Normal Adherent Cells Research Articles

T-lymphocytes phenotype and functions in patients with head and neck cancer.

We studied the phenotype of T-lymphocytes isolated from 18 patients with head and neck cancer, their capacity to express Ia antigens upon activation by lectins in vitro, their capacity to function either as responder or stimulator cells in autologous mixed lymphocyte reaction, and their capacity to cooperate with the normal adherent suppressor cells (NASC). The T-lymphocytes isolated from these patients have several functional defects including an impaired capacity to activate allogeneic lymphocytes in mixed lymphocyte reactions (MLRs), a lack of a proliferative capacity in autologous MLRs, an impaired sensitivity to inhibition by NASC, and an impaired capacity to express Ia antigens upon activation by mitogens in vitro. These data indicate that, in patients with head and neck cancer, immune function is characterized by a defect in T-lymphocytes functions which concerns the process of cell to cell cooperation.

Impaired gamma interferon production by cells from patients with lymphoproliferative disorders of mature T and NK cells.

We investigated interferon (IFN) production by peripheral blood mononuclear cells from four patients with chronic OKT4 T-lymphocytic leukaemia and three patients with abnormal expansions of granular lymphocytes. No spontaneous production of IFN-gamma was found in supernatants of cultures from both patients and normal controls. However, whereas the enzyme galactose oxidase or staphylococcal enterotoxin B was able to induce IFN-gamma production by normal cells, no production could be obtained in the cells under study. The possibility that this lack of production might have been attributed to an excess of N-acetylneuraminic acid masking galactose residues or to a defect of monocyte accessory cells was ruled out either by pre-treating the cells with neuraminidase or by adding normal adherent cells to the cultures, both of which resulted in a lack of production. On the contrary, the calcium ionophore A23187 (considered to act as a second specific step, following oxidation of galactose residues, toward genetic derepression of IFN-gamma) induced considerable IFN-gamma production in all the three tested patients. It can be concluded that, although in rare cases, as previously reported by other authors, cells from patients with T or NK lymphoproliferative disorders may spontaneously produce IFN-gamma, this is not a general mechanism that underlies the disease. In fact, in all our cases a defect of IFN-gamma production was found. This defect seems due to an alteration at the membrane level of the galactose-containing glycoproteins and can be restored by induction with a calcium ionophore.

Defective monocyte function in patients with systemic lupus erythematosus

Peripheral blood adherent cells from patients with systemic lupus erythematosus (SLE) were shown to have markedly reduced phagocytic activity as compared to normal adherent cells or those from non-SLE patients receiving corticosteroid therapy. Both resting and phagocytosing monocytes showed decreased hexose monophosphate shunt and glycolytic activity. Mononuclear cells from SLE patients showed grossly impaired proliferative activity after NaIO 4 activation. Furthermore, addition of SLE adherent cells to normal adherent cell-depleted lymphocytes decreased [ 3H]thymidine incorporation of the latter cells following NaIO 4 treatment. Addition of normal adherent cells to SLE lymphocytes corrected the previous defect, indicating that an adherent abnormality is responsible for the defect in SLE mononuclear cell proliferation to NaIO 4 activation.

Immunoregulatory lymphokines of T hybridomas from AIDS patients: constitutive and inducible suppressor factors.

Supernatants derived from peripheral blood mononuclear cell cultures of certain patients with the acquired immunodeficiency syndrome (AIDS) or its prodromes have the capacity to block T cell-dependent immune reactivity in vitro. T cells derived from a patient positive for antibody to the lymphadenopathy associated virus ( LAV ), and elaborating high titers of these soluble suppressor factors, were fused to a mutagenized clone of the human T lymphoblastoid cell line KE37 . Molecules capable of profoundly depressing T cell-dependent polyclonal antibody production and DNA synthetic responses, either directly or after incubation with normal adherent cells, were isolated from stable hybrid clones.

Role of normal adherent cells in the regulation of the autologous mixed lymphocyte reactions in humans.

The effect of normal adherent suppressor cells on the blastogenesis of human T lymphocytes in the mixed lymphocyte reaction (MLR) was studied in both allogeneic and autologous combinations. Non-T cells and Ia+ T lymphocytes were used as stimulator cells in both allogeneic and autologous MLR. The addition of adherent cells to the stimulators inhibited blastogenesis of T lymphocytes in both types of MLR when the stimulator population was made up of non-T lymphocytes but did not interfere with blastogenesis when Ia+ T lymphocytes were used as stimulator cells. The present data indicate that the T lymphocytes able to respond to Ia+ T cells (in the MLR, autologous or allogeneic) may be different from those which respond to non-T lymphocytes or may be less sensitive to the regulatory function of normal adherent cells.

Monocyte-induced inhibition of lymphocyte response to phytohaemagglutinin in progressive systemic sclerosis.

