Relationship between peptide binding and T cell epitope selection: a study with subtypes of HLA-B27

The effect of HLA-B27 polymorphism on antigen presentation was analysed by comparing the binding of three Epstein-Barr virus-derived peptide epitopes to HLA-B27 subtypes with their immunogenicity and antigenicity in the context of these subtypes. The effect of altering the major anchor residue Arg2 on binding or on recognition by peptide-specific cytotoxic T lymphocytes (CTL) was also examined. The three peptides bound significantly to all the B*2701-B*2706 subtypes. This did not correlate with the peptides being immunogenic or recognized by specific CTL in the context of only particular subtypes. In addition, of the three viral epitopes tested, those that were immunogenic in B*2702- or B*2705-restricted responses bound to these subtypes less efficiently than one peptide that was immunogenic only in the B*2704 context. Thus, among several potentially immunogenic peptides from the same virus, the antiviral response is not necessarily directed against the one that binds best to the restricting subtype. These results indicate that HLA- B27 polymorphism influences antigen presentation in ways other than simply peptide affinity. Synthetic analogues lacking the canonical Arg2 motif of HLA-B27-bound peptides, even when binding much worse to the restricting subtype, were recognized equally by CTL specific for the parental peptide. This indicates that Arg2 is not required to maintain the structure of the epitope. The implications of these results for pathogenetic models of HLA-B27-associated disease are discussed.      

Assessing Coronary Flow Physiology with Intracoronary Doppler Following Coronary Interventions

Although coronary angiography has been the gold standard for assessing coronary artery stenoses, it yields information primarily about the anatomical severity of coronary artery disease, which frequently does not correlate with its physiological severity. Coronary interventions (PTCA, atherectomy, laser, etc.) are performed primarily to improve coronary flow physiology. Coronary flow physiology may be a more important end point than angiography following coronary interventions that were performed to normalize coronary flow physiology. In addition, the physiological significance of angiographically intermediate stenoses should he assessed before proceeding with catheter-based revascularization. Currently, the Doppler guidewire is available for routine clinical assessment of coronary flow physiology in the Cardiac Catheterization Lab. Several Doppler measurements have been used to assess the physiological effect of a stenosis, including the diastolic-systolic velocity ratio, proximal-distal velocity ratio, coronary flow reserve, continuity equation, and the hyperemic diastolic pressure-flow relationship. The Doppler derived coronary flow reserve correlates highly with stress nuclear perfusion images. These Doppler measurements have been made following PTCA, directional atherectomy, rotational atherectomy, and excimer laser. Following coronary interventions, adverse clinical events may be predicted if there is impaired flow physiology or cyclic flow variations. Many of the Doppler measurements used for assessing the lesion severity remain abnormal following successful coronary interventions for reasons unrelated to the lesion. Conversely, normalization of coronary physiology does not guarantee an adequate anatomical result. Further clinical trials will provide a more complete definition of the exact role for coronary flow velocity assessment following coronary interventions. (J Interven Cardiol 1996;9:163–173)     

Molecular Characterization of MPT83: a Seroreactive Antigen of Mycobacterium tuberculosis with Homology to MPT70

The Mycobacterium bovis antigens MPB70 and MPB83 are homologous cross-reactive proteins. It has been reported previously that MPB83 is glycosylated and exists in two forms with apparent molecular masses of 23 kDa and 25 kDa, whereas the apparent molecular mass of MPB70 is 22 kDa. Using a monoclonal antibody, SB10, which recognizes an epitope common to both MPB70 and MPB83, we compared the expression of these proteins in M. bovis BCG, virulent M. bovis and virulent Mycobacterium tuberculosis by Western blotting of bacterial lysates. The previously described pattern of high and low producing substrains of BCG for MPB70 was also applicable for MPB83. Virulent M. bovis was found to express high levels of MPB70 and MPB83. Immunoblotting experiments using sera from Balb/c mice infected with live M. tuberculosis H37Rv revealed that although the MPB83 homologue of M. tuberculosis, MPT83, is expressed at low levels in M. tuberculosis when grown in vitro, the protein is highly immunogenic during infection with live bacteria. A clone from a mycobacterial shuttle cosmid library of M. tuberculosis H37Rv was isolated which expressed both MPT70 and MPT83. Genetic analysis of this cosmid revealed that MPT70 and MPT83 were encoded by separate genes with the gene encoding MPT83 situated 2.4 kb upstream of mpt70. Both genes are transcribed in the same direction. The gene encoding MPT83 was cloned and DNA sequencing revealed an open reading frame of 660 bp encoding a protein with a predicted molecular mass of 22 kDa. Recombinant MPT83 was expressed in Escherichia coli from the native AUG initiation codon by translational coupling. In E. coli MPT83 was expressed as a 23 kDa antigen whereas in the rapid growing mycobacterium Mycobacterium smegmatis the protein was expressed as a 25 kDa protein indicating post-translational modification of the protein by M. smegmatis. In recombinant M. smegmatis MPT83 was predominantly cell associated whereas MPT70 was secreted into the culture medium. Amino acid sequence comparison between MPT83 and MPT70 revealed a 61% identity between the proteins, although little homology was apparent at the amino terminus. In MPT83 this region contained a typical lipoprotein signal peptide cleavage motif and a putative signal motif for O glycosylation. Both these motifs were absent from the amino acid sequence of MPT70.     

Mycotic Aneurysm of the Aortic Arch with Aortoesophageal Fistula

An elderly man presented with an aortoesophageal fistula (AEF) and underwent resection of a mycotic aneurysm of the aortic arch. After an initial success, the patient ultimately succumbed to mediastinal sepsis. The various approaches to combined aortic and esophageal pathology are presented with the recommendation for aortic replacement along with early removal of mediastinal contamination.     

Auditory steady-state response, upper facial EMG, EEG and heart rate as predictors of movement during isoflurane-nitrous oxide anaesthesia

We have studied the relationship between patient movement andchanges in the auditory steady-state evoked potential, upperfacial muscle electromyogram (FEMG), electroencephalographic-zerocrossing frequency (EEG–ZCF) and heart rate during emergencefrom anaesthesia. Twelve healthy patients underwent surgeryduring stable isoflurane-nitrous oxide-oxygen anaesthesia withoutneuromuscular block. After skin closure, anaesthesia was discontinuedabruptly while mechanical ventilation was continued until thepatient moved. The magnitude of change in each physiologicalsignal was evaluated in decibels (dB). Both the auditory steadystate evoked potential and FEMG showed significant increasesin amplitude during the last 5-min period before movement (6.1and 10.7 dB, respectively). EEG-ZCF increased rapidly afteranaesthesia was discontinued (2.5 dB) but there was no furtherincrease in activity before movement. Heart rate did not changebefore movement. The use of the decibel transformation offersa promising method of displaying and interpreting changes inphysiological variables during anaesthesia.