Lymphocyte Blastogenic Response Research Articles

Alternate-day prednisone therapy and human lymphocyte subpopulations.

The mechanisms and kinetics of the immunosuppressive effects of alternate-day prednisone were investigated in a group of patients with a variety of inflammatory diseases receiving a range of alternate-day prednisone doses from 5 to 120 mg. Total circulating lymphocyte and monocyte counts, as well as proportions of lymphocyte subpopulations defined both by surface markers and by in vitro functional capacities, were studied. At 8 a. m. of the day on prednisone, just before drug administration, lymphocyte and monocyte counts, proportions of lymphocyte subpopulations, as well as in vitro lymphocyte blastogenic responses to various mitogenic and antigenic stimuli were normal. 4 h after the administration of prednisone, there was a profound lymphocytopenia and monocytopenia, with a differential depletion of thymus-derived lymphocytes as well as various functionally defined lymphocyte subpopulations. Lymphocyte kinetic studies using a radioactive chromium-labeled autologous lymphocytes showed that the lymphocytopenia was due predominantly to a transient depletion of the recirculating portion of the intravascular lymphocytepool. All these parameters returned to normal by 8 a.m. of the following day (off prednisone) and remained normal throughout the day. This very transient lymphocytopenia and monocytopenia after prednisone, with normal cell numbers, proportions, and functions throughout the remainder of the 2-day cycle, was associated with suppression of disease activity, yet did not affect cutaneous delayed hypersensitivity in these patients nor increase the likelihood of infectious complications. This drug-associated cyclic and transient monocytopenia and selective lymphocytopenia is best explained by a redistribution of recirculating lymphocytes to other body compartments, particularly the bone marrow.

Active specific immunization in malignant melanoma.

The effects of active specific immunization with nonirradiated autologous and irradiated cultured allogeneic melanoma tumor cells (TC) on cell-associated immunity was studied in 11 patients with widespread malignant melanoma receiving chemoimmunotherapy. Immunization with 1 X 10(6)-8 X 10(7) TC was done in the draining area of a BCG scarification on day 7 of the study. The in vitro lymphocyte blastogenic response (BR) to autologous TC was studied in terms of the stimulation index on day 1, 7, 14, and 21. The stimulation index (SI) was the counts per min (cpm) in the TC stimulated culture minus the cpm among the TC alone divided by the cpm in the unstimulated controls. A positive BR was considered as a SI equal to or greater than 3. Six out of 11 patients had a significant increase in BR lasting 7-14 days after immunization. The pre- to post-immunization changes in the SI of the 6 patients were 2.5 to 7.5; 0.7 to 3.2; 0 to 5.2; 0 to 16; 0.1 to 6.6; and 6.5 to 30.0. Nine out of 11 patients showed a delayed local inflammatory reaction at the immunization site. Five out of 11 patients mounted a cutaneous delayed hypersensitivity reaction to autologous tumor cells at a separate test site following immunization. There were no side effects. Active specific immunization in the drainage of a BCG reaction appears to be safe. Since a positive BR to autologous TC correlates with a good prognosis in patients with solid tumors, studies of the immunotherapeutic efficacy of this approach may be warranted.

Serial studies of immunocompetence of patients undergoing chemotherapy for acute leukemia.

Immunocompetence was followed serially for 1 yr from the onset of treatment in 55 adult patients with acute leukemia. The tests used were delayed hypersensitivity responses to a battery of five recall antigens (dermatophytin, dermatophytin 0, candida, streptokinase-streptodornase, and mumps) and in vitro lymphocyte blastogenic responses to phytohemagglutinin and streptolysin 0. There was a strong correlation between immunocompetence at the start of treatment and a good prognosis; 32/39 patients who subsequently entered remission were initially immunocompetent compared to 4/15 who failed to enter remission. In the complete remission group there was a decline in competence starting from 2 to 5 mo after the onset of treatment. In those who remained in remission for 1 yr, competence recovered at 6 mo and remained vigorous thereafter. In those who relapsed before 1 yr, the decline in competence occurred 1 mo before relapse and competence continued to decline progressively during the 1 yr follow-up period. These studies suggest that therapeutic approaches which restore immunocompetence or prevent its decline will improve both the remission rate and the remission duration of patients with acute leukemia.

Potentiation of phytomitogens action by neuraminidase and basic polypeptides.

TREATMENT of lymphocytes with neuraminidase enhances their blastogenic response to some antigens and mitogens1, increases their lectin-induced agglutinability2,3, antigenicity and immunogenicity4,5, alters their homing properties6 and renders the cells highly susceptible to the cytolytic effects of alloantibody and complement7,8. These changes might result from the exposure of new sites on the cell surface2,3,9, a reduction in the net surface charge of the cells or a combination of the two effects. Polycations agglutinate different cell types10–12 and markedly reduce their anodic electrophoretic mobility10. In order to study the effect of cell surface charge on lymphocyte blastogenic response to phytomitogens the effect of polycations on this process was compared with that of neuraminidase.

Serum inhibitory factors in cancer.

