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Biomedical subjects

F Gotch

Publications and source records attributed to F Gotch.

At least 73 records · Page 4Linked to original sources
PubMed1993

HIV-specific cytotoxic T-cell activity in an HIV-exposed but uninfected infant.

The factors necessary for protective immunity against HIV-1 are unknown. Important information about these factors should come from study of people at high risk of HIV infection who have not apparently become infected. Among these are the estimated 60-85% of children who may be exposed in utero or perinatally to HIV-1 but do not become infected. We observed the transient appearance of HIV-specific cytotoxic T-lymphocyte (CTL) activity in a baby born to HIV-1-infected parents, in whom all standard markers of infection remained negative. These findings suggest that HIV-specific CTLs may be a marker for recently exposed, but uninfected, individuals.

CD4-CD8 Ratio↗
PubMed1993

A sequence pattern for peptides presented to cytotoxic T lymphocytes by HLA B8 revealed by analysis of epitopes and eluted peptides.

HLA B8-restricted cytotoxic T lymphocytes (CTL) specific for influenza A virus were generated and shown to recognize the nucleoprotein (NP). The dominant epitope was mapped using recombinant vaccinia viruses that expressed fragments of the NP and then synthetic peptides based on the NP amino acid sequence. The peptide 380-393 was first identified and further refined; it was shown that the glutamic acid at position 380 was essential for recognition by CTL and that the nonamer 380-388 was the optimum peptide. Six HLA B8-positive influenza immune donors that we have tested respond to this peptide as part of their influenza-specific CTL response. The amino acid sequence of the peptide epitope was compared to six other known virus peptides known to be restricted by HLA B8 and a sequence homology was identified, which predicted nonamer and octamer epitope sequences. Probable anchor residues were identified at peptide residues 3 (lysine/arginine), 5 (lysine/arginine) and 9 (leucine/isoleucine). Support for this pattern came from sequencing peptides eluted from purified HLA B8 molecules, where lysines were predominant at positions 3 and 5. One of the predicted epitope peptides was made and shown to be recognized by specific CTL. These and the two others were shown to compete with NP 380-388 for binding to HLA B8. A model was made of the HLA B8 molecule and negatively charged pockets predicted, which could accommodate the positively charged side chains of the peptide anchor residues.

Amino Acid Sequence↗
PubMed1993

CAPD prescription in current clinical practice.

A total of 132 patients from 15 dialysis centers were studied. Analyses were made of each drained dialysate exchange over 24 hours to determine total peritoneal urea clearance (KpT, liters/day), and a 24-hour urine was collected to determine total renal urea clearance (KrT, liters/day) and the sum of KpT+KrT or KprT, liters/day. Body water volume (V, L) was estimated from gender and surface area, and daily fractional urea clearance (KprT/V) was calculated. Normalized protein catabolic rate (PCRN, grams/kilogram/day) was also calculated from the urea data. Major results were the following: KrT comprised 25% of KprT; the mean KprT/V was 0.28, but ranged from 0.10-0.50; an equivalent thrice-weekly hemodialysis KT/V was calculated from the KprT/V values and showed mean KT/V = 1.07, but 67% of values were less than 1.0. In contrast, the Health Care Finance Administration (HCFA) consensus criteria indicated 91% of prescriptions were adequate. These data indicate the need for clinical outcome studies with KprT/V randomized over the range 0.20-0.30 to better define the domain of adequate CAPD.

Creatinine↗
PubMed1993

The measured creatinine generation rate in CAPD suggests only 78% of prescribed dialysis is delivered.

The creatinine generation was measured (GCr) in 132 continuous ambulatory peritoneal dialysis (CAPD) patients from the product of total daily peritoneal plus renal creatinine clearance and plasma creatinine (CpCr) plus the product of estimated gut clearance of 0.036 L/kg/day and CpCr. The creatinine generation was also predicted (PGCr) from gender, age, and weight. The mean ratio of MGCr/PGCr was 1.29. Analysis of these data suggests that on average, only 78% of the prescribed dialysis is actually delivered in CAPD patients. Further studies are suggested for the verification of the assumed gut creatinine clearance.

