Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin

M. A. Casolaro, G. Fells, M. Wewers, J. E. Pierce, F. Ogushi, R. Hubbard, S. Sellers, J. Forstrom, D. Lyons, G. Kawasaki, Ronald Crystal

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

To evaluate the potential use of recombinant DNA-produced α-1-antitrypsin (α-1-AT) to augment the lung antineutrophil elastase defenses in α-1-AT deficiency, we compared the kinetics of intravenously administered recombinant produced α-1-AT (rα-1-AT) and purified normal human plasma α-1-AT (pα-1-AT) in the blood and lung of rhesus monkeys. The rα-1-AT was produced in yeast transformed with an expressing plasmid containing a full-length human α-1-AT complementary deoxyribonucleic acid and purified to > 99% homogeneity. The rα-1-AT has a molecular weight of 45,000, no carbohydrates, and is identical in sequence to normal plasma α-1-AT except for an additional N-terminal acetylmethionine. Despite its lack of carbohydrates, the rα-1-AT inhibited human neutrophil elastase with an association rate constant similar to that of pα-1-AT. Rhesus monkeys were infused intravenously with 120 mg/kg of rα-1-AT (n = 13) or pα-1-AT (n = 12) and the serum, urine, and lung epithelial lining fluid (ELF) concentrations of these molecules quantified at various intervals. Although the initial serum levels of the rα-1-AT and pα-1-AT were both dose dependent, the pα-1-AT remained in the blood for at least 4 days, whereas the rα-1-AT disappared rapidly, such that it was barely detectable at 24 h and strikingly, although no pα-1-AT was detectable in the urine at any time, 38% of the intravenously administered rα-1-AT was excreted within 3 h. Similar to its behavior in humans, the pα-1-AT diffused into the lung such that its concentration in the ELF of the lower respiratory tract 1-4 days after infusion was ~ 10% that in serum. Interestingly, the rα-1-AT molecule also diffused into the lung, with ELF levels at 24 h similar to that of the pα-1-AT. Furthermore, although the rα-1-AT ELF levels declined by 48 and 96 h to below those of the pα-1-AT ELF levels, the rα-1-AT ELF levels exceeded those in blood at the same time points and, like the pα-1-AT, resulted in a significant augmentation of the antineutrophil elastase capacity of the ELF. Thus, in primates, human-based rα-1-AT has very different pharmacokinetics than does human pα-1-AT, likely because of its modified charge and/or conformation. Despite this, however, the rα-1-AT does diffuse into the lung and augment the antineutrophil elastase capacity of the ELF of the lower respiratory tract, suggesting that it has potential as a therapeutic agent in the treatment of disorders such as α-1-AT deficiency.

Original languageEnglish
Pages (from-to)2015-2023
Number of pages9
JournalJournal of Applied Physiology
Volume63
Issue number5
Publication statusPublished - 1 Dec 1987
Externally publishedYes

Fingerprint

Pancreatic Elastase
Lung
Macaca mulatta
Respiratory System
Serum
Carbohydrates
Urine
Leukocyte Elastase
Recombinant DNA
Primates
Plasmids
Pharmacokinetics
Yeasts
Molecular Weight

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Casolaro, M. A., Fells, G., Wewers, M., Pierce, J. E., Ogushi, F., Hubbard, R., ... Crystal, R. (1987). Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin. Journal of Applied Physiology, 63(5), 2015-2023.

Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin. / Casolaro, M. A.; Fells, G.; Wewers, M.; Pierce, J. E.; Ogushi, F.; Hubbard, R.; Sellers, S.; Forstrom, J.; Lyons, D.; Kawasaki, G.; Crystal, Ronald.

In: Journal of Applied Physiology, Vol. 63, No. 5, 01.12.1987, p. 2015-2023.

