Fate of tumor cells injected into left ventricle of heart in BALB/c mice: role of natural killer cells.

The arrest, retention, and elimination (i.e., clearance) of radiolabeled YAC-1 lymphoma cells injected either iv or into the left ventricle (LV) of the heart were studied in male BALB/c mice, with special emphasis on the role of natural killer (NK) cells. After iv injection YAC-1 cells were arrested and, to a large extent, destroyed in the lungs, which contain the first capillary bed that iv injected tumor cells meet. After LV injection the initial distribution of the tumor cells, which depends on the distribution of cardiac output at the time of injection, was estimated by use of radiolabeled microspheres. Using this technique, we have shown that LV-injected tumor cells, in contrast to iv injected tumor cells, were not arrested in the first capillary bed that they encountered but passed viably through the microvasculature of the brain, heart, kidneys, intestinal tract, and to some extent, the bone, skin, and muscle. The only organs that could arrest the LV-injected tumor cells were the lungs and the liver. In the lungs clearance of YAC-1 cells began immediately after the cells were arrested. However, the rate of clearance could be almost abrogated by pretreatment of the recipients with anti-asialo GM1 antiserum, which destroys most of the NK cells in vivo and strongly depresses the in vitro NK cell activity. In contrast, YAC-1 cells arrested in the liver were not cleared from this organ during the first 1-2 hours after arrest. After this delay clearance of the cells commenced. Pretreatment of the recipients with anti-asialo GM1 also strongly depressed the clearance of tumor cells from the liver. Although pretreatment with polyinosinic-polycytidylic acid enhanced in vitro NK cell activity, it could augment only slightly the clearance of YAC-1 cells from the lungs and the liver. Thus these results strongly support the hypothesis that the rapid clearance of tumor cells from both the lungs and the liver depends, at least partially, on the NK cell activity within these organs.

SL12 murine T-lymphoma: a new model for tumor cell heterogeneity.

It has been observed that subclones from the spontaneous murine AKR/J T-lymphoma cell line SL12 with similar in vitro growth characteristics exhibit stable differences in tumorigenicity. The cell line is composed of at least three distinct cloned cell types that are highly, moderately, or poorly tumorigenic in syngeneic host animals. When healthy, young, syngeneic host animals were given iv injections with the same number of viable growth phase cells, each cloned cell type had a different tumor incidence, latent period, and pattern of tumor spread. The unusual stability of the cloned cell lines is shown by a similar incidence, latency, and spread of the tumors when studied after more than 1 year of continuous in vitro culture. The SL12 clones also differ in several phenotypic characteristics commonly used to classify thymocyte maturation, e.g., a) the expression of three of seven surface antigens examined, b) the cellular response to glucocorticoid hormone, and c) the expression of terminal deoxynucleotidyl transferase.

Inhibition of lymphoma invasion and liver metastasis formation by pertussis toxin.

We have examined whether pertussis toxin, an agent known to inhibit entry of normal lymphocytes into tissues, affects invasion and metastasis formation by malignant lymphoma and T-cell hybridoma cells. The toxin reduced invasion in vitro in hepatocyte cultures to 20% of control values. Inhibition was maximal after pretreatment for 2 h with approximately 100 ng/ml. The effect of pretreatment with 1 to 5 micrograms toxin/ml for 4 h persisted for at least 5 days, despite a more than 100-fold increase in cell number. The proliferation rate was not affected. Liver metastasis formation after tail vein injection of TAM2D2 T-cell hybridoma cells in syngeneic AKR mice, measured as liver weight, was reduced to 10 to 25% of controls after pretreatment of the cells for 4 h with 1 microgram pertussis toxin/ml. Metastasis to kidneys, ovaries, and lymph nodes was not, or less evidently, affected. With MB6A lymphosarcoma cells no effect was seen after treatment with 1 microgram/ml, but a significant reduction of the liver tumor burden to approximately 50% of controls was achieved by treatment with at least 5 micrograms toxin/ml. Spleen metastasis by MB6A cells was not affected. These results provide evidence for a similarity in invasion mechanisms of normal and malignant lymphoid cells, and they suggest that invasiveness is an important factor in the formation of lymphoma metastases, particularly in the liver.

Successful high-dose intravenous cytarabine treatment of parenchymal brain involvement from malignant lymphoma.

