Suppressor Cell Precursors Research Articles

CD115− monocytic myeloid-derived suppressor cells are precursors of OLFM4high polymorphonuclear myeloid-derived suppressor cells

Myeloid-derived suppressor cells (MDSCs) consist of monocytic (M-) MDSCs and polymorphonuclear (PMN-) MDSCs that contribute to an immunosuppressive environment in tumor-bearing hosts. However, research on the phenotypic and functional heterogeneity of MDSCs in tumor-bearing hosts and across different disease stage is limited. Here we subdivide M-MDSCs based on CD115 expression and report that CD115− M-MDSCs are functionally distinct from CD115+ M-MDSCs. CD115− M-MDSCs increased in bone marrow and blood as tumors progressed. Transcriptome analysis revealed that CD115− M-MDSCs expressed higher levels of neutrophil-related genes. Moreover, isolated CD115− M-MDSCs had higher potential to be differentiated into PMN-MDSCs compared with CD115+ M-MDSCs. Of note, CD115− M-MDSCs were able to differentiate into both olfactomedin 4 (OLFM4)hi and OLFM4lo PMN-MDSCs, whereas CD115+ M-MDSCs differentiated into a smaller proportion of OLFM4lo PMN-MDSCs. In vivo, M-MDSC to PMN-MDSC differentiation occurred most frequently in bone marrow while M-MDSCs preferentially differentiated into tumor-associated macrophages in the tumor mass. Our study reveals the presence of previously unrecognized subtypes of CD115− M-MDSCs in tumor-bearing hosts and demonstrates their cellular plasticity during tumorigenesis.

Influence of the Ity/Lsh/Bcg gene on the development of suppressor cell precursors in the early phase of the infection of mice with Mycobacterium lepraemurium.

In vitro inducible suppressor cell precursors were detected in the spleen of BALB/c but not in DBA/2 mice infected intraperitoneally with 10(8) Mycobacterium lepraemurium bacilli, thus suggesting that their development is genetically controlled. Two pairs of mouse strains congenic at the Ity/Lsh/Bcg locus (BALB/c-C.D2 and B10.A-B10.A.Bcgr) were used to investigate whether this phenomenon is influenced by this gene known to control the relative susceptibility of mice to Myco. lepraemurium infection. This seems likely, as the detection of culture-induced suppressor activity was delayed for 5-6 weeks in C.D2 and B10.A.Bcgr mice infected intravenously with 10(4) Myco. lepraemurium bacilli. However, despite the retardation in the detection of suppressor cell precursors, the level of in vitro induced suppressor activity at onset in spleen cell suspensions of mice carrying the resistant allele was higher than in cell cultures derived from susceptible mice. As the resistant allele has a different effect when found on BALB/c or DBA/2 background, other genetic factors are apparently involved in the development of suppressor cell precursors. We finally observed that, in spleen cell cultures from intravenously infected Ity/Lsh/Bcg congenic mice on the BALB/c background, adherent and non-adherent cells were required in the inductive phase of suppressor cell development, whereas in vitro induced suppressor activity was found exclusively in the adherent cell fraction. Given these properties, we thus conclude that suppressor cell precursors detected in the spleen of these intravenously infected mice are similar to those previously observed in C3H mice infected intraperitoneally with a thousand times more bacilli.

Cyclophosphamide treatment antagonizes the in vitro development of Mycobacterium lepraemurium-induced suppressor cell precursors.

The in vitro-inducible maturation of splenic suppressor cell precursors detected during the early phase of Mycobacterium lepraemurium infection can be abrogated when a high dose of cyclophosphamide (Cy) is inoculated to infected mice 2 days before assay. The drug does not act directly on adherent suppressor cell precursors, but rather inhibits their activation by a non-adherent cell subset whose phenotype has not yet been elucidated. It was established by flow cytometry analyses, that despite a marked increase in the total number of splenic non-adherent cells following M. lepraemurium infection, the effect of Cy on Ia+, Thy-1+, CD4+ and CD8+ cells in infected mice was comparable to that observed in normal controls. It was not possible to determine the duration of the inhibiting effect of Cy on non-adherent regulatory cells, because the drug was itself inducing suppressor cells from 7 days after inoculation. By the time spleen cell suspensions were totally free of Cy-induced suppressor cells, infection-dependent suppressor cell precursors were once again detected, indicating that Cy treatment did not prevent their in vivo accumulation. Therefore, even though M. lepraemurium-induced adherent suppressor cell precursors are themselves fully resistant to Cy, their development is transiently abrogated by the drug, most probably through the impairment of a non-adherent cell subset regulating their maturation.

