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Post-capillary venules of the mouse lymphatic tissues with special reference to the distribution of their endothelial IgG.

Post-capillary venules (PCV) of the lymph nodes and Peyer's patches were studied in frozen sections stained with an indirect immunoperoxidase technique to demonstrate their endothelium-associated IgG in DBA/2 mice rendered T-lymphocyte depleted with anti-theta- and anti-thymus-globulins. Three types of IgG-distribution (luminal, intraendothelial and basement membrane, types I, II and III, respectively), each confined to one of the three PCV grades (graded on the basis of the endothelial cell height), could be distinguished. The PCV grades, on the other hand, were closely correlated with the degree of T-lymphocyte depletion. The significance of this intimate relationship between the IgG-distribution pattern and the PVC grades was discussed in regard to the T-lymphocyte recirculation. A hypothesis was presented describing the presumed sequence of events involved in the passage of T-lymphocytes through the endothelium of the PCV which is the crucial step in their recirculation from blood of the lymphatic tissue.

Animals

Dicarbocyanine fluorescent probes of membrane potential block lymphocyte capping, deplete cellular ATP and inhibit respiration of isolated mitochondria.

3,3'-Dipropylthiodicarbocyanine iodide, a widely used fluorescent probe of membrane potential, was found to inhibit anti-Ig antibody, induced capping of mouse lymphocytes. The dye also lowered the cell ATP content. Experiments with isolated mitochondria revealed that the probe had a potent inhibitory action at site I of the respiratory chain. This mitochondrial blockade helps to explain the ATP depletion and blockade of capping, and gives cause for caution in the use of this dye as a probe of cell membrane potential. Three related dicarbocyanine dyes had similar toxic effects, but two cyanine dyes with much longer alkyl side chains, which have been used as probes of membrane fluidity, did not.

Adenosine Triphosphate

Mechanism of the establishment of Epstein-Barr virus genome-containing lymphoid cell lines from infectious mononucleosis patients: studies with phosphonoacetate.

A concentration of disodium phosphonoacetate (PA) has been defined which will reduce the synthesis of infectious EB virus in a producer cell line to 1% of control values but which will not affect the growth of EB virus-transformed cells in a 12-week colony-forming assay. When total mononuclear cells or T-lymphocyte-depleted mononuclear cells from the blood of acute IM patients were cultured in the presence of PA at the above concentration, the regular establishment of EB virus genome-containing cell lines seen in control cultures was almost totally abolished. In further experiments, when T-lymphocyte-depleted IM mononuclear cells were co-cultivated with foetal cells of the opposite sex in the presence and absence of PA, cell lines of mixed or of exclusively foetal origin were obtained not only from control co-cultures but also on those rare occasions when transformed foci developed in PA-treated co-cultures. The results suggest that all cell lines derived from the blood of IM patients are initiated in culture by a two-step process of virus release and secondary infection, and argue against the occurrence of any direct outgrowth of IM cells transformed by the virus in vivo.

Antigens, Viral

Activation of human lymphocyte subpopulations by rabbit anti-human beta2-microglobulin and by lipopolysaccharide.

Rabbit anti-human beta2-microglobulin (anti-beta2m) was found to increase DNA synthesis in peripheral blood lymphocytes (PBLs) and in cells from abdominal lymph nodes, spleen, tonsil, adenoid, appendix, and bone marrow. The response to anti-beta2m was highest in cells originating from abdominal lymph node, appendix, and spleen. These organs were shown to contain a high proportion of surface-Ig-positive cells. No response to anti-beta2m was seen in thymus cells or in B-cell-depleted lymphocyte populations. Lipopolysaccharide (LPS) increased DNA synthesis in spleen cells, bone marrow cells, tonsil cells, and, sometimes, in cells from abdominal lymph nodes but weakly or not at all in PBLs. To study whether anti-beta2m and LPS activated the same subpopulation of lymphocytes, cultures were exposed to both mitogens in various concentrations. The effect on DNA synthesis in spleen cells was almost additive. This may indicate that these two polyclonal B-cell activators (PBAs) stimulate mainly distinct subsets of B cells in spleen. On the other hand, these two mitogens have a synergistic effect on DNA synthesis in PBLs. Since anti-beta2m is the first described selective B-cell mitogen activating human PBLs, it might be of clinical importance in the functional characterization of lymphocyte subpopulations.