A patient with systemic recurrence of large-cell malignant lymphoma developed radiologic evidence of parenchymal brain metastasis. Treatment with systemic high-dose cytarabine resulted in complete regression of peripheral adenopathy and complete radiologic response in the central nervous system, with normalization of the brain computed tomographic scan. This case further demonstrates the ability of high-dose cytarabine to penetrate the blood-brain barrier and achieve therapeutic drug concentrations in the central nervous system, an area that might otherwise remain a pharmacologic sanctuary for tumor cells.

Dissemination and proliferation of intravenously injected murine, syngeneic T lymphoma cells.

BALB/c mice were inoculated intravenously with syngeneic T lymphoma cells. Dissemination and proliferation of malignant cells in vitro were determined from time of cell injection to the time of death of the animal, using an agar medium in which only tumor cells grew. Tumor cells were detected in kidney from day 7, and in liver, lungs and ovaries from days 9-12. The myeloid-lymphatic system was only sparsely invaded. Three weeks after injection the animals died of general metastases, and colonies could be grown from all organs. Peripheral blood tumor cells colonies were grown in quantities of 1-2% on injected cells during the first hour. They then disappeared from blood, but reappeared 2-3 weeks later. Within the first 15 min after i.v. infections of vitro labelled tritiated-thymidine tumor cells, 45% of the radioactivity was recovered from lungs and 40% from the liver. After this time the activity gradually declined. Only about 1% of the initial activity of the injected cells was found in the spleen, kidney and brain during the first 24 hours. Blood glow of the organs was examined by means of indicator fractionation. Highest flow rates were measured in liver, kidney, brain and lungs. Thus, the sites of metastases after tumor cell injection were determined neither by the initial deposition of tumor cells, nor by blood flow rates through the organs. This result shows that cellular interaction between neoplastic cells and the immediate environment must be major determinants in the development of metastases.

Prevention of metastatic spread by postoperative immunotherapy with virally modified autologous tumor cells. II. Establishment of specific systemic anti-tumor immunity.

The successful application of a non-oncogenic virus, Newcastle disease virus (NDV), which can be used to modify a highly metastatic tumor to become more immunogenic is reported. Such NDV modified tumor cells were found to be effective as tumor vaccine for anti-metastatic therapy in combination with surgical removal of the primary tumor. The protection in the animals seen after this treatment is paralleled by an establishment of specific systemic anti-tumor immunity. This protective immunity depended on recognition of a distinct tumor antigen. The therapy protocol also worked in animals bearing the plastic adhesive variant ESb-MP. It did not work, however, when using an immune escape variant not expressing a specific tumor antigen.

In vivo and in vitro properties of malignant variants of RAW117 metastatic murine lymphoma/lymphosarcoma.

Using the RAW117 lymphoma/lymphosarcoma system syngeneic to Balb/c strain mice, variant sublines have been selected for enhanced blood-borne liver colonization in vivo or for lack of binding to immobilized lectins in vitro. The kinetic organ distributions of intravenously injected, 3H-thymidine-labelled RAW117 parental cells and a subline sequentially selected ten times for enhanced liver colonization were similar, suggesting that the differences in malignancy between these two cell lines were not due to dramatic differences in organ localization properties. Examination of the malignant properties of the selected sublines and cell clones derived from these in immune-impaired animals indicated that host immune status was important in determining the quantity of experimental metastases in this system. Although impairment of T-cell or NK-mediated anti-tumor responses by using 400 R 60Co-irradiated or Balb/c nude (nu/nu) mice suggested that certain immunologic responses were not effective in preventing experimental metastasis, impairment of macrophage function with chlorine, silica, trypan blue, carrageenan, cyclophosphamide or pristane were effective and resulted in enhanced malignancy of the parental RAW117 line. In contrast, impairment of macrophage function had little or no effect on the experimental metastatic properties of highly malignant RAW117 sublines or clones. In vitro humoral responses or cell-mediated immunologic assays using lymphoid cells from normal or tumor-bearing hosts failed to demonstrate antibody-mediated or antibody-dependent cell-mediated cytotoxicity (ADCC), T-cell or NK-cell responses against RAW117 cells. However, poly I: C activated macrophages were more effective against parental RAW117 cells than against a highly metastatic subline in cytolysis and cytostasis assays suggesting that the highly metastatic RAW117 cells can more readily escape macrophage-mediated host defenses.

