Carcinomatous Pleural Effusions Research Articles

MO234LIFE THREATENING MALIGNANT HYPERCALCEMIA IN BREAST CANCER: CAN THE NEPHROLOGIST CHANGE THE PATIENT DESTINY?

Abstract Background Malignant hypercalcemia is a common complication in cancer patients and can be predictive of poor prognosis and advanced malignancy. Cancer-related mechanisms of hypercalcemia depends either on ectopic hypersecretion of humoral factors (PTH, PTH-like molecules, Vit D) by the tumoral mass, or on osteolysis due to bone invasion. In consideration of the high renal and cardiac impact of severe hypercalcemia, its prompt recognition and treatment can be lifesaving. AKI is frequently associated with malignant hypercalcemia and recognizes a multifactorial pathogenesis (direct renal vasoconstriction, volume depletion, tubule-interstitial damage, etc.). Substitutive renal treatment can be necessary, but not always viable in patients with poor physical performance and is a difficult choice in cancer patients with advanced malignancies. Case report We present the case of a 46 years old woman with an infiltrating non special type (NST) carcinoma of the right breast (luminal B, ER 70%, PgR 60%, Ki67 35%, HER2 1+). Unfortunately, she quickly progressed after a neoadjuvant treatment with Epirubicin/Cyclophosfamide (4 cycles) followed by Paclitaxel (10 cycles). A first line hormonal treatment (Palbociclib, Letrozole and LHRH analogous) was then started, but a further rapid disease progression was observed. A new biopsy showed a muted and more aggressive cancer phenotype (high grade triple negative carcinoma with no PDL1 expression). Due to a rapid worsening of general conditions with cognitive impairment, the patient was admitted to the Oncology department. Blood test showed severe hypercalcemia (corrected calcium: 23.26 mg/dl) and severe hypokalemic metabolic alkalosis (blood pH 7.57; HCO-3 32 mmol/l; K+ 2.6 mmol/l) associated to AKI (creatinine doubled to 1.42 mg/dl; eGFR 45 ml/min/1,73 mq). Alkaline phosphatase and Vitamin D were normal and iPTH was < 2 pg/ml. ECG showed a significant QTc prolongation to 550 msec. Imaging exams revealed lung metastasis, lung carcinomatous lymphangitis and bilateral pleural effusion, while Bone Scan showed a low caption limited to right ribs. Oncologists and Intensive Care Physicians referred to our Nephrology and Dialysis Unit proposing Hemodialysis support. In consideration of the preserved diuresis, the advanced disease stage and the unstable hemodynamics, our advice was instead for a conservative medical approach. An i.v. infusion of 4000 ml NaCl 0.9% + KCl 60 mEq/24h; a continuous i.v. infusion of Furosemide 5mg/h were started and maintained for several days, leading to a progressive and full renal function recovery (creatinine decreased to 0.7 mg/dl) and metabolic alkalosis correction. However, corrected calcium remained largely over target, being 16.3 mg/dl. Sodium bicarbonate 80 mEq, Zoledronic acid 4 mg and Desametasone 4mg/day were administered and at day 16 even calcium was in normal range. We realized that this was a favourable window for acting on the underlaying cause of malignant hypercalcemia, that likely was the aberrant secretion of non-dosable PTH-like molecule(s). Thus, we suggested the Oncologist to start chemotherapy, and they were allowed to treat the patient with Carboplatin/Gemcitabine. The patient was successfully treated with stabilization of normocalcemia and is still alive. Conclusion Malignant hypercalcemia is a life-threatening complication in advanced malignancies. Our case report highlights the key role of the nephrologist in treating a complex and fragile oncologic patient, achieving the full correction of several severe renal disorders, hemodialysis avoidance and survival improvement.

