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Biomedical subjects

C Bird

Publications and source records attributed to C Bird.

At least 55 records · Page 3Linked to original sources
PubMed1991

Homeostatic action of interleukin-4 on endogenous and recombinant interleukin-2-induced activated killer cell function.

Cytokine-secreting, major histocompatibility complex-unrestricted activated killer (AK) cells are toxic to a wide range of virus-infected or malignant target cells and may be generated endogenously, eg, after bone marrow transplantation, or by infusion of cytokines such as recombinant interleukin-2 (rIL-2). Although AK cells secrete cytokines such as gamma-interferon and tumor necrosis factor, which are themselves able to recruit fresh cytokine-secreting AK cells, activation in both settings is short-lived, implying the existence of homeostatic regulatory mechanisms. We now demonstrate one mechanism by which rapid homeostasis is achieved. We show that IL-4 is produced in patients with both endogenously and exogenously generated AK cells. The cytokine was detected in serum after marrow transplantation, and IL-4 transcripts appeared in circulating lymphocytes during rIL-2 infusion. Although IL-4 inhibited the induction phase of AK cell function, it had no significant inhibitory effect on the ability of AK cells from these individuals to respond to restimulation. Nonetheless, neutralization of the IL-4 induced during cell activation doubled the half-life of AK function, once activating stimuli were removed, from 18 to 44 hours and produced a 2-log increase in AK cell secretion of tumor necrosis factor and gamma-interferon. These data suggest that IL-4 induced in vivo during lymphocyte activation abbreviates AK cell responses once the triggering stimuli have been removed. Neutralization of endogenous IL-4 in vivo by appropriate monoclonal antibodies might prolong the duration of AK function.

Bone Marrow Transplantation↗
PubMed1991

Development of immunoassays for human interleukin 3 and interleukin 4, some of which discriminate between different recombinant DNA-derived molecules.

Two panels of hybridomas were produced that secreted monoclonal antibodies (MAbs) against recombinant DNA-derived human interleukin 3 and interleukin 4 (rhIL-3 and rhIL-4). From each panel, sensitive immunoradiometric assays (IRMAs) were developed which were capable of detecting the recombinant molecule used as the immunogen but were unable to recognize natural or other recombinant forms of the same cytokine. Subsequent studies using the MAbs from each panel showed that a number of the MAbs appeared only to recognize that particular recombinant molecule used as immunogen, with little or no binding to other recombinant forms of the molecule. By using MAbs that were found to be unrestricted in their recognition for different recombinant forms of the cytokines, it was possible to develop an IRMA for IL-4 that was capable of detecting natural IL-4 as well as all the recombinant forms equally. An IRMA was also developed for IL-3 but was not of equivalent sensitivity in detecting the different recombinant forms of IL-3 used in the study. The recombinant DNA-derived cytokine molecules used to raise the two panels of MAbs contained amino acid substitutions relative to the natural sequences, and these findings indicate that caution should be exercised when using immunoassays to estimate natural sequence molecules if antibodies raised to modified rDNA-derived molecules are used.

Animals↗
PubMed1991

Cytokines in sera from insulin-dependent diabetic patients at diagnosis.

Cytokines are known to play an important role in autoimmunity and have been suggested to be involved in the pathogenesis of insulin-dependent diabetes (IDDM). In the present study we have measured IL-1, IL-2, IL-4, IL-6, interferon-gamma (IFN-gamma) and tumour necrosis factor (TNF) (using both immunoassays and bioassays) in sera from 50 patients affected by IDDM at the time of clinical diagnosis and 51 age and sex matched controls. Detectable levels of IL-1, IL-2, IL-6 and IFN-gamma were found in the serum of a small percentage of subjects and were not significantly different between patients and controls. IL-4 was detectable in a higher number of both patients and controls and circulating TNF-alpha (greater than 1 U/ml) was found in a percentage of patients (24%) significantly higher than controls (P less than 0.01). Raised levels of TNF-alpha were detectable using an immunoenzymatic assay whereas TNF bioactivity in these samples was negligible. We conclude that the presence of immunoreactive TNF-alpha in the patient's sera may reflect an increased localized production of this cytokine at pancreatic level. However, the measurement in serum of other cytokines does not add information on the role that they may play in the pathogenesis of IDDM.

Cytokines↗
PubMed1990

Expression of human immunodeficiency virus 1 (HIV-1) envelope gene products transcribed from a heterologous promoter. Kinetics of HIV-1 envelope processing in transfected cells.

