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1.
For determination of the localization of lymphoid and erythroid precursor cells in embryos of Xenopus laevis , diploid-triploid chimeras were produced either by joining embryos antero-posteriorly or by orthotopic grafting of various tissues into N ieuwkoop -F aber st. 22–23 tailbud embryos. The sources of the hemopoietic cells were determined in the chimeric animals at various stages by microspectrophotometry of F eulgen -stained cells. Analyses of chimeras produced by joining embryos antero-posteriorly at different levels showed that the precursor cells that contribute to the hemopoietic cells are localized in the posterior half to three quarters. Orthotopic grafting of ventral or dorsal tissues revealed that the precursor cells that contribute to hemopoietic cells in early larvae are mostly localized in the ventral blood island (VBI) mesoderm, whereas those for late larvae and adults are localized both in the dorso-lateral plate (DLP) mesoderm comprising the prospective mesonephros and in the VBI mesoderm. Reciprocal heterotopic grafting of VBI- and DLP mesoderms showed that the two compartments differ in their capacities to differentiate into hemopoietic cells. It is proposed that the VBI-derived cells migrating towards the primary lymphoid organs constitute the transient hemopoietic population of early larvae, and the importance of the mesonephric region for definitive hemopoiesis is pointed out.  相似文献   

2.
In an effort to label very early erythrocyte and lymphocyte populations and to follow their fate in normally developing postmetamorphic frogs and goitrogen-treated permanent larvae, diploid (2N) and triploid (3N) ventral blood island (VBI) mesoderm was exchanged between neurula stage embryos (about 16-22 hr old). Beginning at 15 days of age, half of the 2N or 3N hosts were treated with sodium perchlorate to prevent thyroxine-induced developmental changes. At larval stages 55-59 (41-48 days) and at 1-2 months postmetamorphosis (110-120 days), the untreated control chimeras and age-matched perchlorate-treated chimeras were killed for analysis of the VBI contribution to blood, spleen, and thymus populations by flow cytometry. The data suggest that grafting of ventral blood island mesoderm is an effective way to label an early larval erythrocyte population that declines after metamorphosis. In perchlorate-blocked permanent larvae this early VBI-derived erythrocyte population persists. In contrast, grafting of VBI mesoderm was less useful as a method to label a larvally distinct lymphocyte population in the thymus and spleen. At the late larval stages that we examined, the proportion of VBI-derived cells in thymus and spleen was not different from that observed after metamorphosis. Inhibition of metamorphosis interfered with the thymocyte expansion that normally occurs after metamorphosis, but the proportion of VBI-derived cells in thymus and spleen was not affected. This suggests that lymphopoiesis occurring in late larval life and after metamorphosis uses a stable persisting population of VBI-derived stem cells as well as dorsally derived stem cells.  相似文献   

3.
Hemopoietic stem and progenitor cells ordinarily residing within bone marrow are released into the circulation following G-CSF administration. Such mobilization has a great clinical impact on hemopoietic stem cell transplantation. Underlying mechanisms are incompletely understood, but may involve G-CSF-induced modulation of chemokines, adhesion molecules, and proteolytic enzymes. We studied G-CSF-induced mobilization of CD34+ CD10+ CD19- Lin- and CD34+ CD10+ CD19+ Lin- cells (early B and pro-B cells, respectively). These mobilized lymphoid populations could differentiate only into B/NK cells or B cells equivalent to their marrow counterparts. Mobilized lymphoid progenitors expressed lymphoid- but not myeloid-related genes including the G-CSF receptor gene, and displayed the same pattern of Ig rearrangement status as their bone marrow counterparts. Decreased expression of VLA-4 and CXCR-4 on mobilized lymphoid progenitors as well as multipotent and myeloid progenitors indicated lineage-independent involvement of these molecules in G-CSF-induced mobilization. The results suggest that by acting through multiple trans-acting signals, G-CSF can mobilize not only myeloid-committed populations but a variety of resident marrow cell populations including lymphoid progenitors.  相似文献   