ROLE OF TUMOR MARKERS IN THE DIAGNOSIS OF MALIGNANT PLEURAL EFFUSIONS

As an aid in the differential diagnosis of oxidative pleural effusions tumor markers were investigated. We measured carcinoembrionic antigen (CEA) and a glycoprotein CA 15-3 in the pleural fluid of 33 patients with carcinomatous pleural effusions and of ten patients with tuberculous pleurisy and other causes because we have frequently found those diseases to be associated with exudative pleuritis. CEA and CA 15-3 were significantly higher in carcinomatous pleural fluids than in tuberculous fluids (p value of 0.0287), therefore combined assay of CEA and CA 15-3 may be useful in distinguishing pleural effusions due to malignancies from those of tuberculous origin.

Peritoneal and meningeal relapse from lung adenocarcinoma after a response to gefitinib: A case report

The prognosis of carcinomatous peritonitis and meningitis is poor since ascites and focal neurological dysfunctions usually do not improve despite the available treatment options for this devastating disease. This is a case report of peritoneal and meningeal relapse from lung adenocarcinoma following a response to gefitinib. A 72-year-old woman was diagnosed with adenocarcinoma in the upper lobe of the left lung, accompanied by a massive carcinomatous pleural effusion. Following a cisplati-based chemotherapy, gefitinib was initiated, achieving satisfactory disease control. At 8 months after gefitinib initiation, the patient developed abdominal distension, gait disturbance and visual disorder of the right eye. Adenocarcinoma cells were detected in the ascitic and pleural fluids. The findings of this case report suggest that clinicians should remain vigilant for this type of metastasis, although it is rarely encountered.

Osteoplastic bone metastasis in esophageal squamous cell cancer: report of a case

This report presents a case of esophageal squamous cell cancer with osteoplastic bone metastasis. A 58-year-old male patient underwent multimodality treatment for esophageal cancer. Sclerotic changes resembling bone metastasis from prostate cancer were detected in the 4th thoracic and the 5th lumber vertebral body soon after the adjuvant chemoradiotherapy. Systemic examinations revealed no primary cancer as a cause of osteoplastic bone metastasis and no esophageal cancer recurrence. A needle biopsy revealed metastases of esophageal squamous cell cancer with osteoplastic changes. Multiple sclerotic changes were detected in the systemic bones at that time, and new carcinomatous bilateral pleural effusion developed. The drastic systemic progression of the cancer caused the rapid deterioration of the patient's general condition.

Elevated soluble interleukin-2 receptor and interleukin-2 receptor positive cells in carcinomatous pleural effusions

Soluble interleukin-2 receptor (IL-2R) level and IL-2R positive (IL-2R+) cells were studied in twenty patients with carcinomatous pleural effusions. The mean value of soluble IL-2R level in carcinomatous pleural effusions was 2930 +/- 1722 U/ml and that in sera was 965 +/- 610 U/ml. Soluble IL-2R level in carcinomatous pleural effusions was found to be significantly higher (p less than 0.001) than that in sera of patients with carcinomatous pleural effusions and that in transudates. Serum soluble IL-2R level in patients with carcinomatous pleural effusions was found, to be significantly higher (p less than 0.001) than that in normal controls (264 +/- 70 U/ml). We also studied IL-2R+ cells in pleural fluids and peripheral blood of patients with carcinomatous pleural effusions. The mean percentage of IL-2R+ cells in carcinomatous pleural fluid lymphocytes was 22.8 +/- 17.8%, as compared with 3.0 +/- 2.2% in peripheral blood lymphocytes of normal controls (p less than 0.001). No significant differences were observed among the cell types of lung cancer examined (adenocarcinoma, squamous, small cell and large cell carcinoma) and no correlation among levels of soluble IL-2R and IL-2R+ cell in either pleural fluid or blood. Our results suggest that in patients with carcinomatous pleural effusions, T cell-mediated active immune mechanisms (IL-2/IL-2R system) against cancer cells are more active in pleural fluid than in peripheral blood.

