Size distribution of human lung elastin-derived peptide antigens generated in vitro and in vivo.
The protease-antiprotease hypothesis of emphysema development suggests that degradation of elastin in the lung interstitium may give rise to abnormal quantities of circulating elastin-derived peptides (EDP) during periods of inflammation. Recent studies have shown a relationship between emphysema and high levels of EDP in human plasma. This report characterizes elastin digests on the basis of antigenicity, size, and method of preparation, as well as the size distribution of EDP found in the plasmas of nonsmokers, smokers, and emphysema patients. Gel filtration of elastin digests prepared by hydrolysis of human lung elastin using a low (1:500) ratio of neutrophil elastase to elastin generated a broad protein peak of approximately 70,000 daltons. In contrast, a high (1:25) ratio of neutrophil elastase to human lung elastin gave a broad protein peak, with a size distribution in the 10,000 to 30,000 dalton range. This digest showed distinct immunochemical properties. A polyclonal antibody directed against the low-ratio digest showed a minimum detection of 2 ng/ml for the homologous antigen but required 1,000 ng/ml of the high-ratio digest for detectable inhibition in an indirect ELISA assay. Gel filtration of plasmas from normal nonsmokers and the majority of normal smokers revealed a single immunoreactive EDP fraction of approximately 70,000 daltons. Plasmas from selected normal smokers and emphysema patients with high levels of circulating EDP (greater than 90 ng/ml) fractionated into a complex pattern of peptides in which the 70,000 dalton component represented 50% of the immunoreactive material and several lower molecular weight peptides represented the remaining circulating elastin antigens.(ABSTRACT TRUNCATED AT 250 WORDS)
Kucich
U
U
Graduate Hospital, Department of Medicine, Philadelphia, PA 19146.
Rosenbloom
J
J
Kimbel
P
P
Weinbaum
G
G
Abrams
W R
WR
D000941:Antigens / Q000032:analysis*; D005591:Chemical Fractionation; D004549:Elastin / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005541:Forced Expiratory Volume; D006801:Humans; D008168:Lung / Q000378:metabolism*; D008173:Lung Diseases, Obstructive / Q000276:immunology; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D012016:Reference Values; D012129:Respiratory Function Tests; D012907:Smoking
D000941:Antigens; D010455:Peptides; D004549:Elastin