The cellular response and serum blocking factors of 70 cancer patients were assessed by micro-MLC and PHA-stimulation techniques. Serum level of carcinoembryonic antigen (CEA) was determined by radioimmunoassay, and an attempt was made to correlate the concentration of CEA with the inhibitory effect of each serum on the blastogenic response of lymphocytes from cancer and healthy subjects. A direct correlation was found between the serum inhibitory activity, the CEA level and the extent of the disease. However, CEA negative cancer sera also inhibited transformation of autologous and normal homologous lymphocytes. This indicates that circulating CEA does not account for serum blocking activity.

Effects of surgery, anesthesia and intraoperative blood loss on immunocompetence

The effects of surgery, anesthetic agents, and/or intraoperative blood loss on the immune system were studied by comparing the preoperative, intraoperative, and postoperative peripheral blood lymphocyte blastogenic responses to mitogens and antigens. All anesthetic agents were associated with some degree of immunosuppression, but some were differentially suppressive of either the T-cell or B-cell subpopulation of the peripheral blood lymphocytes. A greater depression of blastogenic responses was associated with ether than with halothane, and the B-cell responses seemed more selectively affected by ether. Cyclopropane anesthesia was more depressive than nitrous oxide, and both T- and B-cells were equally affected. The volume of blood lost during surgery was also correlated with the degree of immunosuppression. This anesthesia and surgery-associated immunosuppression persisted for at least 7 days.

Cellular immunity in man: Correlation of leukocyte migration inhibition factor formation and delayed hypersensitivity

The formation of leukocyte migration inhibition factor (MIF) by the lymphocytes of 13 normal persons immune to the protein antigen keyhole limpet hemocyanin (KLH) has been investigated. KLH-induced MIF formation expressed as percent migration was compared with delayed hypersensitivity, antibody, and in vitro lymphocyte blastogenic responses to this antigen. Individuals were studied 404–840 days (median 540 days) after their last exposure to KLH. Nine persons had delayed hypersensitivity to KLH and 10 had circulating KLH antibody. The lymphocytes of 11 showed an in vitro blastogenic response to KLH stimulation, while the lymphocytes of nine produced MIF after KLH stimulation. The mean percent migration for the subjects with KLH delayed hypersensitivity was 48.2 (range 20.4–70.4) compared with 133 (range 120–161) for the four persons who did not have KLH delayed hypersensitivity ( P < 0.05). The correlation coefficient between the precent migration and delayed hypersensitivity was −0.78 ( P < 0.01). No correlation was demonstrated between migration inhibition and the other parameters of immunity.

Antigen solubilized from human solid tumours: lymphocyte stimulation and cutaneous delayed hypersensitivity.

LYMPHOCYTE blastogenic responses to autologous tumour cells have been demonstrated in some patients with solid tumours1,2, but it is still debatable whether these responses represent a primary immune response similar to the mixed leukocyte reaction or pre-existing immunity to the tumour-associated antigens3.

Auto-immunization with acute leukemia cells: demonstration of increased lymphocyte responsiveness.

AbstractNine patients with adult acute leukemia were autoimmunized with 108 and/or 106 non‐irradiated leukemia cells during remission. Six of nine receiving 108 cells demonstrated a 100% or greater increase in in vivo lymphocyte blastogenic response to their own leukemia cells measured 7 days after immunization. In four responders, increased blastogenesis was noted only in autologous serum, but not in allogeneic serum. Only one of four demonstrated increased lymphocyte blastogenic response in autologous serum after immunization with 106 cells. The increased immune response was always accompanied by significantly increased blastogenic response to phytohemagglutinin (PHA) and/or Streptolysin‐O (SLO). Three patients studied 28–42 days following immunization showed a persistent increase in lymphocyte blastogenic response to leukemia cells, but the response to PHA and SLO had returned to pre‐immunization levels.

Immunoglobulin on tumor cells and tumor-induced lymphocyte blastogenesis in human acute leukemia.

Abstract Combined studies of direct membrane immunofluorescence with anti-immunoglobulin serum and of lymphocyte blastogenic responses to autologous leukemia cells were carried out in 34 adult patients with acute leukemia. Twenty-four of 34 (71 per cent) has positive blastogenic responses to their own leukemia cells. Eight of 19 patients with acute myelogenous leukemia and one of five with acute lymphoblastic leukemia had complete or partial abrogation of this positive blastogenesis when the lymphocytes were cultured in autologous rather than allogeneic serum. Direct membrane immunofluorescence with anti-immunoglobulin serum showed bound IgG on the cells of seven of eight patients with acute myelogenous leukemia and serum inhibition, and one without the serum inhibitory effect. Membrane immunofluorescence was negative in two patients with positive blastogenesis and serum inhibition, 14 of 15 with positive blastogenesis and no serum inhibition and, finally, all 10 patients with negative blastogenic respons...

Suppression of lymphocyte blastogenesis in man following cis-platinous diamminodichloride administration.

