Elaboration of granulocyte-macrophage colony-stimulating factor by human tumor cell lines and normal urothelium.

A total of 72 cell conditioned media (CCM) were screened for their ability to stimulate colony formation by human granulopoietic progenitor cells. Granulocyte-macrophage (GM) colony-stimulating factor(s) (CSF) were found in CCM of nine tumor cell lines, two primary urinary bladder tumors, and three epithelial cell cultures of normal urinary tract. The most active medium came from urinary bladder carcinoma cell line 5637. CSF released by the 5637 cell line induced dose-dependent GM colony formation from human fetal, normal adult, and CML bone marrow (BM) and from mouse BM. Human fetal and normal adult BM formed more colonies when stimulated with 5637 CCM than with peripheral blood leukocyte (PBL) feeder layers, while CML BM produced more colonies with PBL feeder layers. CCM from 5637 was more active in stimulating GM colony formation than human placenta conditioned medium (PCM) and PHA-LCM. 5637 CCM produced in serum-free hormone-supplemented medium was nearly equipotent and can serve as suitable starting material for purification.

Phenomenon of formation of giant fat-containing cells in human bone marrow cultures induced by human serum factor: normal and leukemic patterns.

Normal human sera induce the formation of fat-containing cells (FCC) in human bone marrow cultures. A nearly complete monolayer of FCC is formed after 7-14 days of cultivation with 20% human sera in the medium. FCC-inducing activity (FCCIA) is nondialyzable through 14,900-dalton cutoff membrane and is stable at 56 degrees C for 30 min. Abundant FCCIA was found in 83% of normal human sera but in only 20% of sera from untreated patients with different hemopoietic disorders and in 32% of treated leukemic patients. It is suggested that FCCIA may be involved in regulation of the bone marrow microenvironment an that it varies in normal individuals and in patients with different diseases.

The properties of human fetal bone marrow stromal and hemopoietic cells.

The capacity of fetal bone marrow to form stromal cell colonies, granulocyte-macrophage colonies, stromal cell monolayers and to produce granulocyte-macrophage colony stimulating activity from these monolayers was evaluated in comparison to adult bone marrow. Granulocyte-macrophage colony formation on routine feeder layers was approximately equal in both cases. The fetal bone marrow colonies were larger in size than those from adult precursors. There were at least ten times more stromal cell precursors in fetal bone marrow than in adult bone marrow. The fetal bone marrow stromal cell monolayer formed within 7-12 days whereas adult stromal monolayer formation required 3-4 weeks. Fetal bone marrow stromal cell monolayer produced granulocyte-macrophage colony-stimulating activity (GM-CSA) effective only for fetal GM precursors, whereas adult bone marrow produces GM-CSA effective for fetal and adult normal and CML bone marrow.

Induction of giant fat cells in human bone marrow culture by human serum factor.

Evidence is presented below that normal human sera contain a potent non-dialyzable factor inducing abundant giant fat cells in human bone marrow culture, normal as well as CML. Media with 20% heated (56 degrees C) human serum induce during 7-14 days almost complete monolayer of fat cells on the bottom of the plastic flasks or dishes. Fetal bovine sera do not exhibit this effect and shift cultures to the proliferation of fibroblasts. We are studying the functions of fat cells in hemopoiesis as well as the biochemical nature of fat cell-inducing factor in human sera.

Dependence of gastro-intestinal tumors on gastro-intestinal hormones: pentagastrin stimulates growth of transplanted colon adenocarcinoma in mice.

Gastrin was administered several times to mice with transplantable hepatoma, sarcomas, colon adenocarcinoma and adenocarcinoma of small intestine. Injections of gastrin stimulated the growth of colon adenocarcinoma in mice but not of the other tumors tested. Dependence upon gastro-intestinal hormones of "spontaneous" carcinogenesis in gastro-intestinal tract and the growth of many gastro-intestinal pancreatic and liver tumors is suggested. This hormone dependence could be used for the treatment and prophylaxis of some tumors of the gastro-intestinal tract.

Macrophage-like cell transformation and CFU(c) fluctuations in normal and leukemic human marrow cultures treated by phorbol diester.