In patients with progressive systemic sclerosis (PSS) lymphocyte responses to phytohaemagglutinin (PHA) are abnormal (27.2 +/- 3.5 X 10(-3) counts per minute (cpm) versus 69.8 +/- 4.4 X 10(-3) for normal persons, p less than 0.005). Removal of adherent peripheral blood mononuclear cells improves the response of PSS lymphocytes (42.3 +/- 3.4 X 10(-3) cpm, 155% of control) but diminishes the response of normal lymphocytes (60.3 +/- 5.9 +/- 10(-3), 86% of control). Supernatant fluids from cultures of PSS unfractionated and adherent cells depress normal T cell response to PHA (64% and 55% of control respectively), but supernatant fluids from normal unfractionated and adherent cells do not (104% and 101% of control). Supernatant fluids of PSS and normal adherent cells, cultured in the presence of indomethacin, are not inhibitory to normal T cells (109 +/- 15% and 124 +/- 14% of control respectively). Supernatant fluids from PSS patients are more inhibitory than comparable fluids from patients with systemic lupus erythematosus (60 +/- 8% of control versus 80 +/- 5% of control). These data support the hypothesis that cellular immunity is abnormal in patients with PSS and indicate that adherent mononuclear cells mediate at least one component of the abnormality via an indomethacin-sensitive mechanism.

Abnormalities of immunocompetent cells in primary biliary cirrhosis.

Primary biliary cirrhosis (PBC) is a chronic cholestatic progressive liver disease frequently associated with various immunological abnormalities. We have studied the influence of normal and PBC adherent cells (AC) on peripheral blood lymphocytes (PBL) colony growth both in liquid and soft agar culture. Co-culture experiments with different combinations of AC and PBL of PBC patients and normal donors provided evidence that PBC adherent cells function abnormally. However, this impaired function is not the sole factor involved, since the number of colonies formed by PBC lymphocytes in the presence of normal AC was twice the normal value. The possibility that this increase is correlated with the state of activation of T cells is discussed in the light of the high number of DR-positive T cells found in the PBC patients studied. Finally, none of these immunological abnormalities was related to the stage of the disease.

Murine cytomegalovirus-induced immunosuppression.

The mechanism of murine cytomegalovirus-induced immunosuppression was investigated by examining the roles played by lymphocytes and adherent cells derived from spleens of infected SWR/J mice. As few as 100 infected cells per spleen were correlated with complete abrogation of mitogen responses at 4 and 5 days after infection. In a series of cell mixing experiments it was shown that the deficiency in infected spleens was due partly to the adherent cells, which apparently secreted an immunosuppressive factor, and partly to the infected lymphocytes, which upon exposure to this factor could no longer respond to concanavalin A presented to them by normal adherent cells.

Increased Sensitivity to Normal Adherent Suppressor Cells in Untreated Advanced Hodgkin’s Disease

AbstractMononuclear leukocytes isolated from the peripheral blood of 10 untreated patients with advanced stages of Hodgkin’s disease and 26 normal volunteers were stimulated with allogeneic cells from normal donors in a one-way mixed lymphocyte culture. When the patients’ cells were stimulated by an increased number of normal cells, the baseline mixed lymphocyte culture was suppressed 77.1% ± 4.1%. In contrast, when the normal responder cells were cultured with increased numbers of normal stimulator cells, mean suppression was only 46.6% ± 4.4% (p < 0.001). The removal of phagocytic cells from the high concentration of normal stimulators totally abolished the suppression observed with the patients’ cells, resulting in a mean increase in stimulation of 84% above that found in the baseline culture. The suppression could also be reversed by depletion of adherent cells from stimulator leukocytes. When adherent and nonadherent stimulator cells were recombined, significant suppression of proliferation was again observed. The increased suppression was not caused by the presence of autologous plasma in patients’ cultures, since similar results were obtained utilizing AB serum. The addition of indomethacin to the cultures only partially reversed the suppression observed with the patient’s cells. These studies demonstrate that mononuclear leukocytes from the peripheral blood of untreated patients with advanced stages of Hodgkin’s disease have increased sensitivity to a normal adherent suppressor cell system.

Increased sensitivity to normal adherent suppressor cells in untreated advanced Hodgkin's disease

Mononuclear leukocytes isolated from the peripheral blood of 10 untreated patients with advanced stages of Hodgkin's disease and 26 normal volunteers were stimulated with allogeneic cells from normal donors in a one-way mixed lymphocyte culture. When the patients' cells were stimulated by an increased number of normal cells, the baseline mixed lymphocyte culture was suppressed 77.1% +/- 4.1%. In contrast, when the normal responder cells were cultured with increased numbers of normal stimulator cells, mean suppression was only 46.6% +/- 4.4% (p less than 0.001), The removal of phagocytic cells from the high concentration of normal stimulators totally abolished the suppression observed with the patients' cells, resulting in a mean increase in stimulation of 84% above that found in the baseline culture. The suppression could also be reversed by depletion of adherent cells from stimulator leukocytes. When adherent and nonadherent stimulator cells were recombined, significant suppression of proliferation was again observed. The increased suppression was not caused by the presence of autologous plasma in patients' cultures, since similar results were obtained utilizing AB serum. The addition of indomethacin to the cultures only partially reversed the suppression observed with the patients' cells. These studies demonstrates that mononuclear leukocytes from the peripheral blood of untreated patients with advanced stages of Hodgkin's disease have increased sensitivity to a normal adherent suppressor cell system.

