TBPP_Alpha-1 Antitrypsin (AAT)

inhibition of neutrophil elastase by AAT, conformational change by binding of active serine protease and RCL, elastase inactivation: movement of the upper to the lower pole of the protein, RCL inserted into ?-sheet: prevents release of protease.

1. method: purification AAT using an immunoaffinity column with antibodies, protection of protein: mild elution conditions. 2. method: base fractionation, purification (extraction, filtration, IEC and HIC), concentration, viral inactivation (pasteurization, ultrafiltration), concentration

wash cells (Ca2+/Mg2+ buffer), add trypsin-EDTA, cell incubation (37°C, 5-10 min.), add serum, harvest cells, centrifugation, resuspension, cell counting, seed cells with appropriate density

patients with levels < 70mg/dL, pulmonary: respiratory disease, protease-antiprotease imbalance, inflammation, mechanical damage. Non-pulmonary: liver disease (AAT polymers in the liver), skin disease, vasculitis. Therapy: life-long weekly infusions with plasma-derived AAT

described by Laurrel and Eriksson in 1963, Chronic Obstructive Pulmonary Disease (COPD), autosomal recessive, SERPINA1 gene, 123 mutations described, alleles: M (normal), S, null and Z (pathogenic), Z mutation is the most common (Glu342Lys, ?-sheet is wider, RCL can accept other AAT = accumulation): polymerization and retention of AAT in hepatocytes (liver) leading to liver cirrhosis and decreased AAT in circulation and in the lung, Z-type polymers cause inflammation

interaction with endogenous proteins (cytokines, caspase 3), interactions with cells (leukocytes), interaction with pathogens (viruses), inhibition of TLR mediated cytokine release in human neutrophils

not on the market, AAT without elastase inhibitory capacity, recombinant AAT more potent inhibitor of LPS driven cytokine release, but no receptor for AAT identified yet

islet transplantation, skin transplantation, autoimmune diabetes, cancer