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J Sprent

Publications and source records attributed to J Sprent.

At least 73 records · Page 4Linked to original sources
PubMed1996

Bone marrow-derived cells fail to induce positive selection in thymus reaggregation cultures.

The requirements for inducing positive selection of T cells were examined in thymus reaggregation cultures, a system in which dispersed populations of immature CD4+8+ cells and purified thymic epithelial cells (TEC) are reaggregated in tissue culture. Studies with TEC from mice selectively lacking major histocompatibility complex (MHC) class I (I-II+), class II (I+II-), or both class I and II (I-II-) molecules showed that class II expression was essential for the differentiation of CD4+8+ cells into CD4+8- cells. Unexpectedly, the generation of TCRhi CD4-8+ cells from CD4+8+ cells was apparent with I-II+ TEC but not with I-II- TEC, perhaps reflecting cross-reactive specificity of CD4-8+ cells for class II molecules. Significantly, the failure of I-II- TEC to generate TCRhi CD4+8- or CD4-8+ cells could not be overcome by adding MHC+ bone marrow-derived cells. These findings, together with experiments on purified subsets of TEC, suggest that positive selection in thymus reaggregation cultures is an exclusive property of cortical TEC.

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PubMed1996

Dual function of Drosophila cells as APCs for naive CD8+ T cells: implications for tumor immunotherapy.

With unseparated mouse spleen cells as responders, Drosophila cells expressing MHC class I (L(d)) molecules alone lead to peptide-specific responses of CD8+ cells in the absence of exogenous cytokines. Under these conditions, DNA released from dying cells stimulates the B cells in spleen to up-regulate costimulatory molecules; these activated B cells then provide bystander costimulation for CD8+ cells responding to class I-peptide complexes on the Drosophila APCs. By stimulating B cells and presenting antigen to T cells, Drosophila cells thus serve two different functions in promoting primary responses of CD8+ cells in vitro. With this system, we show that Ld-transfected Drosophila cells are able to induce autologous spleen cells to respond to a tumor-specific peptide in vitro and, after transfer, cause tumor rejection in vivo.

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PubMed1996

The thymus and central tolerance.

Thymic selection programs T lymphocytes to tolerate self antigens but respond to foreign antigens. The mechanisms involved in self tolerance induction are discussed.

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PubMed1995

Antigen-presenting cells. Professionals and amateurs.

Professional antigen-presenting cells, notably dendritic cells, play a key role in stimulating naive T cells--but nonprofessional antigen-presenting cells, such as fibroblasts, may also contribute to this process.

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PubMed1995

Thymic selection and cell division.

Cell division during thymic selection was studied with a system in which purified populations of T cell antigen receptor (TCR)- CD4+8+ (double-positive [DP]) cells and fetal thymic epithelial cells (TEC) were reaggregated in tissue culture. In this system, immature DP cells differentiate into mature single-positive (SP) CD4+8- and CD4-8+ TCRhi cells within 3-4 d, indicative of positive selection. By adding the DNA precursor, bromodeoxyuridine, to the cultures and staining cells for bromodeoxyuridine incorporation, T cell division in reaggregation cultures was found to be high on day 1, low on day 2, and high on days 4-5. Cell separation studies established that cell division on day 1 was restricted to DP blast cells. In the absence of blast cells, small DP cells failed to proliferate and differentiated into SP cells without cell division, thus indicating that proliferation is not an essential component of positive selection. This applied to SP cells generated within the first 2-3 d. Surprisingly, the SP cells generated later in culture showed a high rate of cell division; the proliferating SP cells were TCRhi and included both CD4+8- and CD4-8+ cells. Turnover of TCRhi SP cells was also prominent in the normal neonatal thymus and in TEC reaggregation cultures prepared with adult lymph node T cells. We speculate that division of mature SP cells in the perinatal thymic microenvironment is driven by stimulatory cytokines released from TEC. Such proliferation could be a device to expand the mature T cell repertoire before export to the periphery.

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PubMed1995

Upregulation of surface markers on dying thymocytes.

Using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) to detect cells undergoing early apoptosis, we have defined the surface markers expressed on CD4+CD8+ thymocytes undergoing spontaneous or steroid-induced apoptosis in tissue culture. Some surface markers, e.g., CD4, CD8, and heat stable antigen, are downregulated on apoptotic thymocytes. Surprisingly, however, other markers are upregulated; this applies to T cell receptor beta/CD3, CD69, and CD25 expression. Upregulation of these markers is restricted to a discrete subset of apoptotic cells.

