Serum Antielastase and Neutrophil Elastase Levels in PiM Phenotype Cigarette Smokers with Airflow Obstruction

Abstract

In order to assess blood factors which might explain why some cigarette smokers develop airflow obstruction while others do not, we compared two groups of PiM phenotype volunteers matched for age, sex and total pack-years of cigarette smoking; one group had airflow obstruction and the other did not. Functional levels of alpha-2-macroglobulin (alpha-2-M) and alpha-1-protease inhibitor (alpha-1-PI) were separately assessed by a protease binding procedure. Neutrophils were isolated from blood by counterflow centrifugation, and their elastase content was assayed with ³H-elastin-SDS (sodium dodecyl sulfate). The obstructed and nonobstructed groups were not different with respect to functional or immunoreactive levels of alpha-1-PI and alpha-2-M or elastase levels in their neutrophils. We do not find imbalances of circulating elastase or antielastase levels in PiM phenotype smokers with airflow obstruction.

Defenses of the Hamster Lung against Human Neutrophil and Porcine Pancreatic Elastase

“…The properties of our preparations of HNE were indistinguishable from those reported by others [4,[7][8][9][10]. The optical density at 280 nm (E|'"," 280 nm = 9.85) was used to deter mine the enzyme concentration [9J.…”

“…with its active site protected by the pres ence of a saturating concentration of peptide sub strate, was radiolabeled with IJ5ICI as previously de scribed [8]. The radioiodinated material exhibited a single peak of radioactivity, corresponding to a mo lecular mass of 29,000 daltons, in a 7.5% SDS poly acrylamide gel [8], Elastolytic assays of l!…”

“…with its active site protected by the pres ence of a saturating concentration of peptide sub strate, was radiolabeled with IJ5ICI as previously de scribed [8]. The radioiodinated material exhibited a single peak of radioactivity, corresponding to a mo lecular mass of 29,000 daltons, in a 7.5% SDS poly acrylamide gel [8], Elastolytic assays of l! 'l-HNE with tritiated elastin, after subtracting the contribu tion of radioiodine to the measurement of radioactiv ity in the tritium channel, indicated no loss of activity from the radioiodination procedure [7,13].…”

“…16]. The amount of elastase present in a component was cal culated by correcting the recovered radioactivity for the overall loss on the column and then dividing by the specific radioactivity of the elastase preparation per microgram as previously described [8,16]. Mean recovery of radioactivity from the columns exceeded 85%.…”

“…The amino acid compo sition was similar to published values: the mean devi ation in residues per 1.000 for each amino acid was 3 as compared with that determined by Lo et al [22], Determination of the concentration of this a-l-PI so lution with the serum-calibrated plates yielded a val ue within I SE (±6%) of the value expected based upon the solution's light absorbance. When possible, plasma samples were stored at 4°C and run within 24 h, since the functional efficiency of a-l-PI dimin ishes during prolonged freezing at -20°C [8]. When longer storage was necessary samples were stored at -80°C.…”