Body Weight↗
PubMed1992

Identification of the nonamer peptide from influenza A matrix protein and the role of pockets of HLA-A2 in its recognition by cytotoxic T lymphocytes.

Influenza matrix peptide 58-66 is shown to be the optimal nonamer for binding to HLA-A2 and presentation to cytotoxic T lymphocytes (CTL). If titered out to 2 x 10(-10) - 4 x 10(-10) M in CTL-mediated lysis assays and to 3 x 10(-9) M in an HLA-A2 assembly-stabilization assay in cell lysates. The peptide was shown to make probable contacts with its carboxy terminus close to residue 116 in the floor of the cleft of HLA-A2, close to the F pocket. The side chain of the amino-terminal amino acid was unimportant, but its free amino and carbonyl groups in the A pocket appeared important in optimizing peptide presentation. The B pocket probably accommodates the side chain of residue 2 (isoleucine) and was shown to be critical in peptide presentation.

Amino Acid Sequence↗
PubMed1992

Enhanced peritoneal dialysis delivery with PD-PLUS.

We prospectively studied the effects of enhanced continuous ambulatory peritoneal dialysis (CAPD) on the normalized protein catabolic rate (NPCR) and Kprt/V of two patients with zero residual renal function and marginal or below minimal HCFA values on urea kinetic modeling (Kprt/V = 0.21). Predictable increases in NPCR (from 0.61 to 0.76 and from 0.73 to 0.81 gm/kg/day, respectively) were seen after two weeks of enhanced CAPD achieved by the addition of a fifth nighttime exchange by a portable, easily operable, automated device (PD-PLUS).

Creatinine↗
PubMed1991

Kinetics and quality assurance: prescription therapy through kinetic modeling.

With recent startling statistics showing an increase in mortality risk of dialysis patients in the United States compared to other countries, managers and practitioners have been motivated to carefully examine not only the dialysis prescription, but what is actually delivered to the patient. In October 1990, prior to ANNA's Clinical Concerns in Nephrology meetings in Cincinatti and San Antonio, attendants were invited to participate in an education session examining the monitoring and evaluation of prescription therapy for peritoneal and hemodialysis. The title of the session was "The Nurse's Role in Prescription Therapy--What Does it Really Mean?" The sponsor was Fresenius USA, Inc. This educational offering gave the nephrology nurse the opportunity to learn how to manage the delivery of care based on the theory of kinetic modeling for peritoneal and hemodialysis therapy. Quality assurance indicators, data collection tools, and the latest technology were described for the monitoring and evaluation of the adequacy of therapy. The following article is adopted from the original transcripts of the five-member panel of experts who participated in the session. We appreciate the willingness of the speakers to share this timely and important material with the ANNA Journal readers.

Data Collection↗
PubMed1990

Presentation of viral antigen controlled by a gene in the major histocompatibility complex.

We describe a mutant human cell line (LBL 721.174) that has lost a function required for presentation of intracellular viral antigens with class I molecules of the major histocompatibility complex (MHC), but retains the capacity to present defined epitopes as extracellular peptides. The cell also has a defect in the assembly and expression of class I MHC molecules, which we show can be restored by exposure of the cells to a peptide epitope. This phenotype suggests a defect in the association of intracellular antigen with class I molecules similar to that described for the murine mutant RMA-S (ref. 5), but in the present case the genetic defect can be mapped within the MHC locus on human chromosome 6.

Antigen-Presenting Cells↗
PubMed1989

Anti-HLA-A2 antibody-enhancement of peptide association with HLA-A2 as detected by cytotoxic T lymphocytes.