Research output: Contribution to journalArticle

Casolaro, MA, Fells, G, Wewers, M, Pierce, JE, Ogushi, F, Hubbard, R, Sellers, S, Forstrom, J, Lyons, D, Kawasaki, G & Crystal, R 1987, 'Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin', Journal of Applied Physiology, vol. 63, no. 5, pp. 2015-2023.
Casolaro MA, Fells G, Wewers M, Pierce JE, Ogushi F, Hubbard R et al. Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin. Journal of Applied Physiology. 1987 Dec 1;63(5):2015-2023.
Casolaro, M. A. ; Fells, G. ; Wewers, M. ; Pierce, J. E. ; Ogushi, F. ; Hubbard, R. ; Sellers, S. ; Forstrom, J. ; Lyons, D. ; Kawasaki, G. ; Crystal, Ronald. / Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin. In: Journal of Applied Physiology. 1987 ; Vol. 63, No. 5. pp. 2015-2023.
@article{ccd8ea823ef64af799f133133fa64450,
title = "Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin",
abstract = "To evaluate the potential use of recombinant DNA-produced α-1-antitrypsin (α-1-AT) to augment the lung antineutrophil elastase defenses in α-1-AT deficiency, we compared the kinetics of intravenously administered recombinant produced α-1-AT (rα-1-AT) and purified normal human plasma α-1-AT (pα-1-AT) in the blood and lung of rhesus monkeys. The rα-1-AT was produced in yeast transformed with an expressing plasmid containing a full-length human α-1-AT complementary deoxyribonucleic acid and purified to > 99{\%} homogeneity. The rα-1-AT has a molecular weight of 45,000, no carbohydrates, and is identical in sequence to normal plasma α-1-AT except for an additional N-terminal acetylmethionine. Despite its lack of carbohydrates, the rα-1-AT inhibited human neutrophil elastase with an association rate constant similar to that of pα-1-AT. Rhesus monkeys were infused intravenously with 120 mg/kg of rα-1-AT (n = 13) or pα-1-AT (n = 12) and the serum, urine, and lung epithelial lining fluid (ELF) concentrations of these molecules quantified at various intervals. Although the initial serum levels of the rα-1-AT and pα-1-AT were both dose dependent, the pα-1-AT remained in the blood for at least 4 days, whereas the rα-1-AT disappared rapidly, such that it was barely detectable at 24 h and strikingly, although no pα-1-AT was detectable in the urine at any time, 38{\%} of the intravenously administered rα-1-AT was excreted within 3 h. Similar to its behavior in humans, the pα-1-AT diffused into the lung such that its concentration in the ELF of the lower respiratory tract 1-4 days after infusion was ~ 10{\%} that in serum. Interestingly, the rα-1-AT molecule also diffused into the lung, with ELF levels at 24 h similar to that of the pα-1-AT. Furthermore, although the rα-1-AT ELF levels declined by 48 and 96 h to below those of the pα-1-AT ELF levels, the rα-1-AT ELF levels exceeded those in blood at the same time points and, like the pα-1-AT, resulted in a significant augmentation of the antineutrophil elastase capacity of the ELF. Thus, in primates, human-based rα-1-AT has very different pharmacokinetics than does human pα-1-AT, likely because of its modified charge and/or conformation. Despite this, however, the rα-1-AT does diffuse into the lung and augment the antineutrophil elastase capacity of the ELF of the lower respiratory tract, suggesting that it has potential as a therapeutic agent in the treatment of disorders such as α-1-AT deficiency.",
author = "Casolaro, {M. A.} and G. Fells and M. Wewers and Pierce, {J. E.} and F. Ogushi and R. Hubbard and S. Sellers and J. Forstrom and D. Lyons and G. Kawasaki and Ronald Crystal",
year = "1987",
month = "12",
day = "1",
language = "English",
volume = "63",
pages = "2015--2023",
journal = "Journal of Applied Physiology",
issn = "8750-7587",
publisher = "American Physiological Society",
number = "5",

}

TY - JOUR

T1 - Augmentation of lung antineutrophil elastase capacity with recombinant human α-1-antitrypsin

AU - Casolaro, M. A.

AU - Fells, G.

AU - Wewers, M.

AU - Pierce, J. E.

AU - Ogushi, F.

AU - Hubbard, R.

AU - Sellers, S.

AU - Forstrom, J.

AU - Lyons, D.

AU - Kawasaki, G.

