Increased expression and internalization of the endotoxin coreceptor CD14 in enterocytes occur as an early event in the development of experimental necrotizing enterocolitis

Mollen, Kevin P.; Gribar, Steven C.; Anand, Rahul; Kaczorowski, David J.; Kohler, Jeffrey W.; Branca, Maria F.; Dubowski, Theresa; Sodhi, Chhinder P.; Hackam, David J.

doi:10.1016/j.jpedsurg.2008.02.050

Cited by 23 publications

(15 citation statements)

References 31 publications

(35 reference statements)

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“…However, M.tb infection has been shown to upregulate the expression of CD14 on monocytes (Shams et al, 2003). Given the role of CD14 as the TLR4 co-receptor for LPS binding, the downregulation is likely due to macropinocytosis-mediated internalization upon TLR4 stimulation (Mollen et al, 2008; Poussin et al, 1998). Lower frequencies of CD14 + cells after stimulation may also be due to shedding of CD14 or monocyte death.…”

Section: Discussionmentioning

confidence: 99%

Optimization of a whole blood intracellular cytokine assay for measuring innate cell responses to mycobacteria

Shey¹,

Hughes²,

Kock³

et al. 2012

Journal of Immunological Methods

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Innate cells are essential for host defense against invading pathogens, and the induction and direction of adaptive immune responses to infection. We developed and optimized a flow cytometric assay that allows measurement of intracellular cytokine expression by monocytes, dendritic cells (DC) and granulocytes, as well as cellular uptake of green-fluorescent protein (GFP)-expressing mycobacteria, in very small volumes of peripheral blood. We show that innate cell stimulation resulted in increased granularity of monocytes and mDCs and decreased granulocyte granularity that precluded flow cytometric discernment of granulocytes from monocytes and myeloid DC by forward and side scatter gating. Anti-CD66a/c/e antibody staining allowed reliable identification and exclusion of granulocytes for subsequent delineation of monocytes and myeloid DC. Intracellular cytokine expression by granulocytes, monocytes and mDC was remarkably sensitive to the dose of mycobacterial inoculum. Moreover, activation of monocytes and mDCs with live BCG reduced expression levels of CD14 and CD11c, respectively, necessitating optimization of staining conditions to reliably measure these lineage markers. Finally, we characterized expression of IL-12/23p40, TNF-α, IL-6, and IL-10, by GFP+ and GFP− monocytes and mDC from 25 healthy adults. This assay may be applied to the study of innate cell responses to any GFP-expressing pathogen, and can be performed on blood volumes as low as 200µL per condition, making the assay particularly suitable for pediatric studies.

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Section: Discussionmentioning

confidence: 99%

Optimization of a whole blood intracellular cytokine assay for measuring innate cell responses to mycobacteria

Shey¹,

Hughes²,

Kock³

et al. 2012

Journal of Immunological Methods

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“…For example, the lipopolysaccharide (LPS, endotoxin) co-receptor CD14 and LPS binding protein (LBP, which binds intravascular LPS and facilitates its attachment to CD14) are both increased during neonatal sepsis83–85. Genetic variations in these proteins have been associated with increased risk for sepsis in adults47, 49, 50. Gene polymorphisms in myeloid differentiation-2 (MD-2), a small protein involved in LPS signaling through TLR4, increase the risk for organ dysfunction and sepsis in adults86 but the significance in neonates is unknown.…”

Section: Pathophysiology Of Sepsis and Shock:molecular And Cellular Ementioning

confidence: 99%

Pathophysiology and Treatment of Septic Shock in Neonates

Wynn

Wong

2010

Clinics in Perinatology

198

152

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Neonatal septic shock is a devastating condition associated with high morbidity and mortality. Definitions for the sepsis continuum and treatment algorithms specific for premature neonates are needed to improve studies of septic shock and assess benefit from clinical interventions. Unique features of the immature immune system and pathophysiologic responses to sepsis, particularly those of extremely preterm infants, necessitate that clinical trials consider them as a separate group. Keen clinical suspicion and knowledge of risk factors will help to identify those neonates at greatest risk for development of septic shock. Genomic and proteomic approaches, particularly those that use very small sample volumes, will increase our understanding of the pathophysiology and direct the development of novel agents for prevention and treatment of severe sepsis and shock in the neonate. Although at present antimicrobial therapy and supportive care remain the foundation of treatment, in the future immunomodulatory agents are likely to improve outcomes for this vulnerable population.

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“…In line with the above observations, studies have shown that increased TLR4 expression and up-regulated TLR4 signaling are associated with NEC in mice, rat and premature infants [106, 128–131]. In addition, NEC is associated with increased TLR4 activator HMGB1 [132], co-receptor myeloid differentiation protein-2 (MD-2) [133] and co-receptor cluster of differentiation 14 [134]. Proof that TLR4 is required for the pathogenesis of NEC is found in studies by Jilling et al [128], as well as from our group [106, 135], in which TLR4 mutant mice were protected from the development of NEC as compared to wild-type mice.…”

Section: The Role Of Tlr4 In the Pathogenesis Of Necmentioning

confidence: 83%

Toll-like receptor regulation of intestinal development and inflammation in the pathogenesis of necrotizing enterocolitis

2014

Self Cite

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Toll-like receptors (TLRs) are a structurally related family of molecules that respond to a wide variety of endogenous and exogenous ligands, and which serve as important components of the innate immune system. While TLRs have established roles in host defense, these molecules have also been shown to play important roles in the development of various disease states. A particularly important example of the role of TLRs in disease induction includes necrotizing enterocolitis (NEC), which is the most common gastrointestinal disease in preterm infants, and which is associated with extremely high morbidity and mortality rates. The development of NEC is thought to reflect an abnormal interaction between microorganisms and the immature intestinal epithelium, and emerging evidence has clearly placed the spotlight on an important and exciting role for TLRs, particularly TLR4, in NEC pathogenesis. In premature infants, TLR4 signaling within the small intestinal epithelium regulates apoptosis, proliferation and migration of enterocytes, affects the differentiation of goblet cells, and reduces microcirculatory perfusion, which in combination result in the development of NEC. This review will explore the signaling properties of TLRs on hematopoietic and non-hematopoietic cells, and will examine the role of TLR4 signaling in the development of NEC. In addition, the effects of dampening TLR4 signaling using synthetic and endogenous TLR4 inhibitors and active components from amniotic fluid and human milk on NEC severity will be reviewed. In so doing, we hope to present a balanced approach to the understanding of the role of TLRs in both immunity and disease pathogenesis, and to dissect the precise roles for TLR4 in both the cause and therapeutic intervention of necrotizing enterocolitis.

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Increased expression and internalization of the endotoxin coreceptor CD14 in enterocytes occur as an early event in the development of experimental necrotizing enterocolitis

Cited by 23 publications

References 31 publications

Optimization of a whole blood intracellular cytokine assay for measuring innate cell responses to mycobacteria

Optimization of a whole blood intracellular cytokine assay for measuring innate cell responses to mycobacteria

Pathophysiology and Treatment of Septic Shock in Neonates

Toll-like receptor regulation of intestinal development and inflammation in the pathogenesis of necrotizing enterocolitis

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