4.
Data obtained from studies on the origin and development of hemopoietic cells in several classes of vertebrate embryos argue for two distinct sources of hemopoietic cells, the intraembryonic dorsal lateral plate and the extraembryonic ventral blood island/yolk sac. In the present study, a stage by stage comparison of the hemopoietic potential of both of these regions was made during development of the frog, Rana pipiens. Either dorsal lateral plate or ventral blood island mesoderm was reciprocally transplanted between cytogenetically labeled triploid and diploid embryos. The ratio of donor-derived cells to host-derived cells (labeling index) was determined from Feulgen-stained DNA measurements of cells harvested from hemopoietic organs of young larvae. Blood island transplants consistently resulted in larvae with positive labeling of the circulating blood. Transplanted dorsal mesoderm supplied mesonephric granulocytes and thymocytes, but not circulating erythrocytes to larvae. However, the contribution of dorsal mesoderm to larval hemopoiesis fluctuated with respect to embryonic stage at transplantation.  相似文献   

5.
Integrin alphaIIb is a cell adhesion molecule expressed in association with beta3 by cells of the megakaryocytic lineage, from committed progenitors to platelets. While it is clear that lymphohemopoietic cells differentiating along other lineages do not express this molecule, it has been questioned whether mammalian hemopoietic stem cells (HSC) and various progenitor cells express it. In this study, we detected alphaIIb expression in midgestation embryo in sites of HSC generation, such as the yolk sac blood islands and the hemopoietic clusters lining the walls of the major arteries, and in sites of HSC migration, such as the fetal liver. Since c-Kit, which plays an essential role in the early stages of hemopoiesis, is expressed by HSC, we studied the expression of the alphaIIb antigen in the c-Kit-positive population from fetal liver and adult bone marrow differentiating in vitro and in vivo into erythromyeloid and lymphocyte lineages. Erythroid and myeloid progenitor activities were found in vitro in the c-Kit(+)alphaIIb(+) cell populations from both origins. On the other hand, a T cell developmental potential has never been considered for c-Kit(+)alphaIIb(+) progenitors, except in the avian model. Using organ cultures of embryonic thymus followed by grafting into athymic nude recipients, we demonstrate herein that populations from murine fetal liver and adult bone marrow contain T lymphocyte progenitors. Migration and maturation of T cells occurred, as shown by the development of both CD4(+)CD8- and CD4-CD8(+) peripheral T cells. Multilineage differentiation, including the B lymphoid lineage, of c-Kit(+)alphaIIb(+) progenitor cells was also shown in vivo in an assay using lethally irradiated congenic recipients. Taken together, these data demonstrate that murine c-Kit(+)alphaIIb(+) progenitor cells have several lineage potentialities since erythroid, myeloid, and lymphoid lineages can be generated.  相似文献   

6.
The thymocytes in the early larvae of Xenopus laevis have been shown to be derived from precursor cells immigrating interstitially through the mesenchyme into the organ rudiments at 3-4 days of age (Nieuwkoop and Faber stages 42-45). Orthotopic grafting of diploid tissues onto triploid stage 22 embryos followed by ploidy analyses of their hemopoietic cells revealed that both thymocytes and erythrocytes in early larvae are derived from the ventral blood islands (VBI), whereas those in late larvae and adults come mainly from the dorsolateral plate (DLP). To study how the VBI cells of embryos at stage 22 participate in hemopoiesis, a number of interspecific chimeras were produced in X. laevis and X. borealis embryos. Sections of the chimeras at various developmental stages were examined by employing the unique stainability of X. borealis nuclei to quinacrine as a marker; the results show that the VBI-derived cells enter into the circulation around stage 35/36, and that some of them leave the blood vessels to migrate interstitially through the mesenchyme toward the thymic rudiment during stages 43-45. A minor population of the VBI-derived cells was also found extravascularly in the mesonephric primordia. In contrast to the VBI, the DLP-derived cells contributed to the hemopoietic cell population not in early larvae, but in late ones as a major constituent in the mesonephros, thymus, liver, and peripheral blood.  相似文献   

