PubMed Health⌕ Search

Biomedical subjects

E J Jenkinson

Publications and source records attributed to E J Jenkinson.

At least 19 recordsLinked to original sources
PubMed1992

Apoptosis and T-cell repertoire selection in the thymus.

Thymic tolerance depends on induction of apoptosis (programmed cell death) in immature thymocytes by antigen/MHC complexes on dendritic cells (and possibly other bone marrow-derived APCs). Interactions with antigen/MHC complexes on thymic epithelial cells promote maturation of double-positive thymocytes to single-positive cells. However, the nature of the antigen/MHC complexes on thymic epithelial cells is unknown, and if the stromal cell interaction model as just outlined is correct, then presumably these complexes must be different from those presented on dendritic cells, otherwise all cells signaled for positive selection would be subject to negative selection by similar complexes on APCs. The nature of the signals provided by thymic epithelial cells versus dendritic cells is unknown and may provide a key to understanding the processes of positive and negative selection within the thymus. Clearly the intense selection of the T-cell repertoire within the thymus explains the high level of cell death observed within the immature thymocyte compartment. Such intensive selection shapes the T-cell repertoire in a way that provides an explanation for the genetic basis of immune responsiveness and for the susceptibility of certain individuals to autoimmunity.

Animals↗
PubMed1992

Studies on T cell maturation on defined thymic stromal cell populations in vitro.

We describe an in vitro system in which positive selection of developing T cells takes place on defined stromal cell preparations, which include major histocompatibility complex class II+ epithelial cells but exclude cells of bone marrow origin. In this system, maturation of double-positive T cell receptor negative (TCR-), CD4+8+ thymocytes into single-positive TCR+, CD4+ and CD8+ cells takes place together with the development of functional competence. As in vivo, this maturation is associated with the upregulation of TCR levels as cells progress from double-positive to single-positive status. We also show that class II+ epithelial cells in these cultures are less efficient than dendritic cells in mediating the deletion (negative selection) of V beta 8+ cells by the superantigen staphylococcal enterotoxin B. For the first time, this approach provides a model in which the cellular interactions involved in both positive and negative selection can be studied under controlled in vitro conditions.

Animals↗
PubMed1992

Inositol lipids and phosphates in the proliferation and differentiation of lymphocytes and myeloid cells.

It is established that receptor-stimulated hydrolysis of phosphatidylinositol 4,5-bisphosphate is an essential signalling reaction in the responses of many haemopoietic cells to stimuli: examples include platelet activation, antigen-driven initiation of cell proliferation in mature B and T lymphocytes and histamine release by mast cells, and chemotaxis and oxygen radical generation by neutrophils. However, the roles of inositol lipids and phosphates in the development of haemopoietic and immune cells are less well understood. This paper discusses three such situations: the sequential employment of phosphatidylinositol 4,5-bisphosphate hydrolysis and cyclic AMP accumulation as two signals essential to the action of the B lymphocyte-stimulatory cytokine interleukin 4; the involvement of antigen receptor-triggered inositol lipid hydrolysis in apoptotic elimination of immature anti-self T lymphocytes in the fetal mouse thymus; and the possible role of changes in the levels of abundant inositol polyphosphates in the differentiation of HL-60 promyelocytic cells and of normal human myeloid blast cells.

Bone Marrow Cells↗
PubMed1992

Effects of the thymic microenvironment on the response of thymocytes to stimulation.

We show that, in vitro, the response of thymocytes to certain stimuli, and their survival largely depend on the nature of the culture environment, i.e. whether thymocytes are stimulated within intact thymus lobes or in cell suspension. Exposure of isolated thymocytes to 12-O-tetra-decanoylphorbol 13-acetate (TPA)+ionomycin rapidly abolishes the expression of recombination-activating gene-1 (RAG-1) mRNA (3 h), down-regulates CD4 surface antigen expression (3 h), and enhances apoptosis (24 h). On the other hand, when thymocytes are cultured in intact lobes, TPA plus ionomycin down-regulate rather than abolish RAG-1 mRNA expression (3 h), have little effect on CD4 expression even following 24-h exposure, and only marginally induce apoptosis (24 h). Differences between the culture systems are less pronounced in response to anti-CD3 antibodies. Therefore, it appears that removing thymocytes from their thymic microenvironment makes the cells more susceptible to certain stimuli, possibly by altering their physiological status. In addition, it has been suggested that termination of RAG-1 expression can be linked to thymocyte selection processes. We found that the down-regulation of RAG-1 expression was not dependent on the induction of apoptosis, supporting a proposed link with positive selection.

Animals↗
PubMed1990

Alkaline phosphatase-fast red, a new fluorescent label. Application in double labelling for cell cycle analysis.