Pilot Study on Transdiaphragmatic Thoracoscopic-Assisted Pleural Biopsy and Intrathoracic Washing Cytology for Stage IIIc Ovarian Cancer With Diaphragmatic Metastases

The significance of investigations of thoracic cavities as well as pleural biopsy and intrathoracic washing cytology through transdiaphragmatic thoracoscopy for stage IIIc ovarian cancer with diaphragmatic metastases was assessed as a prospective pilot study. Eligibility criteria were established to include patients with stage IIIc ovarian cancer in whom pleural effusions were not detected preoperationally, but prominent diaphragmatic metastases were observed when the abdomen was opened and those who submitted consent. Transdiaphragmatic thoracoscopy was performed after diaphragm stripping. Then, biopsy of the lesion suspected to be disseminated or the plural membrane of the thoracic opening was performed followed by washing cytology using physiological saline. Ten subjects were enrolled. Disseminated lesions were observed on plural membranes, and positive results were obtained in biopsy and washing cytology in 3 subjects. In addition, positive results were seen with biopsy alone in 2 subjects and with washing cytology alone in 2 subjects. Hence, a total of 7 subjects (70.0%) were up-staged to the level of stage IV. Postoperational complications were not observed in any of these cases. It was suggested that stage IIIc ovarian cancer with prominent diaphragmatic metastasis may advance to the level of stage IV from a clinical point of view even if carcinomatous pleural effusions are not detected pre-operationally. Therefore, it is thought that this operational method is useful in the management of progressive ovarian cancer.

Detection of tumour necrosis factor/cachectin in pleural effusion of patients with lung cancer

We have found that the pleural effusion obtained from a patient with lung cancer (adenocarcinoma) has cytotoxic activity against the patient's lung cancer cells. This finding occurred in the course of establishing a lung cancer cell line from the patient's pleural effusion. The cytotoxic factor was partially purified from the pleural effusion and characterized. It had cytotoxicity against L-929 mouse fibroblasts in the standard 18-h killing assay of tumour necrosis factor (TNF). By molecular sieving chromatography, the activity appeared at molecular weight of 50,000. This activity was completely blocked by a monoclonal antibody to TNF. From these results, we conclude that the cytotoxic factor in the pleural effusion is TNF. The concentration of TNF in the pleural effusion was 34.5 pg/ml by radioimmunoassay. In addition, we detected TNF activity and protein in two other cases of carcinomatous pleural effusion. Therefore, it would appear that in vivo TNF displays cytotoxic activity against cancer cells.

Strategy to augment the efficacy of immunotherapy for refractory breast cancer using trastuzumab combined adoptive cell therapy

3062 Background: Antitumor monoclonal antibody (mAb) is able to make molecular change in cancer cells and may also induce immune response via its Fc domain. CTLs selectively kill autologous tumor cells (AuTCs) is powerful for adoptive cell therapy of cancer. To investigate the immunological activity and anti-cancer effect in the combination therapy using the anti-HER2 Mab (trastuzumab, Tr) with adoptive cell therapy, a pilot clinical trial was performed for patients (pts) with refractory breast cancer. Methods: Fourteen pts with recurrent breast cancer were enrolled, they all had failed the conventional chemoradiotherapy. MLTC was performed using PBMC and primary- cultured AuTCs for 7–14 days. The generated cells were analyzed by Flow Cytometry, and the cytokine production and killing activity were evaluated by the ELISA and 51Cr-release assay. The cells were administrated by intra-peritumoral injection biweekly and Tr (2mg/kg) was infused every week. The treatments were repeated for 6 to 22 injections, and the total numbers of administered T cells were reached to ∼5.6 x 109. To assess the mechanism of Tr-effects, the antigen-loaded dendritic cells (DC) were examined in the phenotype, cytokine productions, and the ability to induce HER2 specific T cells in vitro. Results: After ∼ 2 weeks of MLTC, the derived cells, containing a preponderance of CD3+CD4+ or CD8+ T cells and CD56+ NK cells, showed significant degrees of cytotoxicity and IFN-γ production against AuTCs and HLA-matched allogenic tumor cell lines in 4 cases tested. The clinical response showed PR/SD/PD was 1/6/7 and adverse effect was tolerable. The tumor marker proteins (CEA, etc) were decreased in 5 pts. The carcinomatous pleural effusion was well controlled in 6 pts. In DC experiments, Tr-opsonized antigen-loaded DC enhanced the ability to induce CD8+ T cells specific for HER2-peptides in IFN-γ ELISPOT assay. Conclusions: Clinical responses were observed in pts with refractory breast cancer after receiving the adoptive cell therapy plus Tr. Our results suggested that the efficacy of immunotherapy might be augmented by anti-Her2 mAb, and it might be in part due to the involvement of mAb in the ability of DC cross-presentation followed by the enhancement of antitumor cellular immunity. No significant financial relationships to disclose.

