Differentiation of pDC and mDC from cord blood derived stem cells to study the effect of early nutritional intervention on allergy in vitro

B. Nandanan, D. Kumar, A.J. Nauta, J. Garssen, E. Sandalova

Research output: Contribution to journalMeeting AbstractOther research output

Abstract

Background: Accumulating evidence indicates that commensal microbiota is one of the key drivers of immune system development. Nutritional modulation of the gut microbiota has been identified as a novel approach for the allergy prevention. The ability of DCs to orchestrate allergic immune responses makes them unique targets for functional studies in allergy. DCs may also play a role when probiotic intervention is used to prevent or treat allergic diseases. Recent literature suggests both myeloid dendritic cells (mDCs) and plasmacytoid dendritic cells (pDCs) also play a role in allergic manifestations. Method: As the number of DCs is quite low in blood it become crucial to develop an in vitro methodology to develop both mDC and pDC from single donor blood, to study the mechanism of allergic modulation in individuals' in vitro. In addition, DCs derived from adult PBMCs might not exactly reflect what is happening in an infant, since the immune system in neonates is still naïve. Here we describe a method to differentiate pDC and mDC from cord blood-derived stem cells. CD34+ cells isolated from CBMC were cultured in presence of Human SCF, GM-CSF, IL-4, and Flt3L for mDC's differentiation and Human TPO, Flt3L, and IL-3 for pDCs. Toll-like-Receptor agonists (TLR) were added to the cells for activation and the cells were analysed for expression of activation markers and production of cytokines Results: We show that DCs obtained by this method express DC markers and readily respond to TLR stimulation. The yield of mDC was 1.2E6 cells and of pDC, 9.75E6 from 1.5E6 CD34+ cells. Activated cells showed high expression of cytokines compared to immature cells. Conclusion: Taken together, results of this study indicated that both pDC and mDCs can be derived from cord blood and thereby provides a methodology to study the development and maturation of naïve cells. This in vitro model can provide valuable insights into the influence of early nutritional programming through modulation of gut microbiota on DC development and response.
Original languageEnglish
Article number914
Pages (from-to)346
Number of pages1
JournalAllergy
Volume69
DOIs
Publication statusPublished - 1 Sept 2014

Keywords

  • marker
  • cytokine
  • toll like receptor agonist
  • probiotic agent
  • interleukin 3
  • interleukin 4
  • allergy
  • umbilical cord blood
  • stem cell
  • in vitro study
  • clinical immunology
  • human
  • modulation
  • methodology
  • blood
  • intestine flora
  • immune system
  • laryngeal mask
  • newborn
  • adult
  • donor
  • immune response
  • prevention
  • myeloid dendritic cell
  • infant
  • microflora
  • model
  • stimulation
  • plasmacytoid dendritic cell
  • commensal
  • allergic disease
  • maturation
  • cerebrospinal fluid

Fingerprint

Dive into the research topics of 'Differentiation of pDC and mDC from cord blood derived stem cells to study the effect of early nutritional intervention on allergy in vitro'. Together they form a unique fingerprint.

Cite this