Most cytotoxic T lymphocytes (CTL) not only recognize epitopes of viral or other foreign proteins in association with class I major histocompatibility complex (MHC) molecules, but also recognize target cells sensitized with short synthetic peptides representing the epitopes. There is increasing evidence that these synthetic peptides associate with the class I molecule both at the cell surface and intracellularly. We have now investigated the effect of a monoclonal antibody specific for HLA-A2 and HLA-B17 (B57/58) molecules (antibody MA2.1)3 on the sensitization of target cells with peptide for lysis by HLA-A2-restricted CTL. Previously, anti-HLA class I monoclonal antibodies have been shown to inhibit the recognition of target cells, infected with influenza A virus, by virus-specific CTL. We find, however, that target cells treated with MA2.1 antibody can be sensitized with peptide for CTL lysis much more rapidly than untreated cells, or at greater than 100-fold lower peptide concentration than that required for sensitization of untreated cells. This implies that the antibody, which is believed to bind to one side of the peptide-binding groove, directly affects the binding of peptide to the HLA-A2 molecule at the cell surface.

Amino Acid Sequence↗
PubMed1989

Recognition of influenza virus proteins by cytotoxic T lymphocytes.

Recombinant DNA techniques have been used to express the proteins of influenza virus individually. Target cells expressing single viral proteins were then used to identify the molecules recognized by cytotoxic T lymphocytes (CTLS). Results have shown that, contrary to expectation, the majority of the proteins recognized by class I major histocompatibility complex-restricted CTLS are not transmembrane glycoproteins. Experiments with deletion mutants of the nucleoprotein (NP) gene showed that transport of epitopes to the membrane for recognition by CTLS was independent of a definable signal sequence. In addition, the epitopes recognized were contained within short linear sequences of amino acids, and rapid degradation of large NP fragments within the target cell did not prevent recognition by CTLS. These results led to the suggestion that the epitopes recognized by class-I-restricted CTLS resulted from degradation of viral proteins. If so, the epitopes should, like those for class-II-restricted T cells, be replaceable in vitro with short synthetic peptides. Five different epitopes of NP have now been demonstrated that can be defined with short peptides in vitro. Each peptide is recognized with a specific class I molecule (Db, Kk, Kd and HLA B37). This has been extended to the influenza matrix protein, and a peptide epitope defined that is recognized by human CTLS in association with HLA-A2. The question arose as to whether a similar phenomenon would be found with viral proteins which are naturally inserted in the target cell membrane. A mutant haemagglutinin has been produced that lacks a hydrophobic signal sequence. This protein is expressed as a short-lived, unglycosylated, intracellular protein. However, target cells expressing this molecule were recognized efficiently by CTLS raised to the wild-type haemagglutinin and vice versa. These and more recent results with non-viral glycoproteins are consistent with the existence of a mechanism for degrading viral (and perhaps host) proteins and exposing them at the cell surface for recognition by cytotoxic T cells in association with class I molecules of the major histocompatibility complex.

Animals↗
PubMed1988

Recognition of influenza A matrix protein by HLA-A2-restricted cytotoxic T lymphocytes. Use of analogues to orientate the matrix peptide in the HLA-A2 binding site.

CTL specific for the influenza A virus matrix peptide 57-68 and restricted by HLA-A2 were studied. Their ability to recognize a set of analogue peptides, each of which differed from the natural peptide by a single amino acid, was analyzed. This revealed a core of five amino acids, 61-65, where one or more changes completely abrogated recognition. The glycine at position 61 was the only residue where no substitution was tolerated. Analogue peptides that did not induce CTL-mediated lysis were tested as competitors with the natural peptide; those with substitutions at positions 60, 64, and 65 inhibited, identifying residues that interact with the TCR. Another approach was to test a set of four CTL clones on all of the analogues. Marked differences in recognition by individual CTL clones were observed for several substituted peptides. The data indicate that most of the analogues bind to HLA-A2 with possible differences in fine positioning of the peptide. An alpha helical orientation for the peptide is discussed.

Binding Sites↗
PubMed1987

Lymphocyte proliferation and cytotoxic assays using flat-bed scintillation counting.

