M components and cancer.

Intention of this study was to prove or exclude in patients with different neoplasms some M components which can escape routine electrophoretic analysis or immunoelectrophoretic analysis with polyvalent antiserum. All serum samples from 606 patients were investigated also with monospecific antisera against heavy chains--alpha, gamma, mu and delta--and in suspicion of abnormality also with antisera against light chains. Patterns similar to those found in cases of heavy chain diseases were absent in all patients. M components of IgD class were not found, as well. As to M components of usual types a relatively high incidence was found in lymphoproliferative malignancies excluded myelomas or Waldenstroöm's macroglobulinemias (5.61% in adults and 2.32% in children). On the basis of results the importance of immunoelectrophoretical analysis with monospecific antisera in all these cases is pointed out; not only in attempt to discover additional subjects of heavy chain diseases but also to detect mild "macroglobulinemic" abnormalities which may not be so far detected on electrophoretical pattern.

Chromatographic pattern of DNA isolated from liver tissue during hepatoma development.

DNA was isolated from livers of the rats treated with DAB during various steps of hepatoma development. After histological examination the tumor tissue was separated from the normal liver tissue and used as the source of DNA. Chromatographic fractionation on DEAE and Ecteola celluloses shows characteristic patterns for DNA isolated at various steps of hepatoma development. The largest differences in hepatoma DNA as compared to normal liver DNA were demonstrated in the DNA fraction eluated with 2.0 M-NaCl and NH3, gradient 0.1--1.0 M (m. w. 2--9 x 10(6)), and an increase in the first DNA fraction (m. w. less than 1 x 10(6)) was observed. Differences in the chromatographic patterns are discussed in terms of direct DAB action on DNA.

Hormone dependency of rat mammary carcinoma--a comparison of two assay systems.

Succinic dehydrogenase activity of the DMBA induced tumor explants cultured with or without hormones was assessed histochemically while receptors for estrogen (ER) and progesterone (PgR) were estimated from the cytosol fraction of the tumor tissue. Tumor regression following ovariectomy (OVX) was kept as the end point for determining hormone dependency. By in vitro method positive correlation was observed in 5 of 6 responsive or hormone dependent tumors, and 13 out of 14 independent tumors. Presence of receptors (ER + PgR, PgR) correlated with responsiveness in 4 of 6 tumors while their absence in the non-responsive group correlated in 6 of 14 tumors. Prolactin responsive tumors did not regress following OVX even if ER + PgR or ER/PgR were present. Using the same tumor tissue the results of hormone dependency by the two methods were identical in only 9 of 20 tumors.

Comparison of the excretion of different types of melanogens in the melanoma patients.

Urinary melanogens are indolic and phenolic compounds which are excreted in an elevated amount in the urine of melanoma patients. Urinary melanogens can be divided into two principal groups according to their behavior during Thormählen test--Thormählen positive melanogens (TPM) and Thormählen negative melanogens (TNM). When comparing TPM and TNM (represented by homovanillic and vanillactic acids) urinary excretion in stage III of melanoma patients we have found two significantly different relations for melanogens excretion, i. e. predominating TPM urinary excretion in melanotic forms of melanomas and predominating homovanillic acid and vanillactic acid urinary excretion in amelanotic forms of melanomas. We assumed that it could be an indirect evidence of the different catalytic activity of either cresolase or catecholase activities of tyrosinase.

Individual serum proteins and acute phase reactants in monoclonal immunoglobulinopathies (a study in patients with IgA myeloma and light chain myeloma).

Immunostimulating effect of Listeria monocytogenes, BCG and their biologically active extracts on the model of mouse ascites tumors.

Urinary free cortisol, cortisone, cortisol sulfate, cortisone sulfate and 17-ketosteroids in breast cancer.

Inhibition of cell-mediated cytotoxicity against tumor-associated antigens by suppressor lymph node cells from mice bearing methylcholanthrene-induced sarcomas.

