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Biomedical subjects

T A Davis

Publications and source records attributed to T A Davis.

At least 55 records · Page 3Linked to original sources
PubMed1998

Bone marrow repopulation by human marrow stem cells after long-term expansion culture on a porcine endothelial cell line.

In vitro exposure of murine hematopoietic stem cells (HSCs) to cell cycle-inducing cytokines has been shown to result in a defect in the ability of these cells to engraft. We used a porcine microvascular endothelial cell (PMVEC) line in conjunction with exogenous interleukin (IL)-3, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and stem cell factor (SCF) to expand human HSCs that express the CD34 and Thy-1 antigens but lack lineage-associated markers (CD34+Thy-1+Lin- cells). Ex vivo expansion of hematopoietic cells was evaluated in comparison to stromal cell-free, cytokine-supplemented cultures. Cells expressing the CD34+Thy-1+Lin- phenotype were detectable in both culture systems for up to 3 weeks. These cells were reisolated from the cultures and their ability to engraft human fetal bones implanted into SCID mice (SCID-hu bone) was tested. HSCs expanded in PMVEC coculture were consistently capable of competitive marrow repopulation with multilineage (CD19+ B lymphoid, CD33+ myeloid, and CD34+ cells) progeny present 8 weeks postengraftment. In contrast, grafts composed of cells expanded in stroma-free cultures did not lead to multilineage SCID-hu bone repopulation. Proliferation analysis revealed that by 1 week of culture more than 80% of the cells in the PMVEC cocultures expressing the primitive CD34+CD38- phenotype had undergone cell division. Fewer than 1% of the cells that proliferated in the absence of stromal cells remained CD34+CD38-. These data suggest that the proliferation of HSCs in the presence of IL-3, IL-6, GM-CSF, and SCF without stromal cell support may result in impairment of engraftment capacity, which may be overcome by coculture with PMVECs.

Animals↗
PubMed1997

Human mast cells produce the CD4+ T lymphocyte chemoattractant factor, IL-16.

CD4+ T cell infiltration is known to occur in tissues at sites of mast cell activation. The molecules produced and released by mast cells that account for this lymphocyte accumulation are poorly characterized. Here we report that a CD4+ T cell chemoattractant cytokine, IL-16, is stored preformed in bone marrow-cultured human mast cells and a human mast cell line, HMC-1, as demonstrated by intracytoplasmic cytokine staining and flow cytometry, and in human lung mast cells, as detected by immunohistochemistry. In response to the anaphylatoxin, C5a, or to PMA treatment, IL-16 mRNA transcripts detected by Northern blot analysis in HMC-1 cells increased 6- to 10-fold. HMC-1 cell lysates and activated supernatants contained IL-16 protein, as demonstrated by both ELISA and in vitro lymphocyte chemotaxis assays, the latter of which was blocked 59 to 88% by the addition of neutralizing Ab to recombinant human IL-16. IL-16 bioactivity was detected in the supernatants 2 to 4 h after PMA or C5a activation, and this activity remained elevated through 24 h. The capacity of human mast cells to synthesize and release biologically active IL-16 provides a possible link between mast cell activation and the accumulation of T cells in mast cell-dependent inflammation, thus amplifying the immune response and perpetuating the pathologic process.

CD4-Positive T-Lymphocytes↗
PubMed1997

CTLA4-Ig and anti-CD40 ligand prevent renal allograft rejection in primates.

Selective inhibition of T cell costimulation using the B7-specific fusion protein CTLA4-Ig has been shown to induce long-term allograft survival in rodents. Antibodies preventing the interaction between CD40 and its T cell-based ligand CD154 (CD40L) have been shown in rodents to act synergistically with CTLA4-Ig. It has thus been hypothesized that these agents might be capable of inducing long-term acceptance of allografted tissues in primates. To test this hypothesis in a relevant preclinical model, CTLA4-Ig and the CD40L-specific monoclonal antibody 5C8 were tested in rhesus monkeys. Both agents effectively inhibited rhesus mixed lymphocyte reactions, but the combination was 100 times more effective than either drug alone. Renal allografts were transplanted into nephectomized rhesus monkeys shown to be disparate at major histocompatibility complex class I and class II loci. Control animals rejected in 5-8 days. Brief induction doses of CTLA4-Ig or 5C8 alone significantly prolonged rejection-free survival (20-98 days). Two of four animals treated with both agents experienced extended (>150 days) rejection-free allograft survival. Two animals treated with 5C8 alone and one animal treated with both 5C8 and CTLA4-Ig experienced late, biopsy-proven rejection, but a repeat course of their induction regimen successfully restored normal graft function. Neither drug affected peripheral T cell or B cell counts. There were no clinically evident side effects or rejections during treatment. We conclude that CTLA4-Ig and 5C8 can both prevent and reverse acute allograft rejection, significantly prolonging the survival of major histocompatibility complex-mismatched renal allografts in primates without the need for chronic immunosuppression.

