Evaluation of a new Sebia isoelectrofocusing kit for α1-antitrypsin phenotyping with the Hydrasys® System

Zerimech, Farid; Hennache, G.; Bellon, Franck; Barouh, Guy; Lafitte, J.J.; Porchet, Nicole; Balduyck, Malika

doi:10.1515/cclm.2008.036

Cited by 48 publications

(45 citation statements)

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“…To clarify whether AAT was being produced by the neutrophil, as has been described by others (15), or whether it was being absorbed from the serum, we compared the phenotype of MM and ZZ-AATD neutrophil membrane-associated AAT with that of a ZZ-AATD individual after liver transplantation. Figure 3A shows the different patterns of phenotypes frequently observed for serum of MM and ZZ-AATD individuals (16). The phenotype of the ZZ-AATD serum sample after liver transplantation was entirely MM, a result confirmed by the use of an ELISA specific for the ZZ protein (17,18) (Figure 3B).…”

Section: Figurementioning

confidence: 63%

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α-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8

Bergin¹,

Reeves²,

Meleady³

et al. 2010

J. Clin. Invest.

254

305

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Hereditary deficiency of the protein α-1 antitrypsin (AAT) causes a chronic lung disease in humans that is characterized by excessive mobilization of neutrophils into the lung. However, the reason for the increased neutrophil burden has not been fully elucidated. In this study we have demonstrated using human neutrophils that serum AAT coordinates both CXCR1-and soluble immune complex (sIC) receptor-mediated chemotaxis by divergent pathways. We demonstrated that glycosylated AAT can bind to IL-8 (a ligand for CXCR1) and that AAT-IL-8 complex formation prevented IL-8 interaction with CXCR1. Second, AAT modulated neutrophil chemotaxis in response to sIC by controlling membrane expression of the glycosylphosphatidylinositolanchored (GPI-anchored) Fc receptor FcγRIIIb. This process was mediated through inhibition of ADAM-17 enzymatic activity. Neutrophils isolated from clinically stable AAT-deficient patients were characterized by low membrane expression of FcγRIIIb and increased chemotaxis in response to IL-8 and sIC. Treatment of AATdeficient individuals with AAT augmentation therapy resulted in increased AAT binding to IL-8, increased AAT binding to the neutrophil membrane, decreased FcγRIIIb release from the neutrophil membrane, and normalization of chemotaxis. These results provide new insight into the mechanism underlying the effect of AAT augmentation therapy in the pulmonary disease associated with AAT deficiency.

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Section: Figurementioning

confidence: 63%

“…Controls for Western blotting included recombinant human (Rh) FcγRIIIb (R&D Systems) or serum purified AAT. A Sebia isoelectrofocusing kit was employed for AAT phenotyping with the HYDRASYS system as previously described (16).…”

Section: Methodsmentioning

confidence: 99%

α-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8

Bergin¹,

Reeves²,

Meleady³

et al. 2010

J. Clin. Invest.

254

305

View full text Add to dashboard Cite

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“…Characterization of AAT phenotypes was performed using the Hydrasys electrophoresis platform (Sebia) and the Hydragel 18 A1AT Isofocusing kit (Sebia, Evry, France) (15). AAT levels were measured by nephelometry (Siemens DadeBehring BN II, Tarrytown, NY).…”

Section: Determination Of Aat Phenotypementioning

confidence: 99%

Clarification of the Risk of Chronic Obstructive Pulmonary Disease in α₁-Antitrypsin Deficiency PiMZ Heterozygotes