Suppression of cell-mediated cytotoxicity directed against tumor-associated membrane antigens of methylcholanthrene (MC)-induced sarcomas by lymph node cells (LNC) from tumor-bearing mice was examined by inhibition of the 3H-thymidine incorporation assay. Normal LNC from B10 mice had been sensitized in vitro by 6-day cultivation on layers of irradiated syngeneic sarcoma cells and the effect of the generated cytotoxic lymphocytes was then inhibited by the admixture of suppressor cells present in lymph nodes of tumor-bearing mice. Suppressor cells were detected in lymph nodes from 7 of 16 (44 per cent) individually examined tumor bearers when LNC were added to cytotoxic lymphocytes which had been presensitized in vitro. When the experimental schedule was reverted and the tumor bearers' LNC were admixed to the normal LNC prior to the in vitro sensitization so that they were present in the population of the effector cells throughout the period of sensitization, the suppressor effect was detected in only 1 of 9 (11 per cent) tumor-bearers. The suppressive effect of tumor bearers' LNC was found to be nonspecific, being elicited not only by LNC derived from bearers of the sensitizing tumor, but also by LNC from bearers of an unrelated, immunologically noncross-reacting MC-induced sarcoma.

Investigations on the methods of clinical application of a soluble BCG fraction (F70) in lung cancer.

Several parameters for an optimal treatment scheme of a soluble BCG fraction (F70) were investigated. Among lung cancer patients treated with F70 a restricted selection was made of patients treated with small doses (5--10 U) every second or third month (sparing scheme) and of patients treated every month with sharply increasing doses up to tens of thousands units (aggressive scheme). It was found that the survival rate and the rate of marked X-ray regressions were higher in the former group. As it was previously established for lung cancer patients treated with living BCG, in the group of sparing scheme-treated patients the longest survival period pertained to patients treated once and patients treated twice or three times and an inverse correlation existed between the number of applications of F70 and the mean survival period. It was concluded that, as with living BCG, a sparing approach to the immunotherapy of lung cancer with F70 is to be preferred to an aggressive approach. Illustrative cases treated once, twice and three times are presented.

Oncogenicity in newborn and adult Syrian hamsters of SV40 temperature-sensitive (ts) mutants.

Newborn and adult Syrian hamsters were injected with wild-type SV40 and its temperature-sensitive (ts) mutants A30, A209, A239, B201 and BC210. In contrast to wild-type SV40, ts A30, ts A 239 and ts BC210 were oncogenic in adult hamsters inducing tumors after almost the same latent period as wild type SV40 in newborns but in lower number of animals. Study of TSTA in some tumors (1 generation) induced in newborn and adult hamsters by wild type SV40 and its ts mutants (with the use of immunogenicity and immunosensitivity tests) revealed no significant difference among compared tumors: most of them were immunogenic and immunosensitive. In contrast hamster embryo cells in vitro transformed by SV40 ts A30, ts A239 and ts A209 mutants, studied at different passage levels were all immunoresistant during about 30 in vitro passages and in most cases 10--100 times less immunogenic than hamster embryo cells in vitro transformed by wild type SV40. At higher passage level in some of these lines expression of TSTA improved. The data obtained are discussed in connection with the recently demonstrated [5] significant quantitative difference in tumor specific transplantation antigen activity in hamster cells in vivo and vitro infected, or transformed by wild type SV40 and its ts A mutants.

EBV-specific humoral antibodies in nasopharyngeal carcinoma patients in Cuba.

Sera of 24 patients with nasopharyngeal carcinoma (NPC) and of 60 healthy donors were studied for presence of IgG and IgA antibodies against different antigens induced by Epstein--Barr virus (EBV). The results obtained demonstrate that NPC in Cuba, as in other countries, is accompanied by elevated antibody titers against EBV-specific IgG antibodies, whose levels strongly differ from those of healthy persons. Patients with NPC in Cuba had higher titers of antibody against VCA of EBV in comparison with patients of Caucasian origin in those countries where NPC, the same as in cuba, is rarely found. Cuban NPC patients had high levels of antibody against EA, often higher than in NPC patients in endemic zones of this disease.

Increased susceptibility of leukemia-infected chickens to Marek's disease.

Intramuscular (i.m.) injection of RAV-49 into 1--3 day-old chickens infected with Marek's disease herpes virus (MDHV) by contact led to exceedingly high rate of Marek's disease (MD) incidence. According to the data obtained in three experiments 52 out of 108 (48.1%) chickens fell ill in this group. While in the group uninfected with RAV-49 but contactly infected with MDV-Kekava there were only 11 out of 102, (10.7%) incidences. Simultaneous inoculations of RAV-49 and MDV had no effect on MD morbidity. Increased susceptibility of i.m. RAV-49-infected and contactly MDV-infected chickens to MD was abolished by simultaneous inoculation of RAV-49 and herpes virus of turkeys (HVT).

Studies of the leukemic cell associated antigen(s) in mice with myeloid leukemia (Graffi), using the immunoperoxidase techniques.

