Human Milk Contains Detectable Levels of Immunoreactive Leptin

  • Robert E. Lyle
  • Stephen C. Kincaid
  • Janet C. Bryant
  • Audra M. Prince
  • Robert E. McGeheeJr.
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 501)


Leptin, the recently cloned product of the obese (ob) gene, is a 16 kDa-protein that acts as a circulating satiety factor. It also serves to regulate energy expenditure and may act as a counter regulatory hormone to insulin. Initially thought to be exclusively produced by mature adipocytes, its mRNA has now been identified in significant levels in the placenta as well as the fetus raising speculation regarding its importance as a growth factor. Given studies demonstrating that exclusively breast-fed infants are leaner due to decreased energy intakes than formula-fed infants, we hypothesized that the presence of leptin in human milk could participate in mediating the earlier satiety of those infants fed human milk. We undertook this initial study to qualitatively examine the presence of leptin in human milk utilizing an immunoblot approach. Random milk samples during the first 2 weeks of lactation were available for study from 4 mothers delivering at term. Milk samples were centrifuged, the aqueous layer removed, and the protein content quantitated. One-hundred micrograms of total protein were separated by sodium dodecyl sulfate—polyacrylamide-gel electrophoresis (SDS-PAGE), transferred to nitrocellulose, and immunoblotted with an antileptin antibody. As controls, recombinant human leptin alone and a sample of milk containing added leptin were similarly electrophoresed and immunoblotted. Labeled proteins were visualized by chemiluminescence. Significant amounts of leptin protein were identified in all milk samples examined. No difference in protein detec-tion was identified in fresh milk vs. frozen milk, and little difference was apparent in foremilk samples vs. hindmilk samples. These preliminary data reveal the presence of leptin in term human milk and suggest that further studies to document bioactivity of milk-derived leptin are warranted.