Lymphocyte 51Cr release and [3H]thymidine uptake assays were evaluated with respect to measurement of sample radioactivity using the flat-bed scintillation counter. 51Cr lysates were spotted onto a glass fibre filter sheet while [3H]thymidine-labelled cells were filtered onto a similar sheet using a cell harvester. The 96 samples were rapidly processed for counting, without removal of individual sample areas. Either form of preparation showed good linearity of count rate with the quantity of material on the filter. Reproducibility was good; the coefficient of variation for 96 samples being within 5%. The low background and high efficiency of this counter results in increased assay sensitivity and allows considerable economies in materials to be made. A commercial version of the counter has six counting heads permitting a high rate of sample throughput.

Beta Particles↗
PubMed1987

Identification of viral molecules recognized by influenza-specific human cytotoxic T lymphocytes.

Human cytotoxic T cells specific for influenza A virus were tested for recognition of each of the ten influenza A virus proteins expressed in target cells using recombinant vaccinia viruses. They recognized the matrix M1, polymerase PB2, and nucleoproteins of influenza virus in association with MHC class I antigens. These internal viral proteins were seen by CTL in conjunction with one or more of the available dependent HLA gene products. There was no detectable recognition of influenza virus surface glycoproteins in target cells.

Antigens, Viral↗
PubMed1981

The human cytotoxic T cell response to influenza A vaccination.

The human cytotoxic T lymphocyte (CTL) response to challenge with influenza A vaccine was studied. Six of eight volunteers given killed whole influenza virus A/USSR (H1N1) vaccine showed substantial increases on the level of CTL memory 1 month after immunization. The CTL measured at this time showed complete cross-reactivity in their specificity for influenza A/USSR (H1N1) and A/X31 (H3N2) infected cells and also showed HLA restriction. The level of CTL memory increased in only three out of nine donors given subunit vaccine and showed no change in those not given vaccine. If cytotoxic T cells are important in influenza prophylaxis, killed whole virus vaccine should offer better protection than subunit vaccine.

Antibody Formation↗
PubMed1978

Precise anticoagulation for routine hemodialysis.

A pharmacokinetic model for minimal dose heparinization for chornic hemodialysis patients was recently described by Gotch and Keen. The model requires the determination of two parameters: dose sensitivity (computed for a given heparin dose from increase in WBPTT above a baseline value) and the heparin elimination constant. This study describes the extension of this model to the more precise control of anticoagulation during routine dialysis. S and K were measured in 30 stable chronic dialysis patients and were found to differ markedly (0.015 less than or equal to S 0.08 sec/unit; 0.04 less than or equal to K less than or equal to 1.7 hr-1). The mean S value was 0.041 +/- 0.002 sec/unit (N = 30) and the mean K value was 0.90 +/- 0.06 hr-1 (N = 30). In addition, the mean t 1/2 of heparin obtained in the group of 30 patients from individual rate constants was 0.86 +/- 0.06 hr, in excellent agreement with values obtained in normal subjects given similar doses of the drug. Variations in sensitivity during dialysis were minimal, but variations in elimination rate of up to 50% were encountered during modeling. However, the large variations in K did not affect the applicability of the model of control clotting times during dialysis when infusion requirements were based on mean values of S and K taken over four to five dialyses. In an initial group of five patients, whose heparin requirements were reduced by an average of 38% +/- 20 (range 13% to 65%), there was no significant change in the degree of dialyzer clotting in comparing premodeling and postmodeling heparin therapy. In most of the remaining patients (N = 22) the pattern was similar: a reduction in total heparin administration without increased dialyzer clotting. In three patients (10%) over-all heparin dose had to be moderately increased (3% to 13%). Heparin modeling has been successfully applied to routine anticoahe technique cannot be extended to other clinical procedures involving either intermittent or continuous infusion of heparin. To assist in the application of heparin modeling, nomograms have been developed.

Blood Coagulation Tests↗