The presentation of cell surface alloantigens to T cells.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to T Owens.
Explore the source record for details and available documents.
Both B lymphocytes and cytotoxic T lymphocytes respond to signals from the T helper (Th) compartment, and such signals are mediated by a number of biochemically distinct factors. This raises the question whether help for B cells and T cells is a function of one or several different kinds of Th cell. Here we describe an in vitro and in vivo study of this problem, using a Th clone, designated MTH-1. The clone carries the cell surface markers Thy-1 and L3T4a, but lacks Lyt-2. It recognizes a minor alloantigen shared by DBA/2, B10.D2 and NZB spleen cells, and such recognition is restricted by H-2Ed. Recognition of antigen in vitro is accompanied by secretion of IL-2. In vivo, both primary and secondary CTL responses to multiple minor alloantigens are enhanced by small numbers (less than or equal to 10(4] of MTH-1 cells. Recognition of alloantigen in a T-depleted B cell population results in the polyclonal activation and maturation of the B cells to secrete immunoglobulin; also, antigen-primed B cells are augmented in their in vivo synthesis of specific antibody to the Thy-1 X 1 alloantigen by around 10(5) MTH-1 cells. Taken together, these results suggest a single Th clone can help both B cells and T cells.
Explore the source record for details and available documents.
When splenic lymphocytes from RNC nu/+ mice were enriched for T cells by lectin purification, mixed to constant cell density with splenocytes from syngeneic nude mice, and cultured with ConA, a proliferative response ensued which was greater than that expected from the T cells alone. This was shown both by incorporation of 3H-thymidine after 48 h of culture and by uptake of potassium (measured as 86Rb) after 15 h. Analysis of metaphase chromosomes stained with Hoechst dye 33258 from co-cultured T-enriched and nude lymphocytes (from female and male donors, respectively) mixed in the proportion 1:4 showed that 40% of the mitotic cells came from nude spleen. About half of the blast cells in such mixtures stained strongly with fluorescein-coupled goat anti-mouse immunoglobulin; T-cell blasts did not stain under these conditions. Treatment of the nu/nu cells with anti-Thy 1.2 and complement had no effect on their subsequent proliferation in coculture. B lymphocytes from nude mouse spleen were therefore activated to proliferate in this system. This B-cell activation can be detected by increased potassium uptake 15 h after the initiation of co-culture. Thus the increased monovalent cation flux (previously demonstrated when B and T lymphocytes were separately stimulated) also occurs when B cells are stimulated through cooperation with mitogen-activated T cells, and is also detectable early in culture. T-dependent activation of B cells is therefore detectable considerably earlier than by conventional assays (such as plaque formation).
Low doses (30-84 ergs/mm2, 1 erg = 10(7) J) of ultraviolet radiation (UV) caused severe inhibition of the proliferation of human lymphocytes in vitro. Greatest inhibition was produced when resting cells were irradiated immediately prior to stimulation with concanavalin A (Con A); this was true whether activation was measured by the incorporation of labelled leucine, uridine, or thymidine. If UV was applied at 44 h after culture in presence of Con A, the incorporation of [3H]thymidine measured 4 h later was seen to be inhibited but transcription and translation were scarcely affected. UV, applied after the appearance of the Con A induced thymidine transport system, did not inhibit its function. The disaggregation of chromatin and increase in nuclear volume characteristic of, and essential to, the proliferative response of lymphocytes were completely inhibited if the cells were irradiated before mitogen was added to the cultures, but were unaffected if irradiation occurred after 16 h of culture in presence of Con A. Cells irradiated with 84 ergs/mm2 at the onset of culture with mitogen did not show the early increase of cation pump function which is a characteristic of stimulated lymphocytes, when this was measured by means of 86Rb uptake after 2-4 h culture. The mitogen-stimulated activation of cation pump function has previously been shown to be unaffected by concentrations of cycloheximide and actinomycin D which produce virtually complete inhibition of protein and RNA synthesis, respectively. The major inhibitory effect of UV treatment of lymphocytes at onset of culture with Con A is therefore not on DNA or on DNA synthesis, but on some component(s) of the early activation process, possibly at the cell periphery; inactivation of this component prevents cells from proceeding into later stages of the proliferative pathway.
Explore the source record for details and available documents.
Explore the source record for details and available documents.
Co-culture at constant cell density of nude mouse spleen cells (by themselves unresponsive to the T-cell mitogen concanavalin A (Con A)), with congenic T-enriched lymphocyte suspensions and Con A caused anomalously high activation of K+ transport (measured by 86Rb uptake) and of incorporation of thymidine into DNA; the expected dilution of these two responses by nude spleen cells did not occur. However, if the nude splenocytes were added immediately prior to assay to the enriched T cells that had been precultured in presence of Con A, the expected dilution of the activated T-cell responses occurred; both 86Rb uptake and thymidine incorporation were reduced proportionally to the degree of dilution of the T cells by the nonresponding cells. These data indicate that during co-culture in presence of Con A there is interaction between the T cells, capable of responding to mitogens, and the nude spleen cells. Attempts to demonstrate a diffusible factor in the supernatants of stimulated T cells were unsuccessful. The measured interaction is sufficient to explain our previous paradoxical findings that enrichment of T cells as measured by membrane markers did not cause a corresponding enrichment for either cation transport or for thymidine incorporation, and that depletion of T cells in the B-enriched cultures did not cause a corresponding decrease in these two Con A induced responses.
Quantitative microscopical measurements have been made of the proportion of cell volume occupied by cytoplasm in a cell suspension culture derived from cotyledons of bush bean (cv. Contender). On a 7-day culture cycle, the content of cytoplasm varies from 25% at the time of transfer to 45% at the start of the phase of rapid cell division. If the culture is continued beyond 7 days, the vacuole volume reaches 90% of cell volume by day 12.Attempts to measure relative cytoplasmic volumes by compartmental analysis of nonelectrolyte efflux were unsuccessful. The proportion of cell volume occupied by cytoplasm is roughly correlated with protein content, but shows no correlation with cell size or with intracellular concentrations of K or Na. The most striking observation is that the growth of cytoplasmic volume for the culture as a whole appears to be constant throughout the culture cycle, despite changes in the rate of cell division, cell size, rate of increase in fresh or dry weight, amount of cytoplasm per cell, the cellular concentration or fluxes of Na or K, and the rate of vacuolation. It is suggested that cytoplasmic volume is under the control of its own regulatory mechanism, which operates to give a constant exponential increase in cytoplasmic volume independently of most other observed cellular properties.
Explore the source record for details and available documents.
T-cell-derived cytokines are therefore individually unnecessary and collectively insufficient for microglial response. This somewhat provocative interpretation does not exclude a role for T-cell cytokines in induction of a microglial response in EAE, but it may be easier to show a non-requirement then to prove such a role. The point that emerges is that cytokine production in the CNS parenchyma is itself dependent on the prior infiltration of immune cells, and that without immune cell entry, EAE does not occur. This identifies events at the BBB, and in particular in the perivascular space, as critical immunoregulatory events in development and progression of EAE.