Isolation of cDNA clones coding for human tissue factor: primary structure of the protein and cDNA.

Tissue factor is a membrane-bound procoagulant protein that activates the extrinsic pathway of blood coagulation in the presence of factor VII and calcium. lambda Phage containing the tissue factor gene were isolated from a human placental cDNA library. The amino acid sequence deduced from the nucleotide sequence of the cDNAs indicates that tissue factor is synthesized as a higher molecular weight precursor with a leader sequence of 32 amino acids, while the mature protein is a single polypeptide chain composed of 263 residues. The derived primary structure of tissue factor has been confirmed by comparison to protein and peptide sequence data. The sequence of the mature protein suggests that there are three distinct domains: extracellular, residues 1-219; hydrophobic, residues 220-242; and cytoplasmic, residues 243-263. Three potential N-linked carbohydrate attachment sites occur in the extracellular domain. The amino acid sequence of tissue factor shows no significant homology with the vitamin K-dependent serine proteases, coagulation cofactors, or any other protein in the National Biomedical Research Foundation sequence data bank (Washington, DC).     

Reduced StartReact effect and freezing of gait in Parkinson’s disease: two of a kind?

Freezing of gait (FOG) is a disabling feature of Parkinson’s disease. Emerging evidence suggests that dysfunction of the pedunculopontine nucleus (PPN) and pontomedullary reticular formation (pmRF) plays a role in the causation of FOG. These brainstem structures can be examined by the StartReact paradigm, which utilizes a startling stimulus to accelerate reaction times (StartReact). Here, we examined gait initiation in PD patients with and without FOG using this paradigm. Twenty-six patients with Parkinson’s disease (12 freezers and 14 non-freezers) and 15 controls performed two tasks: rapid gait initiation in response to an imperative ‘go’ signal; and a control condition, involving a simple reaction-time task involving ankle dorsiflexion. During both tasks, a startling acoustic stimulus was combined with the imperative signal in 25 % of trials. In controls, the startle accelerated gait initiation and shortened the onset latency of tibialis anterior responses during ankle dorsiflexion. This acceleration was intact in non-freezers, but was significantly attenuated in the freezers. Independent of the occurrence of a startle, freezers showed a reduced length of the first step compared to non-freezers and controls. The diminished StartReact effect in freezers probably reflects deficient representation or release of motor programs at the brainstem reticular level due to dysfunction of the PPN, the pmRF, or both. These brainstem structures are presumably involved in integrating anticipatory postural adjustments with subsequent stepping movements. We suggest that with time-varying demands, these structures may no longer be able to coordinate the integration of anticipatory postural adjustments with steps, leading to FOG episodes.     

Detection of choroid- and retina-antigen reactive CD8 and CD4 T lymphocytes in the vitreous fluid of patients with birdshot chorioretinopathy

Birdshot chorioretinopathy (BSCR), a progressive form of non-infectious uveitis, is the strongest HLA-associated disease described to date, with >95% of the patients displaying HLA-A29. Since indirect evidence indicates the involvement of T cells in the etiopathology of the disease, we now isolated, cultured and analyzed the vitreous fluid-infiltrating T cells from two BSCR patients with respect to their phenotype, cytokine profile, clonal distribution and antigen specificity. Phenotypic analyses revealed the predominant presence of both CD4⁺ and CD8⁺ T cells in vitreous fluid. Further analyses on short term expanded and cloned T cells suggested that eye-infiltrating T cells generally displayed a Th1 like cytokine profile with secretion of high levels of IFN-γ and TNF-α. In one patient an oligoclonal CD4⁺ and CD8⁺ T cell infiltration, with a moderate to strongly skewed TCR Vβ usage was suggestive for an antigen driven infiltration/expansion. Indeed, a number of intraocular CD4⁺ and CD8⁺ T cells responded to crude retinal and choroidal lysates. These results, which demonstrate for the first time the existence of eye-antigen-specific T cells in the vitreous fluid of BSCR patients, substantiate the current view on the role of eye-antigen specific T cells in the etiopathology of BSCR.     

CT Screening for Pulmonary Pathology in Common Variable Immunodeficiency Disorders and the Correlation with Clinical and Immunological Parameters

Background

Pulmonary disease is common in patients with common variable immunodeficiency disorders (CVID) and involves infections, chronic airway disease and interstitial lung disease. Chronic pulmonary disease is associated with excess morbidity and early mortality and therefore early detection and monitoring of progression is essential.

Methods and Purpose

Thin slice CT scan and pulmonary function were used to determine the prevalence and spectrum of chronic (pre-clinical) pulmonary disease in adult CVID patients regardless of symptoms. CT Scans were scored for airway abnormalities (AD) and interstitial lung disease (ILD). Other CVID related complications and B and T lymphocyte subsets were analyzed to identify patients at risk for pulmonary disease.

Results

Significant pulmonary abnormalities were detected in 24 of the 47 patients (51 %) consisting of AD in 30 % and ILD in 34 % of cases. In only 7 (29 %) of these 24 patients pulmonary function test proved abnormal. The presence of AD was correlated to (recurrent) lower respiratory tract infections despite IgG therapy. The presence of ILD was correlated to autoimmune disease and a reduction in the numbers of CD4?+?T cells, naïve CD4?+?T cells, naïve CD8?+?T cells and memory B cells and lower IgG through levels over time.

