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Howard T Petrie

Publications and source records attributed to Howard T Petrie.

6 recordsLinked to original sources
PubMed2003

Kinetics of steady-state differentiation and mapping of intrathymic-signaling environments by stem cell transplantation in nonirradiated mice.

Upon thymus entry, thymic-homing progenitors undergo distinct phases of differentiation as they migrate through the cortex to the capsule, suggesting that the signals that induce these differentiation steps may be stratified in corresponding cortical regions. To better define these regions, we transplanted purified stem cells into nonirradiated congenic recipients and followed their differentiation with respect to both tissue location and time. The earliest progenitors (DN1) remained confined to a very narrow region of the cortex for about the first 10 d of intrathymic residence; this region virtually overlaps the sites of thymic entry, suggesting that DN1 cells move very little during this lengthy period of proliferation and lineage commitment. Movement out of this region into the deeper cortex is asynchronous, and corresponds to the appearance of DN2 cells. Differentiation to the DN3 stage correlates with movement across the midpoint of the cortex, indicating that stromal signals that induce functions such as TCR gene rearrangement reside mainly in the outer half of the cortex. The minimum time to reach the capsule, and thus transit to the DP stage, is approximately 13 d, with the average time a few days longer. These findings reveal for the first time the kinetics of steady-state progenitor differentiation in the thymus, as well as defining the boundaries of cortical regions that support different phases of the differentiation process. We also show that the first lineage-positive progeny of transplanted stem cells to appear in the thymus are dendritic cells in the medulla, suggesting that each new wave of new T cell production is preceded by a wave of regulatory cells that home to the medulla and ensure efficient tolerance and selection.

Animals↗
PubMed2003

A simple method for detecting up to five immunofluorescent parameters together with DNA staining for cell cycle or viability on a benchtop flow cytometer.

In this manuscript, we describe modifications to a commercial three-laser benchtop flow cytometer, as well as relevant biological methods, that allow analysis of up to five immunofluorescent parameters together with an ultraviolet (UV)-excitable DNA stain. This method allows expanded capacity for multiparameter immunophenotyping of complex mixed cell populations, together with accurate measurements of DNA content (cell cycle) or cell viability, on a stable, end-user operated platform.

Cell Cycle↗
PubMed2002

Stromal cells provide the matrix for migration of early lymphoid progenitors through the thymic cortex.

During steady state lymphopoiesis in the postnatal thymus, migration of precursors outward from the deep cortex toward the capsule is required for normal differentiation. Such migration requires, at a minimum, expression of adhesive receptors on the migrating lymphoid cells, as well as a stable matrix of their ligands persisting throughout the region of migration. In this study, we address the nature of this adhesive matrix. Although some precursor stages bound efficiently to extracellular matrix ligands, a specific requirement for the cell surface ligand VCAM-1 was also found. In situ analysis revealed that early precursors are found in intimate contact with a matrix formed by stromal cells in the cortex, a proportion of which expresses VCAM-1. In vivo administration of an anti-VCAM-1 Ab resulted in decreased thymic size and altered distribution of early precursors within the cortex. These results indicate that precursors migrating outward through the cortex may use a cellular, rather than extracellular, matrix for adhesion, and suggest that the VCAM-1(+) subset of cortical stroma may play a crucial role in supporting the migration of early precursors in the steady state thymus.

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PubMed2002

Access roads for RAG-ged terrains: control of T cell receptor gene rearrangement at multiple levels.

Antigen-specific immune response requires the generation of a diverse antigen (Ag)-receptor repertoire. The primary repertoire is generated through somatic gene rearrangement and molded by subsequent cellular selection. Constraints during gene recombination influence the ultimate shape of the repertoire. One major control mechanism of gene rearrangement, investigated for many years, is exerted through regulated chromosomal accessibility of the recombinase to the antigen receptor loci. More recent studies began to explore the role of interactions between the recombinase and its cognate recognition DNA sequences. The emerging results suggest that formation of the primary repertoire is controlled by two, partially independent factors: chromosomal accessibility and direct recombinase-DNA interactions.

Animals↗
PubMed2002

Role of thymic organ structure and stromal composition in steady-state postnatal T-cell production.

The most conspicuous period of cellular proliferation and differentiation occurs during the embryonic stage of development. In some cell lineages, however, including T lymphocytes, this process must continue postnatally and throughout life. Under ordinary circumstances, postnatal T-cell production occurs in the thymus, and can be separated into five functional phases. The first is recognition of thymus-derived recruitment signals by multilineage progenitors in the bloodstream, followed by their extravasation and entry into the organ. Second is the lymphopoietic process, i.e. the expansion of this small number of blood-derived progenitors to produce the raw materials for all subsequent phases. Third is the screening of these cellular products for the ability of their T-cell receptors to appropriately recognize major histocompatibility/peptide ligands, i.e. positive and negative selection. Fourth is functional maturation, a process that follows but is distinct from positive selection. Finally, cells that successfully undergo all of the prior processes must be induced to leave the thymus and enter the peripheral lymphoid pool. From the above, it can be seen that all the hematopoietic components of the thymus are transient, with uncommitted progenitors entering and lineage committed progeny being exported or removed. This process reveals a subtle but critically important fact about the nature of the thymus, namely that the functional components of the thymus are not the hemato-lymphoid cells, but rather the stable (stromal) elements that induce their differentiation. Understanding the nature of these stromal elements, and the signals they deliver to nascent T lymphocytes, is therefore critical not only for understanding how T lymphocytes are produced normally but, by analogy, what goes wrong in congenital, acquired, or age-associated deficiencies in T-lymphocyte production.

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