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Receptor signals and nuclear events in CD4 and CD8 T cell lineage commitment.

MHC specificity in positive selection is a major determinant in the CD4/CD8 T cell lineage decision. Previous studies support the view that quantitative differences in T cell receptor (TCR) signaling in immature CD4+CD8+ double positive thymocytes leads to an instructive bias in CD4/CD8 T cell lineage commitment that must be re-inforced in subsequent selection steps to ensure that MHC-restricted antigen recognition is linked to appropriate effector functions in mature T cells. Recent work has further defined the TCR signaling pathways involved in this process, but a major effort has been made to identify transcription factors and other regulators of CD4 and CD8 T cell lineage commitment. Methods and screens for detecting changes in gene expression, associated with TCR signaling in positive selection and lineage determination, are starting to provide a better understanding of these complex developmental processes.

Animals↗

Progenitor cells of the biliary epithelial cell lineage.

Stem-like cells have been identified in liver that are able to differentiate in vivo and in culture to biliary epithelial cells (BEC), hepatocytes and oval cells. The growth factors/cytokines and signal pathways required for the differentiation processes are beginning to be evaluated. There is increasing evidence to suggest that these stem-like cells may originate from both the bone marrow population or from a precursor remnant from liver embryogenesis, as they share many of the same markers (CD34, c-kit, CD45). Most recently, it has been shown that a population of progenitor cells can copurify with mesenchymal bone marrow cells and differentiate under specific culture conditions to form both hepatic epithelial and also endothelial cells. The interaction of haemopoietic and mesenchymal stem cells needs further evaluation. The close association of ductular reactive cells and neovessels in end-stage cholestatic liver diseases and the relation to Jagged/Notch signalling pathway may be important in the regulation of stem cells to form both biliary epithelial and endothelial cells.

Biliary Tract↗

Circulating osteoblast-lineage cells in humans.

BACKGROUND: Although current evidence suggests that only a minuscule number of osteoblast-lineage cells are present in peripheral blood, we hypothesized that such cells circulate but that their concentration has been vastly underestimated owing to the use of assays that required adherence to plastic. We further reasoned that the concentration of these cells is elevated during times of increased bone formation, such as during pubertal growth. METHODS: We used flow cytometry with antibodies to bone-specific proteins to identify circulating osteoblast-lineage cells in 11 adolescent males and 11 adult males (mean [+/-SD] age, 14.5+/-0.7 vs. 37.7+/-7.6 years). Gene expression and in vitro and in vivo bone-forming assays were used to establish the osteoblastic lineage of sorted cells. RESULTS: Cells positive for osteocalcin and cells positive for bone-specific alkaline phosphatase were detected in the peripheral blood of adult subjects (1 to 2 percent of mononuclear cells). There were more than five times as many cells positive for osteocalcin in the circulation of adolescent boys (whose markers of bone formation were clearly increased as a result of pubertal growth) as compared with adult subjects (P<0.001). The percentage of cells positive for osteocalcin correlated with markers of bone formation. Sorted osteocalcin-positive cells expressed osteoblastic genes, formed mineralized nodules in vitro, and formed bone in an in vivo transplantation assay. Increased values were also found in three adults with recent fractures. CONCLUSIONS: Osteoblast-lineage cells circulate in physiologically significant numbers, correlate with markers of bone formation, and are markedly higher during pubertal growth; therefore, they may represent a previously unrecognized circulatory component to the process of bone formation.

Adolescent↗

Analysing cell lineage with a recombinant retrovirus.

Analysis of neural cell lineage in vertebrates has been limited by a lack of methods for introducing stable tracers into individual cells at relatively late stages of development. Recent progress in the design of recombinant retroviral vectors provides a novel approach to this problem. When a retrovirus infects a dividing cell, its genome integrates into a chromosome of the infected cell and is inherited by that cell's progeny. For lineage tracing, viral structural genes are replaced by a bacterial beta-galactosidase gene; infected cells are therefore unable to produce new virions, but can produce galactosidase, which is detectable histochemically. By infecting cells and identifying their progeny at appropriate stages, it has been possible to obtain new data on cell lineage in retina, cerebral cortex, optic tectum, and peripheral nerve.

Animals↗

Evolution of cell lineage.

Organisms such as nematodes and mollusks, the development of which involves fixed cell lineages, allow studies of the genetic programming of precision and of evolution of its flexibility. Stochastic specification of cell fate may evolve to a fixed cell lineage and specification by cell interactions to autonomous specification. Comparative developmental studies also shed light on the evolution of asymmetry.

Animals↗

Characteristic distribution density of organelles in the cytoplasm of respective kinds of cell lineages of ascidian embryos.