Radiation-induced changes in regional blood flow in human tumors.

In 43 patients the blood flow in 48 superficial metastatic tumors was measured with the 133xenon wash-out method. In all cases the blood flow was measured before the start of radiotherapy and then one week later during radiotherapy. In 36 cases the blood flow was measured after 2 weeks during radiotherapy, and in six patients the follow-up lasted 5-6 weeks. The blood flow increased during the first week of radiotherapy in the whole series from 20.1 +/- 18.0 ml/min/100g to 31.3 +/- 24.9 ml/min/100g. The increase during the first week was significant (p less than 0.001). During the second week of radiotherapy the blood flow decreased to 27.0 +/- 19.3 ml/min/100g; the decrease was also significant (p less than 0.05). The changes in the different tumor groups during radiotherapy seemed to be in the same direction. In a longer follow-up the gradual decrease in the blood flow seemed to continue.

Brain metastases: results and effects of re-irradiation.

Re-irradiation for recurrent manifestations of brain metastases has been reported to be of benefit by either increasing the duration of survival or improving the quality of life. The records of 455 patients with brain metastases treated by radiation therapy at the University of Colorado Health Sciences Center from 1975 through 1986 were reviewed. Of these, 44 patients (9.7%) were re-irradiated because of suggestive neurological findings and/or imaging studies diagnostic of recurrent disease. The primary site distribution was as follows: lung (non-small cell)--15 (34%), lung (small cell)--9 (20%), melanoma--5 (11%), breast--4 (9%), genitourinary--4 (9%), unknown--4 (9%), lymphoma--2 (4%), and endometrium--1 (2%). Retreated patients received at least two courses of irradiation and one received three. The median interval between the first and second courses was 34 weeks (7.8 months). For the initial course of treatment, all patients were treated to the whole-brain with megavoltage X rays to a dose of 30-36 Gy (median 30 Gy) at 1.5 to 4.0 Gy/fraction (median 3.0 Gy/fraction). Retreatment also consisted of whole-brain irradiation (37/42 patients) to additional doses of 6-36 Gy (median 25 Gy) at 2.0 to 4.0 Gy/fraction (median 3.0 Gy/fraction). The total cumulative doses to the brain, therefore, varied from 38-75 Gy with a median of 60 Gy. Survival data were available for 42 of 44 patients retreated. All patients died with disease. The overall median survival following the initial course of irradiation was 40 weeks (9.2 months) with 10 patients (24%) living beyond 1 year. The median survival following retreatment, however, was only 8 weeks with one patient surviving greater than 1 year. Only 12 patients (27%) showed partial neurological improvement with re-irradiation and over one-half (55%) either failed to respond or deteriorated during or soon following retreatment. Brain necropsies were performed in 8 patients. Three of these had developed brain necrosis and two most likely died as a direct consequence. It is concluded that retreatment of brain metastases is seldom worthwhile. Survival is usually short and most importantly, the quality of survival frequently is not improved.

Incidence, presentation, and outcome of spinal cord disease in children with systemic cancer.

During a 40-month period, in 24 of 643 (4%) newly diagnosed patients with systemic cancer younger than 18 years of age (range: 3 months to 17 years) spinal cord disease developed. Patients with spinal cord disease included 21 children with metastatic spinal cord compression, two with treatment-related transverse myelopathies, and one with an anterior spinal artery stroke. Spinal cord disease occurred in 13 of 102 children (12%) with sarcomas, six of 82 (7%) with neuroblastomas, and four of 94 (4%) with lymphomas. Spinal cord compression occurred as the presenting sign of malignancy in six children (four with sarcomas and two with lymphomas). In the remaining 15 patients, cord compression occurred a median of 13 months after initial diagnosis, and in four patients it occurred at the time of first relapse. Symptoms of metastatic cord compression included back pain in 17 patients (80%), weakness in 14 (67%), sphincter dysfunction in 12 (57%), and sensory abnormalities in three (14%). Findings on plain radiographs of the spine were abnormal in only seven of 20 patients with cord compression, and myelography was needed to differentiate compression from other causes of spinal cord disease. Treatment included high-dose corticosteroids followed by operation (seven patients) or radiotherapy (14 patients). After treatment, nine of 15 nonambulatory patients became ambulatory, and five of 10 incontinent patients regained sphincter control. None of the patients with nonmetastatic spinal cord disease had a satisfactory outcome.(ABSTRACT TRUNCATED AT 250 WORDS)

Emulsion and activated carbon in cancer chemotherapy.

