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Biomedical subjects

J A Culpepper

Publications and source records attributed to J A Culpepper.

12 recordsLinked to original sources

Dendritic cells produce IL-12 and direct the development of Th1 cells from naive CD4+ T cells.

Dendritic cells are APCs that are unique in their potency to stimulate proliferation of primary Ag-specific responses in vitro and in vivo. In this study, we demonstrate that dendritic cells can produce IL-12, a dominant cytokine involved in the development of IFN-gamma-producing T cells. This finding resulted from our observations that dendritic cell-induced Th1 development from total CD4+ T cells upon neutralization of endogenous levels of IL-4 was IL-12-dependent. Furthermore, we demonstrate that dendritic cells can induce the development of Th1 cells from Ag-specific naive LECAM-1bright CD4+ T cells obtained from alpha beta-TCR transgenic mice, provided that CD4+ LECAM-1dull T cells, which produce significant levels of IL-4, are not present in the primary cultures. Production of IL-12 by dendritic cells was confirmed by positive immunofluoresence staining with Abs specific for the inducible IL-12 p40 subunit. This suggests that in addition to inducing proliferation and clonal expansion of naive T cells, dendritic cells, by their production of IL-12, play a direct role in the development of IFN-gamma-producing cells that are important for cell-mediated immune responses.

Animals↗

Interleukin 13, a T-cell-derived cytokine that regulates human monocyte and B-cell function.

We have isolated the human cDNA homologue of a mouse helper T-cell-specific cDNA sequence, called P600, from an activated human T-cell cDNA library. The human cDNA encodes a secreted, mainly unglycosylated, protein with a relative molecular mass of approximately 10,000. We show that the human and mouse proteins cause extensive morphological changes to human monocytes with an associated up-regulation of major histocompatibility complex class II antigens and the low-affinity receptor for immunoglobulin E (Fc epsilon RII or CD23). In addition, they stimulate proliferation of human B cells that have been activated by anti-IgM antibodies or by anti-CD40 monoclonal antibodies presented by a mouse Ltk- cell line transfected with CDw32. Furthermore, the human protein induced considerable levels of IgM and IgG, but no IgA production, in cultures in which highly purified human surface IgD+ or total B cells were cocultured with an activated CD4+ T-cell clone. Based on these findings, we propose that this immunoregulatory protein be designated interleukin 13.

Amino Acid Sequence↗

Identification of Dirofilaria immitis larval antigens with immunoprophylactic potential using sera from immune dogs.

Previous research has demonstrated that dogs that received chemically abbreviated Dirofilaria immitis larval infections were significantly immune to challenge infections. Sera from those immune animals have been effective in passively transferring larval killing and stunting. In the present study, sera from immune and control animals were used to screen various Ag subsets for unique Ag. Through Western blot analysis of larval extracts and excretory-secretory products, and immunoprecipitation of metabolically labeled proteins and larval surface Ag, it was determined that as many as 12 molecules were uniquely recognized by protective immune sera. A 39-kDa molecule was present in both soluble lysates of third- and fourth-stage larvae and larval excretory-secretory products; it was recognized by each of the immune dogs and by none of the infected or uninfected control animals. The 39-kDa molecule appeared to be absent from adults and microfilariae of the parasite. In addition to the unique recognition by immune dog sera, larval stage specificity of this molecule suggests that it may be useful as a vaccine candidate.

Animals↗

Flow resistance of expiratory positive-pressure systems.

We measured the flow-resistance of five commercially available 10 cm H2O expiratory positive-pressure (EPP) valves (n = five per valve type) at bias flows of between 0 and 2,000 ml/s. We found that individual valves of each type and manufacturer functioned similarly. Different valve types, however, functioned differently: with one type, system pressure was higher than rated (p less than 0.05), and with another type, system pressure was significantly flow-dependent (p less than 0.01). The remaining types of valves had no flow-resistive properties and maintained a system pressure of 10 cmH2O. We conclude that system pressure is not similar in all continuous positive airway pressure (CPAP) systems using bias flow and EPP valves. The work of breathing imposed by CPAP circuits will be increased in systems whose EPP valves have flow-dependent properties.

Airway Resistance↗

Effect of mechanical ventilator mode on tendency towards respiratory alkalosis.

The question whether assist-control ventilation (A/C) results in more frequent or more severe respiratory alkalosis than intermittent mandatory ventilation (IMV) is often raised. We prospectively compared the respiratory rates and arterial blood tensions of 18 critically ill patients with respiratory failure of diverse causes who were mechanically ventilated for 1 h with each of these ventilatory modes. Each patient served as his own control. We found that after 1 h of IMV the average pH was 7.42 +/- 0.2 (mean +/- SEM), after 1 h of A/C the pH was 7.45 +/- 0.01 (p less than 0.005), the average PaCO2 during IMV was 40.7 +/- 1.8, the average PaCO2 during A/C was 37.9 +/- 1.6 (p less than 0.001), the average respiratory rate during IMV was 21 +/- 2.0, and the average respiratory rate during A/C was 15 +/- 2.0 (p less than 0.001). One patient became alkalemic (pH 7.55) during A/C. These pH and PaCO2 changes were not associated with any adverse clinical sequelae. We conclude that the responsible physician should be guided by factors other than control of pH in choosing the mode of mechanical ventilation for most patients.

Adult↗

Barotrauma and hypotension resulting from jet ventilation in critically ill patients.

We present the first reports of pneumoperitoneum secondary to jet ventilation, barotrauma secondary to jet ventilation through the suction port of a fiberoptic laryngoscope, and hypotension due to jet ventilation via nasotracheal and orotracheal catheters. We suggest that minimizing airway pressure and using jet catheters with side holes may help decrease the risk of such complications. We cannot, at present, recommend the use of hand-held jet ventilators unless both adequate exhalation space is guaranteed and direct impingement of the catheter's tip on the mucosal surface is avoided.

