Autologous Peripheral Blood Lymphocytes Research Articles

Prednisone-responsive aplastic anemia: a mechanism of glucocorticoid action

We performed serial agar cultures (CFU-C) using marrow cells from a patient with prednisone-responsive aplastic anemia and from five patients with prednisone-resistant aplasia. Colony growth was decreased in all patients. Cortisol (10(-7)-10(-4)M) significantly enhanced colony growth in the prednisone-responsive patient but fauled to enhance colony growth in the remaining five patients. Further studies in the responsive patient indicated that (1) colony growth was enhanced by depleting T lymphocytes from the marrow cells, (2) colon growth T-depleted marrow cells was inhibited by autologous peripheral blood lymphocytes (PBL), (3) cortisol failed to enhance colony growth of T-depleted marrow cells, (4) PBL and PBL-conditioned medium inhibited colony growth of both autologous and allogeneic marrow cells, but neither cortisol-treated PBL nor T-depleted PBL were inhibitory. Serial cultures in the responsive patient showed that colony growth normalized during remission when "suppressor" cells were absent and that colony growth was subnormal during a later relapse when cortisol-resistant "suppressor" cells were present. Therefore, in this prednisone-responsive patient, cortisol-sensitive T lymphocytes suppressed granulopoiesis in vitro. Our observations suggest that aplastic anemia in this patient is immunologically mediated and that prednisone therapy enhanced hemopoiesis in vivo by inhibiting the "suppressor" T lymphocytes.

T-T Interactions in the Induction of Antigen-Specific Human Suppressor T Lymphocytes in Vitro

Abstract We intended to investigate whether the suppression of antigen-induced antibody responses in vitro in man by T suppressor cells required contact of T suppressor cells with target cells or whether this effect was mediated by factors released by T suppressor cells. To this end supernatants of antigen-induced T suppressor cells were tested (by a plaque forming cell assay) for their capacity to suppress antibody responses of autologous and allogeneic human peripheral blood lymphocytes. We have shown that supernatants of antigen-specific T suppressor cells, designated as TsF24: a) can suppress an antibody response of autologous but not allogeneic lymphocytes to the inducing antigen; b) are antigen-specific in their effect; and 3) are produced by radiosensitive T cells. Furthermore, the target of the factor is a radiosensitive T cell. These findings taken together indicate that, in the generation of T-effector suppressor cells in man, T-T interactions occur, and in addition, that cellfree factors may be involved in these interactions.

Suppressor cell function in man: Evidence for altered sensitivity of responder cells with age

Concanavalin A (Con A)-activated suppressor cell function was evaluated in young and elderly adult human donors. Activated suppressor cells reduced the mitogenic response of autologous peripheral blood lymphocytes more markedly in elderly than in young adults (60.2 ± 4.8 vs 35.3 ± 8.9%, P < 0.05). As a measure of the actual amounts of suppressor factors released, we concurrently determined the effect of supernatants from the Con A-activated cells on the proliferative activity of a dividing cell line (L cells). The inhibitory effect of supernatants from activated cells was greater in young than in elderly adults (21.4 ± 3.4 vs 11.4 ± 3.1%, P < 0.05). These results suggest that cultured lymphocytes from elderly donors produce less suppressor factors than do cells from young donors but that the responder cells of the elderly are more susceptible to suppressor influence.

Suppressor cell function in multiple sclerosis: Correlation with clinical disease activity

Concanavalin A (Con A)-activated suppressor cell activity was determined in multiple sclerosis (MS) patients who had been assigned to one of three subgroups, those with active disease, those recovering from a flare-up, and those with stable disease. The level of suppression induced by the Con A-activated suppressor cells on the mitogenic response of autologous peripheral blood lymphocytes was reduced in patients with active disease (3 +/- 8%) compared with stable patients (30 +/- 8%), patients recovering from a flare-up (62 +/- 5%), and controls (40 +/- 5%). As a measure of the actual amounts of suppressor factors released, the effect of supernatants from the Con A-activated cells on the proliferative activity of a dividing cell line (L cells) was determined concurrently. The inhibitory effect of supernatants from activated cells was reduced in active and stable MS patients (7 +/- 3%) compared to controls (21 +/- 4%). Three of 4 with active MS showed mildly elevated immune complex levels as measured by the Raji cell technique; each of these patients had low suppressor activity. Levamisole (1 microgram per milliliter) failed to alter suppressor cell activity in our in vitro system.

Suppressor cells in the pelvic lymph nodes regional to bladder cancer

Regional lymph node cells from some patients with locally advanced bladder carcinoma were found to inhibit the reactivity of normal and autologous peripheral blood lymphocytes in mixed leukocyte culture. The data suggest that suppressor cells are generated in draining lymph nodes of these patients by a high concentration of tumor-derived products and may have suppressive influence on host immunologic defense mechanisms.