Molloy

Hersh²,

Morris³

et al. 2014

Am J Respir Crit Care Med

160

128

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Rationale: Severe a 1 -antitrypsin deficiency (typically PiZZ homozygosity) is associated with a significantly increased risk of airflow obstruction and emphysema but the risk of chronic obstructive pulmonary disease (COPD) in PiMZ heterozygotes remains uncertain.Objectives: This was a family-based study to determine the risk of COPD in PiMZ individuals. (% predicted) (63.84 [38.45-84.35] vs. 72.8 [55.5-97.7]; P = 0.0013) compared with PiMM individuals. This effect was abrogated in never-smoking and accentuated in ever-smoking PiMZ individuals. PiMZ heterozygosity was associated with an adjusted odds ratio for COPD of 5.18 (95% confidence interval, 1.27-21.15; P = 0.02) and this was higher (odds ratio, 10.65; 95% confidence interval, 2.17-52.29; P = 0.004) in eversmoking individuals.Conclusions: These results indicate that PiMZ heterozygotes have significantly more airflow obstruction and COPD than PiMM individuals and cigarette smoke exposure exerts a significant modifier effect.

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“…IEF with or without immunoblotting separates proteins according to their isoelectric points (Figure 3), and enables the user to identify different AAT isoforms [Zerimech et al 2008].…”

Section: Phenotyping and Gene Sequencingmentioning

confidence: 99%

Alpha-1-antitrypsin deficiency: increasing awareness and improving diagnosis

Greulich¹,

Vogelmeier

2015

Ther Adv Respir Dis

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Alpha-1-antitrypsin deficiency (AATD) is a hereditary disorder that is characterized by a low serum level of alpha-1-antitrypsin (AAT). The loss of anti-inflammatory and antiproteolytic functions, together with pro-inflammatory effects of polymerized AAT contribute to protein degradation and increased inflammation resulting in an increased risk of developing chronic obstructive pulmonary disease (COPD) and emphysema, especially in smokers. AATD is a rare disease that is significantly underdiagnosed. According to recent data that are based on extrapolations, in many countries only 5-15% of homozygous individuals have been identified. Furthermore, the diagnostic delay typically exceeds 5 years, resulting in an average age at diagnosis of about 45 years. Although the American Thoracic Society/ European Respiratory Society recommendations state that all symptomatic adults with persistent airway obstruction should be screened, these recommendations are not being followed. Potential reasons for that include missing knowledge about the disease and the appropriate tests, and the low awareness of physicians with regard to the disorder. Once the decision to initiate testing has been made, a screening test (AAT serum level or other) should be performed. Further diagnostic evaluation is based on the following techniques: polymerase chain reaction (PCR) for frequent and clinically important mutations, isoelectric focusing (IEF) with or without immunoblotting, and sequencing of the gene locus coding for AAT. Various diagnostic algorithms have been published for AATD detection (severe deficiency or carrier status). Modern laboratory approaches like the use of serum separator cards, a lateral flow assay to detect the Z-protein, and a broader availability of next-generation sequencing are recent advances, likely to alter existing algorithms.

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Evaluation of a new Sebia isoelectrofocusing kit for α1-antitrypsin phenotyping with the Hydrasys® System

Abstract: This new method is highly specific, rapid and simple to perform. It improves identification of not only the most common but also various rare A1AT phenotypes. It appears to be suitable for routine analysis and screening applications in a clinical laboratory setting.

Cited by 48 publications

References 12 publications

α-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8

α-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8

Clarification of the Risk of Chronic Obstructive Pulmonary Disease in α₁-Antitrypsin Deficiency PiMZ Heterozygotes

Alpha-1-antitrypsin deficiency: increasing awareness and improving diagnosis

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Evaluation of a new Sebia isoelectrofocusing kit for α1-antitrypsin phenotyping with the Hydrasys® System

Abstract: This new method is highly specific, rapid and simple to perform. It improves identification of not only the most common but also various rare A1AT phenotypes. It appears to be suitable for routine analysis and screening applications in a clinical laboratory setting.

Cited by 48 publications

References 12 publications

α-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8

α-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8

Clarification of the Risk of Chronic Obstructive Pulmonary Disease in α1-Antitrypsin Deficiency PiMZ Heterozygotes

Alpha-1-antitrypsin deficiency: increasing awareness and improving diagnosis

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Product

Resources

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Clarification of the Risk of Chronic Obstructive Pulmonary Disease in α₁-Antitrypsin Deficiency PiMZ Heterozygotes