Aged↗

Regulation of IL 3 expression by glucocorticoids in cloned murine T lymphocytes.

We have examined the effects of glucocorticoids on the concanavalin A (Con A)-stimulated production of interleukin 3 (IL 3) by a murine helper T cell clone Cl.Ly-1+2-/9. The addition of Con A to Cl.Ly-1+2-/9 cells leads to the rapid induction of both IL 3 mRNA and activity. A synthetic glucocorticoid, dexamethasone, inhibits the appearance of IL 3 activity in the supernatants of Con A-stimulated cells, as well as the accumulation of IL 3 mRNA in the cells. A similar effect is observed with other Con A-inducible mRNA, such as the mRNA encoding granulocyte-macrophage colony-stimulating factor (GM-CSF), but the hormone effects are not general; the constitutive synthesis of Thy-1 mRNA is not altered by dexamethasone treatment. Several lines of evidence suggest that dexamethasone directly inhibits transcription of the IL 3 gene: the kinetics of dexamethasone action on IL 3 mRNA are rapid; the stability of IL 3 mRNA is not affected by dexamethasone; hormone treatment does not inhibit either mRNA processing or transport from the nucleus. Our findings demonstrate that a set of mitogen-inducible genes in helper T cells can be subject to a second level of regulation mediated by glucocorticoid hormones.

Animals↗

Pretranslational control of tyrosine aminotransferase synthesis by 8-bromo-cyclic AMP in H-4 rat hepatoma cells.

The H-4 rat hepatoma cell line grown in tissue culture was used as a model system to investigate the action of cAMP in tyrosine aminotransferase induction. An immunoprecipitation technique was used to quantitate the amount and the rate of synthesis of tyrosine aminotransferase; the level of mRNA coding for tyrosine aminotransferase was determined by in vitro translation of poly(A)+ RNA isolated from hepatoma cells. Our results demonstrated that 8-bromo-cAMP gave time-dependent and proportionate increases in the tyrosine aminotransferase activity, the amount of immunoprecipitable tyrosine aminotransferase, the rate of synthesis of tyrosine aminotransferase, and the level of mRNATAT in H-4 hepatoma cells. The time course of increase in mRNATAT preceded the increase in synthesis of tyrosine aminotransferase and was dependent on the continuous production of poly(A)+ RNA. Pretreatment of the cells with cordycepin completely abolished the 8-bromo-cAMP-evoked increase in mRNATAT activity. These results provided evidence that the primary action of cAMP in tyrosine aminotransferase induction is the increase of functional mRNATAT and that this increase can completely account for the increase in tyrosine aminotransferase activity.

8-Bromo Cyclic Adenosine Monophosphate↗

Massive hemoptysis and tension pneumothorax following pulmonary artery catheterization.

We report a case of massive hemoptysis and bilateral tension pneumothorax immediately following placement of a fiberoptic pulmonary artery catheter. We postulate air entry into a traumatic communication caused by the catheter, traversing a parenchymal artery, the contiguous airspace and the interstitial space. Dissection of air through the perivascular sheath and into tissue planes of the thorax and abdomen followed. This sequence was rapidly fatal. Tension pneumothorax should be considered if dynamic respiratory system compliance remains poor despite aggressive evacuation of blood from the trachea of a patient with a recently inserted balloon-type catheter.

Catheterization↗

Induction of tyrosine aminotransferase in H-35 hepatoma cells by cAMP captured in phospholipid vesicles.

The uptake, metabolism, and action of cAMP, captured within phospholipid vesicles, in H-35 hepatoma cells were studied. Sonication of lipids in buffer containing cAMP resulted in the formation of 300-A unilamellar lipid vesicles, capturing cAMP in the internal aqueous cavity. Incubation of H-35 hepatoma cells with vesicles containing cAMP (vesicle-cAMP) resulted in rapid incorporation of the vesicle content; apparent saturation of uptake was reached after approximately 30 min of incubation at 37 degrees C. Uptake of vesicle-cAMP was linear over a 10-fold vesicle concentration range. Pretreatment of cells with combined inhibitors of glycolysis and respiration inhibited vesicle uptake by 27%, suggesting vesicle fusion with the cell membrane as a predominant pathway of vesicle uptake. Studies on the metabolism of incorporated cAMP indicated that greater than 50% of the cell-associated radioactivity, derived from vesicle-[3H]cAMP, was preserved as cAMP at the end of a 20-min incubation at 37 degrees C. The incorporation of vesicle-cAMP by H-35 hepatoma cells resulted in increased tyrosine aminotransferase (TAT) activity. The concentration of vesicle-cAMP needed to produce a half-maximal increase in TAT activity was 10 microM, approximately two orders of magnitude lower than that of exogenously added dbcAMP. cAMP was ineffective when added extracellularly. The kinetic relationship of the cAMP-induced increase in TAT activity and the binding of cAMP to its receptor protein, in intact H-35 cells, was examined using vesicle-trapped 8-N3-cAMP, a photoaffinity labeling analogue of cAMP. Incubation of H-35 hepatoma cells with vesicle-8-N3-cAMP resulted in increased TAT activity, preceded by the binding of 8-N3-cAMP to the regulatory subunit of type II cAMP-dependent protein kinase. The use of lipid vesicles provides a means of modulating intracellular cAMP concentration without adding cyclic nucleotide in the millimolar concentration range to the extracellular medium. The increased efficiency of intracellular delivery of cyclic nucleotide with retention of biological activity, provides a useful technique in examining the relationship of occupancy of specific cAMP-receptor protein(s) and the occurrence of a cAMP-mediated biological response in intact cells.

Animals↗