Thymus, spleen and lymph nodes from myeloid leukemic mice (Graffi) were examined by light and electron microscope using immunoperoxidase techniques. As immune sera autologous sera were applied. The percentage of the labeled leukemic cells ranged from 30 to 60. It was established that leukemic cell associated antigen(s) was located mainly on the plasma and nuclear membranes. Occasionally this antigen(s) was observed in some areas of the endoplasmic reticulum, on separate cytoplasmic vesicles, on the budding viruses and on the chromatin. In addition, weakly expressed antigen(s) was located in cytosol. The mature virus particles were not labeled with peroxidase.

Conditions for hormone-stimulated expression of endogenous C57Bl strain-associated mammary tumor virus genome.

Full MMTV-Y gene expression can occur in dexamethasone-insulin-prolactin-stimulated cell cultured derived from C57Bl/10 mammary adenocarcinomas induced by syncarcinogenic action of chemical carcinogens (dimethylbenzanthracene), and mammotropic hormones (estrogen and prolactin). The rate of the hormone-stimulated virus production, as determined by biochemical, immunological and electron microscopical methods, was comparable to the level of spontaneous MMTV production by cells of established mouse mammary cancer cultures (CCL-51 and Mm5mt/cl). On the contrary, no virus production has been detected in hormone-stimulated cultures derived from C57Bl/10 mammary tumors induced by chemical carcinogen alone (urethan).

Carcinoembryonic antigen (CEA) in patients with malignant and non-malignant diseases.

Serum carcinoembryonic antigen (CEA) concentration was found to be raised in 503 of 550 patients (91%) with bladder cancer, lymphoma of intestine, hepatocellular carcinoma, bronchogenic carcinoma, prostate cancer, cirrhosis of liver and bilharziasis. The degree of elevation was moderate in all patients except in 189 patients in whom values more than 20 ng/ml were recorded, of which 53 patients with bladder cancer and 118 patients with bilharziasis. The mean CEA value in the patients with cirrhosis in the non-tumorous liver was slightly higher than that in those without cirrhosis, but the difference did not reach statistical significance (P greater than 0.01). There was no correlation between serum CEA and alph-fetoprotein (AFP) levels in all patients except in patients with bladder carcinoma, hepatoma and bilharziasis.

Virus-specific sequences in nuclear DNA from non-producer hamster Rous sarcoma.

Proviral DNA from non-producer Rous sarcoma in Syrian hamster contains practically all the nucleotide sequences presented in 125I-labeled RNA from Rous sarcoma virus, Carr-Zilber strain. Virus-specific sequences consist of moderately reiterated and unique DNA regions. The amount of Rous sarcoma virus-specific provirus equivalents in hamster Rous sarcoma DNA is equal to 5.2 +/- 0.01. Experiments on transfection show that proviral DNA studied possesses biological activity in respect to cell transformation and virus production. The infectivity of DNA from hamster tumor does not depend on the expression of group-specific (gs) antigen in the recipient cells.

Peripheral blood lymphocytes and EAC-rosette forming cells in B77 tumor bearing rats.

The percentage and absolute numbers of peripheral blood lymphocytes and EAC-rosette forming cells were evaluated in B77 tumor bearing rats. The peripheral blood lymphocytes were in high level even in rats bearing very small tumors; this result suggest the mobilization of lymphoyctes into peripheral blood in tumor bearing rats. The increase of EAC-rosette forming cells was accompanied with the growth of tumors.

Blocking factor in sera of MC1 and B77 tumor bearing rats detected by LAI assay.

A modified form of the leukocyte adherence inhibition assay (LAI assay) has been used to detect the blocking factor in serum of the Lewis (LW) inbred rats and the F1 hybrids of the inbred LW X AVN strains with transplantated syngeneic B77 and MC1 tumors. This study has indicated that the cell mediated antitumor immune response of rats immunized with living tumor cells and the blocking factor in serum of rats with progressively growing tumors may be demonstrated by the LAI assay with incubation time prolonged to 20 hr (LAI assay-IT20). In addition, the specificity of the blocking effect and the disappearance of blocking factor after surgical removal of tumors have also been demonstrated in this paper.

Heterogeneity of neoplastic cells derived from a hamster tumor induced by avian sarcoma virus B77.

