VSSA-2 an VSSA-12 have been estimated to be about 65,000 daltons by SDS-PAGE. Proteolytic activity in the lysates was significantly inhibited by including PMSF or TLC in the lysing buffers. Treatment of the lysate with p-aminobenzamidine-6-aminohexanoic acid-sepharose was equally effective. Characterization of the antigens by classical techniques is now being pursued. Low levels of parasitemia (300 million/ml) were overcome by disturbing the innate resistance of the rodent hosts with cyclophophamide, dexamethasone, prednisolone or anti-neutrophil serum prior to infection. These treatments resulted in infections peaking at greater than 10 to the 9th power/ml. Separation of the trypanosomes from blood cells was accomplished by treating the infected blood with a hemagglutinating lectin from Maclura pomifera. This innovation allowed us to decrease the volume of DEAE-cellulose to only twice that of the blood volume and to run the columns at 4 C. Experiments with the antibiotic tunicamycin have demonstrated that trypanosomes can be inhibited by very low concentrations. Because this drug interferes with glycoprotein biosynthesis, it could be very useful in studying VSSA synthesis and degradation. (Author)
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 1980
- Accession Number
- ADA095616
Entities
People
- Gerald R. Keilman
Organizations
- Colorado State University