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Clinical and Experimental Immunology Jun 1984Human tonsil non-T cells were separated into surface IgD positive (sIgD+), surface IgD negative (sIgD-), sIgM+ and sIgM- fractions by rosetting and gradient...
Human tonsil non-T cells were separated into surface IgD positive (sIgD+), surface IgD negative (sIgD-), sIgM+ and sIgM- fractions by rosetting and gradient centrifugation with ox red blood cells, coated with immunosorbent purified goat anti-human heavy chain antibodies. No activation or suppression was found as a result of the separation technique. The sIg patterns of the isolated B cell fractions showed the effectiveness of the separations; sIgD+ and sIgM+ fractions were to a great extent overlapping populations, but there was a definite population of s mu + delta- cells (about 20% of the sIgD- fraction). The isolated fractions were tested for proliferative responses to Staphylococcus aureus (Sta) and pokeweed mitogen (PWM). The sIgM+ fraction showed the best response to Sta, eight times better than the sIgM- fraction, while the sIgD+ and sIgD- fraction showed equal responses to Sta. The sIgM- fraction responded best to PWM but all fractions responded quite well. Our results indicate that PWM is a mitogen to a whole variety of rather mature B cells while the Sta target B cell is restricted to the relatively immature sIgM+D- and sIgM+D+ cells.
Topics: B-Lymphocytes; Cell Division; Child; Humans; Immunoglobulin Heavy Chains; Immunoglobulin delta-Chains; Immunoglobulin mu-Chains; Palatine Tonsil; Pokeweed Mitogens; Receptors, Antigen, B-Cell; Rosette Formation; Staphylococcus aureus
PubMed: 6430613
DOI: No ID Found -
American Journal of Veterinary Research Aug 2011To determine effects of infection with bovine leukosis virus (BLV) on lymphocyte proliferation and apoptosis in dairy cattle.
OBJECTIVE
To determine effects of infection with bovine leukosis virus (BLV) on lymphocyte proliferation and apoptosis in dairy cattle.
ANIMALS
27 adult Holstein cows.
PROCEDURES
Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood from lactating Holstein cows seronegative for BLV (n = 9 cows), seropositive for BLV and aleukemic (aleukemic; 9), and seropositive for BLV and persistently lymphocytotic (PL; 9). Isolated PBMCs were assayed for mitogen-induced proliferation and were analyzed by means of flow cytometry. The PBMCs from a subset of each group were assayed for apoptosis, caspase-9 activity, and expression of selected genes related to apoptosis.
RESULTS
PL cows had significantly higher total lymphocyte counts and significantly lower proportions of T-lymphocyte populations than did BLV-negative and aleukemic cows. Both groups of BLV-infected cows had significantly higher proportions of B cells and major histocompatibility complex II-expressing cells than did BLV-negative cows. Proliferation with concanavalin A was significantly lower for PL cows, compared with proliferation for BLV-negative cows. Pokeweed mitogen-induced proliferation was significantly higher for aleukemic and PL cows than for BLV-negative cows. Gene expression of apoptosis-inhibitory proteins BCL2 and BCL2L1 was significantly higher for aleukemic cows and expression of BCL2 was significantly higher for PL cows than for BLV-negative cows.
CONCLUSIONS AND CLINICAL RELEVANCE
Cattle infected with BLV had marked changes in PBMC populations accompanied by alterations in proliferation and apoptosis mechanisms. Because the relative distribution and function of lymphocyte populations are critical for immune competence, additional studies are needed to investigate the ability of BLV-infected cattle to respond to infectious challenge.
Topics: Animals; Antibodies, Viral; Apoptosis; B-Lymphocytes; Caspase 9; Cattle; Concanavalin A; Enzootic Bovine Leukosis; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression; Leukemia Virus, Bovine; Leukocyte Count; Leukocytes, Mononuclear; Lymphocyte Activation; Pokeweed Mitogens; Proto-Oncogene Proteins c-bcl-2; T-Lymphocytes
PubMed: 21801063
DOI: 10.2460/ajvr.72.8.1059 -
Immunology Apr 1982The in vitro treatment with Fusarenon-X, a mycotoxin produced by Fusarium nivale Fn 2B, depressed the mitogenic responses of mouse lymphocytes to the T-cell mitogens,...
