ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of total saponins from Sanguisorba officinalis (DYS) on hematopoietic cell proliferation, differentiation and the expression level of IL-3R and c-kit.</p><p><b>METHOD</b>Baf3 and 32D cells were cultured with or without IL-3, then the cells were exposed to DYS in different concentrations of 5, 10, 20, 30 and 40 mg x L(-1) for 24, 48, 72 and 96 hours separately. After that, the cell proliferation and differentiation capacity were determinated by the methods of CCK8 and Giemsa staining separately. The effects of DYS on the expression level of IL-3 receptor in Baf3 cells and the expression level of c-kit in 32D cells were determinated using RT-PCR.</p><p><b>RESULT</b>DYS promotes alone proliferation of Baf3 cells and 32D cells after 48 h. In contrast to control cells, 32D cells containing DYS without IL-3 form many large clusters. DYS also increases the proliferation when cultured with IL-3. High concentration of DYS induce alone the differentiation of 32D cells and increase alone the number of the polyploidy megakaryocyte. Moreover, DYS increases alone the expression level of IL-3R in Baf3 cells and the expression level of c-kit in 32D cells separately.</p><p><b>CONCLUSION</b>Our data shows DYS can promote alone proliferation and differentiation of megakaryocyte progenitor cells. The proliferative and differentiative effect of DYS on megakaryocyte progenitor cells is correlated to the up-regulation of IL-3 receptor and c-kit expression.</p>
Subject(s)
Animals , Mice , Cell Differentiation , Genetics , Cell Line , Cell Proliferation , Dose-Response Relationship, Drug , Gene Expression , Interleukin-3 , Pharmacology , Megakaryocyte Progenitor Cells , Metabolism , Proto-Oncogene Proteins c-kit , Genetics , Receptors, Interleukin-3 , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sanguisorba , Chemistry , Saponins , Pharmacology , Time FactorsABSTRACT
How Th2 immunity develops in vivo remains obscure. Basophils have been considered key innate cells producing IL-4, a cytokine essential for Th2 immunity. Increasing evidence suggests that basophils are dispensable for the initiation of Th2 immunity. In this study, we revisited the role of basophils in Th2 immune responses induced by various types of adjuvants. Mice deficient in IL-3 or IL-3 receptor, in which basophil lymph node recruitment is completely abolished, fully developed wild type level Th2 CD4 T cell responses in response to parasite antigen or papain immunization. Similar finding was also observed in mice where basophils are inducibly ablated. Interestingly, IL-4-derived from non-T cells appeared to be critical for the generation of IL-4-producing CD4 T cells. Other Th2 promoting factors including IL-25 and thymic stromal lymphopoietin (TSLP) were dispensable. Therefore, our results suggest that IL-3- and basophil-independent in vivo Th2 immunity develops with the help of non-T cell-derived IL-4, offering an additional mechanism by which Th2 type immune responses arise in vivo.
Subject(s)
Animals , Mice , Basophils , Immunization , Interleukin-3 , Interleukin-4 , Lymph Nodes , Papain , Parasites , Receptors, Interleukin-3 , T-LymphocytesABSTRACT
The aim of this study was to find new idea for clinical treatment of aplastic anemia. Immune-mediated aplastic anemia mice were developed, IL-3 in the supernatant with PHA stimulating splenic cells was detected by ELISA, semi-quantiting analysis of IL-3R was performed by point hybridization. The results showed that the IL-3 level in the supernatant with PHA stimulating splenic cells of immune-mediated aplastic anemia mice was higher than controls, difference between them was significant (P <0.001), while amount of IL-3 receptor by semi-quantiting analysis was lower than control significantly. In conclusion, the IL-3 receptor expression level is important for pathogenesis and treatment strategy of aplastic anemia.
Subject(s)
Animals , Mice , Anemia, Aplastic , Allergy and Immunology , Pathology , Bone Marrow , Pathology , Interleukin-3 , Mice, Inbred BALB C , Mice, Inbred DBA , RNA, Messenger , Receptors, Interleukin-3 , GeneticsABSTRACT
To investigate the effect of fluoxetine, one of selective serotonin reuptake inhibitors (SSRIs), on the immune system, human peripheral blood mononuclear cells (PBMC) were treated with fluoxetine (10 7 M) for 24 h, and immune-related genes were analyzed by cDNA microarray. Expression of the immune- related genes such as CD107b (LAMP-2), CD47 receptor (thrombospondin receptor), CD5 antigen-like (scavenger receptor cysteine rich family), copine III (CPNE3), interleukin (IL) -18 (interferon-gamma- inducing factor), integrin alpha 4 (CD49d), integrin alpha L subunit (CD11a), IL-3 receptor alpha subunit, L apoferritin, and small inducible cytokine subfamily A (Cys-Cys) member 13 (SCYA13) was induced by fluoxetine. This result suggests that fluoxetine may affect the immune system, and provides fundamental data for the involvement of SSRIs on immunoregulation.