B-Lymphocytes

The effect of thoracic duct drainage on lymphocyte dynamics and clinical symptoms in patients with rheumatoid arthritis.

Thoracic duct drainage (TDD) was performed in 4 patients with severe rheumatoid arthritis. Clinical effects were apparent in all during drainage, but the term of TDD and the cumulative number of lymphocytes drained had no direct relation to the improvement of clinical symptoms. The number of lymphocytes in the peripheral blood increased despite discharge of lymphocytes from the thoracic duct in the very early stage of drainage, suggesting that lymph drainage from thoracic duct accelerates migration of lymphocytes from lymphocyte pools to the blood stream. Biopsy specimens of synovial membranes obtained post-TDD showed marked decrease of mononuclear cell infiltration as compared to the specimens obtained preoperatively. These findings suggest that clinical effectiveness may be due not only to systemic immunosuppression induced by lymphocyte depletion but also to accelerated migration of inflammatory cells from the synovial tissues to the blood stream occurring with dynamic change of lymph flow during TDD.

Adult

Evidence for antibody-dependent cell-mediated cytotoxicity by T cells bearing receptors for IgG.

Human lymphocyte populations comprising T cells, T depleted lymphocytes, and T cells enriched for, or depleted of, IgG Fc receptor-bearing (TG) cells, were separated using rosette techniques. All lymphocytes were assessed for the ability to lyse antibody-coated chicken erythrocytes and SL2 mouse lymphoma cells. Their activity was compared with that of monocytes and neutrophil-enriched preparations. IgG Fc receptor positive cells within the T population were highly active in both cytotoxicity assays; the activity could not be ascribed to contamination by monocytes or neutrophils. The TG cells forming junctions with the target cells possessed a characteristic ultrastructure.

Antibody-Dependent Cell Cytotoxicity

Suppressor cells in mice with murine mammary tumor virus-induced mammary tumors. I. Inhibition of mitogen-induced lymphocyte stimulation.

Suppressor cell activity was present in the glass-adherent fraction of spleen cells from C3H mice bearing murine mammary tumor virus-induced mammary tumors. These cells effectively suppressed the blastogenic response of syngeneic normal lymphocytes to concanavalin A (Con A). Suppression by the spleen cells from mammary tumor-bearing mice was not dependent on DNA synthesis. Removal of the suppressor cells from spleen cell suspensions of tumor-bearing mice was not dependent on DNA synthesis. Removal of the suppressor cells from spleen cell suspensions of tumor-bearing animals (TBA) by passage of the cells on glass wool columns increased the Con A response of the remaining cells by fourfold to eightfold. Characterization of the suppressor population indicated that the cells were also adherent to nylon wool but not to plastic and contained a significantly increased proportion of surface immunoglobulin-bearing and complement receptor-bearing lymphocytes. Depletion of macrophages and T-cells did not remove the suppressive activity from the spleens of the TBA. The results were consistent with the identification of the suppressor cell as a B-cell.

Animals

Thoracic duct drainage and antilymphocyte globulin for renal transplantation in man.

Thoracic duct drainage resulting in a lymphocyte depletion of more than 20 x 10(9) cells was performed during the three months prior to transplantation in 37 patients. Results obtained in this group of patients were compared to those in transplant recipients similarly treated, over the same period, but not subjected to thoracic duct drainage. Both groups received comparable doses of antilymphocyte globulins, azathioprine and corticosteroids. No clear-cut difference in transplantation outcome was found when recipients of kidneys from related living donors (whether HLA identical or HLA haploidentical) were considered. By contrast, an improved transplant survival and a decreased incidence of rejection crises were observed in recipients of kidneys from cadaver donors when a thoracic duct drainage was performed prior to transplantation. The immunosuppressive effect of thoracic duct drainage, probably enhanced by antilymphocyte globulin treatment, is therefore a valuable adjunct to more conventional methods of pretreating human cadaveric transplant recipients.

Antilymphocyte Serum

Clinico-pathological study of heartwater in goats.

The clinico-pathological features of heartwater were studied in goats experimentally infected with a Nigerian isolate of Cowdria ruminantium. Significant drop in haemoglobin values and marked leukopaenia caused by lymphopaenia and neutropaenia and a fall in total serum protein were observed during the course of the disease. A significant increase in the alpha-globulins and an apparent fall in the gamma-globulins also occurred. Marked depletion of lymphocytes in the follicles of spleen and lymph nodes was observed in histological sections. A dramatic rise in blood levels of glucose, pyruvate and lactate, and a drop in blood pH occurred terminally and appeared to contribute to the fatal outcome of the disease.