Autoreactivity between lymphocytes and thymus cells in myasthenia gravis.

Thymus cell preparations from four of five myasthenia gravis patients with thymic hyperplasia and from one additional patient with thymic involution stimulated autologous peripheral blood lymphocytes. No autostimulation was observed in two patients with thymoma. Autostimulation as associated with an increase in the fraction of B lymphocytes in the thymus.

Activation of Human B Lymphocytes

Abstract The kinetics and mechanisms of suppression of the PWM-induced PFC response of human PB lymphocytes by Con A-activated suppressor cells were investigated. It was necessary that Con A suppressor cells be present early in the preocess of activation of human B cells toward antibody synthesis, but maximal suppression of the PFC response occurred later in the culture period. In addition, Con A-activated cells, although suppressing the PFC response to PWM greater than 90% of control, did not significantly suppress the blastogenic response to PWM after 3 or 5 days in culture. On the contrary, after 3 days in culture, background tritiated thymidine incorporation as well as tritiated thymidine incorporation to PWM stimulation was increased when Con A suppressor cells are added to fresh autologous peripheral blood lymphocytes. This increased blastogenic response after three days most likely represented an autologous mixed lymphocyte reaction (MLR) of Con A suppressor cells against fresh autologous non-T cells. The induction of autoreactive cells may be one of several modes of suppression of PFC responses by Con A activated suppressor cells.

Circulating suppressor cells in man as a function of age

Concanavalin A (Con A) was used to activate suppressor cells derived from the peripheral blood of human donors. Activated suppressor cells reduced the mitogenic response of autologous peripheral blood lymphocytes (PBLs) more markedly in elderly than in young adults (72.2 ± 5.0 vs 41.8 ± 7.2%, P < 0.02). The baseline mitogenic response of PBLs from elderly adults was significantly lower than that of PBLs from normal young adults (9497 ± 2609 vs 28,068 ± 3973 cpm, P < 0.01). A group of young multiple sclerosis patients showed poor mitogenic responsiveness (11,017 ± 2679 cpm) and low suppressor activity (36.0 ± 3.0%) suggesting that suppressor influence need not depend on the activity of responder cells. Unconcentrated supernatants from Con A-activated cells did not suppress the mitogenic response of autologous lymphocytes but did suppress a human lymphoblast cell line (Raji).

Plasma cell antigens in human multiple myeloma.

Tumor-specific antigens capable of eliciting a response from autologous lymphocytes have been described in mouse plasmacytoma systems. This paper presents evidence of similar antigens in human myeloma. Myeloma plasma cells isolated from the bone marrow of 27 patients stimulated autologous and allogeneic peripheral blood lymphocytes (PBL) from myeloma patients in mixed leukocyte culture. Plasma cells isolated fromt the bone marrow of normal patients or patients with benign plasmacytosis failed to stimulate PBL in mixed leukocyte culture. Similar results were found in passive cytotoxicity assays with the use of chinken red blood cells (CRBC) coated with 3M KC1 plasma cell extracts from myeloma patients. PBL from myeloma patients caused 30 to 80 percent chromium-51 release from tanned chromium-labeled plasma cell extract-coated CRBC targets, whereas PBL from normal patients or patients with benign plasmacytosis caused only 10 to 25 percent chromium-51 release. This study indicates the presence of material resembling tumor-specific antigens on human myeloma plasma cells. Immune response to suce antigens is elicited in autologous and allogeneic myeloma patients.

Suppression of immunoglobulin synthesis and secretion by peripheral blood lymphocytes from normal donors.

Concanavalin A (Con A)-treated peripheral blood lymphocytes from healthy volunteer donors have been shown to suppress proliferative responses associated with thymus-derived lymphocytes (T-cells). The present investigations demonstrate that peripheral blood lymphocytes incubated with Con A for 48 hr can abrogate pokeweed mitogen-stimulated differentiation of bone-marrow-derived (B) lymphocytes to immunoglobulin-synthesizing and -secreting plasma cells. This effect was manifested when washed Con A-treated peripheral blood lymphocytes were added to pokeweed mitogen-stimulated cocultures containing fresh autologous or allogeneic mononuclear cells, and it did not appear to involve cytotoxicity. Parallel control cultures consisting of mononuclear leukocytes incubated for 48 hr in the absence of Con A also had immunoglobulin suppressor activity in mixing experiments. This effect, however, was most pronounced when preincubated cells were added to fresh autologous pokeweed mitogen-stimulated peripheral blood lymphocytes. Cell mixtures containing peripheral blood lymphocytes demonstrated a spectrum of immunoregulatory effects ranging from suppression to enhancement of pokeweed mitogen-stimulated immunoglobulin synthesis and secretion. Several functional subclasses of suppressor cells that reflect varying levels of specific activity have thus been demonstrated in human beings. Moreover, a degree of genetic identity appears to be required for the expression of "weak" immunoregulatory influences.