Abatacept↗
PubMed1997

Conditioned medium from primary porcine endothelial cells alone promotes the growth of primitive human haematopoietic progenitor cells with a high replating potential: evidence for a novel early haematopoietic activity.

The authors have recently shown that direct contact with primary porcine microvascular endothelial cell monolayers (PMVECs) in combination with haematopoietic growth factors enhances the expansion of primitive human haematopoietic CD34+ bone marrow progenitor cells. It is now demonstrated that serum-free conditioned medium (PMVEC CM, concentrated 70x for proteins >30 kDa) from untreated PMVECs contains haematopoietic growth factor activity that enhances the in vitro proliferation, haematopoietic cell production, and colony cell formation of primitive human haematopoietic progenitor cells. In combination with exogeneously added human growth factors such as interleukin 3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF) and EPO, PMVEC CM enhances the proliferation and colony growth of human haematopoietic CD34+ cells. In contrast, PMVEC CM has no significant synergistic activity on either stem cell factor (SCF) or flt3-ligand-induced CD34+ cell proliferation, cell production or colony formation. Blocking mAbs against the c-kit receptor have no effect on PMVEC CM-induced CD34+ cell proliferation at titres that completely suppress SCF-induced proliferation. Moreover, it is shown that this haematopoietic growth factor supports the proliferation and colony formation of murine, non-human primate, and porcine marrow progenitor cells without any apparent species-specific restrictions in its activity. These finding suggest that PMVEC CM contains a novel early haematopoietic activity.

Animals↗
PubMed1997

Varicella pneumonia in pregnancy.

Pregnant patients with varicella infections have an increased risk of life threatening pulmonary complications. Prompt diagnosis and aggressive early therapy with acyclovir are important aspects of treating varicella pneumonia. The stress of severe hypoxia usually necessitates delivery by cesarean section. Preoperative evaluation of pulmonary function and choice of anesthetic are critical to intraoperative management. This paper presents the successful treatment of varicella pneumonia in the third trimester of pregnancy with survival of both mother and infant.

Journal Article↗
PubMed1997

Mitogenic stimulation of T cells reveals differing contributions for B7-1 (CD80) and B7-2 (CD86) costimulation.

The requirement of accessory cells for concanavalin A (Con A) activation of T cells suggests delivery of a separate costimulatory signal. However, the costimulatory pathways involved have not been identified. These studies assess the role of CD28-B7-mediated costimulation during T-cell activation by Con A. The B7-1/B7-2 binding protein CTLA4-Ig inhibited the proliferative response of primary lymph node cells to either Con A or soluble anti-CD3 mAb. This suppression was dose dependent and could be reversed by CD28 cross-linking. CTLA4-Ig also completely suppressed induction of interleukin-2 (IL-2) mRNA by Con A. CTLA4-Ig-mediated suppression was not due to blockade of the Con A 'receptor(s)' or of the primary activation signal (as measured by the intracellular calcium response). Although both B7-1 and B7-2 were up-regulated following Con A activation, each played a different role in proliferation and cytokine production. Individually, anti-B7-2 Fab partially inhibited the Con A response whereas anti-B7-1 Fab had no effect. However, the combination of anti-B7-1 and anti-B7-2 Fab completely suppressed proliferation and IL-2 production. Therefore, while a part of the Con A response requires B7-2, the remainder of the response can utilize either B7-1 or B7-2. Together, these results demonstrate that Con A activation of T cells requires the delivery of a separate costimulatory signal that is mediated almost entirely by the B7 receptors.