We have observed that the red reaction product of alkaline phosphatase immuno-conjugates and certain substrate preparations produces a brilliant red fluorescence that is visible by fluorescence microscopy using both fluorescein and rhodamine filter combinations. This provides a level of sensitivity greater than that obtained with other commonly used red fluorochromes or by inspection of the reaction product under bright field illumination. Of particular value, the reaction product is unaffected by the denaturing conditions required for the detection of incorporated nuclear BrdU with FITC conjugated anti-BrdU antibody and provides a simple and robust method for the simultaneous detection of cell proliferation and cell surface markers.

Alkaline Phosphatase↗
PubMed1990

Cell growth and gene rearrangement signals during the development of T lymphocytes within the thymus.

The thymus provides signals that control the proliferation and differentiation of T lymphocytes and select the repertoire of T-cell specificities. Antibodies to CD3 molecules inhibit full rearrangement of T-cell receptor beta chain genes in organ cultures of early embryo mouse thymus. Whether this effect is mediated through gamma delta CD3 expressing cells, which are present in small numbers at this stage, or through low amounts of CD3 on alpha beta precursor cells is unclear. A requirement for special gene rearrangement signals within the thymus is supported also by the observations that growth factors such as IL-2 and IL-4, although stimulating proliferation of precursor cells removed from the thymus, do not induce full T-cell receptor gene rearrangements. Recent studies show that newly formed thymic lymphocytes expressing alpha beta CD3 receptors are targets for negative selection (deletion) as a means of removing autoreactive cells. Signalling to immature thymocytes via the alpha beta CD3 complex induces the activation of endogenous endonucleases that cleave DNA into oligonucleosomal fragments. We suggest that the activation of this mechanism is the means by which autoreactive cells are removed.

Animals↗
PubMed1990

Newly generated thymocytes are not refractory to deletion when the alpha/beta component of the T cell receptor is engaged by the superantigen staphylococcal enterotoxin B.

It has been reported that, following the initial expression of the T cell receptor (TcR) alpha/beta, newly generated thymocytes pass through a developmental window characterized by ineffective coupling between the alpha/beta and CD3 components resulting in resistance to deletion (negative selection). However, we now provide evidence that the TcR alpha/beta on developing thymocytes is capable of delivering deletional signals in response to the superantigen staphylococcal enterotoxin B (SEB) as soon as the receptor is expressed. We also show that if TcR+ thymocytes are allowed to mature in organ cultures of embryonic thymus before SEB is added, they respond by proliferation giving rise to blast cells of CD4-CD8-, CD4+CD8- or CD4-CD8+ phenotypes.

Animals↗
PubMed1990

Developmentally regulated fetal thymic and extrathymic T-cell receptor gamma delta gene expression.

The gamma delta T-cell receptor (TCR) is the first TCR to be expressed in ontogeny in all vertebrates in which it has been examined thoroughly. Murine gamma delta cell-surface protein is detected by the fourteenth day of gestation. In this work, the activation of gamma delta RNA has been studied. Data indicate that the first TCR protein to appear in the thymus is encoded by gamma genes that are activated after cells colonize the thymus. However, the sequential appearance of different gamma delta TCR proteins during thymic ontogeny cannot be readily explained by differential temporal activation of V gamma genes in the thymus. There are distinct patterns of gamma and delta gene expression during fetal liver development and in the fetal gut (or tissue associated with it). Cells apparent in the liver of mice at birth express gamma delta cell-surface protein, but they disappear from the liver very soon afterward. One V gamma gene is rearranged and expressed prethymically. In addition, gamma gene expression is detectable in the livers of newborn athymic mice. Together, these observations indicate a thymic-independent pathway of activation of TCR genes.

Animals↗
PubMed1990

T-cell differentiation in thymus organ cultures.

Fetal thymus organ cultures support a full range of T-cell precursor differentiation in vitro, including TCR gene rearrangement and expression. This provides an accessible model system in which the intra-thymic regulation of T-cell development can be investigated. Thymus organ cultures can be manipulated by adding antibodies to block different cell surface components on stromal or lymphoid elements. In addition, it is possible to deplete thymus lobes of their lymphoid cells and recolonize them with T-cell precursors of a different MHC haplotype to produce chimeric lobes. Recolonization can also be achieved with defined numbers or types of precursor cells, including a single micro-manipulated cell. These approaches have been used to obtain information on the signals regulating intra-thymic proliferation, T-cell lineage relationships, antigen receptor diversification within the thymus and the cellular interactions and intracellular mechanisms regulating selection of the antigen receptor repertoire.

Fetus↗
PubMed1989

Apoptosis.

Explore the source record for details and available documents.

Animals↗
PubMed1989

Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures.

The receptors found on most T lymphocytes bind to antigen presented on major histocompatibility complex proteins and consist of dimers of alpha- and beta-polypeptides associated with the invariant CD3 complex. A fully competent immune system requires a diverse array of T-cell antigen receptors (TCRs) with different specificities. This diversity is generated by rearrangement of TCR alpha- and beta-chain gene segments within the thymus where the receptors are first expressed. Any cells carrying self-reactive receptors must be eliminated, suppressed or inactivated so that destructive autoimmunity is avoided. Recently, compelling evidence has shown that one process involved in producing such self-tolerance is clonal deletion of autoreactive cells within the thymus by an as-yet-undefined mechanism. Here we show that engaging the CD3/TCR complex of immature mouse thymocytes with anti-CD3 antibodies produces DNA degradation and cell death through the endogenous pathway of apoptosis. Activation of this process in immature T cells by the binding of the TCR to self-antigens may therefore be the mechanism which produces clonal deletion and consequently self-tolerance.