7.
Characterization of thymic progenitors in adult mouse bone marrow   总被引:5,自引:0,他引:5  
Thymic cellularity is maintained throughout life by progenitor cells originating in the bone marrow. In this study, we describe adult mouse bone cells that exhibit several features characteristic of prothymocytes. These include 1) rapid thymic engraftment kinetics following i.v. transplantation, 2) dramatic expansion of thymic progeny, and 3) limited production of hemopoietic progeny other than thymocytes. The adult mouse bone marrow population that is depleted of cells expressing any of a panel of lineage-specific Ags, stem cell Ag-1 positive, and not expressing the Thy1.1 Ag (Thy1.1(-)) (Thy1.1(-) progenitors) can repopulate the thymus 9 days more rapidly than can hemopoietic stem cells, a rate of thymic repopulation approaching that observed with transplanted thymocytes. Additionally, Thy1.1(-) progenitors expand prolifically to generate thymocyte progeny comparable in absolute numbers to those observed from parallel hemopoietic stem cell transplants, and provide a source of progenitors that spans multiple waves of thymic seeding. Nevertheless, the Thy1.1(-) population yields relatively few B cells and rare myeloid progeny posttransplant. These observations describe the phenotype of an adult mouse bone marrow population highly enriched for rapidly engrafting, long-term thymocyte progenitors. Furthermore, they note disparity in B and T cell expansion from this lymphoid progenitor population and suggest that it contains the progenitor primarily responsible for seeding the thymus throughout life.  相似文献   

8.
An experimental model for human T lymphocyte development from hemopoietic stem cells is necessary to study the complex processes of T cell differentiation in vivo. In this study, we report a newly developed nonobese diabetic (NOD)/Shi-scid, IL-2Rgamma null (NOD/SCID/gamma(c)(null)) mouse model for human T lymphopoiesis. When these mice were transplanted with human cord blood CD34(+) cells, the mice reproductively developed human T cells in their thymus and migrated into peripheral lymphoid organs. Furthermore, these T cells bear polyclonal TCR-alphabeta, and respond not only to mitogenic stimuli, such as PHA and IL-2, but to allogenic human cells. These results indicate that functional human T lymphocytes can be reconstituted from CD34(+) cells in NOD/SCID/gamma(c)(null) mice. This newly developed mouse model is expected to become a useful tool for the analysis of human T lymphopoiesis and immune response, and an animal model for studying T lymphotropic viral infections, such as HIV.  相似文献   

9.
The CD2 family is a growing family of Ig domain-containing cell surface proteins involved in lymphocyte activation. Here we describe the cloning and expression analysis of a novel member of this family, B lymphocyte activator macrophage expressed (BLAME). BLAME shares the structural features of the CD2 family containing an IgV and IgC2 domain and clusters with the other family members on chromosome 1q21. Quantitative PCR and Northern blot analysis show BLAME to be expressed in lymphoid tissue and, more specifically, in some populations of professional APCs, activated monocytes, and DCS: Retroviral forced expression of BLAME in hematopoietic cells of transplanted mice showed an increase in B1 cells in the peripheral blood, spleen, lymph nodes, and, most strikingly, in the peritoneal cavity. These cells do not express CD5 and are CD23(low)Mac1(low), characteristics of the B1b subset. BLAME may therefore play a role in B lineage commitment and/or modulation of signal through the B cell receptor.  相似文献   