The expression of human immunodeficiency virus 1 (HIV-1) envelope glycoprotein products was studied in cells transfected with env gene constructs transcribed from an SV40 promoter. Gene constructs possessing the complete tat, rev (tat+ rev+) and env genes were transiently expressed in COS-1 cells as precursor SU-TM (gp160), SU.TM (gp120 x 41), and nucleolar rev protein. In addition, envelope glycoprotein was detected on the surface of those transfected COS-1 cells expressing abundant levels of env protein. Transfected constructs possessing a mutated tat translational initiation codon (tat-rev+) were expressed in COS-1 cells with at least a 10-fold increase in the level of envelope glycoprotein expression compared to the analogous constructs with an intact tat AUG codon (tat+ rev+). Mutation of the rev initiation codon (tat+ rev-) and (tat-rev-) resulted in no detectable expression of env products but expression of these proteins could be rescued by co-transfection of a cDNA encoding the rev gene. Subgenomic tat/rev transcripts were detected following transfection of all of the gene constructs indicating splicing of the env mRNAs transcribed from a heterologous promoter. Unspliced env transcripts were only detected in the cytoplasm of cells transfected with (rev+) constructs or with the (tat- rev-) construct in the presence of the rev cDNA supplied in trans. In contrast, unspliced transcripts were detected in the nuclear and total cellular RNA of all transfected cells. Expression of rev protein was localized to the nucleolus of transfected COS-1 cells. These results indicate that the export of unspliced env mRNA to the cytoplasm is facilitated by the expression of rev. Following env synthesis, the conversion of SU-TM (gp160) to SU.TM (gp120 x 41) was not quantitative. After a 20-h pulse-chase, only 40% of the SU-TM (gp160) present at the start of the chase period was subsequently accountable as mature SU (gp120), and approximately 30% of the detectable SU (gp120) was found in the culture medium of transfected COS-1 cells. The findings indicate that the surface expression of SU.TM (gp120 x 41) derived from heterologous gene transcripts is modulated (i) the co-expression of rev, (ii) the efficiency of proteolytic processing of SU-TM (gp160), and (iii) the degree of SU (gp120) shedding and/or secretion from the cell.

Animals↗
PubMed1990

The effects of foreign transmembrane domains on the biosynthesis of the influenza virus hemagglutinin.

Eleven chimeric proteins were created in which the transmembrane, the cytoplasmic, or both topological domains of the influenza virus hemagglutinin (HA) were replaced with those from five other glycoproteins. All of the chimeric HAs reached the cell surface but appeared to differ in the degree to which they were stably folded. Comparisons of the rates of folding, passage into the Golgi, and arrival at the plasma membrane of wild-type HA and the chimeric proteins suggest that formation of a stable HA trimer is not an absolute requirement for export from the endoplasmic reticulum. In addition, there appear to be at least two steps at which the rate of transport can be altered during exocytosis, one occurring before and the other after the trimming of oligosaccharides by Golgi mannosidases. Certain of the chimeras differed from HA in their ability to pass through each of these steps. Replacement of the HA transmembrane domain with the analogous sequences from other proteins affected folding and transport of the chimeric HAs in ways that suggest that the HA transmembrane sequences form a specific structure in the membrane that differs from that formed by analogous sequences from the other proteins.

Amino Acid Sequence↗
PubMed1990

Effects of stress and anxiety on the processing of threat stimuli.

Two experiments extended the work of MacLeod and Mathews (1988) and examined whether a cognitive bias for threat information is a function of state or trait anxiety. Color-naming and attention deployment tasks were used to assess the effects of a stress manipulation procedure on attentional responses in high and low trait anxious subjects. Subjects under high stress selectively allocated processing resources toward threat stimuli, irrespective of their trait anxiety level. There was no consistent evidence of a cognitive bias associated with trait anxiety, and the effect of the stress manipulation did not appear to be mediated by state anxiety. It was suggested that trait factors do not modify attentional biases associated with acute stress, but may influence such biases when stress is prolonged.

Adult↗
PubMed1990

Potent inflammatory properties in human skin of interleukin-1 alpha-like material isolated from normal skin.