Increased Susceptibility to Tumor Cell Immunosuppressive Effect in Tumor-Bearing Mice&lt;xref ref-type="fn" rid="FN2"&gt;2&lt;/xref&gt;

Compared to normal hosts, tumor-bearing BALB/c mice were more susceptible to the immunosuppressive effect of tumor cells. At least a tenfold increase was found in the susceptibility mediated by a population of radioresistant spleen adherent cells (AC). The experiments were performed by the study of the suppressive effect of tumor cells on the generation of cytotoxic T-cells in allogeneic mixed lymphocyte culture reactions and allogeneic mixed lymphocyte tumor cell culture reactions. Fewer tumor cells were needed to suppress the T-cell-mediated cytotoxic responses of tumor bearers compared to normal hosts. A normal spleen population could be made to react like the tumor-bearing host by first depletion of its normal macrophages and then reconstitution with spleen AC from tumor bearers. Conversely, reconstitution of the macrophage-depleted tumor bearer's spleen with normal spleen AC made the tumor bearer react like the normal host. Furthermore, tumor cells were needed to trigger the spleen AC of the tumor bearer to fully exert their effect.

Suppressor adherent phagocytic cells in solid tumors: A postulated escape mechanism

A localized graft-versus-host reaction was used to assess the cellular immunity of mice supporting progressive growth of an allogeneic tumor. It was found that early in tumor development anti-tumor cellular-mediated immunity was hyper-responsive, and that adherent phagocytic cells isolated from the neoplasm would inhibit the specific immunological response. Based on these observations and indirect inference it was postulated that a major factor in the success of an immunogenic tumor is the coterminous development of a normal adherent phagocytic cell population which specifically nullifies, in situ , cellular-mediated anti-tumor immunity.

Influence of Malaria Infection on the Elaboration of Soluble Mediators by Adherent Mononuclear Cells

MALARIA RESULTS IN TWO SEEMINGLY PARADOXICAL PERTURBATIONS OF THE IMMUNE RESPONSE: polyclonal B-cell activation and immunosuppression. To determine what immunoregulatory role mediators secreted by adherent cells might play in these alterations, we cultured adherent cells from uninfected mice and from mice at different times during infection with Plasmodium berghei or P. yoelii. Culture supernatants obtained from these cells were tested for their ability to enhance the in vitro proliferative responses of thymocytes to suboptimal concentrations of concanavalin A or to inhibit the mitogen-stimulated proliferation of normal spleen cells. Supernatants obtained from adherent cells of mice early in infection (days 1 to 3) contained significantly elevated levels of enhancing activity which on Bio-Gel P-100 chromatography resembled lymphocyte-activating factor. Later in infection (days 4 and 5), these supernatants contained inhibitory activity. Normal adherent cells, when cocultivated in vitro with parasitized erythrocytes, ingested parasite debris and were stimulated to produce the enhancing factor. At high parasite/adherent-cell ratios, cells elaborated an inhibitory factor. These findings suggest that during malaria, adherent cells are converted from a nonspecific helper role to a nonspecific suppressor role. This modulation in function may be due to the direct interaction between adherent cells and parasitized erythrocytes.

In vitro and in vivo studies of the target cell for dialyzable leukocyte extracts : Evidence for recipient specificity

The effects of dialyzable leukocyte extracts (DLE), assumed to contain the so-called “dialyzable transfer factor,” were tested in vivo and in vitro. In vivo, DLE was used to treat a patient with an apparent monocyte deficiency, before and after treatment with supernatants of buffy-coat cells from normal individuals. In vitro, white blood cells from the same patient and from normal controls were used as target cells for DNA synthesis assays. The normal cells were either cells isolated on a Ficoll-Isopaque gradient (FC) or FC that were nonadherent to cotton after incubation for 45 min at 37°C (NAC). These cells were tested (a) in the presence or absence of DLE and in the presence or absence of antigens to which the donors of the DLE batches were sensitive; (b) in the presence or absence of DLE or antigen, with supernatants from cultures of normal FC (SFC); (c) in the presence or absence of DLE or antigen, with supernatants from nonadherent cells alone (SNAC). DLE was also tested in vitro in a leukocyte migration inhibition factor (LIF) assay with and without antimonocyte serum added to the system. The results, both in vivo and in vitro, suggest that normal adherent cells or a factor(s) released by these cells is required for the effects of DLE. The importance of considering the condition of the patient (recipient specificity) in any theoretical attempt to explain the mechanism and possible specificity of action of DLE is discussed.

