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R C Howe

Publications and source records attributed to R C Howe.

At least 37 records · Page 2Linked to original sources
PubMed1988

CD3-associated alpha/beta and gamma/delta heterodimeric receptors are expressed by distinct populations of CD4- CD8- thymocytes.

Immature double negative (DN) CD4-8- thymocytes expressing the CD3 molecular complex can be subdivided into two distinct subsets based on expression of the B2A2 antigen. Thus, B2A2+ CD3+ DN thymocytes express CD3-associated 35 kDa and 45 kDa polypeptide chains characteristic of a gamma/delta T cell receptor, whereas B2A2- DN thymocytes express predominantly 38 to 43 kDa CD3-associated structures typical of alpha/beta TCR. At the mRNA level, B2A2+ DN thymocytes express full length gamma and delta transcripts but no alpha message and only short (1 kb) B transcripts. In contrast, the B2A2- DN subset expresses full length alpha and beta transcripts and minimal levels of delta transcripts. Interestingly, B2A2- DN thymocytes, unlike mature T cells, also express abundant levels of full length TCR gamma mRNA, perhaps suggesting that these cells represent an early stage after divergence of the gamma/delta and alpha/beta T cell lineages.

Animals↗
PubMed1988

T-cell receptor V beta use predicts reactivity and tolerance to Mlsa-encoded antigens.

T lymphocytes reactive with the product of the Mlsa-allele of the minor lymphocyte stimulating (Mls) locus use a predominant T-cell receptor beta-chain variable gene segment (V beta 6). Such V beta 6-bearing T cells are selectively eliminated in the thymus of Mlsa-bearing mice, consistent with a model in which tolerance to self antigens is achieved by clonal deletion.

Animals↗
PubMed1988

Heterogeneity of immature (Lyt-2-/L3T4-) thymocytes. Identification of four major phenotypically distinct subsets differing in cell cycle status and in vitro activation requirements.

Thymocytes that bear neither Lyt-2 nor L3T4 differentiation Ag (2-4- thymocytes) contain the precursors of mature Lyt-2+ and L3T4+ T cells. In the present study, we have identified four major subpopulations of 2-4- cells in adult C57BL/6 mice that differ in surface phenotype and in situ proliferative status. Two-color immunofluorescence analysis with RL-73 (a mAb recognizing an as yet unidentified activation Ag) and PC-61 (an anti-IL-2R mAb) revealed three distinct subsets of 2-4-thymocytes: RL-73+ IL-2R- (30%), RL-73+/-IL-2R+ (45%), and RL-73- IL-2R- (25%). The RL-73+ IL-2R- subset had the highest percentage of large blasts and cycling cells, whereas the RL-73+/- IL-2R+ and RL-73- IL-2R- subsets had intermediate and low percentages, respectively, indicating that in situ proliferation correlated better with RL-73 intensity than with IL-2R expression. An additional marker, heat-stable Ag (HSA), was found to further subdivide the RL-73- population into RL-73- HSA- (10% of total 2-4-) and RL-73- HSA+ (15%) fractions. The two latter (RL-73-) subsets appeared to be more "mature" than the former since they expressed high levels of Lyt-1 and appeared later during fetal thymus ontogeny. In parallel with the phenotypic analysis, we compared the in vitro activation requirements of each of the four purified 2-4- subsets. All four populations proliferated well to the combination of phorbol ester (PMA), ionomycin, and IL-2. In response to PMA and ionomycin (without added IL-2), only RL-73- HSA-cells proliferated and this proliferation was correlated with IL-2 production. However, if IL-1 was included with PMA and ionomycin then all four populations responded. Finally, a proliferative response to Con A or mitogenic anti-Thy-1 mAb was observed only for RL-73- HSA+ and (to a lesser extent) RL-73- HSA-cells. These data indicate that each of the four phenotypically distinct subpopulations of immature thymocytes can also be distinguished on the basis of their in vitro activation requirements.

Animals↗
PubMed1988

A CD3- subset of CD4-8+ thymocytes: a rapidly cycling intermediate in the generation of CD4+8+ cells.

T lymphocytes with the surface phenotype CD4+8- and CD4-8+ are considered to be representative of functionally mature cells. We show here that adult murine thymus contains a subpopulation of CD4-8+ cells that differ from CD4-8+ cells found in the periphery in that they do not express the T cell receptor-associated CD3 molecular complex. Such CD3-4-8+ thymocytes are cortisone sensitive and rapidly cycling in situ. Furthermore, in contrast to mature T cells, most CD3-4-8+ thymocytes express low levels of CD5 and high levels of the B2A2 antigen. CD3-4-8+ thymocytes fail to respond to a variety of mitogenic stimuli in vitro but do give rise upon short-term culture to CD4+8+ cells. It is suggested that CD3-4-8+ thymocytes represent a transitional stage of thymus differentiation between the CD4-8- and CD4+8+ compartments.