Recombinant human interleukin 1 (IL-1) possesses potent inflammatory properties in both animal and human skin. However, IL-1-like material has been isolated from normal epidermal samples. In view of the uncertainty concerning the structure and biologic properties of human epidermal IL-1, heel stratum corneum, and chamber fluid samples from normal skin have been purified by successive reversed phase and anion exchange high-performance liquid chromatography (HPLC), and aliquots of each HPLC fraction tested for IL-1 activity in an EL-4 NOB-1 assay and for inflammatory activity by intradermal injection of autologous material. The results consistently indicated the presence of inflammatory quantities of IL-1 alpha-like material, which induced persistent erythema lasting at least 24 h, associated in 4-h biopsies with mixed dermal leukocyte infiltrates containing increased numbers of neutrophils, monocytes, and T helper cells. Biologically active quantities of IL-1 beta were not recovered following HPLC purification in most experiments. Analysis of heel stratum corneum extract also showed that the majority of the dilution-related IL-1 activity detected in the EL-4 NOB-1 bioassay was neutralized by IL-1 alpha but not IL-1 beta antiserum. These findings suggest that IL-1 alpha-like material is present in inflammatory amounts in normal human epidermis, and its release may represent a primary inflammatory mechanism in human skin.

Adult↗
PubMed1988

Identification and sequence determination of a cDNA clone for tomato pectin esterase.

Cell wall softening during tomato fruit ripening is brought about through the action of a number of pectolytic enzymes. We have reported previously the cloning and characterisation of the cDNA for the major cell wall softening (degrading) enzyme, polygalacturonase [Grierson, D., Tucker, G. A., Keen, J., Ray, J., Bird, C. R. and Schuch, W. (1986) Nucleic Acids Res. 14, 8595-8603]. We have now isolated a cDNA clone for tomato pectin esterase, an enzyme also implicated in cell wall softening. Here we report the structure of this cDNA and compare it with the structure of pectin esterase derived from amino acid sequence experiments [Markovic, O. and Jornvall, H. (1986) Eur. J. Biochem. 158, 455-462]. We have used the pectin esterase cDNA clone to analyse pectin esterase gene expression during development and ripening of normal and mutant fruit.

Amino Acid Sequence↗
PubMed1988

Clinical validation of an automated thin-film reflectance method for measurement of magnesium in serum and urine.

We evaluated an automated method for measuring total magnesium in serum, plasma, or urine that utilizes dry multilayered thin-film reagents and reflectance photometry for use on the Kodak Ektachem 400 analyzer. Comparison of magnesium results by this method and by atomic absorption revealed the following: the results on samples from a variety of patients agreed closely, the thin-film methodology was not affected by icteric, hemolyzed, or lipemic samples, and over 100 drugs studied did not interfere. Only very high concentrations of citrate and calcium affected magnesium results by this method. Both the accuracy and versatility of the thin-film method are well suited to routine, fast response, and pediatric testing in the clinical laboratory.

Autoanalysis↗
PubMed1987

The primary and secondary cellular immune responses to whole cell Bordetella pertussis vaccine and its components.

The cellular immune responses of Balb/c mice and Wistar rats immunized in hind footpads with intact killed Bordetella pertussis were found to differ from those of similar animals immunized with other bacteria including Bordetella bronchiseptica, Salmonella typhimurium and Escherichia coli. All the bacteria stimulated increases in cell number, proliferation and interleukin 2 (IL-2) production in popliteal lymph nodes which peaked 3-5 days after injection and decreased to resting levels by day 7. However, B. pertussis also caused a second peak in all three parameters at 11 days after immunization. This peak was not seen following injection with any of the other bacteria. Bordetella pertussis also caused systemic effects, increased cellular proliferation in bone marrow and thymus, with similar biphasic kinetics. It possesses a potent toxin, distinguishing it from the closely related B. bronchiseptica. The use of purified materials confirmed that the presence of this pertussis toxin (PT) was responsible for the later peak in stimulation, whereas lipopolysaccharide (LPS) in combination with PT and also the filamentous haemagglutinin (FHA) could mimic the early peak of stimulation. Primary immunization with B. pertussis was also shown to generate lymph node cells which responded in vitro to secondary challenge with B. pertussis cells, FHA or PT. Both proliferation and IL-2 production were enhanced, except with FHA which only increased IL-2 production. Lymph node cells from mice immunized with E. coli showed no such responses.

Animals↗
PubMed1987

The use of monoclonal antibodies raised against a human IgE myeloma paraprotein for the study of allergen extracts and sera from allergic patients.

Murine monoclonal antibodies have been produced against a single human IgE myeloma preparation. A single fusion yielded six different monoclonal antibodies which bound strongly to the immunogen. Two of these failed to react with 'normal' human IgE or with a second IgE paraprotein. The other four antibodies reacted with normal and myeloma-derived IgE and were found to be specific for antibodies of this class. Two of these antibodies were potent reagents for the detection of IgE by two site immunoradiometric assay, immunoblotting, and radioallergosorbent test. The other two antibodies were considerably less potent reagents when used in these assay systems. Our findings suggest that care should be taken in selection of suitable monoclonal antibodies for immunochemical study of allergens and sera from allergic patients.