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PubMed1995

Split tolerance in spleen chimeras.

Transferring small doses of T cells to heavily irradiated F1 mice expressing isolated MHC class I or class II differences invariably leads to rapid death from graft-vs-host disease (GVHD). Paradoxically, GVHD is mild or absent when irradiated F1 mice are reconstituted with large doses of unseparated parental strain spleen cells. This applies when bulk populations of B6 spleen cells are transferred to irradiated class II-different (B6 x bm12)F1 mice or class I-different (B6 x bm1)F1 mice. In this study, we examined whether the donor T cells in long-term spleen chimeras become tolerant to host MHC Ags. On the basis of skin-allograft rejection and induction of GVHD on adoptive transfer, the results show that the donor T cells display strong tolerance to host antigens; this applies to CD4+ cells in class II-different chimeras and to CD8+ cells in class I-different chimeras. In marked contrast to the profound tolerance seen by these in vivo parameters, little or no tolerance is observed in standard in vitro assays. The results illustrate that typical in vitro tests for alloreactivity are an imprecise guide to physiologic tolerance of T cells in vivo.

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PubMed1995

Life span of naive and memory T cells.

The life span of mature T cells is reviewed. Peripheral T lymphocytes are a heterogeneous population and comprise a mixture of naive, effector and memory cells. The recirculating pool of mature T cells is formed during young life through gradual release of naive T cells from the thymus. In adults, the pool of mature T cells is relatively self-sufficient, and input of new T cells from the thymus declines to low levels. Studies on T cell turnover indicate that most peripheral T cells can remain in a resting state for long periods (months in rodents and years in humans). Examination of the phenotype of dividing versus nondividing cells suggests that typical naive T cells are long-lived resting cells whereas the majority of effector and memory T cells have a much more rapid turnover. However, some memory T cells appear to divide very infrequently and eventually return to a resting state. The factors controlling the generation and maintenance of memory T cells are discussed.

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PubMed1995

Lifespan of lymphocytes.

T and B lymphocytes comprise heterogeneous populations of cells at various stages of differentiation and activation. T- and B-cell subsets have different roles in the maintenance of immune homeostasis, and their functional differences are reflected by their respective lifespans. This review briefly summarizes the available data on lymphocyte lifespan, including the kinetics of T- and B-cell development in the primary lymphoid organs and the proliferative behavior of naive, effector and memory lymphocytes in the peripheral lymphoid compartment.

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PubMed1995

Intrathymic and extrathymic clonal deletion of T cells.

Clonal elimination accounts for self-tolerance induction in the thymus and also affects mature T cells responding to exogenous antigens in the periphery. Recent evidence on the microenvironments, cell-cell interactions and signalling requirements for clonal deletion of immature and mature T cells is discussed.

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PubMed1995

Central tolerance of T cells.

The immune system is constructed to tolerate self antigens but give vigorous responses to foreign antigens. How this state of self/nonself discrimination is maintained is controversial. In the case of T cells, many self antigens are transported to the thymus via the bloodstream and induce tolerance (clonal deletion) of self-reactive thymocytes in situ. Although such central tolerance in the thymus is well documented, it is often argued that full induction of tolerance requires peripheral mechanisms such as suppression or induction of anergy. This article proposes that steady-state tolerance of T cells to self components is due solely to central tolerance to circulating self antigens combined with sequestration of tissue-specific antigens. Backup mechanisms for tolerance do exist but such immunoregulation only operates when self tolerance breaks. This scheme allows the immune system to give unrestricted primary responses to foreign antigens.

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PubMed1994

T-cell apoptosis detected in situ during positive and negative selection in the thymus.

Because of positive and negative selection to molecules of the major histocompatibility complex (MHC), only a small proportion of the massive numbers of T cells generated in the thymus are selected for export. Immature thymocytes have a rapid turnover, and it has long been assumed that most thymocytes die in situ, presumably from apoptosis. This has yet to be proved, however, and conventional staining techniques have shown only minimal evidence of cell death in the normal thymus. Using a method for detecting cells with DNA strand breaks, we now present direct evidence for apoptosis in the normal thymus. In sections of thymus from adult mice, apoptotic cells are scattered throughout the cortex and are engulfed locally by F4/80+ macrophages. Apoptosis in the thymic cortex is not reduced in MHC-deficient mice, which suggests that T-cell death is primarily a reflection of lack of positive selection rather than negative selection. Direct evidence for apoptosis due to negative selection was obtained by crossing a V beta 5 transgenic line to I-E+ and I-E- mice: I-E+ mice are known to eliminate V beta 5+ T cells in the thymus whereas I-E- mice do not. In marked contrast to I-E- mice, the medulla of I-E+ V beta 5 transgenic mice contains dense aggregates of apoptotic cells; these cells are engulfed by a distinct population of F4/80- MAC-3+ macrophages. Negative selection of V beta 5+ cells is thus restricted to the medulla.

