Some properties of antisera to serum amyloid A protein (SAA): inhibition of precipitation by complexing of SAA to albumin.
Open Access
- 1 December 1976
- journal article
- research article
- Published by Rockefeller University Press in The Journal of Experimental Medicine
- Vol. 144 (6) , 1679-1682
- https://doi.org/10.1084/jem.144.6.1679
Abstract
Alveolar macrophages (AM) from pathogen-free rabbits were unable to release reactive oxygen intermediates (ROI) unless they were conditioned in serum for 24-48 h before triggering with membrane-active agents. The degree of serum conditioning of AM depended upon the concentration of serum used; optimal ROI release was obtained at or above 7.5% fetal bovine serum (FBS). FBS, autologous rabbit serum, pooled rabbit serum, and pooled human serum were each capable of conditioning AM for release of ROI. Serum conditioning of AM requires synthesis of new protein(s); and the enzyme required for ROI production, NADPH oxidase, was only detectable in serum-conditioned cells. Moreover, serum-conditioned cells lost their ability to release ROI after transfer to serum-free medium, while cells maintained in serum-free medium acquired the capacity to release ROI after their transfer to serum-containing medium, demonstrating the reversibility of the phenomenon. Initial purification data indicate that conditioning is mediated by a discrete serum constituent, which precipitates 40-80% saturated ammonium sulfate, does not bind to Cibacron Blue columns, and has a molecular weight of 30,000 to 50,000, as determined by molecular exclusion chromatography. Unlike gamma interferon, which also enhances ROI release by macrophages, our serum-conditioning factor is not acid labile, retaining 67% of its activity after 120 min incubation at pH 2.0. Moreover, it does not appear to be a contaminating endotoxin, since LPS neither conditioned AM for ROI production, nor triggered ROI production by serum-conditioned AM. We propose that such a conditioning requirement may normally protect the lung against ROI-mediated tissue injury. However, during a pulmonary inflammatory reaction initiated by other mediator systems, the resulting transudation of plasma proteins into the alveolar spaces may condition AM in situ for ROI production.This publication has 11 references indexed in Scilit:
- Isolation and Partial Characterization of SAA—An Amyloid-Related Protein from Human SerumThe Journal of Immunology, 1976
- Amyloid Fibril Protein AA: Purification and Properties of the Antigenically Related Serum Component As Determined by Solid Phase RadioimmunoassayThe Journal of Immunology, 1976
- Isolation and Characterization of Amyloid‐Related Serum Protein SAA as a Low Molecular Weight ProteinScandinavian Journal of Immunology, 1975
- Variation with age and disease of an amyloid A protein-related serum component.Journal of Clinical Investigation, 1975
- Age-associated changes of an amyloid related serum component.1974
- IMMUNOLOGIC STUDIES OF THE MAJOR NONIMMUNOGLOBULIN PROTEIN OF AMYLOIDThe Journal of Experimental Medicine, 1973
- QUANTITATIVE INVESTIGATIONS OF IDIOTYPIC ANTIBODIESThe Journal of Experimental Medicine, 1970
- The effect of hemoglobin on the haptoglobin-anti-haptoglobin reactionImmunochemistry, 1965
- A DIRECT IMMUNOLOGIC ASSAY OF HUMAN SERA FOR BENCE JONES PROTEINS (L-CHAINS)1965
- Diffusion-in-gel methods for immunological analysis.1958