Fractionation of schistosome antigens by high performance electrophoretic chromatography and their screening for the ability to induce Th1 lymphocyte activity.
The technique of high performance electrophoretic chromatography (HPEC) has been used to fractionate soluble antigens from adult Schistosoma mansoni worms on the basis of molecular weight (MW), prior to screening for their ability to stimulate T lymphocyte activity. Approximately 250 micrograms of protein were separated by continuous electrophoresis through an SDS polyacrylamide gel into 30-50 aqueous samples of minimal volume (80 microliters). Each consecutive sample contained a limited number of proteins of progressively greater MW, although the resolution of the fractionation was affected by a number of factors including acrylamide concentration, gel length, gel diameter and electrophoretic current. Following the extraction of SDS using Calbiosorb resin, the aqueous fractions were used directly to stimulate cultures of lymphocytes taken from the lymph nodes of infected or vaccinated mice. The most promising fractions were those containing proteins which induced the release of high levels of interferon-gamma relative to the extent of proliferation. This suggests that these proteins are good inducers of Th1 lymphocyte activity.
Mountford
A P
AP
Department of Biology, University of York, UK.
Wilson
R A
RA
D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology* / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D015801:Helminth Proteins / Q000276:immunology / Q000302:isolation & purification*; D007371:Interferon-gamma / Q000378:metabolism; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D012550:Schistosoma mansoni / Q000737:chemistry / Q000276:immunology*; D012555:Schistosomiasis mansoni / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*
D000947:Antigens, Helminth; D015801:Helminth Proteins; D007371:Interferon-gamma