PubMed per Alfa1AT

[Alpha1-antitrypsin deficiency in Austria: analysis of the Austrian Alpha1-international-registry database.]

Wien Klin Wochenschr. 2010 Jul 14;

Authors: Huber F, Schmid-Scherzer K, Wantke F, Frantal S, Kneussl M

OBJECTIVE: Alpha1-antitrypsin deficiency is a rare hereditary disorder. Deficient patients are at a higher risk to develop lung emphysema at an early age. The alpha1-antitrypsin registry was founded on 1996 to get new insights into the pathogenesis of the disease and to develop new therapeutic strategies. In this study the epidemiological and clinical findings of the Austrian alpha1-antitrypsin deficient patients are presented. MATERIAL AND METHODS: A total of 139 patients with severe alpha1-antitrypsin deficiency, identified by phenol- and genotyping, were evaluated retrospectively by their physicians with the help of a questionnaire. RESULTS: Most patients were smokers or ex-smokers (71.9%) who developed symptoms in their fourth decade. The mean duration between the onset of symptoms and the final diagnosis was 6.5 years. About 25% of the evaluated patients were unable to practice their profession because of their illness. CONCLUSION: Alpha1-antitrypsin deficiency is a rare condition with delayed diagnosis. Because of the benefit of an early diagnosis further effort should be put towards early detection.

PMID: 20623257 [PubMed - as supplied by publisher]

Intradermal alpha1-antitrypsin therapy avoids fatal anaphylaxis, prevents type 1 diabetes and reverses hyperglycaemia in the NOD mouse model of the disease.

Diabetologia. 2010 Jul 1;

Authors: Ma H, Lu Y, Li H, Campbell-Thompson M, Parker M, Wasserfall C, Haller M, Brantly M, Schatz D, Atkinson M, Song S

AIMS/HYPOTHESIS: Human alpha1-antitrypsin (hAAT) gene therapy prevents type 1 diabetes in a NOD mouse model of diabetes. However, repeated i.p. injections of hAAT into NOD mice leads to fatal anaphylaxis. The aim of the study was to determine if an alternative route of administration avoids anaphylaxis and allows evaluation of hAAT's potential for diabetes prevention and reversal. We also sought to determine if the addition of granulocyte colony-stimulating factor (G-CSF), augments hAAT's capacity to prevent or reverse disease in the NOD mice. METHODS: To evaluate hAAT pharmacokinetics, serum hAAT levels were monitored in NOD mice receiving a single dose (2 mg) of hAAT by i.p., s.c. or i.d. injection. For studies of type 1 diabetes prevention and reversal, mice received i.d. hAAT (2 mg/mouse/3 days) for 8 or 10 weeks or hAAT and G-CSF (i.p., 6 mug/day) for 6 weeks. Blood glucose determinations, glucose tolerance testing and insulin tolerance tests were performed. RESULTS: Both i.p. and s.c. injections resulted in fatal anaphylaxis. The i.d. injection avoided anaphylaxis and i.d. injection of hAAT into 11-week-old NOD mice prevented disease (p = 0.005, AAT vs PBS at 40 weeks of age). Treatment of diabetic NOD mice with hAAT or hAAT plus G-CSF provided long-term (at least 100 days) reversal of diabetes in 50% of treated animals. G-CSF did not enhance the reversal rates of hAAT. Glucose tolerance and insulin levels were normalised in mice with hAAT prevention and reversal. CONCLUSIONS/INTERPRETATION: Intradermal hAAT prevents and reverses disease in a NOD mouse model of type 1 diabetes without inducing anaphylaxis.

PMID: 20593162 [PubMed - as supplied by publisher]

Specific detection of gastric alpha1-antitrypsin by immobilized trypsin on polyHEMA films.