Animals↗
PubMed1989

The effects of anti-CD2 antibodies on the differentiation of mouse thymocytes.

Using a rat monoclonal antibody against mouse CD2, we determined the expression of this marker on thymocytes during ontogeny. CD2 expression becomes detectable at day 15 and reaches adult levels (approximately 95% positivity) by day 19. Furthermore, the effect of anti-CD2 antibodies on T cell differentiation was analyzed by addition of antibodies to thymus organ cultures or repeated injection into newborn mice. Anti-CD2 antibodies inhibit CD2 expression in organ cultures and drastically reduce its expression on thymocytes and peripheral lymphocytes in vivo. In either situation, suppression of CD2 expression does not significantly alter the generation of T cells expressing CD3, CD4, CD8 and T cell receptor V beta 8. These results do not support a role for CD2 in early steps of thymocyte differentiation.

Animals↗
PubMed1989

Antigen-induced apoptosis in developing T cells: a mechanism for negative selection of the T cell receptor repertoire.

Herein we have investigated the ability of antigen to induce thymocyte death by apoptosis on the basis that this may be the mechanism for the deletion of autoreactive cells during T cell development. We show that the ability of the bacterial "superantigen" staphylococcal enterotoxin B to cause specific depletion of V beta 8+ cells when added to thymus organ cultures is accompanied by DNA degradation into oligonucleosomal fragments, indicating that depletion involves apoptosis. Our results provide the first direct evidence that antigen-induced apoptosis can be triggered in developing T cells.

Animals↗
PubMed1989

Developmental control of lymphokine gene expression in fetal thymocytes during T-cell ontogeny.

We have used the technique of in situ hybridization to investigate the expression of lymphokine genes by immature thymocytes during intrathymic development. In 13-day fetal thymocytes a population of cells constitutively produces low levels of interleukin 2 (IL-2) and interleukin 4 (IL-4) mRNAs. A second phase of lymphokine gene expression occurs in the majority of 15-day thymocytes, and a population of cells constitutively produces both IL-2 and IL-4 mRNAs. Thymocytes at 14 days of gestation and after 16 days up until birth do not express detectable lymphokine mRNA. By contrast, the population of IL-2 receptor mRNA-producing thymocytes increases progressively up to 15 days of gestation, and expression thereafter decreases up to birth. In addition, thymocytes expressing interferon gamma mRNA were not present until just prior to birth. Our findings indicate developmental control of lymphokine and lymphokine receptor gene expression in fetal thymocytes during ontogeny.

Animals↗
PubMed1989

Regulatory factors in lymphoid development.

The process of T lymphocyte maturation within the thymus involves a complex series of differentiation events involving T cell receptor gene rearrangement and expression. A beginning has now been made at defining the growth and differentiation signals required for these events. An additional level of complexity is provided by the selection of the repertoire of T cell specificities so that the pool of peripheral T cells available for interaction with antigens is 'shaped' by the thymus. The maturation of stem cells to B lymphocytes takes place in association with bone marrow stromal cells. Large numbers of newly-formed B cells can now be generated in vitro on stromal cell layers. Stromal cells capable of supporting B lymphopoiesis have now been cloned and, recently, a growth factor (IL-7) which acts on early B cells has been identified.

Animals↗
PubMed1988

Murine CD2 gene expression during fetal thymus ontogeny.

The expression of the murine CD2 gene and the interleukin 2 receptor IL2R has been investigated during fetal thymus ontogeny. CD2 mRNA was undetectable at embryonic day 14 but was readily detectable at day 15 of gestation. Surface IL2R expression was maximal at day 14 and was clearly detectable at day 13. These results indicate that IL2R expression precedes that of CD2 by at least 2 days.

Animals↗
PubMed1988

Factors affecting the acceptance of deoxyguanosine-treated thymus allografts.

dGuo-treated mouse embryo thymus lobes depleted of lymphoid and dendritic cells are not rejected by normal allogeneic mice despite expression of class I and class II major histocompatibility (MHC) antigens on thymic epithelial cells. Our results suggest that host mice are not tolerant to donor antigens, but treated lobes survive because thymic epithelial cells are not immunogenic. Multiple grafts of dGuo-treated lobes are rejected perhaps because numbers of residual dendritic cells reach a threshold necessary for priming. Mice previously primed by injection of spleen cells reject dGuo-treated thymus lobes, and crossreactions between major and minor histocompatibility antigens can be demonstrated. The acceptance of dGuo-treated thymus allografts by normal mice provides a system for investigating the requirements for priming.

Animals↗