SummaryThe lymphocyte blastogenic response to PHA was studied in patients receiving cis-platinous diamminodichloride for various tumors. This compound produced significant inhibition of blastogenesis (p < .01) in doses ranging from 1 to 4 mg/kg body weight given in different schedules. The inhibition lasted from 18 to 72 hr. A single dose administration produced higher degree of inhibition than divided doses.We thank Mr. Harry Clifford for his technical assistance in carrying out lymphocyte blastogenesis procedure.

In vitro lymphocyte responses to malignant, benign neoplastic and normal tissue extracts.

SummaryTo identify tumor antigens and tumor immunity in man, we have investigated the use of the blastogenic response of lymphocytes to tissue extracts. Extracts of malignant, benign, and normal tissues of 60 patients with cancer were used to stimulate their own lymphocytes, the lymphocytes of other patients with the same tumor and those of normal subjects. Autochthonous extracts of the neoplastic tissue stimulated blas-togenesis in 56% of patients and a linear dose-response curve was observed. This suggested the presence of TSA and specific tumor immunity, even in patients with advanced metastatic disease. However, stimulation of autochthonous lymphocytes by extracts of skin in 50% of patients cast doubt on the tumor specificity of the response to extracts. The release of nonspecific mitogens, or of usually concealed autoantigens, or the presence of tumor antigens in the skin of patients with malignant disease were hypothesized as the possible cause. Because of this lack of specificity it seemed that tum...

Effect of arabinosyl-6-mercaptopurine on the blastogenic responses in vitro of human lymphocytes to mitogenic agents

The effects of arabinosyl-6-mercaptopurine (Ara-6-MP) on lymphocyte blastogenic responses in vitro to mitogens were studied. Ara-6-MP was not degraded in the cultures until after 5 days of incubation. Levels of drug of 30 μg/ml or greater significantly inhibited blastogenic responses to phytohemagglutinin, streptolysin O and allogeneic leukocytes as measured by [ 3H]thymidine incorporation (greater than 50 per cent reduction). Levels over 100–200 μg ml inhibited thymidine incorporation over 90 per cent, but complete inhibition was not achieved at any dose up to 300 μg ml . Morphological examination of parallel replicate cultures revealed that blastogenesis characterized by cell enlargement and the development of cytoplasmic basophilia was not significantly inhibited by the drug. Inhibition, at least at higher drug levels, began within 2 hr after exposure of the cells to the drug. Inhibition was completely reversible if the drug was washed from the cultures after 48 hr of continuous exposure and partially reversible when washed from the cultures after up to 96 hr of continuous exposure. In contrast to thymidine incorporation, uridine and leucine incorporation were only slightly inhibited by Ara-6-MP. These studies suggest that Ara-6-MP is a potent reversible and incomplete specific inhibitor of DNA synthesis among mitogen-stimulated, cultured leukocytes in vitro and that it might be an inhibitor of the proliferative phase of the specific immune response in vitro. In addition, these studies indicate that the morphological events associated with lymphocyte transformation can proceed in the virtual absence of DNA synthesis.

L-glutaminase: suppression of lymphocyte blastogenic responses in vitro.

Continuous exposure to 0.1 international unit or more of Escherichia coli L-glutaminase inhibited responses of human lymphocytes to phytohemagglutinin, streptolysin O, and allogeneic leukocytes. Inhibition was completety reversed by removing the enzyme from the culture or adding L-glutamine but not L-asparagine. Cytoxicity did not occur. L-Glutaminase should be immunosuppressive in vivo.

Patient survival after chemotherapy and its relationship to in vitro lymphocyte blastogenesis.

To study the relationship between chemotherapy immunocompetence and prognosis, in vitro lymphocyte blastogenic responses were studied serially, in 40 patients, receiving intensive chemotherapy for metastatic solid tumors. Suppression of lymphocyte blastogenic responses to phytohemagglutinin and streptolysin “O” was similar in all patients at the end of chemotherapy and was highly significant (PHA: P < 0.01, SLO: P < 0.02). Recovery of such responses occurred with significant overshoot above the pretreatment levels (P < 0.01) for PHA and SLO in those patients whose tumors regressed and survival was long, while such recovery was lacking in those patients whose tumors progressed and whose survival was short (PHA: P < 0.01, SLO: P < 0.05). This shows that the rebound with overshoot of immunologic reactivity after chemotherapy correlates with tumor regression and prognosis, and continued suppression may be an indicator of poor prognosis.

Replication of poliovirus in human lymphocytes stimulated with specific antigens.

Human lymphocytes from donors immunized with poliovirus vaccine and tetanus toxoid supported the replication of poliovirus type 1 when stimulated with these specific antigensin vitro. The blastogenic response of lymphocytes infected with poliovirus proved to be decreased as measured by partial inhibition of DNA synthesis. The inhibition of DNA synthesis was measured by liquid scintillation and radioautographic methods. The inhibition occured when virus was added up to 4 days after initiation of specific antigenic stimulation. Tuberculin-stimulated lymphocytes from Mantoux positive donors supported the replication of measles virus and poliovirus. The blastogenic response of these infected cells was also inhibited.