Regulatory T cells and cytokines in malignant pleural effusions secondary to mesothelioma and carcinoma

Immunotherapy against a variety of malignancies, including pleural-based malignancies, has shown promise in animal models and early human clinical trials, but successful efforts will need to address immuno-suppressive factors of the tumor and host, particularly certain cytokines and CD4+ CD25+ regulatory T cells (Treg). Here, we evaluated the cellular and cytokine components of malignant pleural effusions from 44 patients with previously diagnosed mesothelioma, non-small cell lung cancer (NSCLC), or breast cancer and found significant differences in the immune profile of pleural effusions secondary to mesothelioma vs. carcinoma. Although a high prevalence of functionally suppressive CD4+ CD25+ T cells was found in carcinomatous pleural effusions, mesothelioma pleural effusions contained significantly fewer CD4+ CD25+ T cells. Activated CD8+ T cells in pleural fluid were significantly more prevalent in mesothelioma than carcinoma. However, there is clear patient-to-patient variability and occasional mesothelioma patients with high percentages of CD4+ CD25+ pleural effusion T cells and low percentages of CD8+ CD25+ pleural effusion T cells can be identified. Mesothelioma pleural effusions contained the highest concentrations of the immuno-suppressive cytokine transforming growth factcor (TGF)-beta. Thus, the contribution of cellular and cytokine components of immuno-suppression associated with malignant pleural effusions varies by tumor histology and by the individual patient. These results have implications for the development of immunotherapy directed to the malignant pleural space, and suggest the need to tailor immunotherapy to overcome immunosuppressive mechanisms in tumor environments.

Proinflammatory Cytokine Levels in Patients with Lung Cancer and Carcinomatous Pleurisy

Increased levels of interleukin-6 (IL-6) and interleukin-8 (IL-8) have been reported in various diseases, including lung cancer. The role of the soluble form of the IL-6 receptor (sIL-6R) remains to be explored. We therefore measured IL-6, IL-8 and sIL-6R in effusion fluid and blood serum of 10 lung cancer patients with carcinomatous pleurisy (5 men, 5 women, age 64.3 ± 4.4 years) by enzyme-linked immunosorbent assays. Serum levels of healthy individuals served as control. Concentrations of sIL-6R were much higher in serum compared to pleural effusion fluids of tumor patients (25,698 ± 1,993 vs. 9,438 ± 1,407 pg/ml; p < 0.0001). In contrast, IL-6 and IL-8 were found at high concentrations in carcinomatous pleural effusions in comparison to serum (IL-6: 964 ± 176 vs. 10.2 ± 1.3 pg/ml, p < 0.0001; IL-8: 319 ± 85 vs. 9.6 ± 9.6 pg/ml, p < 0.0001). The serum concentrations of IL-6 were not significantly increased in lung cancer patients (10.2 ± 1.3 pg/ml) in comparison to controls (7.3 ± 1.0 pg/ml). IL-8 was detected in the serum of only 1 patient and in low levels in the serum of controls (8.0 ± 1.5 pg/ml; all values are mean ± SEM). We conclude from this study that decreased levels of sIL-6R, but increased levels of IL-6 and IL-8, are found in pleural effusion fluid of patients with lung cancer and carcinomatous pleurisy. The low sIL-6R levels in the presence of high IL-6 levels in pleural effusions and the high sIL-6R levels in the presence of low IL-6 levels in serum may suggest a downregulation of sIL-6R expression or sIL-6R shedding in the presence of excessive amounts of IL-6.

Procoagulant activity of purified protein derivative-stimulated pleural effusion mononuclear cells in tuberculous pleurisy.