Animals↗
PubMed1997

Colostrum enhances the nutritional stimulation of vital organ protein synthesis in neonatal pigs.

Our objective was to determine the relative importance of the macronutrient components of colostrum in the stimulation of vital organ protein synthesis in neonatal pigs. We studied colostrum-deprived newborn pigs within 4-6 h after birth (unfed) and three groups fed for 24 h mature milk, colostrum, or a formula containing a macronutrient composition comparable to that of colostrum. We measured protein synthesis in vivo using a flooding dose of 3H-phenylalanine. The fractional rates of protein synthesis (Ks) in the brain, heart, lung, kidney and spleen were significantly higher in all fed groups than in the unfed newborns. Among the three fed groups, brain and heart protein synthesis rates were greater in colostrum-fed than in either milk- or formula-fed pigs. Kidney and spleen protein synthesis rates in colostrum- and formula-fed pigs were not significantly different, but both were higher than in milk-fed pigs. The stimulation of kidney protein synthesis in response to feeding was primarily a consequence of greater protein synthetic efficiency; however, protein synthetic capacity in the heart, lung and spleen was generally greater in colostrum- and formula-fed pigs than in unfed newborns. Our results suggest that the predominant stimulus for vital organ protein synthesis in colostrum-fed neonatal pigs is nutrient intake. However, there was a specific stimulation of both brain and heart protein synthesis in colostrum-fed pigs that cannot be attributed to macronutrients.

Amino Acids↗
PubMed1997

Intrauterine growth restriction does not alter response of protein synthesis to feeding in newborn pigs.

This study aimed to determine the effect of intrauterine growth restriction (IUGR) on the acute response of tissue protein synthesis to feeding in newborn pigs. Newborn pigs of sows fed either control or protein-restricted diets throughout gestation were designated C or IUGR, respectively. Both groups were either fasted for 9 h after birth or fed hourly 30 ml colostrum/kg body wt for 2.75 h after a 6-h fast. Fractional rates of tissue protein synthesis (Ks) were measured in vivo with a flooding dose of L-[4-3H]phenylalanine. Birth weight was reduced by 33% in IUGR pigs. IUGR had no effect on Ks in skeletal muscles, heart, liver, jejunum, or pancreas. Feeding stimulated tissue Ks similarly in C and IUGR pigs. Fasting plasma insulin concentrations and their rise with feeding were unaffected by IUGR. Plasma insulin-like growth factor I (IGF-I) concentrations were reduced by 42% in IUGR pigs and were not altered by feeding in either IUGR or C pigs. There were positive nonlinear relationships between tissue Ks and circulating concentrations of insulin. The results indicate that, in newborn pigs, tissue Ks are unaffected by IUGR, despite reduced plasma IGF-I concentrations. The efficiency with which nutrients stimulate tissue Ks is also not altered by IUGR, perhaps because the rise in plasma insulin concentrations with feeding is unaffected by IUGR.

Animals↗
PubMed1997

Insulin-stimulated amino acid utilization during glucose and amino acid clamps decreases with development.

Neonatal animals utilize their dietary amino acids for protein accretion with high efficiency, and this efficiency declines during early life. The factors responsible for this developmental change are unknown. Our objectives were to determine whether amino acid (AA) utilization is stimulated by insulin in the neonate and whether this response changes during the suckling period. Two hyperinsulinemic-euglycemic clamp infusion studies, using 10-2,000 ng insulin.kg-0.66.min-1, were performed in 7- and 26-day-old pigs. In study I, no AA were provided during the infusion, and the resultant decline in plasma AA levels was defined. In study II, plasma AA were clamped at near-fasting levels, and whole body utilization of exogenous AA was determined by measuring the rate of infusion of an AA mixture necessary to maintain basal plasma lysine concentrations. In study I, the half-maximal effective dose (ED50) for the fall in AA concentrations with increasing plasma insulin concentration was lower in 7- than in 26-day-old pigs, and the nadir in AA concentration was achieved by only 20 microU/ml insulin. In study II, the utilization of exogenous AA during hyperinsulinemic-euglycemic AA clamps exhibited a higher maximum response (Rmax) (49 vs. 26 mumol AAtotal.min-1.kg-1) and a lower ED50 (18 vs. 45 microU insulin/ml) in 7- than in 26-day-old pigs. Plasma urea nitrogen concentrations did not rise with increasing insulin and AA infusion rates. These results indicate that insulin stimulates the utilization of exogenous AA in neonatal pigs and that both the insulin sensitivity and responsiveness of AA utilization decline over the suckling period. The infused AA were likely utilized for protein accretion.