Decreased phagocytosis by peritoneal macrophages from BCG-treated mice: Induction of the phagocytic defect in normal macrophages with BCG in vitro

Mice treated up to 31 weeks previously with intraperitoneal BCG yielded peritoneal macrophages with decreased phagocytosis of starch granules, latex beads, graphite dust and formalinized Listeria monocytogenes, with or without opsonin, compared to macrophages from untreated mice. These assays were selected to allow quantitative determinations of the rate or extent of particle uptake under nonrate-limiting conditions. Phagocytosis could be depressed to a similar degree by the prior addition of either starch granules or BCG to normal adherent peritoneal cell cultures in vitro. However, with these two particles, two different mechanisms of inhibition of subsequent phagocytosis appeared to be at work. Inhibition of phagocytosis by prior exposure to large amounts of starch granules appeared to consist largely of mechanical interference, as if through the preemption of intracellular space. In contrast, inhibition by prior uptake of BCG occurred with very small amounts of BCG and appeared gradually with time after uptake of BCG. The ability of a “macrophage activating agent” to inhibit selected functions of parasitized cells may help to explain some of the discordant results obtained by others studying phagocytosis by “activated” macrophages. Such agents may simultaneously enhance some macrophage functions and depress others.

Resistance to Tumor Growth Mediated by Listeria Monocytogenes: Collaborative and Suppressive Macrophage-Lymphocyte Interactions in Vitro

Spleen (SC) and peritoneal cells (PC) from C57BL/6 mice immune to Listeria monocytogenes (LM) have a marked cytotoxic effect on the in vitro growth of B-16 melanoma. Cytotoxicity to B-16 is mediated by the interacting semsitized thymus-derived (T) lymphocytes and macrophages (Mchi) or adherent cells. Separation of immune SC or PC into their adherent and nonadherent components, depletion of the adnerent cells or elimination of the theata-bearing cells from an enriched T cell population all result in the abrogation of cytotoxicity, whereas, recombination of T cells and Mchi re-establishes this effect. Three modes of Mchi/T cell interactions occur: 1) LM immune peritoneal or splenic T cells interact or synergize with their respective adherent cells to become cytotoxic to B-16; 2) combination of peritoneal Mchi and splenic T cells are noncollaborative and therefore noncytotoxic; 3) the cooperative effect of spleen T cells with Mchi are suppressed or inhibited by addition of peritoneal Mchi. Splenic Mchi, however, are neither uncooperative nor inhibitory when interacting with peritoneal T cells. Normal adherent cells can substitute for immune cells in all the cell-cell interactions with similar end results. These results demonstrate that specifically sensitized T cells interact with Mchi for expression of LM-induced nonspecific tumor cell destruction in vitro, however, such interactions may also be noncooperative or suppressive, thus resulting in tumor cell proliferation.

Strong Expression of Fc-Receptors on Leukemic Cells in Hairy Cell Leukemia

The neoplastic cells of a patient with hairy cell leukemia were found to express a high Fe-receptor activity on their surface. When the Fc-receptors were determined quantitatively by measuring the uptake of 125I-labelled aggregated IgG under saturation conditions, it was found that the hairy cells bound approximately 3 times more aggregated IgG than normal adherent mononuclear cells. The bound 125I-Agg. IgG was evenly distributed on the cell surface when the cells were labelled at 4°C and was rapidly redistributed in form of a cap at one cell pole during incubation at 20°C.

The Third Cell Type Required for the Immune Response of Spleen Cells in vitro

Abstract Cells that adhere to glass or plastic surfaces, obtained from spleens of nor mal and immunized mice, were compared for the ability to interact synergistically with nonadherent cells in the in vitro response to sheep erythrocytes. Adherent cells obtained from normal and immunized mice were equally x-ray resistant at several levels of x-irradiation and equally effective in enhancing the response of nonadherent cells. The plaque-forming cell response of cultures that contained dilutions of adherent cells from immunized mice were equivalent to responses of cultures containing corresponding dilutions of normal adherent cells. Adherent cells, therefore, did not appear to increase in number or change in function following in vivo antigenic stimulation. Adherent cells, whether derived from normal or immunized mice, were required for the in vitro response of non-adherent cells obtained from immunized animals. Spleen cell cultures obtained from immunized mice, however, required higher dilutions of non-adherent cells to interact synergistically with adherent cells. The enhanced responsiveness of non-adherent cells from immunized mice was consistent with an increase in number or change in function of non-adherent cells as a result of antigenic stimulation in vivo.