In the early embryos of the ascidian (Halocynthia roretzi), regional cytoplasmic differences arise just after fertilization. We successively studied the characteristic features of the cytoplasm of those regions in the embryonic cells during the entire ontogenic process from the unfertilized egg to the tadpole larva. The embryos and larvae were fixed in a mixture of osmium and glutaraldehyde, and the distribution of the organelles in the entire cytoplasm was observed with light and electron microscopy. According to the analysis of the distribution density of the organelles which occupied a given area of the cytoplasm on the section, we introduced the "organelle region" in the cytoplasm and further calculated the approximate area ratios of the "organelle regions" in the cytoplasm of each cell lineage. As a result, it was demonstrated that the characteristic distribution mode was already present in the cytoplasm of the cell lineages of the 8-cell-stage embryos. The features of the cytoplasm in each embryonic cell lineage, moreover, reflected those of the corresponding larval tissue. Our results not only demonstrate that the structural tissue specificity is expressed at an extremely early stage during the ascidian embryogenesis but also suggest that the corresponding functional differentiation among the cell lineages occurs early.

Animals↗

Developmental patterns and cell lineages of vermiform embryos in dicyemid mesozoans.

Patterns of cell division and cell lineages of the vermiform embryos of dicyemid mesozoans were studied in four species belonging to four genera: Conocyema polymorpha, Dicyema apalachiensis, Microcyema vespa, and Pseudicyema nakaoi. During early development, the following common features were apparent: (1) the first cell division produces prospective cells that generate the anterior peripheral region of the embryo; (2) the second cell division produces prospective cells that generate the posterior peripheral region plus the internal cells of the embryo; (3) in the lineage of prospective internal cells, several divisions ultimately result in cell death of one of the daughter cells. Early developmental processes are almost identical in the vermiform embryos of all four dicyemid genera. The cell lineages appear to be invariant among embryos and are highly conserved among species. Species-specific differences appear during later stages of embryogenesis. The number of terminal divisions determines variations in peripheral cell numbers among genera and species. Thus, the numbers of peripheral cells are fixed and hence species-specific.

Animals↗

Cloning and characterization of a vasa-like gene in rainbow trout and its expression in the germ cell lineage.

The origin of germ cells and the molecular mechanisms of primordial germ cell (PGC) determination in teleosts are unclear. Vasa is a member of the DEAD protein family and plays an indispensable role in germ cell determination in Drosophila and Xenopus species. In this study, we isolated and characterized a rainbow trout vasa cDNA as a first step towards understanding the molecular mechanisms of PGC determination and development and to develop a molecular marker to identify the PGCs in rainbow trout. Cloning of vasa cDNA was performed by degenerate- and RACE-PCR. The predicted amino acid sequence of rainbow trout Vasa contained eight consensus sequences for the DEAD protein family and five arginine-glycine-glycine repeats, a common character of known Vasa homologues. Overall amino acid similarity to the Vasa of Drosophila was 79.2%. Whole-mount in situ hybridization of eyed stage embryos (eighty somite stage) revealed that signals were localized to the putative PGCs. In adult rainbow trout tissues, both ovaries and testes contained large amounts of vasa gene transcripts. A reverse transcription-polymerase chain reaction analysis of unfertilized eggs proved that trout vasa is a maternal factor. Although we have not determined whether rainbow trout vasa functions as a germ cell determinant, its limited expression in the germ cell lineage proved that rainbow trout vasa can be used as a marker molecule for PGCs. This marker will make it possible to identify the PGCs or presumptive PGCs in early trout embryos whose germ cells can not be distinguished by morphological characteristics.

Amino Acid Sequence↗

Life history of cells mediating natural resistance to tumor cells and bone-marrow transplants: the respective roles of cell lineage commitment and host environment in determining strain characteristics of natural resistance to foreign marrow grafts.

A class of cells present in the blood and lymphoid tissues of mammals produces rapid cytolysis of tumor cells on first contact. Abundant evidence suggests that such natural killer (NK) cells play a role in tumor immunosurveillance in vivo. A similarly prompt and spontaneous activity can cause the rejection of foreign marrow transplants. These phenomena are known collectively as natural resistance. The cells mediating natural resistance are lymphoid in morphology, but are neither T nor B lymphocytes. Kinetically, NK cells and cells mediating natural resistance to foreign marrow grafts are themselves nondividing but are rapidly renewed from radiosensitive proliferating precursors in the bone marrow. They appear to have no long-lived (memory) component. Newly formed NK cells have a short residence time in the spleen. Other general properties of the natural-resistance cell lineage, including strain variation, ontogeny, and cell phenotype, are reviewed in this article. The present study aimed to examine the respective roles of cell lineage commitment and of the host environment in determining strain characteristics of natural resistance to foreign marrow grafts. Chimeras produced by inoculating mice of a strain that normally has little or no natural resistance with bone marrow from adult mice of a highly resistant strain develop resistance to a third-party marrow allograft. Such chimeras do not develop the full rejection capacity of the high-resistance strain, however; and chimeras created by inoculating marrow from infant mice develop less resistance than those reconstituted by bone marrow from adult mice. The results demonstrate that the ability to reject foreign marrow grafts develops as an intrinsic property of the natural-resistance cell lineage. The host environment may provide an additional influence, however, particularly in the initial development of natural resistance in early postnatal life.