Cells derived from a hamster tumor induced by avian sarcoma virus Bratislava 77 (B77) in vivo, were cultivated in vitro. After few passages two morphologically different cell lines were isolated from the parental culture (B77/H). One cell line consisted of fibroblastic cells (B77/H/fi and the other from epithelioid cells (B77/H/ep). Cells of both lines were highly tumorigenic in neonatal syngeneic hamsters, B77/H/ep cells were able to form colonies in soft agar and contained complete integrated B77 viral genome. In contrast, the B77/H/fi cells grew poorly in soft agar and did not contained B77 virus genome.

Calf serum- and cell surface proteins released from cultured avian sarcoma virus-transformed and untransformed rat fibroblasts.

Cell surface and calf serum proteins were released in vitro from cultured virus-transformed, chemically transformed and normal rat embryo fibroblasts in 0.2 M urea containing serum-free culture medium. Fibronectin (LETS protein) was found in considerably higher amount in medium from normal rat embryo fibroblasts than in that from transformed cells. Major calf serum protein released from cultured normal and transformed rat cells corresponded by its electrophoretic mobility to calf serum albumin. No substantial differences in electrophoretic patterns of calf serum proteins released from normal and transformed cells into the medium were found. Time course of release of serum proteins from cell surface was measured quantitatively with the aid of radioiodinated Staphylococcus aureus protein A radioimmune assay. More than 50% of cell bound calf serum proteins remained on cell surface after 24 hours of incubation in serum-free medium.

Study of the avian sarcoma virus infection of chemically transformed cells.

BHK-21/13 cells transformed with various chemical carcinogens and mutagens were tested for susceptibility to avian sarcoma virus infection. The chemically transformed BHK cells were highly tumorigenic. The treatment of these cells in vitro with avian sarcoma virus strain Schmidt-Ruppin D resulted in chemically transformed and viral infected cells with different rescuability of the integrated virus genome. The different rescuability is not due to the difference in virus penetration as was shown by vesicular stomatitis virus Schmidt-Ruppin virus pseudotype VSV(SR-D) technique. The sarcoma virus genome in the doubly transformed cells was not inducible by various virus inducers.

Sero-epidemiologic study of the Epstein--Barr virus infectivity in a healthy Cuban population.

The indirect immunofluorescence test was used to study the level of humoral antibodies against viral capsid antigen of the Epstein--Barr virus (VCA-EBV), and early antigen (EA) in the sera of 322 healthy donors of the Havana City province. The age of donors ranged from several months up to 95 years. The results obtained indicate that the healthy population of Cuba is infected by EBV as well as in other countries, even though the spread and the course of infection have its specific characteristics. In the early childhood population group (3--4 years old) the infection is relatively more frequent (73%) than in adults (62--77%), despite the fact that the country is located in the tropical zone. In all the studied age groups the geometric mean titers (GMT) of antibodies were found to be lower in comparison with the other populations studied. The increase of EBV-antibody levels in the group of donors above 55 years of age, together with a large number of seropositive individuals, increase in the number of persons with high anti-VCA-EBV antibody titers (1 : 160), agrees with the results obtained by other investigators. This may be associated with some form of depression of cell-mediated immune response. Infectious mononucleosis-morbidity in the studied age groups was found to be low in 3--4-year- and 15--19-year-old age groups, where higher levels of antibodies against EA of EBV were found.

Serum immunoglobulins in acute myelogenous leukemia.

Forty-five patients with acute myelogenous leukemia have been followed sequentially for 12 months. All these patients had serum immunoglobulin IgA, IgG, IgM and IgE levels measured by immunodiffusion and radioimmunoassay prior to chemotherapy, then every month after chemotherapy. Significant differences were found in IgA and IgG levels prior chemotherapy, while no differences were found in IgM or IgE levels at any time. There was a positive correlation between percentage of blasts and IgA level. There was negative correlation between the percentage of blasts and IgM levels. These findings raise the value of measuring the levels of immunoglobulins in patients with leukemia as a guide to the subclinical relapse of the disease. These results may also support the hypothesis of the role of early immune defect in immunoglobulin metabolism in the pathogenesis of acute leukemia.

An osteogenic sarcoma of the thyroid gland (report of a case and survey of the literature).

An additional case, counted to be the twenty third, of a primary osteosarcoma of the thyroid gland in a 62-year-old woman is described. The literature on the subject is surveyed, and the possible histogenesis of this unusual neoplasm is discussed.

Paracortical activity in the lymph nodes draining female breast carcinoma.

