Non-cultured cell transplantation in an ovine model of non-ischemic heart failure.

OBJECTIVE: Cell therapy may be a promising alternative or adjunct to current treatment modalities for ischemic heart failure. But little is known on the impact of myogenic cell transplantation in large animal models of non-ischemic cardiomyopathy. The aim of the present study was to explore whether an ovine model of toxin-induced heart disease could benefit from non-cultured skeletal muscle cell transplantation. METHODS: Sequential intracoronary injections of doxorubicin (0.75 mg/kg) were carried out every 2 weeks until echocardiographic detection of myocardial dysfunction. Sheep were then randomly assigned to either non-cultured cell transplantation (n=8) or placebo injection (n=5). For the cell therapy group, a skeletal muscle biopsy (about 10 g) was explanted from each animal approximately 3h before grafting. After thoracotomy, 20 epicardial injections were carried out. The animals were assessed one last time before sacrifice, 2 months after the thoracotomy. Cells were tracked with cmDiI (red fluorescence) and characterized with immunohistochemistry with monoclonal antibodies to a fast skeletal isoform of myosin heavy chain. RESULTS: Two months after intramyocardial grafting, tissue Doppler imaging and conventional echocardiographic assessment of the groups showed a marked improvement in the non-cultured cell therapy group. Ejection fraction (EF) (p<0.05) as well as systolic endocardial velocities (p<0.01) improved versus the placebo group. CmDiI and skeletal myosin heavy chain expression was detected in all animals at 2 months after implantation confirming engraftment of skeletal muscle cells. CONCLUSIONS: In conclusion, our data indicate that non-cultured muscle cell transplantation is feasible and may translate into a functional benefit in an ovine model of dilated heart failure.     

Brain magnetic resonance imaging (MRI) and neurological changes after a single high altitude climb.

PURPOSE: Neurological impairment, mental dysfunction, and brain imaging changes caused by severe hypoxia have been described by several authors. However, the occurrence of transitory, long lasting, or permanent brain damage has been debated. Although climbing to 8000 m is reserved to a small number of climbers, there are hundreds of lowlanders spending relatively short holidays climbing peaks up to 6000 m in the Andes or in the Himalayas. They are usually not well acclimated and often suffer from acute mountain sickness (AMS). The aim of this study was to examine the effect of a single high altitude exposure on the changes in brain MRI and neuropsychological testing in climbers. METHODS: Brain MRI, medical history, and a battery of neuropsychological tests were obtained in eight male climbers between 31 and 48 yr of age a few days before and between 5 and 10 d after returning to sea level following ascent to altitudes of over 6000 m without oxygen. RESULTS: The mean AMS symptom score recorded at 5500 m was three in all climbers, headache being the predominant symptom. CONCLUSION: We did not observe the changes in brain imaging and in neuropsychological testing observed by other authors. The residual central nervous system impairment following return from high altitude was not observed in our study, and the good results in neuropsychological testing were well correlated with the unchanged brain MRI imaging.     

Epitope mapping of PR81 anti-MUC1 monoclonal antibody following PEPSCAN and phage display techniques

PR81 is an anti-MUC1 monoclonal antibody (MAb) which was generated against human MUC1 mucin that reacted with breast cancerous tissue, MUC1 positive cell line (MCF-7, BT-20, and T-4 7 D), and synthetic peptide, including the tandem repeat sequence of MUC1. Here we characterized the binding properties of PR81 against the tandem repeat of MUC1 by two different epitope mapping techniques, namely, PEPSCAN and phage display. Epitope mapping of PR81 MAb by PEPSCAN revealed a minimal consensus binding sequence, PDTRP, which is found on MUC1 peptide as the most important epitope. Using the phage display peptide library, we identified the motif PD(T/S/G)RP as an epitope and the motif AVGLSPDGSRGV as a mimotope recognized by PR81. Results of these two methods showed that the two residues, arginine and aspartic acid, have important roles in antibody binding and threonine can be substituted by either glycine or serine. These results may be of importance in tailor making antigens used in immunoassay.     

Autoimmune Cytopenias and Associated Conditions in CVID: a Report From the USIDNET Registry

Purpose

Autoimmune cytopenia is frequently a presenting manifestation of common variable immune deficiency (CVID). Studies characterizing the CVID phenotype associated with autoimmune cytopenias have mostly been limited to large referral centers. Here, we report prevalence of autoimmune cytopenias in CVID from the USIDNET Registry and compare the demographics and clinical features of patients with and without this complication.