Early accumulation of suppressor cell precursors in the spleen of Mycobacterium lepraemurium-infected mice and analysis of their in vitro-induced maturation

Spleen cells harvested from mice infected intraperitoneally with M. lepraemurium 11-17 weeks prior to harvest acquired the capacity to inhibit concanavalin A (Con A) induced proliferation of normal spleen cells when precultured for up to 24 h in mitogen-free medium. The in vivo induced suppressor activity correlated with the length of the preculture period, the time post-infection and the infecting dose. These findings were interpreted as an indication that suppressor cell precursors accumulated in the spleen of infected mice during the early phase of the disease. The interaction of infection-dependent adherent suppressor cell precursors and infection-independent, non-adherent regulatory cells is necessary for the suppressor activity to develop. Both the cells which transmit the inductive signal and the precursor cells which mature into active suppressor cells are radiosensitive, whereas suppressor activity itself is a function of radioresistant adherent cells. Preculture of cells for a short period, before they were cocultured with Con A-stimulated normal spleen cells, allowed the detection of suppressor cells before they were deleterious to the infected host and also turned out to be a relevant in vitro model for characterization of suppressor cell development during M. lepraemurium infection.

Phenotypic characterization of two cell populations involved in the acquisition of suppressor activity by cultured spleen cells from Mycobacterium lepraemurium-infected mice.

The impairment of cellular immunity in mice infected with Mycobacterium lepraemurium was shown to correlate with the development of suppressor cells. We have previously reported that before suppressor activity is detectable in freshly harvested cell suspensions, suppressor cell precursors accumulate in the spleen of infected mice. Upon overnight culture in the presence of a regulatory cell subset, these precursor cells acquire the capacity to impair the concanavalin A (Con A)-induced proliferation of normal spleen cells. The purpose of this study was to determine the phenotype of the cells involved in this phenomenon. This was done by following the development of suppressor activity in spleen cell suspensions depleted of defined cell subsets of the adherent or the non-adherent cell fractions with selected MoAbs and immunomagnetic beads or by in vivo treatment. Our results indicate that the acquisition of suppressor activity requires the interaction of Ia+CD11b+Fc gamma R+IgG- asialo GM1- adherent cells with Thy1-CD4-CD8-IgG-Ia- asialo GM1-Fc gamma R+CD11b+ non-adherent cells. It is also shown that the development of suppressor activity is impaired by preventing cell-cell contact between these two cell subsets through coculture in 'Transwell chambers'. These observations support the conclusion that the in vitro acquisition of suppressor activity is a consequence of the maturation of suppressor cell precursors of the monocytic lineage induced by a receptor-ligand type interaction with a non-adherent cell subset that is clearly distinct from mature T, B and natural killer (NK) cells.

Abnormal t suppressor cell function in juvenile rheumatoid arthritis

The purpose of this study was to analyze T suppressor cell function in juvenile rheumatoid arthritis (JRA). JRA is a chronic inflammatory childhood disease of unknown etiology that is characterized by arthritis and immunoregulatory abnormalities. T suppressor cell precursors (CD8+, CD28-) were purified from the peripheral blood of 24 JRA patients, using a combination of monoclonal antibodies. These cells were treated with histamine or concanavalin A, agents that are known to induce suppressor activity. They were also tested for their ability to inhibit the proliferative response of autologous T cells to phytohemagglutinin. In some experiments, the accumulation of intracellular cAMP following histamine treatment was also measured. Twelve of 13 patients with clinically active JRA showed abnormal histamine-inducible T suppressor cell function, characterized by the failure of CD8+, CD28- T cells to mediate any detectable suppression. The failure of these cells to accumulate intracellular cAMP after histamine treatment was observed in 5 of 5 patients tested who had active disease. In contrast, 11 of 11 patients with clinically inactive JRA, 5 of 5 patients with cystic fibrosis, and 9 of 9 pediatric control subjects had normal histamine- and concanavalin A-inducible T suppressor cell function, and a normal cAMP response to histamine. These results suggest that patients with clinically active JRA have a reversible defect in T suppressor cell function that is associated with a failure of T suppressor cell precursors to accumulate intracellular cAMP following their exposure to selected immune stimuli.