10.
The role of hemopoietic stem cells and other cell types in the induction and maintenance of immunologic tolerance in the thymus was investigated by intravenous injection of Mls-semi-allogeneic cells into newborn mice less than 24 hr after birth. Mls-specific tolerance was induced by inoculation of peritoneal cells and thymus cells, and the tolerant state was compared with that induced by bone marrow cells which had hemopoietic stem cell activity and were able to create a stable chimera in both central and peripheral lymphoid organs. When peritoneal or thymus cells were injected, the level of tolerance attained was proportional to the number of cells injected, though peritoneal cells were 20 times as effective as thymus cells. In vivo functions of tolerance-inducing cells and their immediate precursors were radiosensitive and belonged to a Thy-1-, nylon-wool-nonadherent (probably non-B), weakly Sephadex G-10-adherent cell population. Tolerance induced by peritoneal cell injections was transient, starting to terminate within the first 2 weeks of life, while tolerance caused by bone marrow cell injections persisted through more than 6 weeks. Such transient tolerance induced by the former became long-lasting when followed by an additional injection of bone marrow cells, which did not cause thymic lymphocyte chimerism. All data indicated that bone marrow stem cells were engaged in tolerance induction and maintenance by continuously supplying tolerance-inducing nonlymphocytes.  相似文献   

11.
E I Deriugina 《Ontogenez》1986,17(2):117-137
A review of the origin of stem blood cells in ontogeny of vertebrates is presented. The comparative analysis of the data on laying, determination and migration of the hemopoietic precursor cells during embryogenesis in various taxonomic groups (teleosteans, urodeleans, anurans, avians and mammals) is performed. The change of the hemopoietic site and erythroid cells populations has been described. The data on sources of blood cell precursors and the origin of hemopoietic cells in the primordiums of hemopoietic organs were classified. A conclusion has been reached that in the course of evolution the hemopoietic anlage is gradually divided into two parts: one part migrates to the extraembryonic (ventral) mesoderm and another one remains intraembryonically and gives rice to the predecessors of definitive hemopoietic stem cells.  相似文献   

12.
Stromal cells of hemopoietic origin   总被引:1,自引:0,他引:1  
Hemopoiesis is a multistep process involving stem cell renewal, commitment, differentiation, maturation and consequent positioning of the cells within the tissue. Stromal cells are a major component of the hemopoietic microenvironment. The in vitro culture of cloned stromal cells has enabled detailed analysis of their functions and has provided answers relating to the contribution of stromal cells to the control of hemopoiesis. Cultured stromal cells were found to support the renewal of stem cells through a mechanism that did not seem to involve already known cytokines. Cloned stromal cells from both marrow and thymus supported the in vitro accumulation of myeloid as well as T and B lymphoid cells. Thus, cloned stromal cells had the ability to induce multilineage hemopoiesis, irrespective of the organ from which they were derived. Invariably, stromal cells tended to select in culture for hemopoietic cells at early differentiation stages and restricted the accumulation of mature cells. These functions may be part of the mechanism that protects the stem cell pool from excess differentiation.  相似文献   

13.
The contribution of two embryonic stem cell compartments to the developing thymus in the amphibian Xenopus was examined throughout the larval, postmetamorphic, and adult periods. Hematopoietic chimeras were produced by transplanting either the ventral blood islands (VBI) or the dorsal stem cell compartment (DSC) from diploid donors onto triploid hosts. The DNA content of isolated nuclei harvested from the thymus and circulating E populations was analyzed using propidium iodide staining and flow cytometry. The DNA content of mitotic figures derived from PHA reactive splenocytes was analyzed using the Feulgen reaction and microdensitometry. These data suggested that both the VBI and DSC contribute to the thymocyte populations from the earliest developmental stages examined. Moreover, the contribution of both stem cell compartments was cyclic. However, the periods of these cycles were different. Both VBI- and DSC-derived cells entered the thymus 4 days postfertilization. VBI-derived thymocytes were at a minimum at 28 days postfertilization, reached a maximum at 35 days postfertilization and a second minimum at 42 days postfertilization. However, DSC-derived cells reached a maximum at 28 days, a minimum at 35 days, and a second maximum at 42 days. The PHA-reactive splenocyte population followed a similar temporal pattern. In contrast, the VBI-derived E population was at a maximum during early development and steadily declined throughout the larval period. DSC-derived E were undetectable during early development but steadily increased throughout the larval period. Both VBI- and DSC-derived hematopoietic cells persisted after metamorphosis and contributed to all populations examined in adult frogs. Because of temporal differences in the VBI and DSC contributions to the developing thymus, these data suggest heterogeneity within the thymocyte population associated with the embryonic origin of the colonizing stem cells.  相似文献   