Methods

Investigators obtained demographic, laboratory, and clinical data on CVID patients within the USIDNET Registry. Patients were considered to have autoimmune cytopenia if they had a diagnosis of hemolytic anemia, immune thrombocytopenia (ITP), or autoimmune neutropenia. Baseline characteristics and associated complications of those with autoimmune cytopenia (+AC) and those without (?AC) were compared.

Results

Of 990 CVID patients included in the analysis, 10.2% (N = 101) had a diagnosis consistent with autoimmune cytopenia: ITP was diagnosed in 7.4% (N = 73), hemolytic anemia in 4.5% (N = 45), and autoimmune neutropenia in 1% (N = 10). Age at diagnosis, gender, and baseline Ig values did not differ between the +AC and –AC groups. The +AC group was significantly more likely to have one or more other CVID-associated non-infectious complications (OR = 2.9; 95%-CI: 1.9–4.6, P < 0.001), including lymphoproliferation, granulomatous disease, lymphomas, hepatic disease, interstitial lung diseases, enteropathy, and organ-specific autoimmunity.

Conclusions

Autoimmune cytopenias are a common manifestation in CVID and are likely to be associated with other non-infectious CVID-related conditions. In light of prior studies showing increased morbidity and mortality in CVID patients with such complications, a diagnosis of autoimmune cytopenia may have prognostic significance in CVID.

     

Pharmacological characterization of CXC chemokine receptor 3 ligands and a small molecule antagonist

The CXC chemokine receptor 3 (CXCR3) is predominantly expressed on T helper type 1 (Th1) cells that are involved in inflammatory diseases. The three CXCR3 ligands CXCL9, CXCL10, and CXCL11 are produced at sites of inflammation and elicit migration of pathological Th1 cells. Here, we are the first to characterize the pharmacological potencies and specificity of a CXCR3 antagonist, N-1R-[3-(4-ethoxy-phenyl)-4-oxo-3,4-dihydro-pyrido[2,3-d]pyrimidin-2-yl]-ethyl-N-pyridin-3-ylmethyl-2-(4-fluoro-3-trifluoromethyl-phenyl)-acetamide (NBI-74330), from the T487 small molecule series. NBI-74330 demonstrated potent inhibition of [(125)I]CXCL10 and [(125)I]CXCL11 specific binding (K(i) of 1.5 and 3.2 nM, respectively) and of functional responses mediated by CXCR3, such as ligand-induced guanosine 5'-O-(3-[(35)S]thio)triphosphate ([(35)S]GTPgammaS) binding, calcium mobilization, and cellular chemotaxis (IC(50) of 7 to 18 nM). NBI-74330 was selective for CXCR3 because it showed no significant inhibition of chemotactic responses to other chemokines and did not inhibit radioligand binding to a panel of nonchemokine G-protein coupled receptors. There was a striking difference in potencies among the three CXCR3 ligands, with CXCL11 > CXCL10 > CXCL9. A comparison of the rank order of K(i) values with the rank order of monocyte production levels of these three ligands revealed a precise inverse correlation, suggesting that the weaker receptor affinities of CXCL9 and CXCL10 were physiologically compensated for by an elevated expression, perhaps to maintain effectiveness of each ligand under physiological conditions.     

Treatment of Thyroid Eye Disease

Thyroid eye disease (TED) is an autoimmune disease characterized by varying degrees of proptosis, congestion and inflammation of the extraocular tissues, and eyelid retraction. It is usually seen in the setting of Graves’ disease, but the severity of TED does not necessarily correlate with the level of systemic disease in a given patient. It is very important, nonetheless, to try to achieve a euthyroid state to minimize the chances of exacerbation of TED. Treatment of TED is based on the signs and symptoms displayed by the patient; there is no “one size fits all” approach. Generally, it is advisable to start with conservative measures, such as ocular lubrication with artificial tears, to manage symptoms of chronic irritation and redness. It is also imperative that the patient be advised to quit smoking, because there is a clear link between smoking and disease activity. Medical treatment with systemic oral or pulsed intravenous corticosteroids should be reserved for patients with active inflammation resulting in increased orbital pressure, compressive optic neuropathy, severe periorbital edema, or similar presentations. Once there is significant improvement in the acute inflammation, it is useful to treat patients who have residual inflammation with external beam radiation in order to be able to wean the patient off steroids and avoid their well-known complications.