HLA class II peptide binding specificity and autoimmunity.
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Alleles↗
Biomedical subjects
J Hammer
Publications and source records attributed to J Hammer.
Antigenic peptides and autoimmunity.
MHC class II alleles play a major role in determining resistance or susceptibility to autoimmune disease. Considerable effort is being expended to establish the role polymorphisms play in influencing the binding of antigens, including autoantigens, in the peptide-binding groove. Single amino acid substitutions in the MHC cleft, for instance at DR beta 71 and DQ beta 57, influence peptide binding. Although candidate autoantigenic peptides have been identified which bind to disease-associated MHC molecules, several critical questions remain to be answered before the role of these peptides in the autoimmune disease process can be established.
Animals↗
Absence of peripheral blood T cell responses to "shared epitope' containing peptides in recent onset rheumatoid arthritis.
OBJECTIVES: To determine if peptides containing the 'shared epitope' sequence, QKRAA, from either endogenous, HLA-DR beta 1 (0401), or exogenous, Escherichia coli dnaJ, sources activate T cells in recent onset rheumatoid arthritis (RA). METHODS: Peripheral blood mononuclear cell (PBMC) proliferative and whole blood cytokine responses to shared epitope containing peptides from DR beta 1 (0401) and E coli dnaJ, to control peptides from DR beta 1 (0402) and hsp40 and to the recall antigen, tetanus toxoid, were tested in 20 untreated, recent onset RA subjects, 20 HLA, age, and sex matched healthy controls and 18 other subjects with inflammatory arthritis. PBMC proliferative responses to a second E coli dnaJ peptide (with the shared epitope at the N-terminus) and two peptides from type II collagen with high affinity for DR4(0401) were tested in a further 16 recent onset RA and 17 control subjects. RESULTS: PBMC proliferation and whole blood interferon gamma or interleukin 10 production in response to the shared epitope containing and control peptides were not different between the disease and control groups. On the other hand, compared with controls, RA subjects had significantly higher proliferation to a collagen II (aa 1307-1319) peptide, but significantly lower proliferation and interferon gamma production to tetanus toxoid. CONCLUSION: Recent onset RA subjects had no demonstrable increase in peripheral blood T cell reactivity to shared epitope containing peptides. However, a proportion had increased T cell reactivity to a peptide of similar length from a candidate RA autoantigen, collagen type II. Their impaired responses to tetanus are in keeping with evidence for general T cell hyporesponsiveness in RA.
Acute Disease↗
The distal colon provides reserve storage capacity during colonic fluid overload.
BACKGROUND: In addition to its absorptive function the capacity of the colon to retain fluid might be relevant in compensating for increased fluid loads and prevention of diarrhoea. The distal colon is considered to be mainly a conduit without extensive storage function. AIMS: To evaluate colonic volume capacity in a model of pure osmotic diarrhoea. METHODS: A non-absorbable, iso-osmotic solution (OS) containing polyethylene glycol (500 ml) was infused into the caecum of nine healthy volunteers; the control group (n = 5) received an equal amount of an easily absorbable electrolyte solution (ES). Fluids were radiolabelled with technetium-99m and gamma camera images were obtained for 48 hours. Counts in the proximal and distal colon were measured and regional and overall colonic transit and stool output were quantified. RESULTS: After OS, in contrast to ES, faecal output was increased significantly (p < 0.05), but colonic transit after OS was not different from transit after ES (p > 0.05). This indicates storage of OS in the colon: after OS infusion, counts in the proximal colon decreased linearly while the distal colon stored approximately 30% of radioactivity for the whole 48 hour study period. After OS, stool output correlated with distal (p < 0.01), but not with proximal (p > 0.05), colonic transit. In contrast, after ES, stool output was determined by proximal colonic transit (p < 0.05) but not by transit through the distal colon (p > 0.05). CONCLUSION: The distal colon retains non-absorbable fluid volumes extensively. In our model transit through the distal colon--but not the proximal colon--determined the time at which diarrhoea occurred.
Adult↗
Effect of prone and supine positions on functional residual capacity, oxygenation, and respiratory mechanics in ventilated infants and children.