Subject(s)
Humans , Apoferritins , Cysteine , DNA, Complementary , Fluoxetine , Immune System , Interleukins , Oligonucleotide Array Sequence Analysis , Receptors, Interleukin-3 , Selective Serotonin Reuptake InhibitorsABSTRACT
The hematopoietic system of the mouse arises from extraembryonic mesoderm that migrate through primitive streak to the presumptive yolk sac at day 7.0 of gestation. However, the mechanisms regulating mesoderm commitment to hematopoietic lineages remain poorly understood. Previous studies demonstrated that the development kinetics and growth factor responsiveness of hematopoietic precursors derived from embryonic stem cells (ES cells) is similar to that found in the yolk sac, indicating that the onset of hematopoiesis within the embryoid bodies (EBs) parallels that found in the embryo. Furthermore, in vitro differentiation of ES cells to hematopoietic cells is valuable for establishment of therapeutic clone against a variety of hematological disorders. Despite the identification of multipotential hematopoietic progenitors in EBs, a subset of more primitive progenitors, identical to the high proliferative potential colony-forming cells (HPP-CFC) derived from human and murine hematopoietic tissues, have not been clearly identified regarding particular their replating potential in vitro. HPP-CFC is among the most primitive hematopoietic multipotent precursors cultured in vitro. In this study, our aim was to investigate the in vitro and in vivo hematopoietic capacity of HPP-CFC within the day 12 EBs, rather than the expansion of more committed progenitors. In this study the HPP-CFC could be detected within EBs differentiated for 5 to 14 days of murine ES cells, but the development dynamics of the HPP-CFC differed greatly among distinct serum lots. Qualitatively HPP-CFC is capable of forming secondary colonies. As to our expectation the ES cells-derived HPP-CFC demonstrated similar regeneration capacity to those from yolk sac, giving rise to secondary granulocyte, erythrocyte, macrophage and mast cells, however largely differed from the counterparts of adult bone marrow. In addition, by RT-PCR ES cells-derived HPP-CFC were found to express transcription factors associated closely with stem cell proliferation including SCL, GATA-2 and AML1 as well as various receptors of hematopoietic growth factors such as c-kit, GM-CSF receptor and interleukin 3 receptor et al. Finally, in order to understand the in vivo hematopoietic capacity of the ES cells-derived HPP-CFC, spleen colony-forming unit (CFU-S) assay was performed. Nevertheless, typical CFU-S was not observed after transplantation of the day 12 EB cells or HPP-CFC colonies into lethally irradiated adult murine. In conclusion the HPP-CFC differentiated from murine ES cells displayed robust hematopoietic activity in vitro, however their in vivo reconstitution ability was not detected. The difference between in vitro and in vivo hematopoietic activities of ES cells-derived primitive hematopoietic precursors deserves further investigation.
Subject(s)
Animals , Humans , Mice , Basic Helix-Loop-Helix Transcription Factors , Genetics , Cell Differentiation , Genetics , Physiology , Colony-Forming Units Assay , Core Binding Factor Alpha 2 Subunit , Genetics , Embryonic Stem Cells , Cell Biology , GATA2 Transcription Factor , Genetics , Hematopoietic Stem Cells , Cell Biology , Metabolism , Proto-Oncogene Proteins , Genetics , Proto-Oncogene Proteins c-kit , Genetics , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor , Genetics , Receptors, Interleukin-3 , Genetics , Reverse Transcriptase Polymerase Chain Reaction , T-Cell Acute Lymphocytic Leukemia Protein 1ABSTRACT
La protección del ataque de microorganismos invasores incluye mecanismos de resistencia específicos y no específicos. Los primeros son los responsables de impedir la mayoría de las infecciones, mientras que los segundos participan una vez que los microorganismos o sus productos han entrado a los tejidos. Los mecanismos de resistencia específicos, también conocidos como respuesta inmunológica, incluyen la participación de células y moléculas con capacidad de reconocer y reaccionar específicamente en contra del microorganismo invasor. En la activación de estos mecanismos participan linfocitos y células accesorias que se comunican a través de una compleja red de señales que incluyen moléculas asociadas a la membrana y moléculas solubles. Del tipo de interacciones establecidas de producirá una respuesta mediada por anticuerpos o por células inmunes, en algunos casos estas interacciones también generan una falta de respuesta (anergia) específica