14.
The somitic level of origin of embryonic chick hindlimb muscles   总被引:1,自引:0,他引:1  
Studies of avian chimeras made by transplanting groups of quail somites into chick embryos have consistently shown that the muscle cells of the hindlimb are derived from the adjacent somites, however, the pattern of cell distribution from individual somites to individual hindlimb muscles has not been characterized. I have mapped quail cell distribution in the chick hindlimb after single somite transplantation to determine if cells from an individual somite populate discrete limb muscle regions and if there is a spatial correspondence between a muscle's somitic level of origin and the known spinal cord position of its motoneuron pool. At stages 15-18 single chick somites or equivalent lengths of unsegmented somitic mesoderm adjacent to the prospective hindlimb region were replaced with the corresponding tissue from quail embryos. At stages 28-34, quail cell distribution was mapped within individual thigh muscles and shank muscle regions. A quail-specific antiserum and Feulgen staining were used to identify quail cells. Transplants from somite levels 26-33 each gave rise to consistent quail cell labeling in a unique subset of limb muscles. The anteroposterior positions of these subsets corresponded to that of the transplanted somitic tissue. For example, more anterior or anteromedial thigh muscles contained quail cells when more anterior somitic tissue had been transplanted. For the majority of thigh muscles studied and for shank muscle groups, there was also a clear correlation between somitic level of origin and motoneuron pool position. These data are compatible with the hypothesis that motoneurons and the muscle cells of their targets share axial position labels. The question of whether motoneurons from a specific spinal cord segment recognize and consequently innervate muscle cells derived from the same axial level during early axon outgrowth is addressed in the accompanying paper (C. Lance-Jones, 1988, Dev. Biol. 126, 408-419). Quail cell distribution was also mapped in chick embryos in which quail somites or unsegmented mesoderm had been placed 2-3 somites away from their position of origin. In all cases donor somitic tissues contributed to muscles in accord with their host position. These results indicate that muscle cell precursors within the somites are not specified to migrate to a predetermined target region.  相似文献   

15.
Our previous investigations in 3- to 4-day avian chimeras have revealed that the wall of the aorta is a site from which hemopoietic stem cells can be obtained. In the present work using an in vitro clonal assay, we searched for cells with monocytic potentiality in this location as well as in the remainder of the embryo's body. In each experimental series thoracic segments from 30 chick embryo aortae were dissociated by a pancreatin treatment and plated in agar medium containing chicken serum and fibroblast-conditioned medium. Eighty to 620 macrophage colonies developed when 50,000 cells from 4-day aortae were plated, somewhat fewer when 3-day cells were plated (19-110). By contrast no progenitors were detected when cells were plated from 3- or 4-day embryos after their aorta had been removed. The cell composition and morphology of colonies deriving from aorta cells, their growth requirement and kinetics of development were identical to these of colonies deriving from young chicken bone marrow cells, cultured in the same conditions. The presence of macrophage progenitors in the wall of the 3- or 4-day embryo aorta and their absence in the rest of the embryo argues for a specific role of that region in embryonic hemopoiesis, namely that this is the location where intraembryonic hemopoietic stem cells emerge from the mesoderm at that period of development.  相似文献   