Animals↗

Loss of CD23 is a consequence of B-cell activation. Implications for the analysis of B-cell lineages.

When splenic CD5- B cells are stimulated with antiimmunoglobulin they become CD5+ and have a prolonged in vitro life. Further treatment with IL-6 induces a loss of surface CD23 and IgD; that is, they resemble freshly isolated peritoneal CD5+ cells. These data suggest that the CD5 phenotype is induced after sIg-mediated B-cell activation. Additional support for this view arises from the observation that the loss of CD23 and IgD can be induced by another activation inducer, LPS, although in this case CD5 is not expressed. Thus, activation by anti-Ig plus IL-6 or by LPS induces CD23 loss. Consistent with the hypothesis that the loss of CD23 is a consequence of activation, we now report that the surface expression of CD23 varies inversely with the amount of total cellular RNA. We also find both CD23 positive and negative B cells among freshly isolated splenic CD5- B cells. In young mice a proportion of small splenic CD5+ B cells are CD23+, providing additional evidence that CD23 is present on all B cells prior to activation. A comparison of the features of CD5+ B cells and the antibody responses to thymus-dependent and thymus-independent antigens leads us to hypothesize that the CD5 phenotype arises as a consequence of thymus-independent type 2 (TI-2) stimulation. The relationship of CD5 expression to B-cell lineage (fetal vs. adult bone marrow) is discussed.

Animals↗

Perspective: stem cells react! Cell lineages as complex adaptive systems.

It may be argued that adult stem cell processes or, more precisely, the cell lineages that arise from them, represent complex reactive or adaptive systems. Approaching hematopoietic and other stem cell lineages from this perspective has direct bearing on current debates regarding the plasticity of these lineage systems as well as on interpretation and modeling of clinical data regarding many diseases.

Adaptation, Physiological↗

An interactive computer system for the analysis of cell lineages.

We have developed an interactive computer system for analysing cell lineage data. It can be utilized in studies of cell motility, cell division, cell differentiation, and cell aging. It has enabled us to document the heterogeneity of human foreskin fibroblasts in culture and to propose that loss of proliferative potential may mean that cells enter a state of differentiation which makes them unable to respond to mitotic stimulation. Our method, which enables us to apply immunological and cytochemical probes after recording the history of a cell lineage, should allow us to define precisely features which uniquely distinguish cycling from noncycling cells on an individual cell basis.

Cell Cycle↗

The development of cell lineages: a sequential model.

The concept of cell lineage and the empirical characterization of specific lineages provide valuable insight into the problems of developmental biology. Of central interest is the decision-making process that results in the diversification of cell lines. Studies of the haemopoietic system, in which stem cells can be committed to one of at least six pathways of differentiation, have suggested that the restriction of differentiation potentials is a progressive and stochastic process. We have recently proposed an alternative model which hypothesizes that lineage potentials during haemopoiesis are expressed individually and in a predetermined sequence as progenitor cells mature. The model first arises from experimental studies which show that both normal myeloid progenitor cells and a human promyeloid cell line, which are able to differentiate towards either neutrophils or monocytes, express these potentials sequentially in culture. The close linear relationship between other haemopoietic progenitor cells is inferred from collective data from studies of bipotent progenitor cells and of haemopoietic proliferative disorders. If the development of haemopoietic cell lineages shows a tendency to follow a particular program, such a mechanism is likely to operate throughout development. In this paper we consider the evidence in favour of programmed events within progenitor cells implementing diversification, and the implications of predetermined and restricted pathways of embryonic development.

Animals↗

Cell lineage analysis reveals multipotency of some avian neural crest cells.

A major question in developmental biology is how precursor cells give rise to diverse sets of differentiated cell types. In most systems, it remains unclear whether the precursors can form many or all cell types (multipotent or totipotent), or only a single cell type (predetermined). The question of cell lineage is central to the neural crest because it gives rise to numerous and diverse derivatives including peripheral neurons, glial and Schwann cells, pigment cells, and cartilage. Although the sets of derivatives arising from different populations of neural crest cells have been well-documented, relatively little is known about the developmental potentials of individual neural crest cells. We have iontophoretically microinjected the vital dye, lysinated rhodamine dextran (LRD) into individual dorsal neural tube cells to mark unambiguously their descendants. Many of the resulting labelled clones consisted of multiple cell types, as judged by both their location and morphology. Cells as diverse as sensory neurons, presumptive pigment cells, ganglionic supportive cells, adrenomedullary cells and neural tube cells were found within individual clones. Our results indicate that at least some neural crest cells are multipotent before their departure from the neural tube.