Based on a lipid-absorbing ability of lymphatic capillaries, a fat emulsion containing anticancer agents was applied to selectively deliver more increasing amounts of anticancer agents into regional lymph nodes. The emulsions, in which the drug solution is contained as the innermost phase or as oily soluble drug, yield high drug concentrations in the lymphatic system. Clinical trial of the emulsion method was carried out preoperatively for 180 patients with stomach cancer. As a result, the emulsion enhanced the chemotherapeutic effect of the anticancer agent on lymph node metastasis. About a 20 m mu-sized activated charcoal, in which anticancer agents were absorbed, selectively delivered the anticancer agents to the lymphatic system. The activated charcoal was also excellent carrier material for the lymphatic system, and we have applied it to patients with lymphatic metastasis.

Melanocyte colonization of nodal metastasis from oral carcinoma.

Melanocyte colonization of metastases in cervical lymph nodes is described. The tumor was a squamous carcinoma of the floor of the mouth, metastasizing to cervical lymph nodes. The melanocytes were easily visible on sections of the nodes stained with hematoxylin and eosin and were confirmed by Masson-Fontana staining. Multiple sections through the oral primary tumor failed to show melanocyte colonization of the tumor. This is the second report of this phenomenon and represents only the third known case of this entity.

On the prevention of haematogenous tumor metastases rats. The role of the proteinase inhibitor "Trasylol".

All malignant tumors shed cells into the circulation. The number of circulating tumor cells bears no relation to the extent of secondary growth. This is determined by the number of tumor cells that cross the vessel wall and implant in extravascular sites. The rate of cellular emigration is regulated by the permeability of vessels harbouring tumor emboli, permeability in turn being dependent on the amount of alpha, 2, macro-globulin (AMG) lining vascular endothelium. Agents which inactivate or digest AMG, e.g. proteases, have been shown to promote the dissemination of tumor. Some tumors secrete large amounts of proteases. These enzymes are believed to lyse the AMG layer and so to facilitate the emigration of tumor cells. On this hypothesis, inhibitors of such proteases would have anti-metastatic properties. Present experiments lend support to this view. The proteinase inhibitor Trasylol, when allowed to act on tumor cells, has been shown to impair their capacity of setting up haematogenous metastases. It does not affect their viability or transplantability. It is suggested therefore, that blood-borne tumor dissemination might be inhibited by perfusing the (primary) tumor bed with proteinase inhibitor, possibly in association with AMG.

Clinical presentation of metastatic lymphoma to ureter.

Clinically significant metastatic involvement of the ureter is reported rarely, experience with it is limited, and therefore suspicion is low. A case of metastatic lymphoma serves to demonstrate the delay in diagnosis. A review of the causes of delay is presented.

Quantitative electron microscopy in the diagnosis of mycosis fungoides. A simple analysis of lymphocytic nuclear convolutions.

In the present study we compared the accuracy of three different ultrastructural methods used to confirm clinically proved cases of mycosis fungoides (MF) from skin biopsy specimens. Method 1 is a qualitative method and confirmed the diagnosis of MF in only 44% of the patients with MF. Method 2 involved image analysis of lymphocytic nuclei and confirmed the diagnosis of MF in only 67% of the patients with MF. The most sensitive ultrastructural method for confirming the diagnosis of MF was a simple scoring of the number of sharply angled nuclear invaginations in 100 lymphocytes (method 3). Control biopsy specimens had many more lymphocytes (19% to 55%) with no sharply angled nuclear invaginations compared with those from patients with MF (3% to 15%). The clinical diagnosis of MF was confirmed in 100% of our patients with MF using method 3.

Evaluation of computed tomography and radionuclide scanning in the staging of cutaneous T-cell lymphoma.

