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Prolastin, a pharmaceutical preparation of purified human alpha 1-antitrypsin, blocks endotoxin-mediated cytokine release

Nita, Izabela LU ; Hollander, Camilla LU ; Westin, Ulla LU and Janciauskiene, Sabina LU (2005) In Respiratory Research 6(12).
Abstract
Background: alpha 1-antitrypsin (AAT) serves primarily as an inhibitor of the elastin degrading proteases, neutrophil elastase and proteinase 3. There is ample clinical evidence that inherited severe AAT deficiency predisposes to chronic obstructive pulmonary disease. Augmentation therapy for AAT deficiency has been available for many years, but to date no sufficient data exist to demonstrate its efficacy. There is increasing evidence that AAT is able to exert effects other than protease inhibition. We investigated whether Prolastin, a preparation of purified pooled human AAT used for augmentation therapy, exhibits antibacterial effects. Methods: Human monocytes and neutrophils were isolated from buffy coats or whole peripheral blood by... (More)
Background: alpha 1-antitrypsin (AAT) serves primarily as an inhibitor of the elastin degrading proteases, neutrophil elastase and proteinase 3. There is ample clinical evidence that inherited severe AAT deficiency predisposes to chronic obstructive pulmonary disease. Augmentation therapy for AAT deficiency has been available for many years, but to date no sufficient data exist to demonstrate its efficacy. There is increasing evidence that AAT is able to exert effects other than protease inhibition. We investigated whether Prolastin, a preparation of purified pooled human AAT used for augmentation therapy, exhibits antibacterial effects. Methods: Human monocytes and neutrophils were isolated from buffy coats or whole peripheral blood by the Ficoll-Hypaque procedure. Cells were stimulated with lipopolysaccharide (LPS) or zymosan, either alone or in combination with Prolastin, native AAT or polymerised AAT for 18 h, and analysed to determine the release of TNF alpha, IL-1 beta and IL-8. At 2-week intervals, seven subjects were submitted to a nasal challenge with sterile saline, LPS ( 25 mu g) and LPS-Prolastin combination. The concentration of IL-8 was analysed in nasal lavages performed before, and 2, 6 and 24 h after the challenge. Results: In vitro, Prolastin showed a concentration-dependent (0.5 to 16 mg/ml) inhibition of endotoxin-stimulated TNF alpha and IL-1 beta release from monocytes and IL-8 release from neutrophils. At 8 and 16 mg/ml the inhibitory effects of Prolastin appeared to be maximal for neutrophil IL- 8 release (5.3-fold, p < 0.001 compared to zymosan treated cells) and monocyte TNFa and IL- 1 release (10.7- and 7.3-fold, p < 0.001, respectively, compared to LPS treated cells). Furthermore, Prolastin (2.5 mg per nostril) significantly inhibited nasal IL- 8 release in response to pure LPS challenge. Conclusion: Our data demonstrate for the first time that Prolastin inhibits bacterial endotoxin-induced pro-inflammatory responses in vitro and in vivo, and provide scientific bases to explore new Prolastin-based therapies for individuals with inherited AAT deficiency, but also for other clinical conditions. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Respiratory Research
volume
6
issue
12
publisher
BioMed Central
external identifiers
  • wos:000227573200001
  • pmid:15683545
  • scopus:25644448893
ISSN
1465-9921
DOI
10.1186/1465-9921-6-12
language
English
LU publication?
yes
id
b1deca11-5855-4141-9d88-22fa0c0f150f (old id 248288)
date added to LUP
2007-08-22 13:58:52
date last changed
2017-02-19 03:27:46
@article{b1deca11-5855-4141-9d88-22fa0c0f150f,
  abstract     = {Background: alpha 1-antitrypsin (AAT) serves primarily as an inhibitor of the elastin degrading proteases, neutrophil elastase and proteinase 3. There is ample clinical evidence that inherited severe AAT deficiency predisposes to chronic obstructive pulmonary disease. Augmentation therapy for AAT deficiency has been available for many years, but to date no sufficient data exist to demonstrate its efficacy. There is increasing evidence that AAT is able to exert effects other than protease inhibition. We investigated whether Prolastin, a preparation of purified pooled human AAT used for augmentation therapy, exhibits antibacterial effects. Methods: Human monocytes and neutrophils were isolated from buffy coats or whole peripheral blood by the Ficoll-Hypaque procedure. Cells were stimulated with lipopolysaccharide (LPS) or zymosan, either alone or in combination with Prolastin, native AAT or polymerised AAT for 18 h, and analysed to determine the release of TNF alpha, IL-1 beta and IL-8. At 2-week intervals, seven subjects were submitted to a nasal challenge with sterile saline, LPS ( 25 mu g) and LPS-Prolastin combination. The concentration of IL-8 was analysed in nasal lavages performed before, and 2, 6 and 24 h after the challenge. Results: In vitro, Prolastin showed a concentration-dependent (0.5 to 16 mg/ml) inhibition of endotoxin-stimulated TNF alpha and IL-1 beta release from monocytes and IL-8 release from neutrophils. At 8 and 16 mg/ml the inhibitory effects of Prolastin appeared to be maximal for neutrophil IL- 8 release (5.3-fold, p &lt; 0.001 compared to zymosan treated cells) and monocyte TNFa and IL- 1 release (10.7- and 7.3-fold, p &lt; 0.001, respectively, compared to LPS treated cells). Furthermore, Prolastin (2.5 mg per nostril) significantly inhibited nasal IL- 8 release in response to pure LPS challenge. Conclusion: Our data demonstrate for the first time that Prolastin inhibits bacterial endotoxin-induced pro-inflammatory responses in vitro and in vivo, and provide scientific bases to explore new Prolastin-based therapies for individuals with inherited AAT deficiency, but also for other clinical conditions.},
  author       = {Nita, Izabela and Hollander, Camilla and Westin, Ulla and Janciauskiene, Sabina},
  issn         = {1465-9921},
  language     = {eng},
  number       = {12},
  publisher    = {BioMed Central},
  series       = {Respiratory Research},
  title        = {Prolastin, a pharmaceutical preparation of purified human alpha 1-antitrypsin, blocks endotoxin-mediated cytokine release},
  url          = {http://dx.doi.org/10.1186/1465-9921-6-12},
  volume       = {6},
  year         = {2005},
}