Aging↗
PubMed1997

Food intake alters muscle protein gain with little effect on Na(+)-K(+)-ATPase and myosin isoforms in suckled rats.

Biochemical maturation accompanies the rapid accretion of skeletal muscle in early life. We wished to determine whether changes in muscle protein accretion, induced by variations in food intake, altered the biochemical maturation of the soleus and the extensor digitorum longus (EDL) muscles. Rat pups were suckled in litters of 4, 10, or 16 to induce differences in food intake. At 21 days of age, muscle protein and DNA were quantitated and biochemical maturation was assessed from measurement of [3H]ouabain-binding site abundance and myosin isoform composition. Differences in food intake produced a twofold range in body and muscle weights and protein and DNA contents. Protein accretion was more sensitive to nutrient intake in the soleus than in the EDL. Serum 3-5,3'-triiodothyronine (T3) and insulin concentrations decreased with a reduction in food intake. Total ouabain-binding sites were not altered in either muscle and were independent of muscle size. Differences in myosin isoform composition were more pronounced for the soleus than the EDL, but were relatively small in magnitude. These results demonstrate that, whereas postnatal muscle protein accretion and circulating hormone concentrations are sensitive to food intake, the biochemical maturation is resilient. The immature muscle does not exhibit the fiber type-specific responses to malnutrition typical of mature muscle.

Animals↗
PubMed1997

Exogenous insulin-like growth factor-I increases weight gain in intrauterine growth-retarded neonatal pigs.

Many cases of intrauterine growth retardation (IUGR) are the result of placental insufficiency, suggesting that potential therapies should focus on the neonate rather than the pregnant female. We wished to determine whether IGF-I could be used therapeutically to stimulate normal rates of growth in these neonates. Eight sows received 2.3 kg/d of either a control (13% protein) or protein-restricted (0.5% protein) diet from d 63 of pregnancy to parturition. Litters were reduced to 6 pigs at 3 d of age, and IUGR neonates were fostered onto a control sow. Three pigs/ litter received an osmotic minipump containing either saline or recombinant human IGF-I, delivered at 4 microg/h from d 3 to d 10 of age. Tissue protein synthesis was measured in all pigs using a flooding dose of [3H]phenylalanine. At birth, both body weight (10%) and circulating IGF-I concentration (30%) were significantly lower in IUGR than in control newborns. The infusion of IGF-I to IUGR neonates significantly increased the circulating concentration of IGF-I, growth rate, and protein and fat accretion to control levels. The infusion of IGF-I did not alter concentrations of insulin, glucose, IGF-II, or the thyroid hormones. Our results suggest that IGF-I may be a potential therapy to restore normal growth in IUGR infants.

Analysis of Variance↗
PubMed1997

Role of milk-borne vs endogenous insulin-like growth factor I in neonatal growth.

Neonatal pigs are characterized by a high efficiency of nutrient utilization and rapid growth rate. The utilization of dietary protein for lean tissue growth is particularly efficient in neonatal pigs and is associated with a high rate of skeletal muscle protein synthesis and deposition. In support of these high growth rates, neonatal pigs consume a milk diet that has a high biological value and is abundant in growth factors, including insulin and IGF-I. During the neonatal period, there are developmental changes in the circulating concentrations of, and tissue responsiveness to, hormones, particularly insulin, IGF-I, and growth hormone that play a central role in growth regulation. Our goal has been to characterize the dietary factors and specific aspects of endocrine function that are responsible for the anabolic stimulus that helps to sustain the high rates of protein deposition in neonatal pigs. Our results suggest that, despite the abundance of growth factors in milk and colostrum, the intake of nutrients is the primary anabolic stimulus for protein synthesis and this response declines with age. There is, however, a nonnutritive and as-yet-unidentified component in colostrum that provides a specific anabolic stimulus for skeletal muscle in newborns, but this is probably neither insulin nor IGF-I. Our studies also indicate that circulating concentration of IGF-I are not a primary stimulus of skeletal muscle protein synthesis and that the primary endocrine signal that mediates the response to nutrient intake may be insulin. Future research should address how the local expression of IGF and the function of insulin and IGF receptors affect the responsiveness of anabolic processes to nutrient intake and hence the efficiency of neonatal growth.

