Leukocyte extracts for affecting the immune system
AssigneeEndeavor Corp.
InventorA. Arthur Gottlieb
Life of this patent
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Abstract
A group of substances derived from human leukocytes (white blood cells) are described that amplify, suppress, or otherwise modulate the response of the immune system to reintroduction of antigens. The described materials are generically effective as to antigens rather than specific as to particular antigens, but the described modulators of immune response are effective only in respect of antigens with which the recipient has previously been challenged. The described materials are extracted from leukocyte dialysates by processes involving high pressure liquid chromatography with specified resin/solvent systems. Uses and pharmaceutical compositions for use of the materials are also described.
Claims43
A process of purifying modulator material from an extract of human leukocytes, and of separating said material from substantially all fluorescamine-reactive material and from L-suppressor, comprising the steps of:
- (1) dialyzing said extract through a dialysis membrane having a nominal molecular weight cutoff of about 3500, thereby producing a dialysate;
- (2) fractionating said dialysate by gel filtration, thereby producing a plurality of dialysate-fractions;
- (3) modulator-assaying said dialysate-fractions, thereby ascertaining modulator activity of said dialysate-fractions;
- (4) selecting modulator-assayed dialysate-fractions having substantial modulator activity and pooling them;
- (5) applying said selected and pooled modulator-assayed dialysate-fractions to a reverse-phase high pressure liquid chromatography column, packed with octadecyl silane,
- (6) eluting said column with an ethanol-in water gradient, thereby producing a plurality of effluent-fractions;
- and
- (7) selecting predetermined effluent-fractions on the basis of ethanol concentration therein, and collecting them.
A process of purifying L-suppressor material from an extract of human leukocytes, and of separating said L-suppressor material from amplifiers 1-6, from S-suppressor, and from substantially all fluorescamine-reactive material, comprising the steps of:
- (1) dialyzing said extract through a first dialysis membrane having a nominal molecular weight cutoff of approximately 12,000, thereby producing a first dialysate,
- (2) dialyzing said first dialysate through a second dialysis membrane having a nominal molecular weight cutoff of about 3500, thereby producing a second dialysate and a retentate of the second dialysis,
- (3) applying said retentate of the second dialysis to a hydroxylapatite chromatography column,
- (4) eluting said column with ammonium bicarbonate aqueous solution gradient from approximately 0.08M to approximately 0.2M, and
- (5) collecting the material eluting between approximately 0.1M and 0.15M.
A composition of matter consisting of material which:
- (a) is derived from leukocytes;
- (b) is in purified form, and is purified substantially free of fluorescamine-reactive substances;
- (c) affects a response of the immunity system of an animal or human body, portion of said body, or matter taken therefrom, where said response:
- (1) is to the reintroduction to said body of at least one antigen to which said body has been previously exposed,
- (2) is specifically attributable to a function of the immunity system of said animal or human, and
- (3) occurs following said reintroduction of said antigen;
- and
- (d) is substantially free of other, different material that has an effect on said response opposite to the effect to the effect of said composition on said response--
- said composition of matter being hereinafter referred to as "modulator material."
A method of modulating the immune response of a human body to the reintroduction of antigen to which said human body has previously been exposed, comprising administering to said body human modulator material, in an effective dosage amount, and in a pharmaceutically acceptable vehicle.
Dependencies are inferred from the claim text.