title: Preterm human milk contains a large pool of latent TGF-β, which can be activated by exogenous neuraminidase.
abstract: Human milk contains substantial amounts of transforming growth factor (TGF)-β, particularly the isoform TGF-β2. We previously showed in preclinical models that enterally administered TGF-β2 can protect against necrotizing enterocolitis (NEC), an inflammatory bowel necrosis of premature infants. In this study we hypothesized that premature infants remain at higher risk of NEC than full-term infants, even when they receive their own mother's milk, because preterm human milk contains less bioactive TGF-β than full-term milk. Our objective was to compare TGF-β bioactivity in preterm vs. full-term milk and identify factors that activate milk-borne TGF-β. Mothers who delivered between 23 0/7 and 31 6/7 wk or at ≥37 wk of gestation provided milk samples at serial time points. TGF-β bioactivity and NF-κB signaling were measured using specific reporter cells and in murine intestinal tissue explants. TGF-β1, TGF-β2, TGF-β3, and various TGF-β activators were measured by real-time PCR, enzyme immunoassays, or established enzymatic activity assays. Preterm human milk showed minimal TGF-β bioactivity in the native state but contained a large pool of latent TGF-β. TGF-β2 was the predominant isoform of TGF-β in preterm milk. Using a combination of several in vitro and ex vivo models, we show that neuraminidase is a key regulator of TGF-β bioactivity in human milk. Finally, we show that addition of bacterial neuraminidase to preterm human milk increased TGF-β bioactivity. Preterm milk contains large quantities of TGF-β, but most of it is in an inactive state. Addition of neuraminidase can increase TGF-β bioactivity in preterm milk and enhance its anti-inflammatory effects.
authors.affiliation: Division of Neonatology, Department of Pediatrics, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Kopperuncholan
authors.initials: K
authors.lastname: Namachivayam
authors.forename: Cynthia L
authors.initials: CL
authors.lastname: Blanco
authors.forename: Brandy L
authors.initials: BL
authors.lastname: Frost
authors.forename: Aaron A
authors.initials: AA
authors.lastname: Reeves
authors.forename: Ramasamy
authors.initials: R
authors.lastname: Jagadeeswaran
authors.forename: Krishnan
authors.initials: K
authors.lastname: MohanKumar
authors.forename: Azif
authors.initials: A
authors.lastname: Safarulla
authors.forename: Partha
authors.initials: P
authors.lastname: Mandal
authors.forename: Steven A
authors.initials: SA
authors.lastname: Garzon
authors.forename: J Usha
authors.initials: JU
authors.lastname: Raj
authors.forename: Akhil
authors.initials: A
authors.lastname: Maheshwari
mesh_terms: D000818:Animals; D005260:Female; D015870:Gene Expression; D006801:Humans; D007774:Lactation / Q000378:metabolism; D051379:Mice; D008895:Milk, Human / Q000201:enzymology / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism*; D047928:Premature Birth / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D047929:Term Birth / Q000378:metabolism; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*
chemical_list: D016328:NF-kappa B; D020033:Protein Isoforms; D016212:Transforming Growth Factor beta; D009439:Neuraminidase
keywords: breast milk; inflammation; necrotizing enterocolitis; sialidase; transforming growth factor-β
references.citation: Transfusion. 2002 Nov;42(11):1428-34
references.pmid: 12421215
references.citation: J Allergy Clin Immunol. 2002 Jan;109(1):119-21
references.pmid: 11799376
references.citation: Am J Dis Child. 1960 Aug;100:252-6
references.pmid: 13793167
references.citation: Kidney Int. 1997 May;51(5):1376-82
references.pmid: 9150447
references.citation: Biol Neonate. 1999;75(1):59-64
references.pmid: 9831685
references.citation: J Cell Sci. 2003 Jan 15;116(Pt 2):217-24
references.pmid: 12482908
references.citation: Am J Physiol Lung Cell Mol Physiol. 2007 Feb;292(2):L550-8
references.pmid: 17071719
references.citation: Immunol Cell Biol. 2000 Feb;78(1):74-9
references.pmid: 10651932
references.citation: Adv Carbohydr Chem Biochem. 2010;64:403-79
references.pmid: 20837202
references.citation: Pediatr Res. 2003 Mar;53(3):419-25
references.pmid: 12595589
references.citation: J Endocrinol. 1996 Oct;151(1):77-86
references.pmid: 8943771
references.citation: Am J Physiol Gastrointest Liver Physiol. 2012 Jul;303(1):G93-102
references.pmid: 22538401
references.citation: J Dairy Sci. 2005 Jun;88(6):1986-93
references.pmid: 15905428
references.citation: Prog Neuropsychopharmacol Biol Psychiatry. 2007 Jun 30;31(5):1044-53
references.pmid: 17433516
references.citation: Protein Expr Purif. 2006 Nov;50(1):9-17
references.pmid: 16901717
references.citation: J Biol Chem. 2009 Feb 27;284(9):5945-55
