Impaired cytokine production in whole blood cell cultures of patients with gynaecological carcinomas in different clinical stages

Abstract

Summary The production of the cytokines IFN-gamma, IL-1-alpha, IL-2 and TNF-alpha was investigated in mitogen-stimulated, whole blood cell culturs from 239 untreated patients with primary gynaecological carcinomas (breast, cervix, ovary, endometrium), and 191 healthy female controls.The cytokines were measured in the 4-day post-inductional supernatants by a sensitive enzymoimmunological assay. In the blood cell cultures of all four groups of cancer patients, significantly lower values of IFN-gamma (P < 0.001), IL-2 (P < 0.01) and IL-1 alpha (P < 0.01) were found as compared to the controls, although lymphocyte and monocyte counts were almost identical.Grouping the tumour patients into different clinical stages we could show in the four groups of carcinomas a gradual depression of the cytokine production according to growing tumour burden.The host's response to malignant tumours is usually considered to be primarily a function of the cellular immune system and various tests have been established to measure the integrity of this system in cancer patients, such as hypersensitivity skin tests (Bolton et al., 1975;Evans et al., 1977), or determination of mitogen-induced lymphocyte proliferation in vitro (Dean et al., 1977;Zembala et al., 1977;Fritze & Dystant, 1984).As the cell-mediated immune response is controlled by a series of soluble cytokines, in recent years the measurement of these substances has also been used to determine the cellular immunological potency of tumour patients (Rey et al., 1983;Santos et al., 1985;Elsasser-Beile et al., 1991a Blood samples Ten ml of heparinised blood was taken from each patient and control person between 9 and 11 a.m. The samples, kept at room temperature, were used within 2 h. A 0.5 ml aliquot was removed for total and differential leukocyte counts.Whole blood cell culture (WBCC) Cultures were performed as previously described with a system for which optimal conditions and kinetics of cytokine production were established (Elsasser-Beile et al., 1991b).In brief, heparinised venous blood was diluted 1/10 with RPMI 1640 (Seromed, Berlin FRG), which was supplemented with 50 U ml-' Penicillin (Seromed) and 50 Sg ml-' Streptomycin (Seromed) and distributed in 0.5 ml aliquots in 12 mm polystyrol tubes. For stimulation, 10 ig ml-' phytohaemagglutinin (PHA Wellcome, Burgwedel, FRG) and 0.5-5 g.g ml-' Pokeweed mitogen (PWM, Sigma, Deisenhofen, FRG) were added.Incubation of the cell cultures was performed at 37°C in a humidified atmosphere of 5% CO2. After 4 days of culture without changes of medium 320 ftl of supernatant was removed from each tube to be assayed for cytokine levels.Determination of cytokines Enzyme-linked immunoassays (ELISA) were applied for qualitative and quantitative determinations of cytokines.These tests are based on the sandwich principle and are performed in one step. Since a correlation was found between assay performance of the naturally occurring cytokines and their recombinant counterparts, the latter were used as standards.Interferon-gamma-ELISA IFN-gamma ...

Altered maturation of peripheral blood dendritic cells in patients with breast cancer

“…A perturbation of the cytokine network characterised by low levels of IL-12 and normal or high levels of IL-10 has been previously reported in patients with neoplasms of various organs (O'Hara et al, 1998;Jacobs et al, 1998;Merendino et al, 1999); it has been suggested that this imbalance in IL-12 and IL-10 levels could explain the shift from type 1 to type 2 cytokine production by T lymphocytes known to occur in cancer patients (Elässer-Beile et al, 1993;Clerici et al, 1994;Clerici et al, 1997Clerici et al, , 1998. Actually, we found that T-helper lymphocytes from our cancer patients were polarised towards the Th2 phenotype, suggesting that altered production of regulatory cytokines by DCs may affect adaptive immune responses.…”

High Incidence of Transiently Appearing Complement-Sensitive Bone Marrow Precursor Cells in Patients with Severe Aplastic Anemia - A Possible Role of High Endogenous IL-2 in Their Suppression

“…It is thought to reflect defective tumor immunity in patients with solid tumors [30, 31]. Accordingly, tumor regression has been observed after treatment with IL-2 [32].…”

Selective suppression of cytokine secretion in whole blood cell cultures of patients with colorectal cancer

“…IL-2 and TNF-a are shown in Table 1. Others have pointed out a potential decrease in cytokine levels with an increase in age (Elsasser-Beile et al 1993a). However.…”

“…including our ow-n. have reported selective changes in the cvtokine profile secreted by lymphocytes and monocytes of carcinoma patients (Rev et al 1983: Elsasser-Beile et al 1993a.b: Fischer et al 1995: De Groote et al 1996. Such a reduction might at least in part occur as a consequence of soluble immunosuppressix e factors that are released bv tumour cells (Ebert et al 1990: Fischer et al 1994).…”