Animals↗
PubMed1988

Corporate strategies: organizational structure.

In previous installments of this series, the authors outlined factors that influence corporation information system strategies. The factor that appears to be most significant is centralization vs. decentralization. This article presents examples of organizational structures, roles and responsibilities for either approach.

Centralized Hospital Services↗
PubMed1988

Corporate information system strategies: alternatives. Part 2.

Analyzing various approach alternatives is a complex process. The business goals of the corporation and the various MIS factors must be evaluated in relation to each approach option. The conditions or assumptions under which the corporation is willing to operate will set the stage for the evaluation and selection of the most appropriate approach. Once the corporate MIS approach alternative is determined, an MIS organizational structure can be developed to support the approach. The next article in this series will present various MIS organizational structures, roles, and responsibilities for both large and small multi-hospital groups that employ either a centralized or decentralized MIS approach option.

Centralized Hospital Services↗
PubMed1987

Developmentally regulated expression of T cell receptor beta chain variable domains in immature thymocytes.

A minor subset of immature (CD4-,8-) thymocytes that lack expression of the B2A2 antigen was found to express low levels of surface TCR protein as detected by mAbs F23.1 and KJ16 (reacting with protein products of the V beta 8 gene family). Interestingly, F23.1/KJ16 determinants were expressed on a two- to three-fold higher proportion of B2A2- thymocytes than mature lymph node T cells in four independent haplotypes. When expanded in short-term culture with PMA and calcium ionophore, B2A2- thymocytes retained their overexpression of F23.1/KJ16 determinants and showed a fivefold elevated level (relative to lymph node) of V beta 8-specific mRNA. Taken together, these findings suggest that expression of TCR V beta genes, like Ig genes, is developmentally regulated.

Animals↗
PubMed1987

Distinction of virgin and memory T lymphocytes. Stable acquisition of the Pgp-1 glycoprotein concomitant with antigenic stimulation.

The Pgp-1 glycoprotein was identified on a minor (27%) subset of peripheral Lyt-2+ or L3T4+ T cells. In contrast, mature medullary-type thymocytes (Lyt-2+ L3T4-, Lyt-2- L3T4+) were nearly devoid of cells expressing detectable surface Pgp-1. The appearance of peripheral Pgp-1- T cells was found to be thymus dependent, as demonstrated by the diminished proportion of Pgp-1- T cells after thymectomy and their virtual absence in athymic nude mice. The subsequent acquisition of surface Pgp-1 was found to be a stable differentiation event occurring concomitantly with primary antigenic stimulation; selected Pgp-1- mature T cells from thymus or periphery acquired constitutive expression of Pgp-1 after stimulation in vitro with alloantigen or mitogens. These observations were extended by studies in vivo showing that immunization with various antigens augmented the percentage of Pgp-1+ spleen cells within the Lyt-2+ subset. Furthermore, the frequencies of antigen-specific CTLp, after immunization by any of three different antigens tested, were greatly enriched in the Pgp-1+ compared with the Pgp-1- subpopulations. Peritoneal exudate Lyt-2+ cells, after a localized allograft rejection, demonstrated a particularly prominent Pgp-1+ subpopulation (78%) that contained virtually all the allospecific cytolytic activity. A model consistent with all of these data proposes that mature thymocytes lacking surface Pgp-1 upon emigration to the periphery acquire its expression at the time of primary antigenic stimulation. Hence, expression of Pgp-1 among peripheral T cells is an important differentiation marker for identifying antigen-stimulated memory T cells.

Animals↗
PubMed1987

Anti-Thy-1-induced proliferation of immature thymocytes expressing the CD3-associated gamma/delta heterodimer.

A series of rat monoclonal antibodies (mAb) directed against the Thy-1 molecule have been evaluated for their ability to stimulate immature (CD4-8-) murine thymocytes. CD4-8- thymocytes could be induced to proliferate by several anti-Thy-1 mAb provided that exogenous interleukin 1 or interleukin 2 was provided. Apparently non-stimulatory anti-Thy-1 mAb could be rendered stimulatory if cross-linked by a rabbit anti-rat immunoglobulin reagent. Anti-Thy-1-stimulated CD4-8- thymocytes were found to uniformly express CD3 but did not stain positively with F23.1, a mAb reactive with alpha/beta T cell receptors utilizing the V beta 8 gene family. Immunoprecipitation analysis of such stimulated CD4-8- thymocytes indicated that they expressed a CD3-associated heterodimer of 45 kDa/35 kDa most likely corresponding to the product of the gamma and (putative) delta T cell receptor genes. Taken together, these data demonstrate that anti-Thy-1 mAb selectively activate a subset of CD4-8- thymocytes committed to the gamma/delta T cell receptor lineage.