Mononuclear cells (MNC) generate cell-bound procoagulant activity (PCA) which shortens recalcification time after incubation with an antigen to which the donor has been sensitized. PCA has been demonstrated in various lung diseases, including exudative pleural effusions. To determine the value of measuring cell-bound PCA in the diagnosis of tuberculous pleural effusions we examined pleural effusion MNC of patients with tuberculosis (n = 19), congestive heart failure (n = 7), and carcinoma (n = 7). MNC were isolated, incubated in 0 or 10 micrograms/ml purified protein derivative (PPD) for 15 min and for 20 h, and recalcification time determined. Incubation with thromboplastin was used as control. The recalcification times in serum incubated for 15 min varied within a wide range, the mean values were longest for tuberculous effusion MNC, incubation for 20 h increased variation. Incubation of cells for 15 min with thromboplastin led to a decrease of mean recalcification time in tuberculous (p < 0.001) and heart failure (p < 0.05), and with no significance in carcinomatous effusions. Incubation with PPD led to decrease of recalcification time which was not significant. Comparisons of the mean relative recalcification times after PPD incubation showed that tuberculosis differed from lung cancer (p < 0.001), lung cancer from heart failure (p < 0.05), but not heart failure from tuberculosis. We conclude from our study that pleural effusion MNC express spontaneous PCA in vitro which is strongest in carcinomatous pleural effusions. Incubation of MNC with thromboplastin and less discernable with PPD leads to an increase in PCA which is more pronounced in tuberculous pleural effusions. However, due to substantial intersubject variability and overlap between the study groups, this test does not allow reliable differentiation of tuberculous from other MNC rich pleural effusions.

Elevated progastrin-releasing peptide(31-98) concentrations in pleural effusions due to small-cell lung carcinoma.

Progastrin-releasing peptide (ProGRP) is a specific and actively secreted product from small-cell lung carcinoma (SCLC) cells. Recently, an enzyme-linked immunosorbent assay, which uses monoclonal and polyclonal antibodies to recombinant proGRP(31-98), a common region of ProGRP, was established. We measured concentrations of ProGRP (31-98), neuron-specific enolase (NSE) and carcinoembryonic antigen (CEA) in carcinomatous and infectious pleural effusions. Significantly increased ProGRP(31-98) and NSE values were found in carcinomatous pleurisy due to SCLC compared to the other carcinomatous pleurisy and infectious pleurisy. CEA values were significantly increased in carcinomatous pleural effusions compared with those in infectious effusions. The ProGRP(31-98) values were not correlated to NSE values in carcinomatous pleurisy due to SCLC. The determination of ProGRP(31-98) in pleural effusions will be helpful for diagnosing carcinomatous pleurisy due to SCLC.

Immunocytochemical characterization of malignant mesothelioma and carcinoma metastatic to the pleura: IOB3--a new tumor marker.

We have tried to find a reliable panel of markers that would allow distinction between mesotheliomas and carcinomas metastatic to the pleura. In a prospective study, we evaluated 54 pleural effusions: In 27 of the patients, a diagnosis of histologically proven metastatic carcinoma was subsequently established, 7 patients had biopsy-proven malignant mesotheliomas and 20 had benign, reactive effusions whose benign etiologies were established by more than 2 years clinical follow-up. The MAb (monoclonal antibody) IOB3 proved to be diagnostic for carcinomas in all 27 cases (100%), whereas CEA (carcinoembryonic antigen) expression was found in only 22 out of 27 (81%). None of the malignant mesotheliomas, nor benign reactive mesothelial cells reacted with these two markers. All carcinomas, as well as one malignant mesothelioma, reacted with the MAb HEA125. Antibodies against 12 single cytokeratins, vimentin, and EMA (epithelial membrane antigen) were not helpful in the differentiation between malignant mesotheliomas and malignant carcinomatous pleural effusions. We conclude that adding the antibody IOB3 to the CEA assay should allow a reliable differentiation between these two entities.