The in vitro treatment with Fusarenon-X, a mycotoxin produced by Fusarium nivale Fn 2B, depressed the mitogenic responses of mouse lymphocytes to the T-cell mitogens, phytohaemagglutinin (PHA) and concanavalin A (Con A), but to a lesser extent to B-cell mitogen, a bacterial lypopolysaccharide (LPS). The in vitro treatment of mice with Fusarenon-X also decreased the responsiveness of splenic lymphocytes to the T-cell mitogen, Con A. Administration of Fusarenon-X to BALB/c mice before immunization significantly reduced anti-DNP IgE and IgG1 responses, while the treatment of mice after immunization was less effective. The number of spleen cells of the treated mice was increased with a reduction of T lymphocytes and an increase of granulocytic elements. In vitro antibody responses of spleen cells from Fusarenon-X treated mice by pokeweed mitogen (PWM) or LPS were also suppressed. Reconstruction experiments using sIg positive and negative spleen cells from normal and Fusarenon-X treated mice showed that this inhibitory effect resided in the sIg negative cell population.
Topics: Animals; Antibody-Producing Cells; Concanavalin A; Immunoglobulin E; Immunoglobulin G; Immunosuppressive Agents; Lipopolysaccharides; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Mycotoxins; Phytohemagglutinins; Pokeweed Mitogens; Rats; Rats, Inbred Strains; Receptors, Antigen, B-Cell; Sesquiterpenes; Trichothecenes
PubMed: 7200080
DOI: No ID Found -
Proceedings of the National Academy of... Jun 1981The fate of two recently described human B lymphocyte-specific antigens (B1 and B2) was studied after B-cell activation in vivo and in vitro. Whereas both B1 and B2 were...
The fate of two recently described human B lymphocyte-specific antigens (B1 and B2) was studied after B-cell activation in vivo and in vitro. Whereas both B1 and B2 were present on virtually all B cells from normal lymph nodes, B2 was absent from approximately 50% of B cells from hyperplastic lymph nodes. When B cells from spleen, tonsil, or peripheral blood were stimulated in vitro with pokeweed mitogen, activated cells were found to lose B2 (days 4-5) and subsequently B1 (days 6-7). Temporally, B2 loss was accompanied by loss of surface IgD, expression of T10, and the development of intracytoplasmic IgM; B1 loss was correlated with the acquisition of surface IgG and the appearance of intracytoplasmic IgG. Peripheral blood B cells, on which B2 is normally only weakly expressed (B1++++B2+) in contrast to B cells from secondary lymphoid organs (B1++++B2++), exhibited a transitory increase in B2 expression to the B1++++B2++ phenotype prior to B2 disappearance during activation. Taken together with other findings, this observation suggests that peripheral blood may contain a relatively immature subpopulation of B cells.
Topics: Antibody-Producing Cells; Antigens, Surface; B-Lymphocytes; Cell Differentiation; Cytoplasm; Humans; Immunoglobulins; Lymphocyte Activation; Lymphoid Tissue; Phenotype; Pokeweed Mitogens; Tissue Distribution
PubMed: 6973760
DOI: 10.1073/pnas.78.6.3848 -
The Journal of Clinical Investigation May 1983The immunoglobulin-synthesizing activities of peripheral blood mononuclear cells from 57 untreated patients with Hodgkin's disease and 47 normal subjects were compared....
The immunoglobulin-synthesizing activities of peripheral blood mononuclear cells from 57 untreated patients with Hodgkin's disease and 47 normal subjects were compared. Cumulative amounts of IgM and IgG synthesized and secreted by unstimulated and pokeweed mitogen-stimulated cells over a 7-d period were determined in a solid-phase radioimmunoassay. Synthesis of IgM in unstimulated cultures and of both IgM and IgG in cultures stimulated with pokeweed mitogen was markedly reduced in patients with Hodgkin's disease, whereas the mean level of the spontaneous IgG synthesis was enhanced. The degree and frequency of in vitro abnormalities were not influenced by disease stage or histology. Depression of pokeweed mitogen-induced immunoglobulin synthesis did not correlate with excessive number of monocytes and it was unaffected by removal of phagocytic cells or addition to the cultures of monocytes from normal individuals. On the other hand, monocytes isolated from blood of patients with Hodgkin's disease were even more effective than normal monocytes in supporting pokeweed mitogen-induced immunoglobulin synthesis by normal phagocyte-depleted mononuclear cells. Synthesis of both IgM and IgG induced by pokeweed mitogen remained subnormal after addition to patient B cell cultures of autologous irradiated T cells or allogeneic normal T lymphocytes. T cells from patients with Hodgkin's disease appeared at least as effective as normal T cells in helping pokeweed mitogen-induced immunoglobulin production by normal B cells. However, when normal T cells were co-cultured with B cells from patients with Hodgkin's disease, spontaneous IgG synthesis declined, whereas the addition of patient T cells to normal B cells resulted in an increase of spontaneous IgG synthesis. In patients showing depression of pokeweed mitogen-induced immunoglobulin synthesis the lymphoproliferative response and immunoglobulin synthesis stimulated by Staphylococcus aureus bacteria of the Cowan first strain, a T cell independent B cell mitogen, were also markedly reduced. These studies demonstrate impairment of immunoglobulin synthesis by cultured lymphocytes from untreated patients with Hodgkin's disease after stimulation with polyclonal B cell activators and suggest that the in vitro abnormalities may be, at least in part, the result of a preexisting in vivo activation of lymphocytes in Hodgkin's disease patients.