references.pmid: 19117939
references.citation: J Cell Biol. 1981 Jun;89(3):485-94
references.pmid: 7019216
references.citation: J Allergy Clin Immunol. 1999 Nov;104(5):1093-8
references.pmid: 10550758
references.citation: J Pediatr. 2010 Feb;156(2 Suppl):S21-5
references.pmid: 20105660
references.citation: Lipids. 2001 Oct;36(10):1179-81
references.pmid: 11768164
references.citation: J Cell Biol. 1991 Aug;114(4):827-39
references.pmid: 1869589
references.citation: Pediatr Res. 1999 Aug;46(2):194-9
references.pmid: 10447115
references.citation: J Pediatr Gastroenterol Nutr. 2012 May;54(5):630-8
references.pmid: 22067113
references.citation: Acta Biochim Pol. 2009;56(3):411-4
references.pmid: 19753334
references.citation: Cytokine. 2002 Feb 21;17(4):182-6
references.pmid: 11991670
references.citation: J Pediatr. 2010 Feb;156(2 Suppl):S16-20
references.pmid: 20105659
references.citation: Microbes Infect. 1999 Dec;1(15):1297-304
references.pmid: 10611758
references.citation: Dev Growth Differ. 2008 Jan;50(1):41-8
references.pmid: 17986262
references.citation: J Pediatr. 2010 Apr;156(4):562-7.e1
references.pmid: 20036378
references.citation: Clin Exp Allergy. 2006 May;36(5):614-8
references.pmid: 16650046
references.citation: Am J Obstet Gynecol. 1992 May;166(5):1515-28
references.pmid: 1595807
references.citation: J Biol Chem. 1999 May 7;274(19):13586-93
references.pmid: 10224129
references.citation: Cancer Cytopathol. 2011 Apr 25;119(2):92-101
references.pmid: 21433001
references.citation: Am J Physiol Gastrointest Liver Physiol. 2013 Jan 15;304(2):G167-80
references.pmid: 23154975
references.citation: Pediatr Res. 2008 Jan;63(1):26-32
references.pmid: 18043506
references.citation: J Allergy Clin Immunol. 1999 Dec;104(6):1251-7
references.pmid: 10589009
references.citation: Cell Immunol. 1994 Nov;159(1):77-84
references.pmid: 7954842
references.citation: Pediatr Allergy Immunol. 2011 Dec;22(8):853-61
references.pmid: 21929601
references.citation: Cancer Surv. 1992;12:81-103
references.pmid: 1638549
references.citation: Eur J Biochem. 1991 Apr 23;197(2):353-8
references.pmid: 2026157
references.citation: Pediatrics. 2006 Aug;118(2):e299-308
references.pmid: 16831894
references.citation: Neonatology. 2011;99(3):192-201
references.pmid: 20881435
references.citation: J Affect Disord. 2010 Jul;124(1-2):68-75
references.pmid: 19900711
references.citation: Science. 1994 Jun 24;264(5167):1936-8
references.pmid: 8009224
references.citation: Adv Exp Med Biol. 2001;501:559-66
references.pmid: 11787729
references.citation: Pediatr Allergy Immunol. 2008 Sep;19(6):497-504
references.pmid: 18221472
references.citation: Pediatr Res. 2003 Apr;53(4):642-7
references.pmid: 12612204
references.citation: Cold Spring Harb Perspect Biol. 2011 Nov 01;3(11):a005017
references.pmid: 21900405
references.citation: PLoS Pathog. 2010 Oct 07;6(10):e1001136
references.pmid: 20949074
references.citation: Clin Exp Allergy. 2011 Jun;41(6):783-93
references.pmid: 21492269
references.citation: J Nutr Biochem. 2001 Oct;12(10):559-564
references.pmid: 12031261
references.citation: Clin Exp Immunol. 1993 Oct;94(1):220-4
references.pmid: 8403511
references.citation: Clin Chem. 2007 May;53(5):955-62
references.pmid: 17363422
references.citation: Pediatr Res. 2000 Jan;47(1):157-62
references.pmid: 10625097
references.citation: J Nutr. 2009 Aug;139(8):1463-8
references.pmid: 19515744
references.citation: Pediatr Nephrol. 2004 Mar;19(3):317-21
references.pmid: 14714171
references.citation: Early Hum Dev. 2004 Apr;77(1-2):67-75
references.pmid: 15113633
references.citation: Clin Exp Allergy. 2008 Oct;38(10):1606-14
references.pmid: 18631345
references.citation: Int J Biochem Cell Biol. 2008;40(6-7):1068-78
references.pmid: 18243766
references.citation: Immunology. 1999 Aug;97(4):595-600
references.pmid: 10457212
references.citation: J Immunoassay. 1998 Feb;19(1):23-37
references.pmid: 9530609
references.citation: Cochrane Database Syst Rev. 2007 Oct 17;(4):CD002971
references.pmid: 17943776
references.citation: Am J Physiol Regul Integr Comp Physiol. 2009 Apr;296(4):R1053-62
references.pmid: 19158412
references.citation: Cytokine. 2009 Apr;46(1):12-6
references.pmid: 19232500
references.citation: Gastroenterology. 2011 Jan;140(1):242-53
references.pmid: 20875417
references.citation: Nature. 1989 Mar 9;338(6211):158-60
references.pmid: 2493139
references.citation: Res Commun Mol Pathol Pharmacol. 1996 Sep;93(3):263-87
references.pmid: 8896040
references.citation: Pediatr Res. 2010 Oct;68(4):330-4
references.pmid: 20581738
references.citation: Am J Reprod Immunol. 1997 Aug;38(2):121-8
references.pmid: 9272211
references.citation: J Nutr. 2009 Jan;139(1):69-75
references.pmid: 19056655
references.citation: Pediatr Res. 1998 Oct;44(4):524-31
references.pmid: 9773841