title: Formula diet alters small intestine morphology, microbial abundance and reduces VE-cadherin and IL-10 expression in neonatal porcine model.
abstract: BACKGROUND
  Breastfeeding is associated with a variety of positive health outcomes in children and is recommended exclusively for the first 6 months of life; however, 50-70 % of infants in the US are formula-fed. To test the hypothesis that immune system development and function in neonates and infants are significantly influenced by diet, 2-day old piglets were fed soy or milk formula (n = 6/group/gender) until day 21 and compared to a sow-fed group (n = 6/gender).
  
  
  METHODS
  Histomorphometric analyses of ileum, jejunum and Peyer's patches were carried out, to determine the inflammation status, mRNA and protein expression of pro-inflammatory, anti-inflammatory and growth-related chemokines and cytokines.
  
  
  RESULTS
  In formula-fed animals, increases in ileum and jejunum villus height and crypt depth were observed in comparison to sow-fed animals (jejunum, p < 0.01 villus height, p < 0.04 crypt depth; ileum p < 0.001 villus height, p < 0.002 crypt depth). In formula-fed the lymphoid follicle size (p < 0.01) and germinal centers (p < 0.01) with in the Peyer's patch were significantly decreased in comparison to sow-fed, indicating less immune education. In ileum, formula diet induced significant up-regulation of AMCFII, IL-8, IL-15, VEGFA, LIF, FASL, CXCL11, CCL4, CCL25 and down-regulation of IL-6, IL-9, IL-10, IL-27, IFNA4, CSF3, LOC100152038, and LOC100736831 at the transcript level. We have confirmed some of the mRNA data by measuring protein, and significant down-regulation of anti-inflammatory molecule IL-10 in comparison to sow-fed piglets was observed. To further determine the membrane protein expression in the ileum, VE-cadherin, occludin, and claudin-3, Western blot analyses were conducted. Sow fed piglets showed significantly more VE-Cadherin, which associated with levels of calcium, and putrescine measured. It is possible that differences in GI tract and immune development are related to shifts in the microbiome; notably, there were 5-fold higher amounts of Lactobacillaceae spp and 3 fold higher Clostridia spp in the sow fed group in comparison to milk formula-fed piglets, whereas in milk formula-fed pigs Enterobacteriaceae spp was 5-fold higher.
  
  
  CONCLUSIONS
  In conclusion, formula diet alters GI morphology, microbial abundance, intestinal barrier protein VE-cadherin and anti-inflammatory molecule IL-10 expression. Further characterization of formula effects could lead to modification of infant formula to improve immune function, reduce inflammation and prevent conditions such as allergies and infections.
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA. vlyeruva@uams.edu.|Arkansas Children's Hospital Research Institute, Little Rock, USA. vlyeruva@uams.edu.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA. vlyeruva@uams.edu.
authors.forename: Laxmi
authors.initials: L
authors.lastname: Yeruva
authors.affiliation: Arkansas Children's Hospital Research Institute, Little Rock, USA.
authors.forename: Nicole E
authors.initials: NE
authors.lastname: Spencer
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Manish K
authors.initials: MK
authors.lastname: Saraf
authors.affiliation: Department of Pathology, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Leah
authors.initials: L
authors.lastname: Hennings
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Anne K
authors.initials: AK
authors.lastname: Bowlin
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Mario A
authors.initials: MA
authors.lastname: Cleves
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Kelly
authors.initials: K
authors.lastname: Mercer
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Sree V
authors.initials: SV
authors.lastname: Chintapalli
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Kartik
authors.initials: K
authors.lastname: Shankar
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Roger G
authors.initials: RG
authors.lastname: Rank
authors.affiliation: Arkansas Children's Nutrition Center, 15 Children's Way, Little Rock, AR, 72202, USA.|Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Badger
authors.affiliation: Department of Pharmacology & Experimental Therapeutics, Louisiana State University Health Sciences Center, New Orleans, LA, USA.
authors.forename: Martin J J
authors.initials: MJ
authors.lastname: Ronis
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D016207:Cytokines / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D004032:Diet; D015536:Down-Regulation; D053222:Fas Ligand Protein / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000069196:Gastrointestinal Microbiome / Q000187:drug effects*; D006801:Humans; D007082:Ileum / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D041943:Infant Formula / Q000494:pharmacology*; D007231:Infant, Newborn; D016898:Interferon-alpha / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D019409:Interleukin-15 / Q000235:genetics / Q000378:metabolism; D064094:Interleukin-27 / Q000235:genetics / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D016209:Interleukin-8 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D016906:Interleukin-9 / Q000235:genetics / Q000378:metabolism; D007421:Intestine, Small / Q000187:drug effects* / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D007583:Jejunum / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology / Q000473:pathology; D053605:Leukemia Inhibitory Factor / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D008892:Milk; D010581:Peyer's Patches / Q000187:drug effects / Q000276:immunology; D012333:RNA, Messenger / Q000187:drug effects* / Q000378:metabolism; D045730:Soy Foods; D013552:Swine; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D002118:Calcium; D016207:Cytokines; D053222:Fas Ligand Protein; D016898:Interferon-alpha; D016753:Interleukin-10; D019409:Interleukin-15; D064094:Interleukin-27; D015850:Interleukin-6; D016209:Interleukin-8; D016906:Interleukin-9; D053605:Leukemia Inhibitory Factor; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D000099152:Cadherin 5
keywords: GALT; Ileum; Jejunum; Nutrition; Peyer’s patches