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Small intestinal intraepithelial TCRγδ+ T lymphocytes are present in the premature intestine but selectively reduced in surgical necrotizing enterocolitis.

Weitkamp JH, Rosen MJ, Zhao Z, Koyama T, Geem D, Denning TL, Rock MT, Moore DJ, Halpern MD, Matta P, Denning PW - PLoS ONE (2014)

Bottom Line: In human preterm infants, the proportion of IEL was reduced by 66% in 11 NEC ileum resections compared to 30 non-NEC controls (p<0.001).While γδ IEL dominated over conventional αβ IEL early in gestation in mice and in humans, γδ IEL were preferential decreased in the ileum of surgical NEC patients compared to non-NEC controls (50% reduction, p<0.05).Loss of IEL in human NEC was associated with downregulation of the Th17 transcription factor retinoic acid-related orphan nuclear hormone receptor C (RORC, p<0.001).

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Vanderbilt University School of Medicine and Monroe Carell Jr. Children's Hospital at Vanderbilt, Nashville, Tennessee, United States of America.

ABSTRACT

Background: Gastrointestinal barrier immaturity predisposes preterm infants to necrotizing enterocolitis (NEC). Intraepithelial lymphocytes (IEL) bearing the unconventional T cell receptor (TCR) γδ (γδ IEL) maintain intestinal integrity and prevent bacterial translocation in part through production of interleukin (IL) 17.

Objective: We sought to study the development of γδ IEL in the ileum of human infants and examine their role in NEC pathogenesis. We defined the ontogeny of γδ IEL proportions in murine and human intestine and subjected tcrδ-/- mice to experimental gut injury. In addition, we used polychromatic flow cytometry to calculate percentages of viable IEL (defined as CD3+ CD8+ CD103+ lymphocytes) and the fraction of γδ IEL in surgically resected tissue from infants with NEC and gestational age matched non-NEC surgical controls.

Results: In human preterm infants, the proportion of IEL was reduced by 66% in 11 NEC ileum resections compared to 30 non-NEC controls (p<0.001). While γδ IEL dominated over conventional αβ IEL early in gestation in mice and in humans, γδ IEL were preferential decreased in the ileum of surgical NEC patients compared to non-NEC controls (50% reduction, p<0.05). Loss of IEL in human NEC was associated with downregulation of the Th17 transcription factor retinoic acid-related orphan nuclear hormone receptor C (RORC, p<0.001). TCRδ-deficient mice showed increased severity of experimental gut injury (p<0.05) with higher TNFα expression but downregulation of IL17A.

Conclusion: Complimentary mouse and human data suggest a role of γδ IEL in IL17 production and intestinal barrier production early in life. Specific loss of the γδ IEL fraction may contribute to NEC pathogenesis. Nutritional or pharmacological interventions to support γδ IEL maintenance in the developing small intestine could serve as novel strategies for NEC prevention.

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Related in: MedlinePlus

Occludin gene expression in human NEC versus control mucosa.(A) We measured gene expression levels of occluding by quantitative RT-PCR in 16 NEC tissue sections and 13 controls by quantitative RT-PCR array. Occludin gene expression was significantly decreased in NEC samples versus controls (p<0.0001). Relative level of mRNA expression of occludin in each sample was normalized to the expression level of reference gene GAPDH.
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pone-0099042-g009: Occludin gene expression in human NEC versus control mucosa.(A) We measured gene expression levels of occluding by quantitative RT-PCR in 16 NEC tissue sections and 13 controls by quantitative RT-PCR array. Occludin gene expression was significantly decreased in NEC samples versus controls (p<0.0001). Relative level of mRNA expression of occludin in each sample was normalized to the expression level of reference gene GAPDH.

Mentions: Occludin forms rings at sites of γδ IEL/epithelial contact and promotes γδ IEL migration into epithelial monolayers [40]. Enterocytes internalized occludin in experimental NEC but expression in human NEC was unchanged in the small intestine by immunohistochemistry [16]. We sought to determine occludin expression in human NEC tissue to test the possibility that reduced expression may inhibit migration of γδ IEL into the intraepithelial compartment. We found statistically significant reduction in occludin gene expression in by quantitative RT-PCR in 16 NEC tissue sections compared to 13 controls (p<0.0001, Figure 9).