Agammaglobulinemia

Optics-free spatial genomics for mapping mammalian brain aging by IRISeq.

Spatial transcriptomics has emerged as a transformative approach for in situ mapping of cellular heterogeneity and interactions, yet existing methods often compromise throughput, cost and tissue coverage. Here we introduce Imaging Reconstruction using Indexed Sequencing (IRISeq): an optics-free, cost-effective platform that leverages spatial interaction mapping by indexed sequencing to profile tissues at adjustable sizes and resolutions (5-50 µm). We applied IRISeq to map gene expression across more than 70 coronal sections from both adult and aged mouse brains, including wild-type and two lymphocyte-deficient models (Rag1 and Prkdc mutants) and generated more than 460,000 spatial transcriptome profiles. Our integrated analysis with 783,264 single-cell transcriptomes revealed region-specific aging signatures that are lymphocyte dependent, notably a downregulation of interferon signaling and inflammation in ventricular regions upon lymphocyte depletion, alongside mutant-specific upregulation of senescence pathways. Furthermore, lymphocyte deficiency was linked to preserved abundance of ependymal cells that line the brain's ventricles and to distinct microglial state dynamics, highlighting a key role for lymphocytes in driving inflammatory processes during brain aging. Overall, IRISeq provides a high-throughput and cost-effective solution for spatially resolved transcriptomic profiling, opening new avenues for elucidating region-specific cellular mechanisms underlying aging and identifying potential therapeutic targets to preserve brain homeostasis.

Animals

Hodgkin's disease. An immunodepleting and immunosuppressive disorder.

Irradiated leukocytes or mononuclear leukocytes, from 16 out of 30 patients with Hodgkin's disease and from one patient with the Sézary syndrome, stimulated in culture subnormal (3H)thymidine incorporation by allogeneic lymphocytes from normal individuals. This abnormality was not demonstrated in any of 30 other patients with non-Hodgkin's lymphomas. Subnormal mixed leukocyte culture reaction activation was caused by suppression of the mixed leukocyte reaction by patients' cells. Inhibition of the reaction by patient mononuclear leukocytes was corrected when adherent cells were removed or when protein synthesis was inhibited with cycloheximide. The inhibitory cells were probably lymphocytes since selective removal of phagocytic cells did not remove the inhibition by other patient mononuclear leukocytes. The presence in culture of as few as 2,500 granulocytes per mm3 also reduced responses when target cells were from patients with Hodgkin's disease. Patient cells no longer suppressed the mixed leukocyte reaction after patients entered clinical remission which suggests that suppression is a reversible, disease-related abnormality. Thus, the immune deficiency with advance Hodgkin's disease caused by ly lymphocyte depletion may be compounded by a relative excess of suppressor lymphocytes. The overall immunodeficiency may be further compounded by suppression of immune response by granulocytes at even physiologic concentrations.

Dermatitis, Exfoliative

Aryl hydrocarbon hydroxylase activity in subpopulations of peripheral blood mononuclear cells.

Peripheral blood mononuclear cells (PMC), isolated by density gradient techniques with Ficoll-Hypaque, contain T- and B-lymphocytes and monocytes. Aryl hydrocarbon hydroxylase (AHH) activity was measured in PMC subfractions consisting of T-lymphocyte-enriched, T-lymphocyte-depleted, and monocyte-depleted populations. The T-cell-enriched populations consistently showed enhancement of AHH activity with both the fluorometric and radiometric technique when compared to the total PMC population. This enhanced AHH activity was observed when T-cell-enriched populations were isolated either before or after 96 hr of lymphocyte culture, by the sheep red blood cell rosette method, or by the nylon wool column technique before lymphocyte culture. T-cell-depleted populations (B-cell enriched) obtained by sheep red blood cell rosette method had diminished AHH activity. Monocytes were shown to contribute to the total PMC AHH activity through an indirect technique by first depleting the monocytes from PMC with the carbonyl iron method. The monocyte-depleted populations had less AHH activity than did the total PMC population after both 24 and 96 hr of culture. The greatest amount of AHH activity was present in PMC populations with their native number of monocytes when cultured for 96 hr in the presence of mitogens.

Aryl Hydrocarbon Hydroxylases

Hodgkin's disease stage I and II. A comparison between two different treatment policies.