Topics: Adolescent; Adult; Aged; B-Lymphocytes; Cells, Cultured; Child; Female; Hodgkin Disease; Humans; Immunoglobulin G; Immunoglobulin M; Lymphocytes; Male; Middle Aged; Monocytes; Pokeweed Mitogens; Staphylococcus aureus; T-Lymphocytes
PubMed: 6602150
DOI: 10.1172/jci110890 -
Poultry Science Jun 2006Fumonisin B1 (FB1) is a mycotoxin produced by the fungus Fusarium verticillioides (formerly Fusarium moniliforme) and is found in diverse crops such as corn, wheat, and...
Fumonisin B1 (FB1) is a mycotoxin produced by the fungus Fusarium verticillioides (formerly Fusarium moniliforme) and is found in diverse crops such as corn, wheat, and barley. Many diseases linked to FB1, such as porcine pulmonary edema, rat hepatic cancer, and equine leukoencephalomalacia, indicate a compromised immune system. The purpose of this study was to determine whether FB1 altered immunological responses in various cell populations of Single Comb White Leghorn chicks. Cells collected for this study were obtained from those immunological organs with well-defined responses (i.e., spleen, thymus, and blood). Cell populations were exposed to 5 to 50 microg/mL FB1 in vitro for 24 to 72 h, and viability and mitogenic response were evaluated. The effects of FB1 on the mitogenic response were evaluated in cell populations from the spleen and blood stimulated with the mitogens, lipopolysaccharide, concanavalin A, and pokeweed mitogen and in thymocytes stimulated with concanavalin A. The 3-(4,5-dimethylthazol-2-yl)-diphenyl-2H-tetrazolium bromide (MTT) reduction assay was used to assess viability and mitogenic response. Fumonisin B1 decreased spleen cell viability and mitogenic response, albeit the degree of decrease varied with mitogen and time of exposure. Fumonisin B1 increased number of viable thymic cells at 50 microg/mL but had no effect on the mitogenic response of thymocytes. Fumonisin B1 had no effect on blood lymphocyte viability or mitogenic response.
Topics: Animals; Cell Survival; Chickens; Concanavalin A; Fumonisins; Lipopolysaccharides; Lymphocyte Activation; Lymphocytes; Mitogens; Mitosis; Pokeweed Mitogens; Spleen; Thymus Gland
PubMed: 16776470
DOI: 10.1093/ps/85.6.1020 -
Cellular & Molecular Biology Letters 2007The edeines analogs were tested in several in vitro and in vivo assays using the mouse model, with edeine B (peptide W1) and cyclosporine A as reference compounds. The...
The edeines analogs were tested in several in vitro and in vivo assays using the mouse model, with edeine B (peptide W1) and cyclosporine A as reference compounds. The peptides displayed moderate, stimulatory effects on concanavalin A-induced (ConA-induced) splenocyte proliferation, whereas their effects on pokeweed mitogen-induced (PWM-induced) splenocyte proliferation were inhibitory. The peptides inhibited lipopolysacharide-induced (LPS-induced) tumor necrosis factor alpha production but had little effect on interleukin 6 production. In the model of the humoral immune response in vitro to sheep red blood cells, peptide 1 was distinctly stimulatory in the investigated concentrations (1-100 microg/ml), whereas peptides 3 and 4 only stimulated the number of antibody-forming cells at the highest concentration (100 microg/ml). In the model of the delayed type hypersensitivity in vivo to ovalbumin, the peptides were moderately suppressive (3 being the most active). The reference peptide W1 stimulated ConA-induced cell proliferation at 1-10 microg/ml but was inhibitory at 100 microg/ml. It also inhibited PWM-induced cell proliferation in a dose-dependent manner. This peptide had no effect on the humoral immune response in vitro or on cytokine production, but inhibited DTH reaction in vivo. The relationship between structure and activity, and a possible mode of action of the peptides, is discussed in this paper.