Usefulness of Soluble Interleukin 2 Receptor in Differentiating Tuberculous and Carcinomatous Pleural Effusions

To evaluate the usefulness of soluble interleukin 2 receptor (sIL-2R) in differentiating tuberculous and carcinomatous pleural effusions. Levels of sIL-2R were measured simultaneously in plasma and pleural fluid in 111 patients with pleural effusions of unknown causes. The causes of pleural effusions were tuberculosis in 42 cases, carcinoma in 41 cases, pneumonia in 18 cases, and heart failure in 10 cases. In all groups of patients, sIL-2R levels were significantly higher in pleural fluid than in plasma. Plasma and effusion levels of sIL-2R were highest in the patients with tuberculosis, followed by those with carcinoma, pneumonia, and heart failure. Levels were significantly higher in the tuberculous group than in the carcinomatous group. To differentiate tuberculous from carcinomatous effusions, the highest plasma and effusion sIL-2R values obtained from the carcinomatous group were chosen as cutoff points (test specificity, 100%). The sensitivities of plasma and effusion sIL-2R levels in differentiating tuberculous and carcinomatous pleural effusions were 50% (21 of 42 samples) and 81% (34 of 42 samples), respectively. Although increased plasma and pleural fluid levels of sIL-2R should not be viewed as a diagnostic test specific for tuberculous pleural effusion, sIL-2R level appears to be clinically useful as a biochemical marker to differentiate tuberculous and carcinomatous pleural effusions.

Usefulness of soluble interleukin 2 receptor in differentiating tuberculous and carcinomatous pleural effusions

Usefulness of soluble interleukin 2 receptor in differentiating tuberculous and carcinomatous pleural effusions

Carcinomatous and tuberculous pleural effusion usefulness of cell markers quantitation

Carcinomatous and tuberculous pleural effusion usefulness of cell markers quantitation

Antipurified-Protein-Derivative Antibody in Tuberculous Pleural Effusions

Using ELISA, we studied anti-PPD antibody values in 31 tuberculous and 39 carcinomatous pleural effusions. Mean ODI values of anti-PPD IgG, IgA and IgM antibodies in tuberculous pleural effusions were higher than those in carcinomatous pleural effusions (p less than 0.01 in IgG and IgA, p less than 0.05 in IgM antibodies). We also analyzed the detected antigens in PPD and BCG whole cell fraction recognized by IgG antibody in tuberculous pleural effusions. Among heteromolecular components, the antigen band from 25 to 40 Kd region was most frequently stained. This antigen was heat-stable, contained sugar components, and included no disulfide bonds.

Discrimination of tuberculous from carcinomatous pleural effusion by biochemical markers: adenosine deaminase, lysozyme, fibronectin and carcinoembryonic antigen.

We measured adenosine deaminase (ADA), lysozyme, fibronectin and carcinoembryonic antigen (CEA) in the pleural fluid of tuberculous and carcinomatous pleural effusion in order to discriminate these two groups. Tuberculous pleural effusion had significantly higher levels of ADA and lysozyme than did carcinomatous effusion. When ADA activity of more than 33 IU/l is considered, diagnostic tests of tuberculous effusions showed a sensitivity of 100%, specificity of 95% and accuracy of 96%. A pleural fluid/serum ADA ratio (pl-ADA/s-ADA) above 1.1 was found in 100% of tuberculous and in 53% of carcinomatous effusions (sensitivity 100%, specificity 47%, diagnostic accuracy 70%). A lysozyme level above 12 micrograms/ml, selected as the discriminating limit, was found in 100% of tuberculous and in 17% of carcinomatous effusions (sensitivity 100%, specificity 83%, diagnostic accuracy 88%). Pleural fluid/serum lysozyme ratio (PL/SL) was also valuable in the discrimination of these two groups. When the cut-off level of 1.2 was considered, diagnostic tests of tuberculous effusions showed a sensitivity of 100%, specificity of 88% and accuracy of 93%, respectively. The mean fibronectin concentration in pleural fluid with tuberculous effusion was significantly higher than that in carcinomatous effusion, but there was a marked overlap between these two groups. On the other hand, CEA was significantly higher in carcinomatous effusions than in tuberculous effusions. At a cut-off level of 5 ng/ml, 53% of patients with carcinomatous effusion showed elevated pleural fluid CEA levels, while none of the tuberculous effusion did (sensitivity 53%, specificity 100%, diagnostic accuracy 65%).(ABSTRACT TRUNCATED AT 250 WORDS)

Generation of cytotoxic factor by human large granular lymphocytes during interaction with autologous tumor cells: lysis of fresh human tumor cells.