Animals↗
PubMed1986

Role of interleukin 1 in early T cell development: Lyt-2-L3T4- thymocytes bind and respond in vitro to recombinant IL 1.

Thymocytes that bear neither Lyt-2 nor L3T4 differentiation antigens (2-4- thymocytes) contain the precursors for mature L3T4+Lyt-2- and Lyt-2-L3T4+ T cells. In the present study we determined the capacity of 2-4- cells to respond to recombinant interleukin 1 (rIL 1) in vitro. The presence of rIL 1 enhanced IL 2-dependent proliferation to the lectins Con A and PHA by threefold to eightfold. In a second assay, rIL 1 enhanced proliferation and IL 2 production by 2-4- cells in response to phorbol myristate acetate (PMA) and the calcium ionophore, ionomycin. Using a direct IL 1 binding assay, we were able to detect both high-affinity (Kd approximately 5 pM) and low-affinity (Kd approximately 200 pM) classes of IL 1 receptors on freshly isolated 2-4- cells. Bound IL 1 was rapidly internalized, suggesting that such receptors were functional. These results are compatible with a role for interleukin 1 during thymocyte maturation.

Animals↗
PubMed1986

Functional status of interleukin 2 receptors expressed by immature (Lyt-2-/L3T4-) thymocytes.

A subpopulation of phenotypically immature (Lyt-2-/L3T4-) thymocytes express receptors for the polypeptide hormone interleukin 2 (IL 2); however, these cells do not proliferate in vitro in response to IL 2. In investigating this phenomenon in greater detail, we observed that the IL 2 receptors (IL 2-R) on freshly isolated immature thymocytes bound IL 2 with about fivefold lower affinity (Kd approximately 100 pM) than IL 2-R on activated mature T cells and T cell lines (Kd approximately 20 pM). Furthermore, in contrast to activated T cells, Lyt-2-/L3T4- thymocytes did not endocytose bound IL 2. When stimulated in short-term culture with a combination of phorbol ester (PMA) and calcium ionophore, Lyt-2-/L3T4- thymocytes proliferated in a largely IL 2-dependent fashion. IL 2-R expression on these activated cells initially disappeared (at 24 hr) and subsequently reappeared (at 48 to 72 hr). Reexpressed IL 2-R on activated thymocytes resembled those on mature T lymphocytes in that they bound IL 2 with high affinity (Kd = 15 to 25 pM) and were capable of endocytosing IL 2. Taken together, these data place certain constraints on the putative physiologic role of IL 2 in intrathymic growth regulation.

Animals↗
PubMed1985

Interleukin 2-mediated induction of lytic activity in a cloned murine CTL line.

We have analyzed the ability of interleukin 2 (IL 2) to induce lytic activity within a cloned murine H-2Dd-specific CTL line. Weakly lytic CTL harvested 6 to 7 days after previous stimulation with irradiated DBA/2J spleen cells and conditioned medium from secondary MLC (MLC SN) could be reactivated to high antigen-specific lytic activity with highly purified gibbon IL 2 or E. coli-produced human recombinant DNA IL 2. Dose-response curves with IL 2 and MLC SN suggest that IL 2 may be the principal detectable activity in MLC SN that is active on these CTL. Doses of IL 2 or MLC SN that were saturating for the induction of lytic activity were suboptimal for the expression of DNA synthesis measured by 3HTdR incorporation. This is consistent with a mechanism in which different threshold IL 2 concentrations are required to induce these two biologic responses. Finally, we show that IFN-gamma has little effect on the expression of lytic activity either alone or in combination with IL 2 in this bioassay.

Absorption↗
PubMed1985

Inhibition of cytotoxic T lymphocyte-mediated lysis by ETYA: effect independent of arachidonic acid metabolism.

Cytotoxic T lymphocyte (CTL)-mediated lysis of target cells was inhibited by 5,8,11,14-eicosatetraynoic acid (ETYA) and other inhibitors of the lipoxygenase pathway at concentrations that inhibited arachidonic acid metabolism in mixed lymphocyte cultures. Inhibition was reversible and selective for the "lethal hit" stage in the CTL-target interaction. Studies to define CTL-specific arachidonic acid metabolites demonstrated that cloned CTL populations have little or no capacity to metabolize arachidonic acid. Therefore, inhibitor actions appear to be independent of the effects on CTL arachidonic acid metabolism. Alternative explanations for inhibitory effects are discussed.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