Three hundred and two female breast carcinomas were assessed histologically with special attention focused on the nuclear grade of the tumor, the stromal lymphocyte reaction and the morphology of the paracortical areas of the regional lymph nodes. These morphologic parameters were correlated with the 5-year survival data of the patients. Nuclear grade of the primary tumor was directly positively related to the 5-year survival as was the paracortical activity of the regional lymph nodes. The paracortical activity was inversely related to the frequency of nodal metastases which were a sign of poor prognosis. The value of the morphology of the regional lymph node paracortex in evaluating the criteria of host resistance in association with breast carcinoma is emphasized.

The pattern of human lymph node involvement by the non-Hodgkin lymphomas of B-cell lineage.

A total of 93 lymph nodes from 24 patients with a primary malignant non-Hodgkin lymphoma of B-cell lineage was assessed histologically with special reference to the presence or absence of neoplastic B-cells in the lymph node cortex and medulla (B-cell areas) as well as in the paracortical (T-cell area) area, and in the lumen of the post-capillary venules (PVC). The B-cell origin of the lymphomas was verified by using an indirect immunoperoxidase technique for the demonstration of the cell surface immunoglobulins. The B-cell areas of the node were involved in all the lymph nodes studied, and the T-cell areas in 95.7 per cent of the nodes. Neoplastic cells were present inside the PCV in 80.6 per cent of the nodes. Furthermore, 15 lymph nodes were seen where the paracortical area was only partially involved despite the total replacement of the B-cell areas by the neoplastic cells. The results suggest that the nodal involvement by the B-cell lymphomas starts from the B-cell areas, a fact that could be used as an aid in the diagnosis of these lymphomas as well as in the study of the dynamics of B-lymphocytes in general.

Suppression of immune anti-tumor lymphocytes by macrophages of advanced tumor bearing rats, across a cell impermeable membrane.

In experiments using double and triple chamber cultures it was demonstrated that suppressive macrophages from advanced T8-Guérin tumor (diameter 5--6.5 cm) bearing rats produced a dialysable factor which suppressed the killer activity of lymphocytes from non-advanced T8-Guérin tumor (diameter 0.5--0.7 cm) bearing rats, as well as from nonadvanced h 18R tumor bearing rats and from Ehrlich ascites bearing mice, against T8-Guérin ascitic cells and, respectively, against h 18R ascitic and Ehrlich ascitic cells. The dialysable suppressive factor inhibits immune lymphocytes but has no effect on the lymphotokin itself already produced.

Experimental data concerning the question of the synergic action of the influenza virus and of some chemical carcinogens in the pathogenesis of lung cancer.

The importance of new discoveries in the field of molecular biology concerning the interaction of oncogenic material in cell leading to a synthesis of knowledge in the field of viral and physico-chemical blastomogenesis is underlined. A concise reference to earlier studies and to the present state of knowledge is presented, with a subsequent review of the results of experimental studies dealing with the problem of the synergic action of influenza viruses (type A2, A/PRS, B, Sendai) and of chemical carcinogens (urethane, cancerogenic hydrocarbons, diethylnitrosamines, cigarette smoke) in the pathogenesis of lung cancer. Both the suggested and already demonstrated mechanisms of the joint action of viruses and chemical carcinogens, illustrating the interrelations between the two factors in the process of malignant transformation, are discussed.

Splenic and thymic histamine forming enzyme activity and weights following organ allograft and tumor cell transplantation.

Kinetic of the increasing activity of histamine forming enzyme--histidine decarboxylase in host spleen and thymus, following kidney and heart allografting and after tumor cells transplantation is presented.

Antitumor effect of sublethal irradiation caused by its tolerance abrogating capacity.

After the resection of methylcholanthrene-induced fibrosarcomas, representing 1--2% of total body weight, "R" rats were immunized with glutaraldehyde-fixed tumor cells, irradiated sublethally [0.1419 C kg-1 (550 R)], and restored immunologically by thymus, spleen and bone marrow cells. Afterwards, 87.5% of them were able to reject a viable challenge cell graft of 1 X 10(5) cells. Sublethal irradiation alone had the same effect, reflected by 90% of rats rejecting the grafts. Five control lots were run. They showed that none of the treatments, applied to "tumor-resected" animals could provide normal animals with the same defense capacity. Results point to the decisive role of the transient presence of the tumor in the host and of the sublethal irradiation in restoration of its defense capacity. Role of partial tolerance, in producing the host's immune inhibition, and of the capacity of irradiation to abrogating it, are discussed.