Although numerous reports have described the improvement in PAO2 in patients in the prone position, the underlying mechanism has yet to be determined. Some authors have suggested this phenomenon may be related to an increase in functional residual capacity (FRC); however, no previous studies have described positional changes in FRC in children with severe lung disease or in those under neuromuscular blockade. We measured arterial blood gases, FRC, Rrs, and Crs in supine and prone positions in 30 patients under neuromuscular blockade with lung disorders including moderately severe restrictive (n = 10) and obstructive (n = 10) disease and control subjects without significant lung disease (n = 10). Prone positioning was not associated with a significant increase in FRC in the cohort of 30 patients, nor in any of the subgroups. Although individual patients demonstrated large improvements in oxygenation, a statistically significant (but clinically insignificant) increase in AaPO2 ratio was observed only in the subgroup of patients with obstructive disease (0.35+/-0.03 to 0.38+/-0.04, p = 0.027). There was no correlation between changes in FRC and changes in AaPO2 (r = 0.225, p = 0.23). A significant improvement in Rrs occurred in the prone position compared to supine in patients with obstructive lung disease, decreasing from 0.264+/-0.024 to 0.216+/-0.021 cm H2O/ml/s, p = 0.009. No significant changes in Crs were seen in the prone position. We conclude that prone positioning has no effect on FRC and in this series of 30 patients significantly improved oxygenation only in patients with obstructive airway disease. A significant decrease in Rrs in patients with obstructive lung disease was also observed.
Blood Gas Analysis↗
Influence of endotracheal tube diameter on forced deflation flow-volume curves in rhesus monkeys.
The forced deflation (FD) technique is the recommended gold standard to generate forced expiratory vital capacity (FVC) curves and to measure maximum expiratory flow-volume (MEFV) relationships in intubated infants and children. However, the influence of the endotracheal tube (ETT) on the site of flow limitation, the shape and the analysis of the resultant MEFV curves have not been defined. Nine anaesthetized (thiopentone, 8 mg x kg(-1) x h(-1)) rhesus monkeys (mean weight (+/-SEM) 10+/-1 kg) were intubated consecutively with ETTs of different internal diameters (ID 3.0-5.5 mm, at intervals of 0.5 mm); the largest representing the appropriate ETT size for the animal. Quadruplicate MEFV curves were generated by FD using the standard +40 cmH2Oinsp/-40 cmH2Oexp pressures and recorded at each ETT diameter. The effect of the different ETT diameters on the FD flow-volume curve was analysed by comparing maximum expiratory flows at isovolume points at 50, 25 and 10% FVC (MEF50, MEF25 and MEF10, respectively). The shape of the resulting MEFV curves could be divided into an initial horizontal part (tube-dependent), and a final descending slope (airway-dependent). No difference in FVC occurred irrespective of the ETT size (p>0.05 by analysis of variance (ANOVA)). MEF50 increased with increasing tube diameter (p<0.05). MEF25 remained unchanged using ETTs which were less than 1.5 mm smaller than the largest ETT. Smaller ETTs resulted in significantly reduced MEF25 (p<0.05). MEF10 was not influenced by the ETT size. We conclude that maximum expiratory flows measured by the forced deflation technique are not influenced by an appropriately sized endotracheal tube at lung volumes below 25% forced vital capacity in our monkey model with normal lungs. We postulate that the effect of endotracheal tubes on maximum expiratory flow volume curves in intubated infants might be of similar or even smaller magnitude, which remains to be established.
Animals↗
The effects of bronchodilators on spontaneous ventilation and oxygen consumption in rhesus monkeys.
The effects of breathing normal saline, salmeterol, fenoterol, ipratropium bromide, or formoterol, and of i.v. infusion of theophylline on oxygen consumption (VO2), carbon dioxide production (VCO2), minute ventilation (VE), heart and respiratory rates, and end-tidal carbon dioxide tension (P(ET)CO2) have been defined in 10 anesthetized, intubated rhesus monkeys (mean age 7.0 y, weight 10.2 kg). VO2 increased over control by + 17.1% after salmeterol (p < 0.001), +33.3% after fenoterol (p < 0.001), +23.7% after formoterol (p < 0.001), +3.9% after theophylline (p < 0.01), but did not change after ipratropium bromide and normal saline. VE increased by 63.0% after fenoterol (p < 0.001), 49.8% after formoterol (p < 0.001), 31.7% after salmeterol (p < 0.01), and 29.7% after theophylline (p < 0.001), but not after ipratropium bromide or normal saline. Heart rate response was greatest after fenoterol, formoterol, and salmeterol, respectively. P(ET)CO2 dropped dramatically after theophylline (-15.7%, p < 0.001), but not at all with any of the inhaled beta2-adrenoceptor agonists. In seven animals, salbutamol (albuterol) caused an increase in V(E) and VO2 of 50.1% and 45.9%, respectively, whereas in the presence of a beta2-adrenoceptor antagonist [racemic or (+/-)-propranolol (0.1 mg/kg i.v.)], inhaled salbutamol (2.5 mg/mL for 10 min) could not increase V(E) (+6.2%, p > 0.05) and VO2 (+1.6%, p > 0.05). The increase in VO2 and V(E) after administration of beta2-agonists may be partly the result of direct stimulation of the respiratory center and partly a response to increased metabolic rate. The dramatic increase in VO2 and V(E) after salbutamol was suppressed in the presence of propranolol, which is consistent with a beta-receptor-mediated mechanism.
