Antonella Alberti

Uptake and degradation of Curosurf® after tracheal administration to newborn and adult rabbits

This paper examines the removal from the airways of Curosurf, a commercial surfactant derived from porcine lungs, administered at pharmacological concentrations to newborn or adult animals. Curosurf was labelled by the addition of radioactive dipalmitoyl phosphatidylcholine (DPPC) and administered intratracheally to newborn and adult rabbits at a dose of 200 mg x kg(-1) body weight. The disappearance of DPPC from the airways and its appearance in alveolar macrophages, lung parenchyma, lamellar bodies, serum, liver, kidneys and brain was then studied for 24-48 h. The in vitro degradation of Curosurf DPPC by alveolar macrophages was also studied. During the first 3 h after instillation, large amounts of Curosurf left the airways and became associated with tissue, indicating that it mixed rapidly with the endogenous pools of surfactant. A fraction of administered DPPC became associated with the lamellar bodies, suggesting that Curosurf can be recycled. Curosurf administration did not stop the secretion of endogenous surfactant. Very little intact radioactive DPPC could be recovered at any time in alveolar macrophages, however, macrophages have the ability, in vitro, to degrade Curosurf. Newborn rabbits lose Curosurf from the lungs at a slower rate than adult rabbits. One and two days after instillation, organic extracts from the liver, kidney, brain and serum contained small but measurable amounts of radioactivity. These results indicate that Curosurf rapidly enters the pathways of surfactant metabolism and that alveolar macrophages may play an important role in the catabolism of Curosurf.

Surfactant apoprotein A modulates interleukin-8 and monocyte chemotactic peptide-1 production

Previous studies have shown that surfactant apoprotein A (SP-A) and natural or synthetic surfactant can modulate the release of pro-inflammatory cytokines from alveolar mononuclear phagocytes. The aim of this study was to assess whether SP-A or Surfactant (Surf) from patients with pulmonary alveolar proteinosis (PAP) can affect the release of two chemokines (interleukin (IL)-8 and monocyte chemtactic peptide (MCP)-1) from human monocytes and rat lung type-II cells. In addition IL-8 and MCP-1 levels were assessed in the brochoalveolar lavage fluid (BALF) of seven patients with PAP and compared with those in a group of control subjects (n=5). SP-A, tested over a wide range of concentrations, significantly increased IL-8 and MCP-1 release from monocytes. SP-A retained its activity after collagenase digestion, but was not active after heat treatment. The release of IL-8 by monocytes was also stimulated by Surf. Finally, median BALF IL-8 and MCP-1 levels in PAP patients were significantly higher than in controls (9.50 and 9.51 pg·mL−1 in controls versus 151.95 and 563.70 pg·mL−1 in PAP, respectively) and significantly correlated with SP-A concentrations in BALF. Overall the results of this study support the view that the high content of alveolar surfactant apoprotein A may contribute to the upregulation of chemokine release in pulmonary alveolar proteinosis, thus contributing to airway inflammation.

In chyloptysis, SP-A affects the clearance of serum lipoproteins entering the airways

Serum lipoproteins may enter the airways and appear in sputum (chyloptysis) when the lymphatic circulation is impaired by inflammation, neoplasia, or an abnormal proliferation of smooth muscle cells. While analyzing the bronchoalveolar lavage fluid of a patient with chyloptysis, we noticed that surfactant could not be separated from contaminating serum lipoproteins and speculated that lipoproteins might interact with surfactant components. To clarify this point we immobilized surfactant protein (SP) A on microtiter wells and incubated it with 125I-labeled very low density lipoproteins (VLDLs), low-density lipoproteins, and high-density lipoproteins. We found that SP-A binds lipoproteins. Studying in greater detail the interaction of SP-A with VLDLs, we found that the binding is time and concentration dependent; is inhibited by unlabeled lipoproteins, phospholipids, and antibodies to SP-A; is increased by Ca2+; and is unaffected by methyl α-d-mannopyranoside. Whole surfactant is a potent inhibitor of binding. Furthermore, we found that SP-A increases the degradation of VLDLs by alveolar macrophages and favors the association of VLDLs with alveolar surfactant. We conclude that SP-A influences the disposal of serum lipoproteins entering the airways and speculate that binding to alveolar surfactant might represent an important step in the interaction between exogenous substances and the lung.Serum lipoproteins may enter the airways and appear in sputum (chyloptysis) when the lymphatic circulation is impaired by inflammation, neoplasia, or an abnormal proliferation of smooth muscle cells. While analyzing the bronchoalveolar lavage fluid of a patient with chyloptysis, we noticed that surfactant could not be separated from contaminating serum lipoproteins and speculated that lipoproteins might interact with surfactant components. To clarify this point we immobilized surfactant protein (SP) A on microtiter wells and incubated it with 125I-labeled very low density lipoproteins (VLDLs), low-density lipoproteins, and high-density lipoproteins. We found that SP-A binds lipoproteins. Studying in greater detail the interaction of SP-A with VLDLs, we found that the binding is time and concentration dependent; is inhibited by unlabeled lipoproteins, phospholipids, and antibodies to SP-A; is increased by Ca2+; and is unaffected by methyl alpha-D-mannopyranoside. Whole surfactant is a potent inhibitor of binding. Furthermore, we found that SP-A increases the degradation of VLDLs by alveolar macrophages and favors the association of VLDLs with alveolar surfactant. We conclude that SP-A influences the disposal of serum lipoproteins entering the airways and speculate that binding to alveolar surfactant might represent an important step in the interaction between exogenous substances and the lung.

LOCALIZATION OF SYNTHETIC SURFACTANT PROTEIN SP-B IN PHOSPHOLIPID VESICLES.|[dagger]| 1947

Surfactant subtypes have been identified by gradient centrifugation and characterized by differences in protein composition, vesicle size, and surface activity. The presence of surfactant proteins in each subtype is thought to be critical to the formation of aggregates and their function (Baritussio, Am J Physiol 1994;266:L436). To evaluate whether synthetic surfactant peptides interact with lipids in a manner comparable to native surfactant proteins, we prepared surfactant vesicles containing synthetic 125I-SP-B1-78 and phospholipids (DPPC:PG:Palmitic acid, 7:2:1) [PL], with and without palmitoylated synthetic SP-C1-34. We separated the aggregates by continuous sucrose gradient (0.1-0.8 M; 100,000 g for 48 h), collecting 50 fractions, and analyzed the PL and peptide profile. The PL peak appeared in fraction 17 and > 80% of PL was recovered in fractions 15-17, at a density of 1.07 g/mL, irrespective of the presence of SP-C. In the SP-B/PL aggregates, SP-B peaked at 51% in fraction 15 and > 95% of SP-B appeared in fractions 11-17. In the SP-B&SP-C/PL aggregates, 17% of the SP-B was detected at the bottom of the gradient with <5% PL and ≈78% was recovered in fractions 5-15 with a peak of 28% in fraction 13. In both aggregates, small amounts of PL, but no SP-B, were recovered at the top of the gradient, at a density of about 1.010 g/mL. In summary, synthetic SP-B associates with phospholipid aggregates at a density of about 1.07 g/mL, as shown by the overlapping peaks. The presence of SP-C does not affect the phospholipid distribution, but changes the SP-B profile significantly. These data are consistent with those obtained for term rabbit surfactant and suggest that the insertion of synthetic surfactant proteins in a simplified phospholipid dispersion reproduces important characteristics of native surfactant.