Immunological Consequences of Mineral Dust Inhalation

Abstract Our experiments have indicated that in vitro lymphocyte function is a sensitive marker capable of detecting low levels of exposure to mineral dusts. Syngeneic PVG rats were exposed to airborne mineral dusts at a range of mean respirable mass concentrations for periods ranging from two weeks to three months. At predetermined times, splenic lymphocyte function was assessed by measuring the in vitro proliferation response to the mitogens phytohaemagglutinin and concanavalin A. Inhalation of chrysotile A asbestos and quartz had profound but different effects on the immunological status of the rats: chrysotile induced a marked suppression of lymphocyte mitogenic activity, whereas quartz was associated with a marked enhancement of response. In contrast, inhalation of titanium dioxide had no significant effect on lymphocyte reactivity over a wide range of dust concentration. Soluble factors released from lymphoid cells taken from dust exposed rats were capable of inducing immune suppression when transferred to syngeneic naive recipients whereas transfer to normal lymphocyte products had no effect. Moreover, the immunomodulatory effect of intraperitoneal dust inoculation was reduced by pretreatment of rats with cyclophosphamide to inhibit the development of suppressor cell precursors.

Lymphocyte function in human bone marrow. II. Characterization of an interleukin 2-sensitive T precursor-cell population.

In the present study of human bone marrow lymphocytes, we analyze a newly recognized population of T suppressor-cell precursors which are found in marrow only and which have the potential to inhibit immunoglobulin (Ig) production in vitro. Following exposure to interleukin 2 (IL2), suppressor precursors acquire E receptor, T3 determinants, suppressor function, and lectin responsiveness. To distinguish this population within the framework of T-cell ontogeny, it was compared to a previously described population of thymus-dependent helper T-cell precursors which express helper function following exposure to thymus-derived mediators. The two populations are completely distinct and can be separated on density gradients. Suppressor precursors expressed T8 and TAC (IL2-receptor) antigens prior to in vitro induction with IL2. The thymic hormone-dependent cells expressed T4 but not T8 or TAC determinants. In two patients with severe combined immunodeficiency disease (SCID), IL2-responsive precursor cells appeared only late after thymus epithelium transplantation, perhaps best explained by a model in which thymus-dependent differentiation pathways precede, induce, or seed pathways of extra-thymic T-cell differentiation. The large pool size of over 10(11) suppressor and helper precursor cells present in adult bone marrow suggests that these populations may play an important role in immune homeostasis.

A method for analysing the clonal precursors of concanavalin A-induced suppressor cells

Spleen cells from 3 different strains of mice (C57 (H-2 b), CBA (H-2 k) and BALB/c (H-2 d)) were stimulated in vitro with different concentrations of concanavalin A (CA) for 48 h. This resulted in the production of cells capable of inhibiting the generation of alloantigen-specific cytotoxic T lymphocytes (CTL) in a mixed lymphocyte culture (MLC). 1 μg/ml was an effective concentration of CA to induce C57 and BALB/c suppressor cells (SC), but 5 μg/ml CA was required to induce CBA SC. SC precursors (SC-P) were shown to be radiosensitive and the results suggest that SC themselves may be radiosensitive. SC were effective in the presence of added interleukin-2 (IL-2). SC were then induced at limit dilution in microwells in a volume of 25 μl. A MLC (200 μl) was then (after 48 h) added to each microwell. This resulted in a dilution of the concentration of CA to a level below which it was effective at inducing suppression. Cytotoxicity was then assessed 7 days later. It was thus possible to analyse SC-P at the clonal level and estimate their frequency. The frequency of C57 splenic SC-P (active against a C57 anti-BALB/c MLC) was 14.4 × 10 −6, the frequency of CBA splenic SC-P (active against a CBA anti-BALB/c MLC) was 92.3 × 10 −6, and the frequency of BALB/c splenic SC-P (active against a BALB/c anti-CBA MLC) was 15.8 × 10 −6. It was possible to analyse SC-P at a clonal level whether or not the MLC contained added IL-2. SC and SC-P were shown to be sensitive to anti-Thy-1 and complement.

Functional and phenotypic studies of Japanese adult T cell leukemia cells.