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PubMed1994

Lymphocyte life-span and memory.

Differentiation of immature T and B cells in the primary lymphoid organs gives rise to a pool of long-lived lymphocytes that recirculate through the secondary lymphoid tissues. On the basis of their surface markers, T and B cells comprise a mixture of naïve and memory cells with differing life-spans. Immunization (and vaccination) causes naïve lymphocytes to proliferate and differentiate into effector cells and memory cells. Whether the survival of memory cells is innate or requires persistent contact with residual antigen is controversial. Resolving this issue may be crucial for designing optimal vaccines.

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PubMed1994

Profound atrophy of the bone marrow reflecting major histocompatibility complex class II-restricted destruction of stem cells by CD4+ cells.

The effector functions of CD4+ cells in vivo are presumed to reflect a combination of lymphokine-mediated bystander reactions and direct cytotoxic T lymphocyte activity. To assess the relative importance of these two mechanisms, we studied the effects of transferring small doses of purified unprimed CD4+ cells to lightly irradiated (600 cGy) recipients expressing major histocompatibility complex class II (Ia) differences. Within the first week after transfer, the host marrow was rapidly repopulated with hemopoietic cells. Thereafter, however, the donor CD4+ cells caused massive destruction of hemopoietic cells, both in marrow and spleen. Marrow aplasia did not affect stromal cells and was prevented by coinjecting donor but not host bone marrow. The use of allotypic markers and fluorescence-activated cell sorter analysis indicated that the destructive effects of CD4+ cells were directed selectively to host Ia+ hemopoietic cells, including stem cells; donor hemopoietic cells and Ia- host T cells were spared. No evidence could be found that the ongoing destruction of host cells impaired the capacity of donor stem cells to repopulate marrow, spleen, or thymus. Moreover, CD4+ cells failed to destroy host-type hemopoietic cells from Ia-deficient mice. Tissue destruction by CD4+ cells thus did not seem to reflect a bystander reaction. We conclude that, under defined conditions, CD4+ cells can manifest extremely potent Ia-restricted CTL activity in vivo, probably through recognition of covert Ia expression on stem cells and/or their immediate progeny.

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PubMed1994

Resting and activated T cells display different requirements for CD8 molecules.

Clonotype-positive (1B2+) T cells from 2C T cell receptor (TCR) transgenic mice were used to define the role of CD8 molecules in the induction phase vs. the effector phase of the primary response to class I alloantigens. Three main findings are reported. First, in the presence of exogenous lymphokines, resting CD8+ 2C cells gave strong proliferative responses to two alloantigens, Ld and Kbm11. In the absence of added lymphokines, however, CD8+ 2C cells responded only to Ld and not to Kbm11; Ld stimulated both interleukin 2 (IL-2) and IL-2 receptor (R) synthesis, whereas Kbm11 elicited only IL-2R synthesis. The primary response of CD8+ 2C cells was thus helper-independent (HI) to Ld but helper-dependent (HD) to Kbm11, presumably reflecting that Ld is a stronger antigen than Kbm11. Second, in contrast to CD8+ cells, CD8- 2C cells mounted only an HD and not an HI response to the strong Ld antigen; conversely, selecting for a minor subset of CD8hi cells enabled 2C cells to mount an HI response to the weak Kbm11 antigen. These findings, together with experiments with heterozygous vs. homozygous stimulator cells, suggest that HI and HD responses reflect differences in the overall avidity of T antigen presenting cell (APC) interaction: high-avidity interaction leads to strong intracellular signaling and an HI response, whereas low-avidity interaction causes weak signaling and an HD response; high-avidity T/APC interaction is heavily dependent on CD8 expression. Third, CD8 expression was found to be less important for CTL activity than for primary proliferative responses. Thus, in contrast to HI proliferative responses, CTL responses of 2C cells to Ld were CD8 independent. However, 2C lysis of Ld targets became strongly CD8 dependent in the presence of limiting doses of anti-TCR (1B2) antibody. Collectively, the data suggest that, both for T cell induction and the expression of effector function, CD8 molecules play a decisive role in augmenting TCR-mediated signaling, either by promoting TCR contact with antigen or delivering kinases (p56lck) to the TCR/CD3 complex, or both.

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