Computed tomography (CT) of the abdomen and pelvis was performed on 30 patients with cutaneous T-cell lymphoma (CTCL) as part of their pretreatment staging evaluation. Twenty-two patients also had liver-spleen radionuclide scans. Physical examination revealed limited cutaneous plaque disease in five patients, extensive plaque disease in 11 patients, cutaneous tumors in six patients, and exfoliative erythroderma in eight patients. Generalized palpable adenopathy was detected in 11 patients, localized palpable adenopathy in ten patients, and no adenopathy in nine patients. Peripheral lymph node biopsy specimens showed CTCL in seven patients and dermatopathic lymphadenitis or sinus histiocytosis in 17 patients. Two patients were at disease stage Ia, five were at stage Ib, seven were at stage IIa, two were at stage IIb, seven were at stage III, and seven were at stage IVa. The CT did not reveal intra-abdominal, retroperitoneal, or pelvic adenopathy, or hepatic or splenic abnormalities in any patient. Radionuclide scans demonstrated nonspecific abnormalities in five patients, but did not appear to reflect disease involvement. Computed tomography of the abdomen and liver-spleen radionuclide scans should not be routine staging procedures for CTCL.

Pulmonary manifestations of renal disease.

Many renal diseases have varied pulmonary manifestations. A pattern-recognition approach coupled with pertinent clinical findings can often lead to diagnosis of a specific disease entity. In this article, an attempt is made to categorize pulmonary manifestations of renal disease and to assign them to groups of customarily used recognition patterns as presented on chest radiographs.

Treatment of extraneural metastatic medulloblastoma with a combination of cyclophosphamide, adriamycin, and vincristine.

Seven patients with extraneural metastases from medulloblastoma were treated with a combination of cytoxan, adriamycin, and vincristine (CAV). None of the patients had evidence of active neural axis disease. All patients with bone metastases responded with a reduction in bone pain and improvement on the radionuclide bone scan. One patient presenting with lymph node metastases showed initial reduction in the size of the palpable nodes. In this group, the median time to the development of extraneural metastasis was 18 months from the time of original diagnosis of central nervous system medulloblastoma. The median duration of response to CAV, after extraneural metastasis, was 17 months (4-65 months). Four of seven patients died of disease-related causes, one patient presumed well was lost to follow-up, and two of seven are still without evidence of active disease at 37 and 65 months. The combination of CAV is well tolerated and provides reasonably good palliation for extraneural medulloblastoma.

Ultrastructural analysis of experimentally induced invasion in the rat lung by tumor cells metastasizing lymphatically.

The colonization of the lung by the rat tumor cells BSp73 ASML which have the ability to metastasize via the lymphatic system was studied at the ultrastructural level. Tumor cells arriving in the lung after i.v. injection become transiently embolized; within hours, however, they begin to extravasate from the blood capillaries. Swelling cellular protrusions open a limited area between endothelium and basal lamina through which tumor cells erupt. Tumor cells then form metastases in the interstitial tissue and, in an apparently lymphotropic action, intravasate the lymphatic vessels in a similar manner to a reverse diapedesis-like process. Within the lympatic system they settle, spread, and build up extensive tumor foci particularly in the subpleural region.

Flow cytometric measurement of ploidy and proliferative activity of carcinomas of the oropharyngeal mucosa.

The most important indicator of ploidy and cell cycle stage in ortho- and pathologic tissues is the nuclear DNA content. To study this parameter of malignancy in squamous carcinomas of the oral, pharyngeal, or laryngeal mucosa we analyzed DNA histograms by aid of flow cytophotometry after using pepsin digestion of tumor specimens for cell dispersal and fluorochromation with combined ethidium bromide and mithramycin. In 30 of 36 tumor tissue specimens aneuploid states of nuclear DNA content (83%) were recognized, in some cases only by comparing tumor cells with admixed diploid human reference cells. All but two tumors (with hypodiploid DNA pattern) exhibited hyperdiploid stem line abnormalities, one specimen even exhibited them with triclonal DNA distribution pattern. The degree of ploidy (DNA index), defined as the ratio of peak modal channel number for the G1/0 proportion of tumor cells to that of normal cells, ranged from 0.59 to 3.24 (mean 1.58). Cell cycle analysis of untreated squamous carcinomas, calculated by the relative DNA distribution pattern in histograms, also offered a considerable variation in proliferative activity.