The cell surface marker profile and functional analysis of peripheral blood lymphocytes from 11 Japanese adult T cell leukemia patients were studied. The phenotypic analysis of Japanese adult T cell leukemia (ATL) cells by a series of 13 monoclonal antibodies showed that all ATL cells are anti-T4 reactive but some differ in their expression of T3, T11, and T12 antigens. Thus, considerable phenotypic heterogeneity exists in these populations of leukemia cells. When analyzed in functional assays, ATL cells were suppressive when added to a pokeweed mitogen- (PWM) driven Ig synthesis system. However, the suppression mechanism seemed to be more complex than originally conceived. ATL cells examined in this study seem to function mainly as an inducer of suppressor cells, and as such, activate normal T8 precursors of suppressor cells rather than function as suppressor effector cells. In addition, no evidence was obtained to suggest that suppression of PWM-stimulated IgG synthesis was mediated by natural killer (NK) activity of ATL cells. Rather, ATL cells seem to be markedly deficient in NK activity. These studies suggest that the majority of ATL cells tested are representative of and seem to be the leukemic counterparts of the T4+ suppressor inducer subset.

Alloproliferative human T cell clones primed and cultured in vitro lose proliferative and gain suppressive activity with age

Alloreactive human T lymphocyte clones were found to lose antigen-stimulated proliferative capacity and their ability to secrete interleukin 2 (IL 2) after a critical period in tissue culture. Instead, they gained the previously absent capacity to suppress lymphoproliferative (LP) responses in the presence or absence of exogenous IL 2. Such “ex-PLT” suppressive clones continued to grow perfectly well, retaining IL 2 and filler cell dependency, apparently normal karyotypes, and their OKT4+, OKT8− phenotypes. At least two suppressive mechanisms were demonstrated: (1) the “induction” of suppressive effectors in normal peripheral lymphocytes, and (2) a direct suppressive activity on lymphocyte proliferation shown by their ability to inhibit restimulation of cloned lymphocytes lacking suppressor cell precursors. The consistent “differentiation” from IL 2-secreting “helper” status to nonspecific suppressive status may represent a novel immunoregulatory phase in the long-term differentiation of normal human T cells.

31P NMR study of erythrocytes from a patient with hereditary pyrimidine-5′-nucleotidase deficiency

In our previous study, thymus cells were shown to be responsible for enhancing the growth of the allogeneic sarcoma 180 (S180) in AKR mice that had been injected with goat anti-Th-B antibody reagent (antiserum raised in goats against Balb/c myeloma MOPC 104E cells and purified). We suggested that the cells producing enhancement are suppressor T cells. We now show that the cells responsible for tumor enhancement are indeed T cells, since they carry the Thy-1 antigen on their surface. Treatment of the cells in vitro with anti-Thy-1 plus complement completely eliminates their ability to enhance tumor growth. The thymocytes responsible for tumor enhancement do not carry the Th-B determinant. Treating thymocytes in vitro with goat anti-Th-B antibody reagent plus complement does not abrogate their tumor-enhancing activity. This suggests that the suppressor T cells involved in tumor enhancement are generated by the interaction of anti-Th-B antibodies with precursor suppressor cells which do carry Th-B. Once generated, the active suppressor cells lose the Th-B antigen. This suggestion is supported by our finding that the thymic precursors of Con A-inducible suppressor cells bear Th-B, since they are killed by anti-Th-B plus complement, whereas active suppressor cells induced by Con A do not carry Th-B, since they are not killed by anti-Th-B plus complement. Neither splenic precursors of Con A-inducible suppressor cells nor the active suppressor cells thus induced carry Th-B since neither is killed by anti-Th-B plus complement. We have also found that there are apparently nonthymic suppressor cell precursors which can also be activated by anti-Th-B, since spleen cells from thymectomized mice bearing S180 and treated with anti-Th-B can transfer the tumor-enhancing effect. We conclude that precursors of suppressor cells carry the Th-B determinant. These precursors differentiate to active suppressor cells when stimulated by anti-Th-B antibodies. This process can take place either outside the thymus or in the thymus. Once differentiated, the mature suppressor cells no longer bear the Th-B marker and migrate from their sites of induction. Such cells can suppress immune mechanisms responsible for allogeneic tumor graft rejection and thus cause tumor enhancement.