Conclusion

Preclinical signs of AD and ILD are common in CVID patients despite Ig therapy and do not correlate to pulmonary function testing. Patients at risk for ILD might be identified by the presence of autoimmunity or a deranged T cell pattern. Larger studies are needed to confirm these findings and to determine thresholds for the T lymphocyte subsets.     

Human immune response to group A streptococcal carbohydrate (A-CHO). III. Comparison of the efficiencies of anti-idiotopic antibody and of nominal antigen in the induction of IgM anti-A-CHO-producing B cells

In this study the efficiencies of a monoclonal anti-idiotopic (Id) antibody (anti-Id498) and of various preparations of nominal antigen in the induction of an antigen-specific human B cell response in vitro were compared. Anti-Id498 recognizes a recurrent, binding site-related Id present on IgM antibodies with specificity for N-acetyl-D-glucosamine, the immunodominant group of streptococcal group A carbohydrate (A-CHO). We have previously shown that anti-Id498 induces IgM anti-A-CHO secretion from B cells of donors that possess Id-498+ antibodies in their serum. A-CHO was presented to B cells either in soluble or insoluble form, i.e. coupled to beads or as intact bacteria. Purified blood B cell populations from three Id+ healthy donors with high numbers of circulating anti-A-CHO B cells were used and antibody-producing B cells are enumerated in a single-cell assay (spot ELISA). The data show that anti-Id498 was superior in the induction of IgM anti-A-CHO-secreting B cells in two donors (factor 4.6 and 13.5 as compared to the most efficient antigenic stimulation). In the third donor antigen stimulation was slightly more efficient than anti-Id but only with Sepharose-bound A-CHO and not with soluble A-CHO or intact bacteria. The increase of specific B cells induced after stimulation with anti-Id498 could be abolished after addition of autologous T cells in two donors. On the contrary, an enhancement of the specific response was observed after addition of autologous T cells in antigen-stimulated cultures. Neither suppression nor enhancement were induced by addition of irradiated T cells.     

Saturating Nonlinearities of Contrast Response in Human Visual Cortex

Response nonlinearities are ubiquitous throughout the brain, especially within sensory cortices where changes in stimulus intensity typically produce compressed responses. Although this relationship is well established in electrophysiological measurements, it remains controversial whether the same nonlinearities hold for population-based measurements obtained with human fMRI. We propose that these purported disparities are not contingent on measurement type and are instead largely dependent on the visual system state at the time of interrogation. We show that deploying a contrast adaptation paradigm permits reliable measurements of saturating sigmoidal contrast response functions (10 participants, 7 female). When not controlling the adaptation state, our results coincide with previous fMRI studies, yielding nonsaturating, largely linear contrast responses. These findings highlight the important role of adaptation in manifesting measurable nonlinear responses within human visual cortex, reconciling discrepancies reported in vision neuroscience, re-establishing the qualitative relationship between stimulus intensity and response across different neural measures and the concerted study of cortical gain control.SIGNIFICANCE STATEMENT Nonlinear stimulus–response relationships govern many essential brain functions, ranging from the sensory to cognitive level. Certain core response properties previously shown to be nonlinear with nonhuman electrophysiology recordings have yet to be reliably measured with human neuroimaging, prompting uncertainty and reconsideration. The results of this study stand to reconcile these incongruencies in the vision neurosciences, demonstrating the profound impact adaptation can have on brain activation throughout the early visual cortex. Moving forward, these findings facilitate the study of modulatory influences on sensory processing (i.e., arousal and attention) and help establish a closer link between neural recordings in animals and hemodynamic measurements from human fMRI, resuming a concerted effort to understand operations in the mammalian cortex.     

Magnetic resonance imaging of renal transplants: its value in the differentiation of acute rejection and cyclosporin A nephrotoxicity

The purpose of this study was to determine the value of magnetic resonance imaging (MRI) in the differentiation of acute rejection and cyclosporin A nephrotoxicity in renal transplant kidneys. Fifty-six magnetic resonance examinations in 46 patients were prospectively and independently evaluated by two radiologists. MRI was performed with a 0.5 T superconducting scanner (Gyroscan S5, Philips) applying both T1 and T2 weighted pulse sequences. Biopsies were performed in 22 cases and histology was reviewed. Fifteen normally functioning transplant kidneys and 41 kidneys with graft dysfunction due to cyclosporin A nephrotoxicity, acute rejection, chronic rejection or acute tubular necrosis were studied. Absence or reduction in cortico-medullary demarcation proved to be a sensitive, but non-specific indicator of parenchymal disease. In cases of cyclosporin A nephrotoxicity the allograft was diagnosed as being normal in 90%. The magnetic resonance appearance of acute rejection may be very similar to that of the combination of acute rejection and cyclosporin A nephrotoxicity, chronic rejection or acute tubular necrosis. However differentiation between acute rejection and cyclosporin A nephrotoxicity was possible according to the following statistical data: sensitivity 100%, specificity 75%, positive predictive value 86%, negative predictive value 100%, accuracy 90%.