Bone marrow from normal and chronic myeloid leukemia donors was grown in liquid cultures without feeder layers and with and without 12-u-tetradecanoyl-phorbol-13-acetate (TPA). In 24-96 hours most of the cells (60-70%) cultured with 10(-7) M and 10(-8) M TPA stuck to the bottom of the flasks and had a peculiar shape resembling macrophages possessing strong phagocytizing activity and surface markers of monocyte-macrophage lineage of differentiation. 10(-7) M and 10(-8) M TPA fully inhibited CFU(c) in cultures of normal marrow as well as of chronic myeloid leukemia (CML) patients; 10(-9) M and 10(-10) M exhibited individually varied partial suppression. Cultivation of bone marrow with 10(-11) M to 10(-13) M TPA led in some cases to statistically significant increase of CFU(c) on day 4 and day 7.

Nonimmune and immune surveillance. II. Effect of recipient's age, tumor immunogenicity, and neonatal thymectomy on tumor growth inhibition.

After a suspension of tumor pieces was grafted into newborn and adult (CBA X C57BL/6J)F1 and BALB/c mice, the growth of Lewis lung adenocarcinoma and mammary gland adenocarcinoma was inhibited in newborn mice, whereas sarcoma of the rectum (SR-1-75) grew at the same rate in newborn and adult recipients. Neonatal thymectomy stimulated the growth of hepatoma (H-2-73) in newborns. The degree of tumor growth inhibition was age-dependent: The maximum inhibition was observed in 1- to 6-day-old recipients, but later it gradually decreased. The hepatoma (H-2-73) and ovarian carcinoma (OC-1-75) with inhibited growth in newborns and the tumor (SR-1-75) with uninhibited growth had equally low immunogenicity. The data suggested that newborns possess factors that inhibit tumor growth but these factors disappear with increased age of recipients.

Body temperature and tumor virus infection. I. Tumorogenicity of Rous sarcoma virus for reptiles.

Rous sarcoma virus (RSV) was oncogenic for the following nine species of reptiles representing 6 families from Chelonia and Squamata orders: family of Testudinidae: 1. Testudo horsfieldi, family Agamidae: 2. Agama sanguinolenta; 3. Agama erythrogastra, family Lacertidae: 4. Eremias persica; 5. Eremias velox; 6. Eremias grammica, family Scincidae: 7. Eumeces taeniolatus, family Boidea: 8. Erix tataricus, 9. Ancistrodom blomhoffi. RSV did not induce tumors in 13 studied species of reptiles. Histologically 26 reptile tumors studied were polymorphous sarcomas with spindle-shaped (fibroblast-like), round and polygonal macrophage-like cells and sometimes peculiar giant polynuclear cells. Chromosomal analysis showed that reptile tumors arose out of reptile cells. RSV was pathogenic for adult reptiles. Reptile tumors did not contain a mature infectious virus. The tumors of 2 snakes were virogenic. The effect of increased temperature at the body level on the transformation of a symptomless viral infection into a viral disease is discussed in the evolutionary aspect.

Rapidly acquired cytotoxicity of lymphoid cells from ice inoculated with allogeneic spleen cells.

Spleen cells from C57BL/6J or CBA mice inoculated iv with spleen cells from BALB/c mice produced a strong nonspecific cytotoxic effect on target cells (mouse L-cells). Lymph node cells from CBA or C57BL/6J mice inoculated sc with BALB/c spleen cells also destroyed L-cells. Lymph node cells from mice inoculated with syngeneic spleen cells were not cytotoxic. The cytotoxic effect was observed ion of allogeneic but not syngeneic spleen cells. This effect was considerably reduced or completely suppressed after partial or total removal of plastic-adherent cells.

Specific xenogenous antiserum to cells of chronic myeloid leukemia (CML) in blastic crisis.

The antiserum was obtained from horses immunized with cells from patients with blastic crisis of CML and completely absorbed with normal white blood cells (WBC). The absorbed antiserum remained cytotoxic to blast cells from nearly half of the patients in blastic crisis and did not react with WBC from patients with Acute Myeloid Leukemia (AML), Acute Lymphoid Leukemia (ALL), Chronic Lymphoid Leukemia (CLL) and Chronic Myeloid Leukemia (CML) in its chronic phase as well as with cells of human normal bone marrow or fetal liver.