AU - Crystal, Ronald

PY - 1987/12/1

Y1 - 1987/12/1

N2 - To evaluate the potential use of recombinant DNA-produced α-1-antitrypsin (α-1-AT) to augment the lung antineutrophil elastase defenses in α-1-AT deficiency, we compared the kinetics of intravenously administered recombinant produced α-1-AT (rα-1-AT) and purified normal human plasma α-1-AT (pα-1-AT) in the blood and lung of rhesus monkeys. The rα-1-AT was produced in yeast transformed with an expressing plasmid containing a full-length human α-1-AT complementary deoxyribonucleic acid and purified to > 99% homogeneity. The rα-1-AT has a molecular weight of 45,000, no carbohydrates, and is identical in sequence to normal plasma α-1-AT except for an additional N-terminal acetylmethionine. Despite its lack of carbohydrates, the rα-1-AT inhibited human neutrophil elastase with an association rate constant similar to that of pα-1-AT. Rhesus monkeys were infused intravenously with 120 mg/kg of rα-1-AT (n = 13) or pα-1-AT (n = 12) and the serum, urine, and lung epithelial lining fluid (ELF) concentrations of these molecules quantified at various intervals. Although the initial serum levels of the rα-1-AT and pα-1-AT were both dose dependent, the pα-1-AT remained in the blood for at least 4 days, whereas the rα-1-AT disappared rapidly, such that it was barely detectable at 24 h and strikingly, although no pα-1-AT was detectable in the urine at any time, 38% of the intravenously administered rα-1-AT was excreted within 3 h. Similar to its behavior in humans, the pα-1-AT diffused into the lung such that its concentration in the ELF of the lower respiratory tract 1-4 days after infusion was ~ 10% that in serum. Interestingly, the rα-1-AT molecule also diffused into the lung, with ELF levels at 24 h similar to that of the pα-1-AT. Furthermore, although the rα-1-AT ELF levels declined by 48 and 96 h to below those of the pα-1-AT ELF levels, the rα-1-AT ELF levels exceeded those in blood at the same time points and, like the pα-1-AT, resulted in a significant augmentation of the antineutrophil elastase capacity of the ELF. Thus, in primates, human-based rα-1-AT has very different pharmacokinetics than does human pα-1-AT, likely because of its modified charge and/or conformation. Despite this, however, the rα-1-AT does diffuse into the lung and augment the antineutrophil elastase capacity of the ELF of the lower respiratory tract, suggesting that it has potential as a therapeutic agent in the treatment of disorders such as α-1-AT deficiency.

AB - To evaluate the potential use of recombinant DNA-produced α-1-antitrypsin (α-1-AT) to augment the lung antineutrophil elastase defenses in α-1-AT deficiency, we compared the kinetics of intravenously administered recombinant produced α-1-AT (rα-1-AT) and purified normal human plasma α-1-AT (pα-1-AT) in the blood and lung of rhesus monkeys. The rα-1-AT was produced in yeast transformed with an expressing plasmid containing a full-length human α-1-AT complementary deoxyribonucleic acid and purified to > 99% homogeneity. The rα-1-AT has a molecular weight of 45,000, no carbohydrates, and is identical in sequence to normal plasma α-1-AT except for an additional N-terminal acetylmethionine. Despite its lack of carbohydrates, the rα-1-AT inhibited human neutrophil elastase with an association rate constant similar to that of pα-1-AT. Rhesus monkeys were infused intravenously with 120 mg/kg of rα-1-AT (n = 13) or pα-1-AT (n = 12) and the serum, urine, and lung epithelial lining fluid (ELF) concentrations of these molecules quantified at various intervals. Although the initial serum levels of the rα-1-AT and pα-1-AT were both dose dependent, the pα-1-AT remained in the blood for at least 4 days, whereas the rα-1-AT disappared rapidly, such that it was barely detectable at 24 h and strikingly, although no pα-1-AT was detectable in the urine at any time, 38% of the intravenously administered rα-1-AT was excreted within 3 h. Similar to its behavior in humans, the pα-1-AT diffused into the lung such that its concentration in the ELF of the lower respiratory tract 1-4 days after infusion was ~ 10% that in serum. Interestingly, the rα-1-AT molecule also diffused into the lung, with ELF levels at 24 h similar to that of the pα-1-AT. Furthermore, although the rα-1-AT ELF levels declined by 48 and 96 h to below those of the pα-1-AT ELF levels, the rα-1-AT ELF levels exceeded those in blood at the same time points and, like the pα-1-AT, resulted in a significant augmentation of the antineutrophil elastase capacity of the ELF. Thus, in primates, human-based rα-1-AT has very different pharmacokinetics than does human pα-1-AT, likely because of its modified charge and/or conformation. Despite this, however, the rα-1-AT does diffuse into the lung and augment the antineutrophil elastase capacity of the ELF of the lower respiratory tract, suggesting that it has potential as a therapeutic agent in the treatment of disorders such as α-1-AT deficiency.

UR - http://www.scopus.com/inward/record.url?scp=0023588232&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023588232&partnerID=8YFLogxK

M3 - Article

VL - 63

SP - 2015

EP - 2023

JO - Journal of Applied Physiology

JF - Journal of Applied Physiology

SN - 8750-7587

IS - 5

ER -