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From the Department of Microbiology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
Abstract
The molecular dimensions of guinea pig lymphotoxin were studied by means of gel filtration, sucrose density gradient ultracentrifugation, and CsCl isopycnic ultracentrifugation. Lymphotoxin was produced by stimulating immune lymph node cells with the specific antigen, ovalbumin, and was assayed by means of a Coulter counter microassay. The D20,w of guinea pig lymphotoxin was estimated by gel filtration on Sephadex G-100 to be (8.0 ± 0.2) x 10-7 cm2/sec, and the apparent S20,w of guinea pig lymphotoxin was estimated to be 4.2 ± 0.1S by sucrose density gradient ultracentrifugation. The buoyant density of guinea pig lymphotoxin was determined to be 1.302 ± 0.002 g/ml by CsCl isopycnic ultracentrifugation. From these values the molecular weight of guinea pig lymphotoxin was calculated to be 45,000, and the frictional ratio was calculated to be 1.1. No evidence was found to indicate that more than a single factor was involved in producing the cytotoxic effect attributed to lymphotoxin.
Footnotes
1 This work was supported in part by a Contract with the Office of Naval Research N00014-67-A-0163-0014, United States Public Health Service Training Grant M 5 T01 AI00282-09, United States Public Health Service Research Grant 5 R01 AI-02566-14, and National Science Foundation Grant GB7406X3.
2 Recipient of Pre-Doctoral Fellowship on United States Public Health Service Training Grant M 5 T01 AI00282-09.
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