Suppressor cells in levamisole-treated mice: A possible role of T-cell-mediated feedback suppression in the drug-induced suppression of the humoral immune response

The suppressive effect of levamisole (LMS) on the primary IgM plaque-forming cell (PFC) response to sheep erythrocytes in mice was studied. The suppressive effect of LMS on day 4 PFC response was found to be modified by dose, the timing of drug administration, and the amount of antigen injected. The experiments, using an inhibitory dose of 10mg/kg LMS, showed that: 1) T cells play an essential role in LMS-induced suppression; 2) antigen-specific suppressor cells are induced in the spleen of LMS-treated mice; 3) the spleen cells from LMS-treated donors either suppress or enhance the PFC response in the recipients, depending on the amount of antigen injected and the number of spleen cells transferred; and 4) a dose of 10 mg/kg LMS, which is inhibitory when assessed on day 4 PFC response, actually enhances the response three and a half days after the immunization. These results suggest that LMS induces precursors of both helper and suppressor cells, and their differentiation and/or maturation to LMS-primed antigen-specific suppressor T cells are modulated, at least in part, by T-cell-mediated feedback suppression.

Lymphocyte subsets involved in tumor-activated nonspecific feedback suppression

Cells from the spleen, lymph nodes, and peritoneum of DBA/2 mice bearing a subcutaneous tumor mediate nonspecific suppression of an in vitro antibody response to sheep red blood cells (SRBC) when cocultured with a normal T-cell subset(s). The spleen cells from the tumor-bearing mouse required for the suppression bear the Lyt 1 and Ala 1 surface markers characteristic of “inducer” T cells and activated cells, respectively. The activity of this cell population is also sensitive to irradiation. The normal T-cell subset which cooperates in the suppression bears the Qa-1 surface antigen which has been associated with suppressor cell precursors in several systems but lacks detectable surface Lyt 1 and 2 markers. Suppression of anti-SRBC antibody responses in spleen cell cultures from tumor-bearing mice alone could also be elicited, but only when increased numbers of cells were cultured. These data are consistent with the theory that a tumor-activated, Lyt 1 + T-cell subset has the capacity to nonspecifically suppress immune responses by activating a Qa-1 + subset(s) of T suppressor cells, perhaps via feedback signals.

Phenotypes of T lymphocytes in systemic lupus erythematosus: Decreased cytotoxic/suppressor subpopulation is associated with deficient allogeneic cytotoxic responses rather than with concanavalin A-induced suppressor cells

Patients with systemic lupus erythematosus (SLE) have decreased precursors of cytotoxic/suppressor T lymphocytes in their peripheral blood, as determined by monoclonal antibodies. To determine whether decrease of the cytotoxic or the suppressor parts (or both) of this subpopulation of T lymphocytes is being reflected by this peripheral mononuclear cell (MNC) abnormality, a series of experiments was conducted in which both the suppressive function (concanavalin A induced and spontaneous) and the generation of cytotoxic responses against alloantigens were tested. Cytotoxic responses were consistently diminished while suppressor capacity of MNC from patients with SLE (measured on several assays of normal T- and B-lymphocyte functions) was comparable to that of MNC from normal individuals. The defect in cytotoxic responses to alloantigens by MNC from SLE patients persisted following secondary stimulation in mixedleukocyte cultures; the cytotoxic responses were not amplified and remained well below the responses of normal MNC. These experiments indicate that the decreased peripheral population of cytotoxic/suppressor lymphocytes in SLE patients represents low or absent precursors of cytotoxic cells rather than of precursors of suppressor cells.

Suppressor T cells activated in a primary in vitro response to non-major histocompatibility alloantigens.

Normal mouse spleen cells are not capable of mounting a primary cytotoxic T lymphocyte (Tc) response to non-H-2 alloantigens in vitro, although a good secondary H-2-restricted response is observable after in vivo immunization of the responder animals. Suppressor cells are generated in such a primary responses provided a Mls incompatibility exists between the responder and stimulator. These suppressors are not antigen specific, are Thy-1+, Lyt-1+, 2-, I-J-, and are highly radiosensitive. The suppressor cell precursors in normal spleen express the same phenotype. These suppressor cells are probably implicated in the lack of a primary Tc response in a primary mixed lymphocyte reaction across non-H-2 incompatibilities that include an Mls difference.

Mechanisms involved in age-dependent decline of immune responsiveness and apparent resistance against tolerance induction in C57BL/6 mice.