Large granular lymphocytes (LGL) obtained by centrifugation (Percoll gradient) of blood from patients with carcinomatous pleural effusions and solid tumors lysed autologous, freshly isolated tumor cells in a 4-hour 51Cr-release assay. When cocultured with autologous tumor cells, LGL released soluble cytotoxic factor(s), large granular lymphocyte-derived cytotoxic factor (LGL-CF). In contrast, small T lymphocytes were unable to kill autologous tumor cells or to produce a cytotoxic factor. The LGL-CF demonstrated cytotoxicity against autologous fresh tumor cells but also against allogeneic fresh tumor cells in a 48-hour microcytotoxicity assay and in an 18-hour 51Cr-release assay in the presence of dactinomycin. Binding of LGL-CF to autologous tumor cells occurred within 2 hours and reached the maximum by 6 hours; this binding was sufficient to cause subsequent lysis of the target cells without the continued presence of LGL-CF. Neither the supernatants produced by culture of LGL alone nor the lysates of LGL had detectable cytolytic activity. In addition, treatment of LGL with cytochalasin A inhibited both direct cell-mediated autologous tumor lysis and generation of LGL-CF. Production of LGL-CF required active cell metabolism and protein and RNA syntheses in LGL, but not DNA synthesis. Addition of dactinomycin to LGL-CF in assays augmented the lysis. LGL-CF was stable at 56 degrees C, but it was reduced at 70 degrees C and destroyed at 100 degrees C. Treatment of LGL-CF with trypsin or proteinase K reduced or abrogated the lytic effect, respectively, while the lytic effect was not affected by papain, catalase, or superoxide dismutase. These results indicate that during interaction with autologous tumor cells, LGL release a soluble cytotoxic factor that mediates lysis of fresh human tumor cells.

Autologous mixed lymphocyte-tumor reaction and autologous mixed lymphocyte reaction. II. Generation of specific and non-specific killer T cells capable of lysing autologous tumor.

The specific and non-specific nature of autotumor cytotoxicity induced in autologous mixed lymphocyte-tumor culture (AMLTC) and autologous mixed lymphocyte culture (AMLC) was studied in patients with carcinomatous pleural effusions. Small- and medium-sized blood lymphocytes that were isolated by centrifugation on discontinuous Percoll gradients did not lyse autologous, freshly isolated effusion tumor cells. In vitro activation of the small lymphocytes, but not of the medium lymphocytes, with autologous tumor cells generated cytotoxic potential restricted to autologous tumor. When stimulated with autologous non-malignant non-T cells, the medium lymphocytes, but not small lymphocytes, were triggered to cytotoxicity that acted not only on autologous tumor cells but also on allogeneic tumor cells, T blasts, and tumor cell lines. Experiments using monoclonal antibodies (MAb) and complement (C') showed that both types of killer cells were CD2+ CD3+ CD16- T cells. Autotumor cytotoxicity developed in AMLTC was mediated by the CD4- CD8+ T cell subset in 6 of 9 cases and the CD4+ CD8- subset in the other 3 cases. In contrast, cytotoxicity induced in AMLC was exerted exclusively by the CD8+ subset. The enrichment of blasts from cultured T cells on discontinuous density gradients enhanced autotumor killing activity, with no reactivity recorded for blast-depleted, resting T cells. Addition of mitomycin-C-treated large granular lymphocytes (LGL) to AMLTC abolished the induction of autotumor killer cells, whereas non-specific killer cells were generated in AMLC irrespective of the presence of LGL. These results indicate that stimulation of autoreactive T cells in AMLTC and in AMLC could induce 2 distinct types of autotumor killer cells.