16.
The frequencies and potentialities of hematopoietic stem cells from 20-hr-old Xenopus embryos were examined by transplanting cytogenetically distinct ventral blood island tissue from diploid to triploid embryos. Thirty-five-day-old larvae were examined for the presence of donor-derived cells in their erythrocyte, thymocyte, and B lymphocyte populations by analyzing DNA content using flow cytometry. These experiments demonstrated that B lymphocytes, as well as erythrocytes and thymocytes, were derived from the ventral blood island. Data obtained by transplanting graded sized pieces of ventral blood island suggested that restricted erythroid precursors were present within the region by 20 hr postfertilization. Differentiation of both B- and T-lymphoid precursors from small pieces of ventral blood island was markedly enhanced when this tissue was grafted onto peripheral areas within the blood island region. Analysis of these data using repopulation statistics suggested that circulating larval erythrocytes of ventral blood island origin were derived from six or seven precursors. Each lobe of the thymus was colonized by three precursors, one of which was ventral blood island derived.  相似文献   

17.
The lymphoid tissue provides the most productive conditions to the application of immunohistochemical techniques. This tissue is composed of many B and T phenotypically different cell populations ranging from stem cells to completely differentiated lymphocytes which can be identified by using monoclonal antibodies. These techniques also are of great help in classifying the malignant lymphocyte counterparts and, thus, in defining diagnostic and prognostic characters of different lymphomas/leukaemias.  相似文献   

18.
Maggi Allan  Paul Harrison 《Cell》1980,19(2):437-447
We have studied the regulation of differentiation within the hemopoietic system by fusing mouse Friend cells (which can be induced to undergo red blood cell differentiation) to various mouse lymphomas and myelomas which express characteristic T and B lymphocyte surface antigens. Our results show that both erythroid and lymphoid differentiation markers can be co-expressed within the same cell. To determine whether this result applies to other differentiation states, we fused suspension Friend cells to three adherent fibroblast cell lines, and isolated both adherent and suspension hybrids. In fact, suspension hybrid clones were inducible for hemoglobin, whereas adherent clones were not. No obvious differences in overall chromosome balance were evident between the adherent and suspension hybrids. A similar correlation between suspension morphology and inducibility of hemoglobin was found in hybrids between suspension Friend cells and an adherent lymphoma line. These results show that different developmental programs can be coexpressed within the same hybrid cell; but the strongly adherent type of morphology is inconsistent with expression of the red blood cell phenotype, both in hybrid cells derived entirely from hemopoietic parental cells and in cells from widely different lineages.  相似文献   

19.
Human embryonic stem cells (hESCs) provide a unique resource to analyze early stages of human hematopoiesis. However, little is known about the ability to use hESCs to evaluate lymphocyte development. In the present study, we use a two-step culture method to demonstrate efficient generation of functional NK cells from hESCs. The CD56(+)CD45(+) hESC-derived lymphocytes express inhibitory and activating receptors typical of mature NK cells, including killer cell Ig-like receptors, natural cytotoxicity receptors, and CD16. Limiting dilution analysis suggests that these cells can be produced from hESC-derived hemopoietic progenitors at a clonal frequency similar to CD34(+) cells isolated from cord blood. The hESC-derived NK cells acquire the ability to lyse human tumor cells by both direct cell-mediated cytotoxicity and Ab-dependent cellular cytotoxicity. Additionally, activated hESC-derived NK cells up-regulate cytokine production. hESC-derived lymphoid progenitors provide a novel means to characterize specific cellular and molecular mechanisms that lead to development of specific human lymphocyte populations. These cells may also provide a source for innovative cellular immune therapies.  相似文献   

20.
Cyclophosphamide-treated 18-day-old chick embryos were transplanted with cells from 7-day intraembryonic mesenchyme; the recipients and donors were identical at the major histocompatibility locus. At the age of 35 days, the cell recipients were studied to determine the reconstitution capacity of the transplanted cells. The transplantation resulted in a complete restoration of IgM and IgG class antibody production against human gammaglobulin and Brucella abortus, and of microscopic morphology of the bursa of Fabricius and of the germinal center formation in the spleen. These findings demonstrate that 7-day intraembryonic mesenchyme of the chick embryo harbor prebursal stem cells. These findings confirm our previous observations in the yolk sac-embryo chimeras indicating that lymphoid stem cells originate in the intraembryonic hematopoietic sites.  相似文献   

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