A retrospective analysis was performed on 145 patients with Stage I and II Hodgkin's disease treated over an 11-year period. Sixty-two patients (Group I) received a mantle field without systematic irradiation of the para-aortic lymph nodes. Eight-three patients (Group II) received radiotherapy according to the folowing policy: all Stage IB and IIB and all mixed cellularity and lymphocytic depletion types received total nodal irradiation while stage IA and IIA nodular sclerosing and lymphocytic predominance cases received irradiation to a mantle field and to the para-aortic lymph nodes. The characteristics of the two groups were roughly comparable in age range, sex, staging, histopathologic subtypes and total irradiation doses. All patients had lymphangiograms although not all underwent staging laparotomies. The staging laparotomy did not appear to have an influence within each group. The extent of irradiation did significantly affect both the incidence of further manifestation of disease as well as survival rates. The frequency of lymph node extension, organ extension and local recurrence for Group I was 24%, 14%, and 3%, while for Group II it was 4%, 6%, and 6%, respectively. The seven-year absolute survival rate for Group I was 57% while for Group II it was 93%.

Adolescent

Immunological characterization of mononuclear cells and morphological findings in patients with mammary carcinoma.

Mononuclear cells from peripheral blood and draining lymph nodes of 40 patients with invasive mammary carcinoma were examined for various immunological cell surface markers including surface membrane immunoglobulins and rosetting properties (E, EA, EAC). No significant relationship could be established to anyone of the following criteria for which the literature reports varying prognostic values: Clinical staging of the disease , histological tumor type, grading, nuclear differentiation, round cell infiltration, perivenous infiltration, sinus histiocytosis, and lymph node reaction patterns (lymphocyte predominance, germinal center predominance, lymphocyte depletion, unstimulated nodes). From the reported results it is concluded that the analysis of lymphocyte cell surface markers in mammary carcinoma is not a suitable parameter for supporting the existence of specific or unspecific anti-tumor immune reactions which may be suspected from certain histological reaction patterns.

Breast Neoplasms

Ecotaxis: the principle and its application to the study of Hodgkin's disease.

A study of the function, characterization and distribution of T and B lymphocytes in five children with Hodgkin's disease is presented. The results, indicating that lymphocyte depletion in the peripheral blood does not necessarily reflect an overall lack of circulating lymphocytes, are presented to demonstrate that failure of ecotaxis (normal lymphocyte migration and distribution) can occur in man. The underlying reasons for such failure and their relevance to the pathogenesis of Hodgkin's disease are discussed.

Adolescent

Human T lymphocyte receptors for IgM: control by IgG-binding lymphocytes.

The capacity of human peripheral blood lymphocytes to bind antigen-IgG or antigen-IgM-antibody complexes was investigated using a rosette technique with ox erythrocytes (E) coated with rabbit IgG (AG) or IgM (AM) antibodies. EAM rosette formation was achieved only in suspensions pre-incubated for 24 h at 37 degrees C. Addition of either EAM or EAG complexes to the culture medium was shown to prevent the formation of EAM rosettes. The inhibition was reversible, it was not due to trace IgM contaminants in the IgG antibody fraction. It was not observed when lymphocytes depleted of EAG-rosetting cells were incubated with EAG complexes. Inhibition of the expression of lymphocyte receptors for IgM can be regarded as a consequence of the modulation of surface receptors for IgG and involves an interaction between the two lymphocyte subsets bearing surface receptors for IgG and IgM, respectively. However, these experiments do not exclude the possibility that a few cells which bind EAG may lose their receptors by modulation and then express a receptor for IgM.

Antigen-Antibody Complex

The thymus in patients with allogeneic bone marrow transplants.

The thymus glands from 11 patients with aplastic anemia or acute leukemia who received allogeneic bone marrow transplants were studied at autopsy. All showed marked cortical involution. In the short-term survivors the medulla and perivascular spaces were lymphocyte-depleted and the epithelial cells formed pseudorosettes. In those surviving over 2 months, increasing numbers of small lymphocytes were present, presumably reconstituted with donor lymphocytes. Phagocytosis of cellular debris was frequent, especially in patients with graft-versus-host reaction (GVHR) or treated with anithymocyte globulin (ATG). Plasma cells were numerous in perilobular tissue and were occasionally found within the medulla. The findings are compatible with the concept that the thymus plays an important role in the immune deficiency experienced after allogeneic bone marrow transplantation and in the subsequent lymphoid reconstitution.

Adolescent