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K A Wilkinson

Publications and source records attributed to K A Wilkinson.

28 records · Page 2Linked to original sources
PubMed1998

Peptide-specific T cell response to Mycobacterium tuberculosis: clinical spectrum, compartmentalization, and effect of chemotherapy.

The T cell repertoire of 59 patients with untreated tuberculosis was compared with that of 46 bacille Calmette-Guérin-vaccinated controls by assaying the proliferative responses to six permissively recognized peptides from the 16-, 19-, and 38-kDa molecules of Mycobacterium tuberculosis. A trend from higher to lower reactivity following this order: vaccinated controls > lymph node disease > localized extrapulmonary > pulmonary > pleural was seen for 4 of the peptides (P < .03). The decreased response of blood lymphocytes from patients with pleural tuberculosis was partially accounted for by sequestration of peptide-responsive cells within the pleural fluid. Chemotherapy "reversed" the depressed proliferative responses of patients with pulmonary and pleural tuberculosis depending on the peptide origin, being greatest for peptides of 16 kDa, transient for those of 19 kDa, and least for those of 38 kDa. These data demonstrate antigen specificity in the decreased responsiveness of patients with tuberculosis.

Adolescent↗
PubMed1997

Modulation of peptide specific T cell responses by non-native flanking regions.

The deduced core (75RYPNVTI81) from a T-cell stimulatory epitope of the 38 kDa protein of M. tuberculosis was studied to identify the structural elements required for the creation of a synthetic peptide antigen from an epitope core, which alone was not capable of inducing CD4+ T-cell responses. Peptides were prepared with extensions composed of native and/or non-native sequences to clarify the role of the flanking regions adjacent to the epitope core. Their binding to isolated H-2-Ab MHC glycoprotein as well as T-cell stimulatory capacity were assayed using a specific murine hybridoma T-cell line [38.H6], lymph node cells from the native 20-mer peptide primed C57BL/10 mice and human PBMCs from sensitised individuals. Elongation of the epitope core by four alanines at both N- and C-terminals resulted in a 15-mer peptide A4-75-81-A4 which was stimulatory for hybridoma T-cells and showed a small decrease in H-2-Ab binding. Substitution of one Ala by Ser in the N-terminal flank had pronounced effect and peptide A2SA-75-81-A4 proved to be more effective than the native 20-mer sequence in the hybridoma as well as in the LN cell proliferation assays. The binding of this peptide and that of the native one were similar. Testing in human PBMC cultures from eight PPD positive individuals showed that in 50% of the donors' cells responded to the 'artificial' A2SA-75-81-A4 peptide. These results suggest that it is possible to construct simple, synthetic CD4+ T-cell stimulatory peptides of high potency from a non-stimulatory, 'silent' epitope core by addition of flanking residues not part of the native sequence.

Amino Acid Sequence↗
PubMed1995

Gastric emptying in patients the day after cardiac surgery.

Fourteen adults underwent cardiac surgery with a standard anaesthetic technique. Prior to surgery and the day after surgery, gastric emptying was determined using the paracetamol absorption technique. Results from 13 patients were available for analysis. The mean time to reach the maximum plasma concentration was 14.1 min (SEM 2.1) pre-operatively and 225.4 min (SEM 42.3) postoperatively. The mean maximum concentration was 23.7 mg.l-1 (SEM 1.9) pre-operatively and 5.1 mg.l-1 (SEM 0.8) postoperatively. The area under the curve (0-60 min) was 892 mg min.l-1 (SEM 57) pre-operatively and 131 mg min.l-1 (SEM 25) postoperatively. The differences between pre- and postoperative values were highly significant for all these three measurements. We conclude that gastric emptying is markedly delayed the day after elective cardiac surgery.

Acetaminophen↗
PubMed1991

Assessment of a hygroscopic heat and moisture exchanger for paediatric use.

A laboratory study of a widely available heat and moisture exchanger marketed for paediatric use was undertaken. The deadspace, measured by volume displacement, was 12 ml, similar to that of a standard catheter mount for paediatric use. Pressure drop across the device was measured at several different flows in five samples of the device in both the dry and wet state. Calculated resistance proved to be markedly lower when compared with that of other anaesthetic equipment such as tracheal tubes, and with similar humidification devices for paediatric use.

Anesthesiology↗
PubMedDate not supplied

Determination of paracetamol (acetaminophen) in blood and plasma using high performance liquid chromatography with dual electrode coulometric quantification in the redox mode.

A simple, but sensitive, micro-assay for paracetamol in blood and plasma is described. Samples were prepared by precipitation with trichloroacetic assay solution and the drug quantified by reversed phase liquid chromatography with dual electrode electrochemical detection in the redox mode (oxidation at +0.25 V followed by reduction at -0.15 V). The limit of detection was 0.1 mg/L for a 10 microL sample of blood or plasma, the coefficient of variation at this concentration being 11.4%. The method was applied to a study of gastric emptying in patients before and after cardiac surgery.

Acetaminophen↗
PubMedDate not supplied

Synthesis and in vitro T-cell immunogenicity of conjugates with dual specificity: attachment of epitope peptides of 16 and 38 kDa proteins from Mycobacterium tuberculosis to branched polypeptide.

T-cell epitope containing peptides covalently attached to macromolecular carriers can be considered as synthetic immunogens for the development of skin-test diagnostics and of vaccines. As a carrier, an amphoteric branched chain polypeptide, poly[Lys-(Glui-DL-Alam)] (EAK) with poly(L-lysine) backbone has been used. This polypeptide with free alpha-amino and gamma-carboxyl groups at the end of the side chains was conjugated with peptides representing two immunodominant regions of the 16 and 38 kDa proteins of Mycobacterium tuberculosis, respectively. Peptide C91SEFAYGSFVRTVSLPVGADE110 was elongated by Cys at the N-terminal and attached to the carrier containing protected SH groups to form disulfide bridges. Peptide 65FNLWGPAFHERYPNVTITA83 was conjugated to the 3-(2-pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP) modified and acetylated EAK by introducing amide bond between the free alpha-amino group of peptide and the free gamma-COOH group of Glu at the terminal position of the branches. This strategy lead to chemically well-defined synthetic immunogens that contain two different epitopes in multiple copies covalently linked to a synthetic branched polypeptide carrier. In vitro T-cell immunogenicity of a prototype conjugate was studied using T-cell hybridomas, lymph node cells from 38 kDa protein immunized mice, and human peripheral blood mononuclear cell (PBMC) cultures from sensitized individuals. These data document that the specific T-cell stimulatory effect of each mycobacterial epitope was maintained in this conjugate. Taken together, these findings suggest that it is feasible to use a biodegradable polymeric polypeptide for producing macromolecular bioconjugates for the stimulation of T-cell responses.

Amino Acid Sequence↗