Adrenergic beta-Agonists↗
Subcellular localization of myosin V in nerve growth cones and outgrowth from dilute-lethal neurons.
Myosin V-null mice (dilute-lethal mutants) exhibit apparent neurological defects that worsen from birth until death in the third postnatal week. Although myosin V is enriched in brain, the neuronal function of myosin V is unclear and the underlying cause of the neurological defects in these mice is unknown. To aide in understanding myosin V function, we examined the distribution of myosin V in the rodent superior cervical ganglion (SCG) growth cone, a well characterized neuronal structure in which myosin V is concentrated. Using affinity purified, myosin V-specific antibodies in immunofluorescence and immunoelectron microscopy, we observed that myosin V is concentrated in organelle-rich regions of the growth cone. Myosin V is present on a distinct population of small (50-100 nm) organelles, and on actin filaments and the plasma membrane. Myosin V-associated organelles are present on both microtubules and actin filaments. These results indicate that myosin V may be carried as a passenger on organelles that are transported along microtubules, and that these organelles may also be capable of movement along actin filaments. In addition, we found no abnormalities in outgrowth, morphology, or cytoskeletal organization of SCG growth cones from dilute-lethal mice. These results indicate that myosin V is not necessary for the traction force needed for growth cone locomotion, for organization of the actin cytoskeleton, or for filopodial dynamics.
Actin Cytoskeleton↗
Isolation of scFv fragments to polymorphic and monomorphic HLA-DR epitopes from a synthetic phage library.
The possibility of producing human recombinant antibodies by the phage display libraries technology has opened up new perspectives in the fields of immunological research and therapeutic applications. Despite the interest for a potential use of this technology in the HLA field, no information is so far available about selection and screening of libraries with HLA antigens. In this study we report our first experience with expression and characterization of human single-chain antibody fragments (scFvs) against HLA-DR epitopes selected from a synthetic phage library.
Bacteriophages↗
Editing of the HLA-DR-peptide repertoire by HLA-DM.
Antigenic peptide loading of classical major histocompatibility complex (MHC) class II molecules requires the exchange of the endogenous invariant chain fragment CLIP (class II associated Ii peptide) for peptides derived from antigenic proteins. This process is facilitated by the non-classical MHC class II molecule HLA-DM (DM) which catalyzes the removal of CLIP. Up to now it has been unclear whether DM releases self-peptides other than CLIP and thereby modifies the peptide repertoire presented to T cells. Here we report that DM can release a variety of peptides from HLA-DR molecules. DR molecules isolated from lymphoblastoid cell lines were found to carry a sizeable fraction of self-peptides that are sensitive to the action of DM. The structural basis for this DM sensitivity was elucidated by high-performance size exclusion chromatography and a novel mass spectrometry binding assay. The results demonstrate that the overall kinetic stability of a peptide bound to DR determines its sensitivity to removal by DM. We show that DM removes preferentially those peptides that contain at least one suboptimal side chain at one of their anchor positions or those that are shorter than 11 residues. These findings provide a rationale for the previously described ligand motifs and the minimal length requirements of naturally processed DR-associated self-peptides. Thus, in endosomal compartments, where peptide loading takes place, DM can function as a versatile peptide editor that selects for high-stability MHC class II-peptide complexes by kinetic proofreading before these complexes are presented to T cells.
Amino Acid Sequence↗
Secondary structure and location of a magainin analogue in synthetic phospholipid bilayers.