Mol Pharm. 2010 Jun 28;

Authors: Rubinstein A, Khazanov E, Emanuel N, Azab AK, Yechezkel B, Yavin E

Early diagnosis of gastric carcinoma is crucial for maximizing medical treatment efficacy. For the purpose of real time diagnosis ("virtual biopsy") of stomach-malignancy we developed a polyHEMA platform capable of capturing human alpha1- antitrypsin precursor (A1AT), a model proteinaceous luminal biomarker. Its specific attachment to the polymeric platform was accomplished by immobilized trypsin, which was linked to the surface of the polyHEMA film by a series of PEG-based spacers. Recognition was enabled by adapting an ELISA-like methodology, using rabbit anti-A1AT and HRP-conjugated anti-rabbit IgG as a secondary antibody. Since this A1AT-sensing platform was designed to be detected by endoscopic means such as a video capsule, its physical stability was tested after casting on top of a polycarbonate surface. It was found that, in contrast to classical ELISA analysis, performed on polystyrene plates, A1AT detection was possible only when spacer arms were used to immobilize the capturing moiety, trypsin, with a 7-fold increase in the optical signal and a linear dependency upon the concentration of the A1AT biomarker.

PMID: 20583815 [PubMed - as supplied by publisher]

Ex Vivo Transduction and Transplantation of Bone Marrow Cells for Liver Gene Delivery of alpha1-Antitrypsin.

Mol Ther. 2010 Jun 15;

Authors: Li H, Lu Y, Witek RP, Chang LJ, Campbell-Thompson M, Jorgensen M, Petersen B, Song S

Adult stem cell-based gene therapy holds several unique advantages including avoidance of germline or other undesirable cell transductions. We have previously shown that liver progenitor (oval) cells can be used as a platform for liver gene delivery of human alpha1-antitrypsin (hAAT). However, this cell source cannot be used in humans for autologous transplantation. In the present study, we tested the feasibility of bone marrow (BM) cell-based liver gene delivery of hAAT. In vitro studies showed that BM cells can be transduced by lentiviral vector (Lenti-CB-hAAT) and recombinant adeno-associated viral vectors (rAAV1-CB-hAAT, and rAAV8-CB-hAAT). Transplantation studies showed that transplanted BM cells homed into liver, differentiated into hepatocytes and expressed hAAT in the liver. Importantly, we showed that transplantation of rAAV8-CB-hAAT vector-transduced BM cells resulted in sustained levels of hAAT in the systemic circulation of recipient mice. These results demonstrated that rAAV vector-mediated BM cell-based liver gene therapy is feasible for the treatment of AAT deficiency and implies a novel therapy for the treatment of liver diseases.

PMID: 20551917 [PubMed - as supplied by publisher]

Role of the lectin VIP36 in post-ER quality control of human alpha1-antitrypsin.

Traffic. 2010 May 7;

Authors: Reiterer V, Nyfeler B, Hauri HP

The L-type lectin VIP36 localizes to the Golgi apparatus and cycles early in the secretory pathway. In vitro, VIP36 binds high mannose glycans with a pH optimum of 6.5, a value similar to the luminal pH of the Golgi apparatus. Although the sugar binding properties of VIP36 in vitro have been characterized in detail, the function of VIP36 in the intact cell remains unclear since no convincing glycoprotein cargo has been identified. Here we used yellow fluorescent protein (YFP) fragment complementation to identify luminal interaction partners of VIP36. By screening a human liver cDNA library, we identified the glycoprotein alpha-1-antitrypsin (alpha1-AT) as a cargo of VIP36. The VIP36/ alpha1-AT complex localized to Golgi and ER. In the living cell, VIP36 bound exclusively to the high-mannose form of alpha1-AT. The binding was increased when complex glycosylation was prevented by kifunensine and abolished when the glycosylation sites of alpha1-AT were inactivated by mutagenesis. Silencing VIP36 accelerated alpha1-AT transport, arguing against a role of VIP36 in anterograde traffic. The complex formed by VIP36 and alpha1-AT in the Golgi recycled back to the ER. The combined data are most consistent with a function of VIP36 in post-ER quality control of alpha1-AT.

PMID: 20477988 [PubMed - as supplied by publisher]