Nonimmune and immune surveillance. I. Growth of tumors and normal fetal tissues grafted into newborn mice.

Growth of various fetal tissues and transplantable tumors in syngeneic newborn and adult mice [BALB/c, DBA/2, and (CBA X C57BL/6J)F1] was compared. Fetal skin, a mixture of all fetal tissues, and tumors were transplanted. The tumors arose spontaneously [hepatomas, mammary gland adenocarcinoma (MGAC)] or resulted from malignant conversion of ectopic transplants either of fetal tissues (urinary bladder carcinoma, adenocarcinoma of small intestine, stomach sarcoma) or of adult animal tissues (ovarian carcinoma) in the syngeneic system. The growth of fetal skin transplants and teratomas, which developed after transplantation of minced tissue from 18- to 20-day and 12- to 14-day fetuses, was considerably inferior in newborn syngeneic recipients, as compared with similar transplants in adults. Inhibition of tumor growth observed in newborn animals was manifested in prolongation of latent period before tumor node appearance and in slowing of growth rate of developed tumors. One of six tumors studied (MGAC) grew at the same rate in newborn and adult recipients. It was suggested that a special type of cellular and/or humoral mechanisms controlling tumor growth exists in newborns. The activity of such factors was conceivably based on fetal tumor antigens as targets. We assumed that weakly antigenic and strongly antigenic tumors behaved differently in respect to nonimmune and immune surveillance mechanisms.

The effect of Trichinella spiralis on graft-versus-host reaction, transplantation immunity and antibody formation.

The effect of Trichinella spiralis in different phases of infection on transplantation immunity, the ability of lymphocytes to induce graft-versus-host reaction, on antibody and plaque-forming cell production was studied. In certain phases of Trichinella spiralis beginhing from the first days of infection and during 40 days significant suppression of transplantation immunity was observed. Thus, on the 24th day of infection skin allograft necrosis occurred much later (26.2 days) as compared with non-infected mice (12.5 days). The spleen cells of C57BL/6j mice infected with Trichinella spiralis on the 22, 29, 36, 42, 56, 72 days either induced slight graft-versus-host reaction in (CBA X C57B1/6j) F1 hybrid mice or did not induce it at all (42nd day of infection). The amount of antibody forming cells in Trichinella-infected mice significantly decreased on the 20, 25, 47, 55th days of infection. Hemagglutinin production to sheep red blood cells was significantly inhibited on the 25th day of Trichinella spiralis infection. The concept of immunodepressors being released from helminths is briefly discussed.

An attempt of treatment of malignant melanoma with bovine pineal tissue.

Seven incurable patients with disseminated malignant melanoma were injected (subcutaneously, sometimes intravenously) with homogenate of bovine pineal tissue. The daily injections of large quantities of pineal tissue appeared to be harmless for patients. Many-fold injections (during 9.5 months) of large quantities of pineal tissue (up to 3950 g) in 1 of 7 cases produced certain effect on subcutaneous metastases of malignant melanoma, i.e. causing reduction and even disappearance.

Inhibition of tumor and fetal tissue growth in newborn recipients.

A comparative study of growth of a variety of fetal tissues and transplantable tumors in syngeneic newborn and adult mice was carried out. Tumors used in the experiments arose spontaneously (hepatomas, mammary gland adenocarcinoma) or resulted from malignant conversion of ectopic transplants either of fetal tissues (urinary bladder carcinoma, adenocarcinoma of small intestine, stomach sarcoma) or of adult animal tissues (ovary carcinoma) in syngeneic system. The growth of "teratomas" developed after transplantation of minced tissues of 18-20-day fetuses was considerably inferior in newborn syngeneic recipients as compared to analogous transplants in adults. Inhibition of tumor growth was also observed in newborn animals. It was manifested in prolongation of latent period before tumor node appearance as well as in slowing down of growth rate of developed tumors. Only one tumor, mammary gland adenocarcinoma, proved to be an exception, its growth being equally progressive in newborn and adult recipients. At transplantation of tumor cells mixed with lymphocytes of adult mouse spleen, stimulation of tumor growth in newborns and inhibition of growth in adult recipients was observed. It is suggested that there exists a special type of cellular or humoral mechanism controlling tumor growth in newborns. The activity of such factors is conceivably based on fetal antigens as targets.