Magainins are cationic, membrane-active peptides which show broad-spectrum antimicrobial activity. We have investigated the secondary structure and location of an analogue of magainin 2 in synthetic phospholipid bilayers using a combination of Fourier transform infrared (FTIR) spectroscopy and solid-state nuclear magnetic resonance (NMR) spectroscopy. Ala19-magainin 2 amide exhibits both alpha-helix and beta-sheet secondary structures in lipid bilayers containing either dipalmitoylphosphatidylglycerol (DPPG) or a 1:1 molar mixture of DPPG and dipalmitoylphosphatidylcholine (DPPC). The combination of FTIR and solid-state NMR results suggests that there are two populations of peptide. The secondary structure of one population is alpha-helix while that of the other population is beta-sheet. We demonstrate that the solid-state NMR technique, rotational-echo double resonance (REDOR), can be used to measure both intra- and intermolecular dipole-dipole interactions in membrane-bound peptides. Our REDOR experiments indicate that alpha-helical Ala19-magainin 2 amide is bound near the phospholipid head groups.
Amino Acid Sequence↗
HLA-DR4-IE chimeric class II transgenic, murine class II-deficient mice are susceptible to experimental allergic encephalomyelitis.
To investigate the development of HLA-DR-associated autoimmune diseases, we generated transgenic (Tg) mice with HLA-DRA-IE alpha and HLA-DRB1*0401-IE beta chimeric genes. The transgene-encoded proteins consisted of antigen-binding domains from HLA-DRA and HLA-DRB1*0401 molecules and the remaining domains from the IE(d)-alpha and IE(d)-beta chains. The chimeric molecules showed the same antigen-binding specificity as HLA-DRB1*0401 molecules, and were functional in presenting antigens to T cells. The Tg mice were backcrossed to MHC class II-deficient (IA beta-, IE alpha-) mice to eliminate any effect of endogenous MHC class II genes on the development of autoimmune diseases. As expected, IA alpha beta or IE alpha beta molecules were not expressed in Tg mice. Moreover, cell-surface expression of endogenous IE beta associated with HLA-DRA-IE alpha was not detectable in several Tg mouse lines by flow cytometric analysis. The HLA-DRA-IE alpha/HLA-DRB1*0401-IE beta molecules rescued the development of CD4+ T cells in MHC class II-deficient mice, but T cells expressing V beta 5, V beta 11, and V beta 12 were specifically deleted. Tg mice were immunized with peptides, myelin basic protein (MBP) 87-106 and proteolipid protein (PLP) 175-192, that are considered to be immunodominant epitopes in HLA-DR4 individuals. PLP175-192 provoked a strong proliferative response of lymph node T cells from Tg mice, and caused inflammatory lesions in white matter of the CNS and symptoms of experimental allergic encephalomyelitis (EAE). Immunization with MBP87-106 elicited a very weak proliferative T cell response and caused mild EAE. Non-Tg mice immunized with either PLP175-192 or MBP87-106 did not develop EAE. These results demonstrated that a human MHC class II binding site alone can confer susceptibility to an experimentally induced murine autoimmune disease.
Amino Acid Sequence↗
[Misleading advertising on a new NSAID].
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Advertising↗
Compound osteosynthesis of pathological fractures of the proximal femoral shaft. Experimental studies on the biomechanical effects of additional intramedullary splinting.
The biomechanical stability of two compound osteosynthesis techniques with and without intramedullary splinting were compared in 20 paired cadaver femora. Group A was stabilised using a contoured 12-hole DC plate and polymethylmethacrylate cement. In group B an additional intramedullary semi-diameter osteosynthesis plate was inserted parallel to the outer plate and transfixed with the same screws. The specimens were tested in a universal testing machine using a force applied to the specimen at 45 degrees to the load to provoke the most unfavourable condition, consisting of a bending moment and torsion. Both force and extension were recorded. The compound osteosynthesis with an additional intramedullary plate (group B) reached an average higher load stability. This increase of stability was not significant statistically.
Aged↗
Minimal chronic inactive gastritis: indicator of pre-existing helicobacter pylori gastritis?