The plaque-forming antibody response of C57BL/6 mice to rabbit gamma globulin (RGG) decreases as a function of ae. RGG in tolerogenic form induces tolerance of young mice but sensitizes older animals. If antigen is administered together with lipopolysaccharide, the age-dependent decline in immune responsiveness is not observed, nor are older animals sensitized by tolerogen. The age-dependent decline in immune responsiveness is due to a loss of T helper capacity; sensitization by tolerogen is attributable to a subpopulation of B cells which becomes sensitized by the tolerogen. Older animals, treated with tolerogen, show a degree of central tolerance of T cells and a relatively slight, age-dependent diminution in colchicine- or cyclophosphamide-sensitive precursors of suppressor cells.

The effect of protecting guinea-pigs against EAE on the meningeal inflammatory cell-infiltrate.

Guinea-pigs were found to be fully protected from the clinical signs of EAE when they were given spinal cord in FIA either 3, 7. or 14 days immunization with spinal cord in FCA. If the animals were protected by the recruitment of suppressor cells then cyclophosphamide (CY) might break this protection. CY at 300 mg/kg and 20 mg/kg was given either before the protective inoculum to try to eliminate suppressor cell precursors or before immunization to eliminate the suppressors cells themselves. No clinical signs of EAE developed in any of these groups of guinea-pigs. It was found, however, that large numbers of meningeal inflammatory cells could be isolated from the brains of guinea-pigs protected from EAE when the protective inoculum was given only 1 week before immunization. These cell numbers were further significantly increased by giving CY in addition to the protective inoculum. The meningeal inflammatory cells and peritoneal exudate cells (PECs) from guinea-pigs protected against EAE were compared to those isolated from animals with acute EAE. These cells from guinea-pigs with acute EAE responded, in the lymphocyte transformation test, to the mitogen. The PECs also responded to the brain antigen, myelin basic protein (MBP) and to tuberculin (PPD). The meningeal inflammatory cells did not respond to either antigen. The PECs from animals protected against EAE responded to mitogen and PPD, but not to MBP, but the meningeal inflammatory cells did not respond to antigen or mitogen. The percentage of T lymphocytes in the meningeal inflammatory infiltrates of protected and unprotected guinea-pigs was similar, but that of phagocytic cells was significantly depressed in the inflammatory infiltrates of animals protected against EAE.

Dissection of distinct human immunoregulatory T-cell subsets by a monoclonal antibody recognizing a cell surface antigen with wide tissue distribution.

A monoclonal antibody, PVR-11, was obtained after hybridization of X63Ag8.653 murine myeloma cells with spleen cells from a mouse immunized with human lymphocytes. It recognizes a 175,000- to 185,000-dalton surface antigen present on approximately 80% of normal human peripheral T lymphocytes, 50% of non-T non-B cells, and less than 10% of B cells as determined by complement-dependent microcytotoxicity. It is also present on various leukemia T cells, on some but not all T lymphoblastoid cell lines, and on a small fraction of some B lymphoblastoid cell lines. Some B-cell chronic lymphocytic leukemia cells also express the PVR-11 antigen. Functional analysis of normal human T lymphocytes demonstrated that the PVR-11-depleted T-cell subset contains the precursors of both cytotoxic and suppressor cells but lacks helper cells. On the other hand, cytotoxic effector T cells express the PVR-11 antigen. These results demonstrate that antigenic determinants with relatively wide tissue distribution can dissect functionally distinct human immunoregulatory T-cell subsets.

Concanavalin A-inducible suppressor cells in pleural effusions and peripheral blood of cancer patients

Carcinomatous pleural effusions of 18 of 20 patients with lung cancer contained suppressor cell precursors that could be activated by concanavalin A (Con A) to suppress the proliferative responses of autologous and allogeneic lymphocytes to phytohemagglutinin and Con A. However, pleural effusion cells showed no suppressor function without prior activation by Con A. In contrast, the peripheral blood of the cancer patients exhibiting impaired mitogenic response contained nonspecific spontaneous suppressor cells capable of inhibiting the lymphoproliferative response to mitogens without prior activation by Con A, but these cells were not able to show further suppressor function even after activation by Con A. The maximum suppression was observed after 48-h treatment of lymphocytes with optimally mitogenic doses of Con A. The Con A-inducible suppressor cells of the pleural effusion and spontaneous suppressor cells of the peripheral blood of cancer patients had the same characteristics with regard to the capacity to suppress the mitogenic responses of autologous and allogeneic lymphocytes, belonging to the group of nylon wool-nonadherent T cells and being sensitive to in vitro culture and resistant to treatment with mitomycin C.