Mononuclear-cell infiltration in ovarian cancer. I. Inflammatory-cell infiltrates from tumour and ascites material

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Summary.-Macrophages have been isolated from ascitic and collagenase -dispersed tumours from patients undergoing surgery for ovarian cancer. Macrophages were present in varying proportions in both sites, though the ratio of macrophages to tumour cells was higher in ascites. Marked variation in size (as detected by sedimentation velocity) and cytochemical markers in the macrophages was noted. Highly enriched macrophage fractions were isolated from the ascites and collagenasedispersed solid tumours by a combination of sedimentation velocity and selective EA RFC or adherence techniques. Suppressor activity in the PHA assay was detected in tumour macrophages (4/10 giving>50% inhibition), ascitic macrophages (1/15) and blood monocytes (2/7). Lymphocyte fractions from tumours were unresponsive to PHA and failed to suppress the blood response. Suppressor activity was also present in the purified tumour-cell fraction of 6/14 patients.ADCC activity was tested in a few patients. When the activity was determined against the SB target cells, tumour-derived macrophages were inactive, whereas the ascitic fraction showed low but significant activity which averaged much lower than patient blood values. The ADCC assays carried out with the CRC target cell indicated activity within the range of patient blood values in 4/4 ascites and 2/4 tumour macrophage fractions.Cytotoxicity was also assessed against co-purified autologous tumour cells. Although activity was detected in many of the tests, the results seemed to reflect target cell sensitivity. There appeared to be a correlation between cytotoxicity with test macrophages and normal blood mononuclear cells.The results indicate that the cytochemical heterogeneity and the variation in size between macrophage fractions is associated with a spectrum of activities.WE HAVE INITIATED a study of immune competence of the various inflammatory cell types infiltrating primary solid and ascitic ovarian tumours. In the first paper (Haskill et al., 1982a) we outlined the methods used to isolate these cells and characterized the cell markers associated with infiltrating cells from these tumours. Two classes were characterized; 1 sedimented at < 6 mm/h and was similar in size to most blood mononuclear cells; the other was composed of larger, strongly adherent macrophages distinct from blood monocytes, which sedimented with the tumour cells. In the second communication (Haskill et al. 1982b) we investigated effector-cell functions (PHA, ADCC and NK) associated with blood and bloodequivalent inflammatory cells present in both ascitic and solid tumours. The data indicated that all tumour-derived effector-cell tests were markedly depres-

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Mononuclear-cell infiltration in ovarian cancer. III. Suppressor-cell and ADCC activity of macrophages from ascitic and solid ovarian tumours

Haskill¹,

Koren²,

Becker³

et al. 1982

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“…Because similar dose-response rela- Tumour (TuL) and ascites (AscL) mononuclear cell fractions were obtained by sedimentation-velocity separation (Haskill et al, 1982). The mean E RFC values for each cell source are given in Fig.…”

Section: Mitogen Responsementioning

confidence: 99%

“…In the accompanying paper (Haskill et al, 1982) we outline the methods of isolating these cells, and their cell markers. Two classes were characterized: those sedimenting less than 6mm/h and similar in size to most of the blood cells and a series of larger macrophages cytochemically distinct from blood monocytes, which sediment with tumour cells.…”

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confidence: 99%

Mononuclear-cell infiltration in ovarian cancer. II. Immune function of tumour and ascites-derived inflammatory cells

Haskill¹,

Koren²,

Becker³

et al. 1982

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Summary.-Mononuclear cell fractions were isolated from blood, ascites and solid tumours of patients undergoing surgery for Stages III and IV PHA responses of patient blood and ascites fractions were about half that of normal blood. Tumour-infiltrating lymphocytes (TIL) were less than 10% as responsive as normal blood. The depressed PHA responses of the TIL were not due to the presence of a suppressor cell population. NK activity of patient blood was less than that of normal blood, but not as much as the ascites or TIL cells. The activity of the ascites-derived lymphocytes was enhanced by treatment with interferon. ADCC activity against both CRBC and SB cells was normal or higher than controls in patient blood, and depressed in the ascites-derived fractions. TIL responded to <10% of the patient blood values.The results indicate a lack of response by ascitic and TIL cells in assays dependent on FcR -bearing effector cells and a greater loss of PHA-reactive cells from the tumour than from blood and ascites. These data could result from intratumour inactivation, or a failure of the particular subset to localize either In the ascites or the tumour site.

“…Methods such as cell electrophoresis or centrifugal elutration (Meistrich et al, 1977) require specialized equipment not accessible to all laboratories. Velocity sedimentation at 1 g requires special equipment, a large volume of cells and reagents, and is time consuming (Haskill et al, 1982). Density gradient separation has proven to be a relatively simple, effective method for separating malignant from stromal cells.…”

mentioning

confidence: 99%

Percoll density gradient separation of cells from human malignant effusions

Hamburger¹,

Dunn²,

White³

1985

Summary A simple method is described for the separation of cells derived from effusions of patients with adenocarcinomas in discontinuous density gradients of Percoll. After separation, cells from different fractions were analyzed by morphologic, histochemical and immunologic criteria. Total cell recovery from 27 experiments was 67+4%. Macrophages (82%) were recovered in the intermediate density fraction (1.056-1.067gml-1) with a purity of 90%. Recovered lymphocytes (98%) were found in the high density fraction (1.067-1.077gml-1) with a purity of 92%. The majority of the lymphocytes recovered were T cells. Malignant adenocarcinoma cells (90%) were recovered in the lowest density fractions (up to 1.056gml-1) with a purity of 79%. Use of effective cell separation procedures should facilitate the analysis of the functional capacities of both normal and neoplastic cells derived from human malignant effusions.