Search

Angiogenic and poorly angiogenic tumor variants were obtained by an intraperitoneal inoculation of cells from clones of polyoma-virus transformed BALB c 3T3 cells into syngeneic mice. The angiogenic tumor cells expressed a higher tumorigenicity phenotype and a higher capacity to produce artificial pulmonary metastases than cells from the poorly angiogenic tumors. The former cells expressed also significantly higher levels of the lymphocyte activation protein Ly-6E.1 than the former cells. The two types of cells did not differ in expression levels of CD44 and of a polyoma-virus specific membrane antigen. These results raise the possibility that the angiogenic phenotype is coregulated with Ly-6. The effect on Ly-6 expression of signal transduction through TNF receptors, functioning as pivotal regulators of angiogenesis was therefore studied. It was found that TNFα and more so antibodies against Fas down-regulate expression levels of Ly-6. This down-regulation seemed to be selective as expression levels of CD44 were not affected by this treatment.

Nonadherent Fc gamma receptors (Fc gamma R) expressing cells from SEYF-a tumors that form rosettes with sheep erythrocytes coated with IgG antibodies (EA) were isolated by Percoll density gradients. The EA-IgG rosette-forming cells were characterized by the parameters of 1) binding of IgG immune complexes; 2) binding of purified monoclonal antibodies against mouse FcR; 3) sensitivity to complement-dependent lysis mediated by syngeneic anti-SEYF-a antibodies; 4) expression of parental H-2 antigen when grown in F1 hybrids; 5) incorporation of [125I]5-iodo-2'-deoxyuridine; and 6) growth in syngeneic mice. The nonadherent EA-IgG rosette-forming cell population was found to be composed of both host lymphocytes as well as of tumor cells. Tumor-seeking lymphocytes then were removed from SEYF-a tumors by velocity sedimentation on Percoll. The remaining cell population was tumorigenic and expressed FcR, as well as tumor antigens. These tumor EA-IgG rosette-forming cells exhibited a very low rate of DNA synthesis compared with that of non-rosetting tumor cells.

Making use of 125 I-labelled monoclonal rat antibodies against mouse Fc receptor we have developed a solid-phase radioimmunoassay for cell-free antigenic mouse Fc receptor (FcR). An 80% acetone solution was used for fixation of relatively large amounts of soluble proteins on PVC microtiter plates. As a result of this treatment FcR practically lost its activity to bind the Fc portion of an IgG molecule but retained its antigenicity, the binding of specific anti-FcR (aFcR) antibody being increased following acetone fixation. Concentrations of cell-free FcR in NP-40 extracts of FcR-expressing cells were calculated from the linear part of a standard curve and expressed in units of antigenic activity, 1 unit being the amount of antigenic FcR capable of binding 1 μg of 125 I-aFcR. The method may be used for detecting cell-free FcR as a minor constituent in a mixture of proteins.

Serum from young normal BALB/c mice was found to contain IgM antibodies able tomediate complement-dependent lysis of certain syngeneic or allogeneic tumor target cells. The titer of such naturally occurring antitumor antibodies (NATA) was found to increase with aging. A longitudinal serological study comparing the cytotoxicity potential of NATA fromnormal and from urethan-treated BALB/c mice was performed. It was found that urethan-treated mice that did not develop primary lung-adenomas within the duration of the experi-ment had significantly lower NATA titers, against one out of 4 target cells assayed, than urethan-treated animals that developed lung adenomas. This difference was evident in two independent experiments. The results suggested that the lower NATA activity of the urethan-treated mice that did not develop tumors existed even before exposure to the carcinogenic insult. This raises the possibility that certain populations could be segregated according to their natural antibody profile into those individuals which will develop primary tumors within a certain period if exposed to a subthreshold amount of carcinogen, and those which will not.