title: Potential immunoregulatory role of heme oxygenase-1 in human milk: a combined biochemical and molecular modeling approach.
abstract: Human milk contains biological factors that are involved in a newborn's growth and immune system regulation. By integrating standard biochemical experimental protocols with computational methods, the present study investigates the presence of heme oxygenase-1 (HO-1), a cytoprotective enzyme, in human milk at different levels of maturation and in milk formulae. Furthermore, we evaluated cytokine and glutathione S-transferase (GSH) levels. Samples were collected from colostrum (on Day 1 after birth), from transition milk (on Postdelivery Days 7 and 14) and from mature milk (on Day 30 after delivery) in 14 healthy women. HO-1 protein, GSH and cytokines levels were measured using enzyme-linked immunosorbent assay and flow cytometry. HO-1 protein levels were significantly higher in colostrum (1.33 ng/ml; 5th centile 0.92; 95th centile 2.38) and in transition milk at 14 days (0.97 ng/ml; 5th centile 0.87; 95th centile 1.45) than in mature milk (0.9 ng/ml; 5th centile 0.8; 95th centile 1.38). Levels of HO-1 in milk formulae were similar to those in colostrum. No significant differences in GSH content were observed in mature milk, transition milk and colostrum, whereas significantly higher GSH levels were observed in milk formulae. No significant levels of cytokines, with the exception of interleukin-8, were found. Computational studies on the possible interactions between HO-1 and CD91 were carried out by a battery of softwares, namely, GRAMM (version 1.03), DALI, CLUSTALW (version 2.0), PatchDock and FireDock, mutually counterchecking and validating each other. The computational results, the strong convergence (to the same "solution") of which finally leads to an "experimental-like" character, showed that HO-1 may bind to CD91, thus suggesting its major role as a new chaperokine in immune response regulation. These findings, which connect and integrate biochemical data and computational data interpretation, represent a synergistic and powerful means of conducting biological research.
authors.affiliation: Department of Biological Chemistry, Medical Chemistry and Molecular Biology, University of Catania, Catania, Italy. livolti@unict.it
authors.forename: Giovanni
authors.initials: G
authors.lastname: Li Volti
authors.forename: Fabio
authors.initials: F
authors.lastname: Galvano
authors.forename: Alessandro
authors.initials: A
authors.lastname: Frigiola
authors.forename: Salvatore
authors.initials: S
authors.lastname: Guccione
authors.forename: Claudia
authors.initials: C
authors.lastname: Di Giacomo
authors.forename: Stefano
authors.initials: S
authors.lastname: Forte
authors.forename: Giovanni
authors.initials: G
authors.lastname: Tringali
authors.forename: Massimo
authors.initials: M
authors.lastname: Caruso
authors.forename: Olayiwola Adedotun
authors.initials: OA
authors.lastname: Adekoya
authors.forename: Diego
authors.initials: D
authors.lastname: Gazzolo
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000378:metabolism*; D003126:Colostrum / Q000276:immunology; D019295:Computational Biology; D005260:Female; D005978:Glutathione / Q000378:metabolism; D051547:Heme Oxygenase-1 / Q000502:physiology*; D006801:Humans; D016209:Interleukin-8 / Q000032:analysis; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008895:Milk, Human / Q000276:immunology*; D008958:Models, Molecular; D011247:Pregnancy
chemical_list: D015703:Antigens, CD; D016209:Interleukin-8; C475476:LRP1 protein, human; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D051547:Heme Oxygenase-1; D005978:Glutathione