Small intestinal intraepithelial TCRγδ+ T lymphocytes are present in the premature intestine but selectively reduced in surgical necrotizing enterocolitis.

Weitkamp JH, Rosen MJ, Zhao Z, Koyama T, Geem D, Denning TL, Rock MT, Moore DJ, Halpern MD, Matta P, Denning PW - PLoS ONE (2014)

Occludin gene expression in human NEC versus control mucosa.(A) We measured gene expression levels of occluding by quantitative RT-PCR in 16 NEC tissue sections and 13 controls by quantitative RT-PCR array. Occludin gene expression was significantly decreased in NEC samples versus controls (p<0.0001). Relative level of mRNA expression of occludin in each sample was normalized to the expression level of reference gene GAPDH.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4048281&req=5

pone-0099042-g009: Occludin gene expression in human NEC versus control mucosa.(A) We measured gene expression levels of occluding by quantitative RT-PCR in 16 NEC tissue sections and 13 controls by quantitative RT-PCR array. Occludin gene expression was significantly decreased in NEC samples versus controls (p<0.0001). Relative level of mRNA expression of occludin in each sample was normalized to the expression level of reference gene GAPDH.
Mentions: Occludin forms rings at sites of γδ IEL/epithelial contact and promotes γδ IEL migration into epithelial monolayers [40]. Enterocytes internalized occludin in experimental NEC but expression in human NEC was unchanged in the small intestine by immunohistochemistry [16]. We sought to determine occludin expression in human NEC tissue to test the possibility that reduced expression may inhibit migration of γδ IEL into the intraepithelial compartment. We found statistically significant reduction in occludin gene expression in by quantitative RT-PCR in 16 NEC tissue sections compared to 13 controls (p<0.0001, Figure 9).

Bottom Line: In human preterm infants, the proportion of IEL was reduced by 66% in 11 NEC ileum resections compared to 30 non-NEC controls (p<0.001).While γδ IEL dominated over conventional αβ IEL early in gestation in mice and in humans, γδ IEL were preferential decreased in the ileum of surgical NEC patients compared to non-NEC controls (50% reduction, p<0.05).Loss of IEL in human NEC was associated with downregulation of the Th17 transcription factor retinoic acid-related orphan nuclear hormone receptor C (RORC, p<0.001).

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Vanderbilt University School of Medicine and Monroe Carell Jr. Children's Hospital at Vanderbilt, Nashville, Tennessee, United States of America.

ABSTRACT

Background: Gastrointestinal barrier immaturity predisposes preterm infants to necrotizing enterocolitis (NEC). Intraepithelial lymphocytes (IEL) bearing the unconventional T cell receptor (TCR) γδ (γδ IEL) maintain intestinal integrity and prevent bacterial translocation in part through production of interleukin (IL) 17.

Objective: We sought to study the development of γδ IEL in the ileum of human infants and examine their role in NEC pathogenesis. We defined the ontogeny of γδ IEL proportions in murine and human intestine and subjected tcrδ-/- mice to experimental gut injury. In addition, we used polychromatic flow cytometry to calculate percentages of viable IEL (defined as CD3+ CD8+ CD103+ lymphocytes) and the fraction of γδ IEL in surgically resected tissue from infants with NEC and gestational age matched non-NEC surgical controls.

Results: In human preterm infants, the proportion of IEL was reduced by 66% in 11 NEC ileum resections compared to 30 non-NEC controls (p<0.001). While γδ IEL dominated over conventional αβ IEL early in gestation in mice and in humans, γδ IEL were preferential decreased in the ileum of surgical NEC patients compared to non-NEC controls (50% reduction, p<0.05). Loss of IEL in human NEC was associated with downregulation of the Th17 transcription factor retinoic acid-related orphan nuclear hormone receptor C (RORC, p<0.001). TCRδ-deficient mice showed increased severity of experimental gut injury (p<0.05) with higher TNFα expression but downregulation of IL17A.

Conclusion: Complimentary mouse and human data suggest a role of γδ IEL in IL17 production and intestinal barrier production early in life. Specific loss of the γδ IEL fraction may contribute to NEC pathogenesis. Nutritional or pharmacological interventions to support γδ IEL maintenance in the developing small intestine could serve as novel strategies for NEC prevention.

Show MeSH
Related in: MedlinePlus