Thymic lymphocytes in myasthenia gravis.

Lymphocytes from the thymus glands of myasthenic patients and controls were characterized in terms of their percentages of B- and T-cells and their ability to stimulate the proliferation of autologous peripheral blood lymphocytes. Percentages of B- and T-cells were the same in the thymuses of myasthenic and control subjects. Autologous peripheral lymphocyte stimulation by thymocytes was observed in only 1 of 3 myasthenic patients, and this was with a thymus gland containing a thymoma. These results thus differ from those of other investigators.

Induction of peripheral blood lymphocyte transformation by autologous synovial fluid lymphocytes and synovial fluid.

Synovial fluid lymphocytes and synovial fluid from most patients with rheumatoid arthritis induced blastogenesis of autologous peripheral blood lymphocytes. In vitro transfer of the mitogenic activity of the synovial fluid to peripheral blood lymphocytes was not accomplished, but diminished response of the peripheral blood lymphocytes to phytohemagglutinin stimulation after incubation in synovial fluid was demonstrated. These findings suggest that a similar blastogenic response in vivo may induce the lymphoid hyperplasia regularly observed in rheumatoid arthritis.

Search for antigen in rheumatoid synovial macrophages.

Peripheral blood monocytes and synovial macrophages obtained from 21 patients with rheumatoid arthritis and from 13 controls were cultured with autologous peripheral blood lymphocytes and stimulation measured by thymidine uptake. Rheumatoid macrophages and monocytes induced a small but significant degree of lymphocyte transformation, and those from the controls did not. The degree of stimulation appears to be more consistent with the nonspecific effect of lymphocyte activating factor, rather than with the presence of a specific synovial antigen.

The thymus in myasthenia gravis. Evidence for altered cell populations.

Abstract Comparisons of several types of immunologic reactivity were made in thymic cells from six patients with myasthenia gravis and thymic hyperplasia, four patients with myasthenia gravis and thymoma and six age-matched control cardiac-surgery patients. In mixed leukocyte reactions, thymic cells from the subjects with hyperplasia were capable of stimulating autologous peripheral blood lymphocytes. Such reactivity was not seen in thymocyte-blood lymphocyte mixtures from the other two groups. There was an increased number of B cells in the thymic-cell populations from the myasthenic patients as compared to that in the control group, carrying predominantly IgM receptors. The thymic cells from myasthenic patients also responded more vigorously to pokeweed mitogen. These findings suggest altered populations of lymphoid cells in the thymus of myasthenic patients that react with autologous lymphocytes from other cell compartments. The pathogenic implications of these findings remain to be determined. (N Engl...

In vitro peripheral blood and synovial fluid lymphocyte interactions.

AbstractIn vitro interaction of autologous peripheral blood and synovial fluid lymphocytes was studied using cells isolated from patients with various forms of arthritis. Addition of mitomycin‐treated peripheral blood lymphocytes from patients with inflammatory arthritis to in vitro cultures of autologous synovial fluid lymphocytes caused an increased incorporation of tritiated thymidine by the synovial fluid lymphocytes. However when the reverse situation was tested, no stimulation of peripheral blood lymphocytes by mitomycin‐treated synovial cells was recorded. In some instances synovial fluid lymphocytes were observed to exert an apparent cytotoxic or inhibitory effect on autologous peripheral blood lymphocytes. The latter response pattern was found to occur to a much greater degree with lymphocyte preparations from patients with rheumatoid arthritis than in those with other forms of inflammatory joint disease.

Lymphotoxin formation by lymphocytes and muscle in polymyositis.

Muscle pieces from 11 patients with dermatomyositis or polymyositis were incubated with autologous peripheral blood lymphocytes and the supernates examined for the production of lymphotoxin, a mediator of delayed hypersensitivity, using human fetal muscle monolayers as the target cell. In the case of all 10 active patients, production of lymphotoxin was demonstrated. This mediator was also demonstrated when muscle alone was incubated from two patients with extensive cellular infiltration. Lymphotoxic activity was not found in supernates obtained by incubation of muscle from nine control subjects with their autologous peripheral blood lymphocytes. Addition of methyl prednisolone to active cultures inhibited the action of lymphotoxin on the muscle monolayers. Lymphotoxin was not demonstrated when breast tumor tissue from a patient with dermatomyositis was incubated with autologous lymphocytes. The lymphotoxic agent in the active supernates had similar chromatographic properties to those of a sample of purified lymphotoxin. These findings suggest that muscle injury in polymyositis is a result of a cellular immune response to an antigen present in involved muscle tissue.

The Effect of Human Tumor Extracts Upon Autologous Peripheral Blood Lymphocytes.

Excerpt The induction of lymphocyte transformation by various antigens has been shown to be a direct reflection of the degree of lymphocytic or delayedtype immunity in the host. Since specific lymp...