Transforming growth factor–β2 is sequestered in preterm human milk by chondroitin sulfate proteoglycans

Kopperuncholan Namachivayam, Hayley P. Coffing, Nehru Viji Sankaranarayanan, Yingzi Jin, Mohan Krishnan, Brandy L. Frost, Cynthia L. Blanco, Aloka L. Patel, Paula P. Meier, Steven A. Garzon, Umesh R. Desai, Akhil Maheshwari

Research output: Contribution to journalArticle

Abstract

Human milk contains biologically important amounts of transforming growth factor-β2 isoform (TGF-β2), which is presumed to protect against inflammatory gut mucosal injury in the neonate. In preclinical models, enterally administered TGF-β2 can protect against experimental necrotizing enterocolitis, an inflammatory bowel necrosis of premature infants. In this study, we investigated whether TGF-β2 bioactivity in human preterm milk could be enhanced for therapeutic purposes by adding recombinant TGF-β2 (rTGF-β2) to milk prior to feeding. Milk-borne TGF-β2 bioactivity was measured by established luciferase reporter assays. Molecular interactions of TGF-β2 were investigated by nondenaturing gel electrophoresis and immunoblots, computational molecular modeling, and affinity capillary electrophoresis. Addition of rTGF-β2 (20–40 nM) to human preterm milk samples failed to increase TGF-β bioactivity in milk. Milk-borne TGF-β2 was bound to chondroitin sulfate (CS) containing proteoglycan(s) such as biglycan, which are expressed in high concentrations in milk. Chondroitinase treatment of milk increased the bioactivity of both endogenous and rTGF-β2, and consequently, enhanced the ability of preterm milk to suppress LPS-induced NF-κB activation in macrophages. These findings provide a mechanism for the normally low bioavailability of milk-borne TGF-β2 and identify chondroitinase digestion of milk as a potential therapeutic strategy to enhance the anti-inflammatory effects of preterm milk.

Original languageEnglish (US)
Pages (from-to)G171-G180
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume309
Issue number3
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

Fingerprint

Chondroitin Sulfate Proteoglycans
Human Milk
Transforming Growth Factors
Milk
Protein Isoforms
Growth
Chondroitinases and Chondroitin Lyases
Biglycan
Necrotizing Enterocolitis
Macrophage Activation
Capillary Electrophoresis
Luciferases
Premature Infants
Biological Availability
Electrophoresis
Digestion
Anti-Inflammatory Agents
Necrosis
Therapeutics
Gels

Keywords

  • Breast milk
  • Chondroitinase
  • Inflammation
  • Necrotizing enterocolitis
  • TGF-β<inf>2</inf>

ASJC Scopus subject areas

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

Cite this

Transforming growth factor–β2 is sequestered in preterm human milk by chondroitin sulfate proteoglycans. / Namachivayam, Kopperuncholan; Coffing, Hayley P.; Sankaranarayanan, Nehru Viji; Jin, Yingzi; Krishnan, Mohan; Frost, Brandy L.; Blanco, Cynthia L.; Patel, Aloka L.; Meier, Paula P.; Garzon, Steven A.; Desai, Umesh R.; Maheshwari, Akhil.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 309, No. 3, 01.01.2015, p. G171-G180.

Research output: Contribution to journalArticle

Namachivayam, Kopperuncholan ; Coffing, Hayley P. ; Sankaranarayanan, Nehru Viji ; Jin, Yingzi ; Krishnan, Mohan ; Frost, Brandy L. ; Blanco, Cynthia L. ; Patel, Aloka L. ; Meier, Paula P. ; Garzon, Steven A. ; Desai, Umesh R. ; Maheshwari, Akhil. / Transforming growth factor–β2 is sequestered in preterm human milk by chondroitin sulfate proteoglycans. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2015 ; Vol. 309, No. 3. pp. G171-G180.
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AU - Krishnan, Mohan

AU - Frost, Brandy L.

AU - Blanco, Cynthia L.

AU - Patel, Aloka L.

AU - Meier, Paula P.

AU - Garzon, Steven A.

AU - Desai, Umesh R.

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