Antibodies in human sera to oncorna virus-like proteins from normal or leukemia marrow cell cultures.

Abstract

Some human marrows in culture release particles with oncornavirus-like properties. This study was designed to examine the immunological properties of similar particles in human marrow culture supernates. Leukemic and nonleukemic marrows were cultured for 5-7 days in the presence of [14C]uridine and [3H]leucine or [3H]glucosamine. Labeled supernatant components banding in sucrose gradient densities of 1.20-1.24 g/ml were used as antigen in a double antibody immunoprecipitation assay. The assay was validated by end point titrations and competition with unlabeled antigen; purified myeloma proteins were used as negative controls. Cross-reactivity with mammalian oncornaviruses, as judged by competitive inhibition of precipitation by these viruses, was slight and at the border of the sensitivity of the method. Precipitated antigens analyzed by SDS polyacrylamide gel electrophoresis contained three distinct polypeptides of about 70,000, 45,000 and 30,000 mol wt; these comigrated with the gp 70, pg 45, and p 30 of a murine leukemia virus. Similar polypeptides were obtained from both leukemic and nonleukemic marrow culture supernates. As determined by the radioimmunoprecipitation assay, 32 of 45 leukemic sera (71%), 36 of 45 normal sera (80%), 15 of 19 sera from family contacts of leukemic patients (79%), 14 of 21 cord blood specimens (67%), and 21 of 23 sera (91%) from patients with systemic lupus erythematosus had detectable antibody activity.

Specificity of human antibodies to oncovirus glycoproteins: recognition of antigen by natural antibodies directed against carbohydrate structures.

“…In the absence of a clearly defined oncovirus of human origin, a number of investigators have approached this problem by investigating human sera for naturally occurring antibodies to oncoviruses of other species. The results obtained have varied from positive to negative or equivocal (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19). Attempts to resolve discrepancies in the published data have centered around discussions of the lability of certain viral protein probes (14) and the use of special reaction conditions influencing human antibody binding to viral antigens (13,14).…”

“…In the absence of a clearly defined oncovirus of human origin, a number of investigators have approached this problem by investigating human sera for naturally occurring antibodies to oncoviruses of other species. The results obtained have varied from positive to negative or equivocal (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19) For preparation of soluble radiolabeled viral 70,000-dalton glycoprotein (gp7O), purified virions were subjected to detergent lysis in a buffer containing 0.05 M NaCI, 0.02 M Tris-HCl (pH 7.4), 0.5% Nonidet P40, 0.5% sodium deoxycholate, and 10% Traysylol protease inhibitor (10,000 kallikrein inactivator units per ml) for 30 min at 370C. After the viral lysates were subjected to centrifugation at 100,000 X g for 60 min in a type 50 rotor, the supernatant fractions containing soluble viral proteins, including radiolabeled gp7O, were collected and stored at -70'C until used.…”

Heterophile binding of human antibodies to glycoproteins of retroviruses

“…It is largely on this basis that Epstein-Barr virus has been identified, not only as the cause of infectious mononucleosis (Niederman et al, 1970) but also as a likely cause of Burkitt's lymphoma (de and nasopharyngeal carcinoma (Henle & Henle, 1976). This type of evidence has also been important in detecting endogenous and exogenous Type C virus activity in animals (Charman et al, 1975;Jhle et al, 1976;Nowinski & Kaehler, 1974).…”

“…The widespread presence of anti-Type C virus antibodies in the human population (healthy and cancer patients) has been described (Aoki et al, 1976;Caldwell et al, 1975: Kurth et al, 1977Kurth & Mikschy, 1978;Loui et al, 1976;Snyder et al, 1976), but also questioned (Gardner et al, 1977; Krakower & Aaronson, 1978: Stephenson & Aaronson, 1976. In the absence of a Type C virus of indisputable origin, proteins from well characterized mammalian Type C viruses have been used in the above studies.…”