Antigen processing: where tumor-specific T-cell responses begin

It is well established that tumor-specific CD8+ T cells have the capacity to prevent and cure malignancies in animals under experimental conditions. This has raised expectations that it will prove possible to achieve similar successes with human cancers. CD8+ T cells recognize peptides of 8-10 residues derived from cytosolic proteins that are bound to the class I molecules of the major histocompatibility complex. To most effectively manipulate the T-cell response to tumor cells, it is essential to understand the means by which the peptide-class I complex is created in cells. An overview of this process is provided with an emphasis toward the recent findings made by our laboratory.     

The comparative role of haemoglobinaemia and hypoxia in the development of canine babesial nephropathy

Renal pathology associated with haemoglobinaemia resulting from Babesia canis infection is ascribed to haemoglobinuria, with or without a contribution from anaemic hypoxia. This study was undertaken to investigate the relative roles of haemoglobinaemia and hypoxia in renal function and pathology in the dog. Three groups of 6 dogs each were used over a 4-day period. The dogs in the 1st group were infused with homologous canine haemoglobin, anaemic hypoxia was induced in the 2nd group, and both treatments were applied in the 3rd group. Full urinalyses, serum urea and creatinine concentrations, fractional clearance of sodium and the activity of urine enzymes, were assessed daily. At the end of the trial period, the glomerular filtration rate (GFR) was determined and kidney specimens collected for light and electron microscopy. In the group with hypoxia only, the urine sediment contained more casts and a greater number of renal tubular epithelial (RTE) cells than in either of the other groups. Hypoxia resulted in greater enzymuria, suggestive of RTE cell pathology, whereas haemoglobinuria did not appear to have any effect on urine enzyme activity. Hypoxia resulted in a decreased GFR. Histological examination revealed a mild, single-cell tubular necrosis in the majority of the animals (all 3 groups), with granular casts in the hypoxic groups. There appeared to be a large individual variation in the ability of the kidney to handle infused haemoglobin. It was concluded that severe haemoglobinaemia did not induce a significant nephropathy, anaemic hypoxia appeared to cause a very mild nephropathy, and the combination of haemoglobinaemia and anaemic hypoxia did not exacerbate this change. These lesions were very different from those described in canine babesiosis.     

Identification and differential subcellular localization of the neuronal class C and class D L-type calcium channel alpha 1 subunits

To identify and localize the protein products of genes encoding distinct L-type calcium channels in central neurons, anti-peptide antibodies specific for the class C and class D alpha 1 subunits were produced. Anti-CNC1 directed against class C immunoprecipitated 75% of the L-type channels solubilized from rat cerebral cortex and hippocampus. Anti-CND1 directed against class D immunoprecipitated only 20% of the L-type calcium channels. Immunoblotting revealed two size forms of the class C L-type alpha 1 subunit, LC1 and LC2, and two size forms of the class D L-type alpha 1 subunit, LD1 and LD2. The larger isoforms had apparent molecular masses of approximately 200-210 kD while the smaller isoforms were 180-190 kD, as estimated from electrophoresis in gels polymerized from 5% acrylamide. Immunocytochemical studies using CNC1 and CND1 antibodies revealed that the alpha 1 subunits of both L-type calcium channel subtypes are localized mainly in neuronal cell bodies and proximal dendrites. Relatively dense labeling was observed at the base of major dendrites in many neurons. Staining in more distal dendritic regions was faint or undetectable with CND1, while a more significant level of staining of distal dendrites was observed with CNC1, particularly in the dentate gyrus and the CA2 and CA3 areas of the hippocampus. Class C calcium channels were concentrated in clusters, while class D calcium channels were generally distributed in the cell surface membrane of cell bodies and proximal dendrites. Our results demonstrate multiple size forms and differential localization of two subtypes of L-type calcium channels in the cell bodies and proximal dendrites of central neurons. The differential localization and multiple size forms may allow these two channel subtypes to participate in distinct aspects of electrical signal integration and intracellular calcium signaling in neuronal cell bodies. The preferential localization of these calcium channels in cell bodies and proximal dendrites implies their involvement in regulation of calcium-dependent functions occurring in those cellular compartments such as protein phosphorylation, enzyme activity, and gene expression.     

Evaluation of a new chromogenic substrate assay for the measurement of the prothrombin time

Since the ingestion studies by Marshall and Morris, Helicobacter pylori has been known to cause both acute and chronic infection in the human stomach activating both the cellular and the humoral immune system. It is of little or no value to evaluate the causative relationship of an infectious agent using Koch's criteria. The more recent criteria for causative relationships used in the science of epidemiology are more useful. These criteria include: (i) the characteristic of the association which is fulfilled for most cases of both duodenal and gastric ulcer; (ii) the temporal relationship which is fulfilled for duodenal ulcer and has not been investigated for gastric ulcer; (iii) the biological gradient which has been fulfilled for duodenal ulcer in a few studies but not investigated for gastric ulcer; (iv) the biological plausibility which is easily fulfilled for both duodenal and gastric ulcer; (v) the effect of an intervention which has been fulfilled for duodenal ulcer and in a few studies for gastric ulcer; and (vi) the coherence of these data with what is known about the disease which is fulfilled for both duodenal and gastric ulcer. Even though there is no need for all criteria to be fulfilled, further studies are necessary to confirm the temporal relationship between H. pylori and peptic ulcer, and the biological gradient of H. pylori in relation to the gastric ulcer. Even so, there is a strong indication that most of the peptic ulcers, apart from those caused by non-steroid anti-inflammatory drugs and Zollinger-Ellison-like syndromes, are caused by H. pylori infection.     

Expression of a phosphorylation-resistant eukaryotic initiation factor 2 alpha-subunit mitigates heat shock inhibition of protein synthesis

Protein synthesis is dramatically reduced upon exposure of cells to elevated temperature. Concordant with this inhibition, multiple phosphorylation and dephosphorylation reactions occur on specific eukaryotic initiation factors that are required for protein synthesis. Most notably, phosphorylation of the alpha-subunit of eukaryotic initiation factor-2 (eIF-2 alpha) on serine residue 51 occurs. To identify the importance of phosphorylation in control of protein synthesis, we have evaluated the effects of expression of a mutant eIF-2 alpha which is resistant to phosphorylation. Expression of a serine to alanine mutant at residue 51 of eIF-2 alpha partially protected cells from the inhibition of protein synthesis in response to heat treatment. The overexpressed serine to alanine 51 mutant subunit was incorporated into the eIF-2 heterotrimer and was resistant to phosphorylation. These results are consistent with the hypothesis that heat shock inhibition of translation is mediated in part through phosphorylation of eIF-2 alpha. Expression of the wild type or mutant eIF-2 alpha did not affect cell survival or induction of hsp70 mRNA upon heat shock, indicating that although eIF-2 alpha is a heat shock-induced protein, its increased synthesis during heat shock does not alter the heat-shock response.     

Estimation of in vivo proton intrinsic and cross-relaxation rate in human brain

By using on-resonance binomial pulse saturation of the immobile component in tissues and by monitoring the time development of the longitudinal decay of the free water magnetization, it has been shown that it is possible to estimate physical model parameters not directly measurable by conventional methods. Since pulsed saturation is easy and safe to implement in a clinical MRI machine, it should be possible to estimate these parameters in vivo by the same means. In this article, we report the results of such estimates of the parameters of the principal components (gray and white matter) in brains of two normal volunteers.