Milk Sample Human Milk Fresh Milk Counter Regulatory Hormone Lower Energy Intake 
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  1. Cunningham AS. Morbidity in breast-fed and artificially fed infants II. J Pediatr 1979;95:685–689.PubMedCrossRefGoogle Scholar
  2. Dewey KG, Heinig MJ, Nommsen LA, Persson JM, Lonnerdal B. Breast-fed infants are leaner than formula-fed infants at 1 year of age: the DARLING study. Am J Clin Nutr 1993;57:140–145.PubMedGoogle Scholar
  3. Dewey KG, Peerson JM, Brown KH, Krebs NF, Michaelsen KF, Persson LA, Salmenpera L, Whitehead RG, Yeung DL, WHO Working Group on Infant Growth. Growth of breast-fed infants deviates from current reference data: A pooled analysis of US, Canadian, and European data sets. Pediatrics 1995;86:495–503.Google Scholar
  4. Drewett RF. Returning to the suckled breast: a further test of Hall’s hypothesis. Early Hum Dev 1982;6:161–163.PubMedCrossRefGoogle Scholar
  5. Elmquist JK, Ahima RS, Maratos-Flier E, Flier JS, Saper CB. Leptin activates neurons in the ventrobasal hypothalamus and brainstem. Endocrinology 1997;138:839–842.PubMedCrossRefGoogle Scholar
  6. Goldman AS. The immune system of human milk: antimicrobial, antiinflammatory and immunomodulating properties. Pediatr Infect Dis J 1993;12:664–671.PubMedCrossRefGoogle Scholar
  7. Hall B. Changing composition of human milk and early development of an appetite control. Lancet 1975;1:779–781.PubMedCrossRefGoogle Scholar
  8. Hamosh M. Does infant nutrition affect adiposity and cholesterol levels in the adult? J Pediatr Gastroenterol Nutr 1988;7:10–16.PubMedCrossRefGoogle Scholar
  9. Harigaya A, Nagashima K, Nako Y, Morikawa A. Relationship between concentration of serum leptin and fetal growth. J Clin Endocrinol Metab 1997;82:3281–3284.PubMedCrossRefGoogle Scholar
  10. Hassink SG, de Lancey E, Sheslow DV, Smith-Kirwin SM, O’Connor DM, Considine RV, pentanova I, Dostal K, Spear ML, Leef K, Ash M, Spitzer AR, Funanage VL. Placental leptin: an important new growth factor in intrauterine and neonatal development? Pediatrics I997;100:El.Google Scholar
  11. Heinig MJ, Nommsen LA, Peerson JM, Lonnerdal B, Dewey KG. Energy and protein intakes of breast-fed and formula-fed infants during the first year of life and their association with growth velocity: the DARLING study. Am J Clin Nutr 1993;58:152–161.PubMedGoogle Scholar
  12. Hoggard N, Hunter L, Duncan JS, Williams LM, Trayhurn P, Mercer JG. Leptin and leptin receptor mRNA and protein expression in the murine fetus and placenta. Proc Natl Acad Sci USA 1997a;94: 11073–11078.CrossRefGoogle Scholar
  13. Hoggard N, Mercer JG, Rayner DV, Moar K, Trayhurn P, Williams LM. Localization of leptin receptor splice variants in murine peripheral tissues by RT-PCR and in situ hybridization. Biochem Biophys Res Comm 1997b;232:383–387.CrossRefGoogle Scholar
  14. Ingalls AM, Dickie MD, Snell GD. Obese, a new mutation in the mouse. J Hered 1950;41:317–318. Knittle JL, Hirsch J. Effect of early nutrition on the development of rat epididymal fat pads: cellularity and metabolism. J Clin Invest 1968;47:2091–2098.Google Scholar
  15. Koistinen HA, Koivisto VA, Andersson S, Karonen SL, Kontula K, Oksanen L, Teramo KA. Leptin concentration in cord blood correlates with intrauterine growth. J Clin Endocrinol Metab 1997;82:3328–3330.PubMedCrossRefGoogle Scholar
  16. Kuczmarski R, Flegal KM, Campbell SM, Johnson CL. Increasing prevalence of overweight among US adults: the National Health Nutrition Examination Surveys, 1960 to 1991. JAMA 1994;272:205–211.PubMedCrossRefGoogle Scholar
  17. Lewis DS, Bertrand HA, McMahan A, McGill HC, Carey KD, Masoro EJ. Preweaning food intake influ-ences the adiposity of young adult baboons. J Clin Invest 1986;78:899–904.PubMedCrossRefGoogle Scholar
  18. Lyle RE, Habener JF, McGehee RE Jr. Antisense oligonucleotides to Differentiation-Specific Element Binding Protein (DSEB) mRNA inhibit adipocyte differentiation. Biochem Biophys Res Comm 1996;226:709–715.CrossRefGoogle Scholar
  19. Mikhail AA, Beck EX, Shafer A, Barut B, Gbur JS, Zupanic TJ, Schweitzer AC, Cioffi JA, Lacaud G, Ouyang B, Keller G, Snodgrass HR. Leptin stimulates fetal and adult erythroid and myeloid development. Blood 1997;89:1507–1512.PubMedGoogle Scholar
  20. Nysenbaum AN, Smart JL. Suckling behavior and milk intakes of neonates in relation to milk fat content. Early Hum Dev 1982;6:205–213.PubMedCrossRefGoogle Scholar
  21. Pelleymounter MA, Cullen MJ, Baker MB, Hecht R, Winters D, Boone T, Collins F. Effects of the obese gene product on body weight regulation inob/obmice. Science 1995;269:546–549.CrossRefGoogle Scholar
  22. Rath EA, Thenen SW. Use of tritiated water for measurement of 24-hour milk intake in suckling lean and genetically obese(oblob)mice. J Nutr 1979;109:840–847.PubMedGoogle Scholar
  23. Schubring C, Kiess W, Englaro P, Rascher W, Dotsch J, Hanitsch S, Attanasio A, Blum WE Levels of leptin in maternal serum, amniotic fluid and arterial and venous cord blood: relation to neonatal and placental weight. J Clin Endocrinol Metab 1997;82:1480–1483.PubMedCrossRefGoogle Scholar
  24. Sivan E, Lin M, Homko CJ, Reece A, Boden G. Leptin is present in human cord blood. Diabetes 1997;46:917–919.PubMedCrossRefGoogle Scholar
  25. Tartaglia LA, Dembski M, Weng XU, Deng N, Culpepper J, Devos R, Richards GJ, Campfield LA, Clark FT, Deeds J. Identification and expression cloning of a leptin receptor, OB-R. Cell 1995;83:1263–1271.Google Scholar
  26. Troianno RP, Flegal KM, Kuczmarski RJ, Campbell SM, Johnson CL. Overweight prevalence and trends for children and adolescents: the National Health and Nutrition Examination Surveys, 1963 to 1991. Arch Pediatr Adolesc Med 1995;149:1085–1091.CrossRefGoogle Scholar
  27. Wang MY, Zhou YT, Newgard CB, Unger RH. A novel leptin receptor isoform in rat. FEBS Lett 1996;392:87–90.PubMedCrossRefGoogle Scholar
  28. Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM. Positional cloning of the mouse obese gene and its human homologue. Nature 1994;372:425–432.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2001

Authors and Affiliations

  • Robert E. Lyle
    • 1
  • Stephen C. Kincaid
    • 1
  • Janet C. Bryant
    • 1
  • Audra M. Prince
    • 1
  • Robert E. McGeheeJr.
    • 1
  1. 1.Division of Neonatology Department of Pediatrics and Lactation Support DepartmentUniversity of Arkansas for Medical Sciences and Arkansas Children’s HospitalLittle RockUSA

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