Animals↗

Essential requirement of antigen presentation by monocyte lineage cells for the activation of primary human gamma delta T cells by aminobisphosphonate antigen.

Human gammadelta T cells respond to nonpeptide Ags such as pyrophosphomonoesters and alkylamines in a gammadelta TCR-dependent manner in the absence of other APCS: Recently, aminobisphosphonates such as pamidronate have also been shown to activate human gammadelta T cells. In the present study, we indicate that activation of primary gammadelta T cells by pamidronate strictly depends on the presence of monocyte-lineage cells, unlike that by pyrophosphomonoesters. Thus, although pamidronate induced cell clustering, proliferation, and IFN-gamma production of gammadelta T cells in the culture of PBMC, it failed to induce any of these activities in the culture of purified primary gammadelta T cells. By adding back the purified monocytes, however, both cell clustering and IFN-gamma production of gammadelta T cells by pamidronate could be restored. The pamidronate-pulsed, but not untreated, myelomonocytic line, THP-1, was capable of activating the purified gammadelta T cells to produce IFN-gamma, which was associated with the down-regulation of gammadelta TCR. Furthermore, pamidronate-pulsed THP-1 cells were significantly more susceptible to gammadelta T cell-mediated cytotoxicity than untreated THP-1. Also, TCR-defective Jurkat T cells transfected with gammadelta TCR genes produced a significant level of IL-2 in response to the pamidronate-pulsed THP-1 cells. These results have suggested strongly that human gammadelta T cells are functionally activated via gammadelta TCR by aminobisphosphonate Ag presented on the surface of monocyte lineage cells rather than directly by its free form.

Antigen Presentation↗

The bifurcating autoregression model in cell lineage studies.

A model for cell lineage data is presented and analysed. The model is an extension of the classical first-order autoregression, used in time-series studies, to bifurcating data trees of general size and shape. Maximum likelihood theory is developed and compared with an extensive simulation study. Some properties of moment estimators are also presented.

Analysis of Variance↗

Characteristics of mitotic cells in developing and adult testes with observations on cell lineages.

This report describes characteristics of dividing cells, primarily in developing (10-40 day) rat testis and relates the structure of the dividing cells to the structure of interphase cells. Mitotic cells were characterized in seven zones. Dividing Sertoli cells were seen prior to day 15 and possessed distinct characteristics as compared with dividing germ cells. Myoid cells showed morphological characteristics of precursor myoid cells; 'clear cells' self-replicated in the myoid cell layer; adult-type Leydig cells, some containing lipid, differentiated early (10th-15th postnatal days) from fibroblast-like cells of the multilayered tubule wall and later (15th-25th postnatal days) from dividing differentiated and semi-differentiated Leydig cells within the lymphatic space; fibroblastic cells arose from cells with similar morphological characteristics; semi-differentiated Leydig cells divided, and differentiated Leydig cells in the lymphatic space self-renewed; undifferentiated perivascular cells most likely gave rise to Leydig cells, pericytes; arteriolar smooth muscle cells and vascular endothelial cells arose from division of the pre-existing respective cell types. Fetal Leydig cells appeared to remain but, with time, they appeared to lose their lipid. The data suggest that (1) early recruitment of Leydig cells from undifferentiated peritubular fibroblast-like cells, (2) later mitosis of differentiated and semi-differentiated Leydig cells primarily in the interstitium but also in the perivascular region, and (3) the continued presence of pre-existing Leydig cells from the fetus constitute the adult population. Leydig cell division in the adult mouse was documented. This study provides the necessary information for the recognition of cell divisions to study of cell lineages among testis cells.

Animals↗

The zinc finger transcription factor Th-POK regulates CD4 versus CD8 T-cell lineage commitment.

Development of immature T-cell precursors (thymocytes) to either the CD4 helper or CD8 killer T-cell lineages correlates precisely with their T-cell receptor specificity for major histocompatibility complex class II or class I molecules, respectively, indicating that the process is carefully regulated. Although intensively studied owing to its importance in determining the composition of the mature T-cell compartment and as a general model of binary lineage decisions, the underlying molecular pathways remain obscure. We have previously reported a spontaneous mouse mutant (HD (helper deficient) mice) in which lineage commitment is specifically perturbed without affecting positive selection. Here we show that a point mutation in the zinc finger transcription factor Th-POK (T-helper-inducing POZ/Krüppel-like factor) is responsible for redirection of class-II-restricted thymocytes to the CD8 lineage in HD mice. Furthermore, we demonstrate that constitutive expression of this factor during thymic development leads to redirection of class-I-restricted thymocytes to the CD4 lineage, indicating that Th-POK is a master regulator of lineage commitment.

Amino Acid Sequence↗