Minimal chronic inactive gastritis is regularly observed in routine histopathology. Presently, it is not clear whether this type of gastritis should be regarded as a histopathological entity or a normal variant. The similarity to lesions observed after H.pylori eradication prompted us to look for an association between minimal chronic inactive gastritis and status post H.pylori eradication. In a prospective study of 110 consecutive patients undergoing upper gastrointestinal endoscopy, at least two mucosal biopsies were taken from the gastric antrum and body. Gastritis was classified according to the Sydney System. Antibodies to H.pylori were determined serologically by immunofluorescence test, ELISA, and complement binding reaction. A status post eradication of H.pylori was revealed by medical history and/or positive serology; H.pylori gastritis was found in 39.1%, reactive gastritis in 12.7%, and minimal chronic inactive gastritis in 29.1%. In 19.1% a combination of reactive/ minimal chronic gastritis was diagnosed according to morphology. Status post eradication was observed significantly more often in cases with minimal chronic inactive gastritis (43.8%) than in cases with reactive gastritis (7.1%, p < 0.004). Furthermore, positive ELISA and/or status after eradication was found in 50% of the cases with minimal chronic inactive gastritis (p < 0.005 vs reactive gastritis), in 42.9% of the cases with mixed reactive/chronic inactive gastritis (p < 0.03 vs reactive gastritis), and in 7.1% of the cases with reactive gastritis. Lymphoid aggregates, considered another sign of former H.pylori presence, were found significantly more often in minimal chronic inactive gastritis than in reactive gastritis (50% versus 7.1%, p < 0.005). Minimal chronic inactive gastritis is significantly associated with both positive H.pylori serology and status post eradication and is, therefore, an indicator of pre-existing H.pylori gastritis.
Biomarkers↗
Effort and volume dependence of forced-deflation flow-volume relationships in intubated infants.
The application of negative pressure to the airway opening [called the forced-deflation (FD) technique] allows the examination of maximal expiratory flow-volume curves in intubated infants who are unable to generate a voluntary maximal expiratory maneuver. We explored the questions of effort and volume dependence of flows generated by FD in 18 intubated, sedated, and paralyzed infants [age 10.6 +/- 2.0 (SE) mo; weight 7.2 +/- 0.7 kg] with normal lungs. Effort dependence was assessed by isovolume pressure-flow curves that were constructed in 10 infants from repeated FD maneuvers from total lung capacity (defined as +40 cmH2O) by varying airway opening pressures from 0 (barometric pressure) to -100 cmH2O at intervals of 20 cmH2O. The effect of volume history was assessed by initiating FD maneuvers from different inspiratory volumes delineated by the inspiratory pressures +10, +20, +30, and +40 cmH2O. We compared maximal expiratory flows at isovolume points [50, 25, and 10% forced vital capacity (FVC) of the standard +40/-40 cmH2O FD maneuver] and found that flow limitation consistently occurred in all infants at and below 25% FVC with -40 cmH2O or greater airway opening pressure. We found no significant influence of volume history on maximal flows at and below 25% FVC. Under well-controlled study conditions, we demonstrated excellent reproducibility of maximal expiratory flows at low lung volumes, analogous to those of voluntary forced expiratory maneuvers in adults and older children. This information may be helpful in setting standards for performance and interpretation of FD maneuvers in intubated infants.
Humans↗
Flow limitation in anesthetized rhesus monkeys: a comparison of rapid thoracoabdominal compression and forced deflation techniques.
Rapid thoracoabdominal compression (RTC) and forced deflation (FD) allow the examination of maximal expiratory flow-volume (MEFV) curves in animals and human infants unable to generate a voluntary maximal expiratory maneuver. We compared flows generated by these two techniques and by constructing isovolume pressure-flow (IVPF) curves examined each method's potential to produce flow limitation. RTC and FD were performed in seven intubated rhesus monkeys from +40 cm H2O inspiratory pressure with deflation pressures ranging from -20 to -100 cm H2O and jacket pressures from +20 to +100 cm H2O (arms outside jacket). We also performed RTC maneuvers in all monkeys with the arms inside the jacket at +100 cm H2O jacket pressure. Maximal expiratory flows achieved by FD and RTC were analyzed at isovolume points. Both techniques produced flow limitation conditions over the last 25% of the MEFV curve. Individual IVPF curves generated by the two techniques demonstrated remarkable differences in shape and flow rates. Maximal transpulmonary pressures achieved with FD were higher than those with RTC in this experiment. Negative effort dependence was observed with higher deflation pressures in the majority of the monkeys. Flows achieved by RTC with the arms in were lower than those achieved with the arms out when compared at identical pressure gradients. We conclude that the difference in expiratory flow rates at isovolume-pressure points is most likely related to different effects on the behavior of intrapulmonary airways and choke point characteristics by the two techniques.
Animals↗
[Crohn disease gastritis: histomorphology and immunohistochemical characterization of inflammatory cell infiltrates].
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Antigens, CD↗