Animals↗
PubMed1996

Molecular cloning and characterization of CFT1, a developmentally regulated avian alpha(1,3)-fucosyltransferase gene.

Although coordinate expression of carbohydrate epitopes during development is well described, mechanisms which regulate this expression remain largely unknown. In this study we demonstrate that developing chicken B cells express the LewisX terminal oligosaccharide structure in a stage-specific manner. To examine regulation of this expression, we have cloned and expressed the chicken alpha(1,3)-fucosyltransferase gene involved in LewisX biosynthesis, naming it chicken fucosyltransferase 1 (CFT1). CFT1 is characterized by a single long open reading frame of 356 amino acids encoding a type II transmembrane glycoprotein. The domain structure and predicted amino acid sequence are highly conserved between CFT1 and mammalian FucTIV genes (52.8% and 46.3% identity to mouse and human respectively). In vitro CFT1 fucosyltransferase activity utilizes LacNAc > 3'sialyl-LacNAc acceptors with almost no utilization of other neutral type II (lactose, 2-fucosyllactose), or type I (lacto-N-biose I) acceptors. CFT1-transfected cells make cell surface LewisX (COS-7) and LewisX + VIM-2 structures (Chinese hamster ovary). CFT1 gene expression is tissue-specific and includes embryonic thymus and bursa. Furthermore, expression of the CFT1 gene and cell surface LewisX structures are closely linked during B cell development. These findings reveal the evolutionary conservation between nonmammalian and mammalian alpha(1,3)-fucosyltransferase genes and demonstrate a role for fucosyltransferase gene regulation in the developmental expression of oligosaccharide structures.

Amino Acid Sequence↗
PubMed1996

Spectrophometric assay for measuring branched-chain amino acid concentrations: application for measuring the sensitivity of protein metabolism to insulin.

Plasma amino acid concentrations fall during insulin infusion. Amino acid concentrations can be maintained using an infusion of amino acids if their plasma concentration can be determined within a few minutes. We developed a spectrophometric assay which determines the total concentration of all three branched-chain amino acids in plasma within 1 min. The enzyme leucine dehydrogenase oxidatively deaminates leucine, isoleucine, and valine, with stoichiometric reduction of NAD that is measured using a spectrophotometer. The assay was developed in both a kinetic and end-point format. For the kinetic assay the buffer conditions were formulated to obtain equivalent rates with all three amino acids so that it could be used in samples containing unknown mixtures. For the end-point assay additional enzyme was added so that an end-point could be reached within 1 min. The application of the kinetic assay for "clamping" the branched-chain amino acids during hyperinsulinemic euglycemic clamps in humans is demonstrated.

Amino Acid Oxidoreductases↗
PubMed1996

CTLA-4 blockade enhances clinical disease and cytokine production during experimental allergic encephalomyelitis.

The B7 family of cell surface molecules expressed on APC provides accessory signals to T cells via either CD28 or CTLA-4. However, while CD28 transduces a costimulatory signal that is required for an optimal immune response, CTLA-4 transmits a negative signal. These studies use an anti-CTLA-4 mAb to directly address the role of this T cell surface molecule in experimental allergic encephalomyelitis (EAE). CTLA-4 regulation of disease was assessed during initial immune cell interactions and during the effector stage of the encephalitogenic immune response. The effects of anti-CTLA-4 treatment were schedule dependent. CTLA-4 blockade during the onset of clinical symptoms markedly exacerbated disease, enhancing mortality. Disease exacerbation was associated with enhanced production of the encephalitogenic cytokines TNF-alpha, IFN-gamma and IL-2. Hence, CTLA-4 regulates the intensity of the autoimmune response in EAE, attenuating inflammatory cytokine production and clinical disease manifestations.

Abatacept↗