The use of protein as a carrier of methotrexate for experimental cancer chemotherapy. III. Human serum albumin-methotrexate derivative, its preparation and basic testing.

The preparation and a more detailed characterization of human serum albumin-methotrexate derivative (HSA-MTX) is described. The synthesis of the derivative was performed by means of 1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide (in methoiodide form) (WSC). Results of many experiments showed that, on the average, about 26 molecules of methotrexate (MTX) were coupled to one molecule of human serum albumin (HSA).

Organ distribution of fibrinogen-bound methotrexate in mice with Gardner lymphosarcoma.

The distribution of fibrinogen-bound 3H methotrexate was investigated in Gardner lymphosarcoma bearing mice. 3H labeled methotrexate (3H MTX) was covalently bound by means of aminopropyl carbodiimide to bovine and mouse fibrinogen (FBG). The preparations as well as the free 3H MTX were applied i.

The use of protein as a carrier of methotrexate for experimental cancer chemotherapy. II. Chemotherapy of Gardner lymphosarcoma with pea seed lectin-methotrexate derivative.

It was demonstrated that the tumorigenicity of tumor cells preincubated in low concentration of free methotrexate (MTX) has not been changed. On the other hand the preincubation of these cells with pea seed lectin (PL), MTX and PL mixture and especially pea seed lectin-methotrexate derivative (PL-MTX) influenced markedly the tumor cells tumorigenicity. The chemotherapy of Gardner lymphosarcoma (LSG) bearing mice with PL-MTX derivative was performed.

Immunogenicity of the Gardner lymphosarcoma for the mice of the strain C3H (H-2k). II. Alteration of the growth and dissemination of solid Gardner lymphosarcoma following immunization by an extract of the ascitic tumor.

Aqueous 1-butanol extracts were prepared from the ascites form of Gardner lymphosarcoma (LSG) maintained in C3H (H-2k) mice and from the solid Lewis lung carcinoma (LLCa) transplanted in B6 mice. C3H mice primed at least two times with cell surface antigens extracted from Gardner tumor cells (LSG-extract) lived longer than untreated controls if challenged with the solid Gardner lymphosarcoma cells. Tumors growing in immunized C3H mice differed from those growing in controls by shape, necrotization and inhibition of dissemination.

Immunogenicity of the Gardner lymphosarcoma for the mice of the strain C3H (H-2k). I. The effect of the 60Co-irradiation of recipients and of the interval between the immunization and transplantation of the tumor on the antitumor resistance of immunized mice.

The C3H (H-2k) mice were immunized by 60Co-irradiation-inactivated Gardner lymphosarcoma (LSG) cells. The degree of resistance of mice with transplanted tumors was determined by difference in survival curves of the immunized and nonimmunized mice. When the tumor was transplanted during 26 weeks after the last of the three immunizing injections the mean survival time of immunized mice was always prolonged over that of nonpretreated controls.

Treatment of C3H mice bearing solid Gardner lymphosarcoma with vaccine prepared from Corynebacterium parvum combined with methotrexate and Alexan.

C3H mice after transplantation of solid Gardner lymphosarcoma were injected with Methotrexate (MTX) or Alexan. The prolonged survival time of mice treated with antimetabolites was in some cases evidently reduced after an additional injection of Corynebacterium parvum (C.p.

Effect of vaccine prepared from Corynebacterium parvum on the growth of Gardner lymphosarcoma in C3H strain mice.

The present paper discusses the effect of a vaccine prepared with formol-killed Corynebacterium parvum (alias Propionibacterium acnes) on the growth of solid Gardner lymphosarcoma transplanted on C3H (H-2k) mice. The vaccine was injected at various time intervals prior to, simultaneously with, or following transplantation of the tumor to intact mice or mice immunized with 60Co-irradiated Gardner tumor cells. The effectivity was evaluated in terms of numbers of mice surviving without tumor 60-day observation period or prolongation of survival time.

Effect of free and bovine fibrinogen-bound methotrexate and dibromoaminopterin on the dissemination of solid Gardner lymphosarcoma.

Solid Gardner lymphosarcoma transplanted subcutaneously to mice of strain C3H/Sumice does not grow solely at the site of inoculation, but disseminates also to remote tissues. An i. v.

Localized chemotherapy of Gardner's lymphosarcoma of C3H mice using combinations of carrier-sorbed antifolics and detoxicating tetrahydrofolates.

The effect of aminohexyl-2-hydroxyethylmethacrylate polymer (HEMA-Hex) with sorbed methotrexate (MTX) or 3',5'-dibromoaminopterin (BrAP) on the survival of C3H mice with Gardner lymphosarcoma was studied. The measured bits of HEMA-Hex-MTX or HEMA-Hex-BrAP were implanted into the solid tumor growing 4 to 8 8 days. The doses of sorbed antimetabolites amounting in MTX 4.

Changes in hemopoiesis of mice of the C3H strain following transplantation of Gardner lymphosarcoma and infection with LDH-virus. III. Blood proteins.

Inoculation of bone marrow and spleen cells of C3H/Sumice strain mice demonstrated that on the ninth day of growth of Gardner lymphosarcoma these tissues were invaded with tumor cells. The weights of mice with advanced tumors increased. Yet the deterioration of health status of mice is characterized by decreased food and water consumption, as well as by lower concentrations of proteins in mouse sera and plasma.

Changes in hemopoiesis of mice of the C3H strain following transplantation of Gardner lymphosarcoma and infection with LDH-virus. II. Spleen and bone marrow.

Cytological examination of spleens of mice with growing Gardner lymphosarcoma contaminated with LDH virus indicates the augmentation of erythropoiesis and granulopoiesis, as well as the decrease of lymphopoiesis. Similar changes were observed after infection with LDH-virus. The counts from the bone marrow smear showed more pronounced changes only in tumorous mice, in which repeatedly increased granulopoiesis was found.

The chemotherapy of C3H mice bearing Gardner lymphosarcoma with bovine fibrinogen-methotrexate derivative.

Derivative of bovine fibrinogen (FBG) containing chemically bound methotrexate (MTX) has been prepared by action of ethyldimethylaminopropyl carbodiimide. The derivative retained its solubility and clotability. Intraperitoneally administered FBG-MTX derivative one day after transplantation of the ascitic Gardner lymphosarcoma prolonged distinctly the survival of C3H mice.