Allergens↗
PubMed1986

Immunochemical characterization of a new platelet specific monoclonal antibody and its use to demonstrate the cytoskeletal association of the platelet glycoprotein IIb-IIIa complex.

We describe the production and characterization of a monoclonal antibody specific for platelets. This antibody reacts strongly with human and primate platelets, but does not recognise human monocytes, polymorphonuclear leucocytes, lymphocytes, erythrocytes, leukaemic nor fibroblast cell lines, nor rodent platelets. Immunoprecipitation studies using radiolabelled platelet membrane proteins showed that the monoclonal antibody binds to the platelet membrane glycoprotein IIb-IIIa complex. Affinity chromatography using immobilized monoclonal antibody allows purification of the antigen, but also co-purifies the cytoskeletal proteins actin and myosin. Our results demonstrate immunochemically that although the GP IIb-IIIa complex is an external structure, it is connected through the cell membrane to the microfilament system.

Actins↗
PubMed1986

Aqueous and serum-based materials compared for use as simulated calibrators for three ionized calcium analyzers.

To determine if bias between different ionized calcium analyzers could be decreased, we analyzed 10 control fluids during a study in which ionized calcium was measured in more than 150 serum and whole-blood samples. After calibrating three ionized calcium analyzers (Radiometer ICA 1, Nova 8, and AVL 980) with the manufacturers' respective calibrators, we used the between-instrument differences of the control fluids to simulate recalibration of the analyzers during each analytical run. A filtered human serum pool containing ionized calcium at 1 mmol/L concentration, with CO2 removed and having no added buffer, was the only material that consistently decreased between-analyzer bias of both serum and whole blood. Another human serum pool containing about 1.3 mmol of ionized calcium and about 10 mmol of bicarbonate per liter was even better at minimizing analyzer biases for serum samples, but was not as effective for whole-blood samples. Some additives used to buffer pH apparently adversely affected both the accuracy and precision of some, but not other, calcium ion electrodes. We conclude that if a reference material is developed for calibration of ionized calcium analyzers, it should be tested on several analyzers for use with both serum and whole blood, and it should be at least as effective as a human serum material, such as that used here.

Calcium↗
PubMed1985

Human B cell proliferation is stimulated by interleukin 2.

The proliferation of human B cells was studied for response to interleukin 2 (IL-2) produced in Escherichia coli using recombinant DNA technology. The IL-2 was found to be an homogenous preparation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using the anti-IL-2 monoclonal antibody DMS-1. IL-2 was found to stimulate B cell proliferation. Activation of the B cells using anti-IgM antibodies increased this response. Resting T cells from the same donors were found to be less reactive to IL-2. The results suggest that human B cell proliferation can be stimulated by IL-2 alone.

Antibodies, Anti-Idiotypic↗
PubMed1984

Anti-androgens do not alter androgen-dependent characteristics of acid phosphatase in the rat ventral prostate.

When administered to intact adult male rats, cyproterone acetate (10 mg/day), flutamide (15 mg/day), or Compound I (1 mg/day) caused a significant decrease in the organ weight to body weight ratios, with a concomitant rise in the specific activity of prostatic acid phosphatase. These compounds do not affect two other markers of androgen activity in the prostate. Neither the percentage inhibition of acid phosphatase activity by tartrate nor the appearance of the secretory band of acid phosphatase on polyacrylamide gels was altered by the administration of anti-androgens. When administered to castrated rats given doses of 5 alpha-dihydrotestosterone (250-750 micrograms/day), flutamide (15 mg/day) was unable to alter the percentage inhibition of acid phosphatase activity by tartrate, or the pattern of activity on polyacrylamide gels. These results suggest that these anti-androgens affect only some of the androgen-dependent functions of the prostate.

Acid Phosphatase↗
PubMed1981

Spleen scanning with 99Tcm-labelled red blood cells after splenectomy.

In order to correlate the haematological changes which occur after splenectomy, with the presence or absence of residual splenic tissue, spleen scans using 99Tcm-labelled red blood cells were performed in 36 patients who had had a splenectomy. Positive spleen scans were found in 44 per cent (8 out of 18) of patients who had undergone splenectomy for trauma and in 17 per cent (3 out of 18) of patients who had undergone elective splenectomy. No relationship was found between the presence of Howell-Jolly bodies, platelet counts, the levels of IgG, IgM and IgA and the scan result. It is concluded that these findings are due to the presence of splenunculi, whose incidence is more common than the 12 per cent usually quoted.

Adolescent↗