Topics: Animals; Cell Proliferation; Concanavalin A; Edeine; Hypersensitivity; Immunity; Interleukin-6; Lipopolysaccharides; Mice; Ovalbumin; Pokeweed Mitogens; Sheep; Spleen; Tumor Necrosis Factor-alpha
PubMed: 17149559
DOI: 10.2478/s11658-006-0061-z -
Clinical and Experimental Immunology Nov 1972Mouse lymphocytes were cultured with phytohaemagglutinin, pokeweed mitogen, Concanavalin A or allogeneic cells. The expression of theta, -2 and Ig determinants on the...
Lymphocyte activation. I. Expression of theta, H-2 and immunoglobulin determinants on lymphocytes stimulated by phytohaemagglutinin, pokeweed mitogen, concanavalin A or histocompatibility antigen.
Mouse lymphocytes were cultured with phytohaemagglutinin, pokeweed mitogen, Concanavalin A or allogeneic cells. The expression of theta, -2 and Ig determinants on the blasts and surviving lymphocytes was assessed by indirect autoradiography using unlabelled mouse antisera and iodinated rabbit anti-mouse light chain. The great majority of the responding cells carried the θ-antigen (82–100%) and few carried Ig determinants (0–12%). There was no difference in the concentration of antiserum required to demonstrate -2 or θ on fresh lymphocytes, cultured lymphocytes or PHA-induced blasts. In mixed leucocyte cultures between parental and F1 cells most of the blasts were derived from the parent (90%) whereas in cultures of two parental strains both strains contributed equally to the response. These data support the view that thymus-derived cells predominate in the response to non-specific mitogens and histocompatibility antigen, and that parental/F1 cultures exhibit a unidirectional response.
Topics: Animals; Autoradiography; B-Lymphocytes; Cells, Cultured; Concanavalin A; Epitopes; Histocompatibility Antigens; Immunoglobulin Fragments; Iodine Isotopes; Lectins; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred BALB C; Mice, Inbred CBA; Rabbits; Spleen; Stimulation, Chemical; T-Lymphocytes
PubMed: 4118608
DOI: No ID Found -
Clinical and Experimental Immunology Dec 1989The effect of human lymphokine-activated killer (LAK) cells on pokeweed mitogen (PWM) induced immunoglobulin synthesis by autologous peripheral blood mononuclear cells...
The effect of human lymphokine-activated killer (LAK) cells on pokeweed mitogen (PWM) induced immunoglobulin synthesis by autologous peripheral blood mononuclear cells (PBMC) was studied. LAK cells induced by the in vitro culture with recombinant human interleukin-2 (IL-2) lysed PWM-activated autologous T cells and B cells, but did not lyse unstimulated lymphocytes. These effector cells which are capable of killing lymphoblasts were shown to express either CD16 surface markers. When CD8(+)-and CD16(+)-enriched cells isolated from the culture with IL-2 were added to cultures containing autologous PBMC and PWM, marked suppression of the IgG production was observed. In contrast, the control CD8(+)-and CD16(+)-enriched cells isolated from the culture without IL-2 showed a weak suppressive effect on PWM-induced IgG synthesis. These results suggest that LAK cells suppress immunoglobulin synthesis by the cytotoxic elimination of activated T cells and B cells.
Topics: B-Lymphocytes; Humans; Immunoglobulin G; Killer Cells, Lymphokine-Activated; Lymphocyte Activation; Pokeweed Mitogens; T-Lymphocytes; T-Lymphocytes, Regulatory
PubMed: 2532992
DOI: No ID Found -
Hereditas 1988
Topics: Environmental Exposure; Humans; Lymphocytes; Micronucleus Tests; Phytohemagglutinins; Piperazine; Piperazines; Pokeweed Mitogens
PubMed: 3192423
DOI: 10.1111/j.1601-5223.1988.tb00194.x