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Co-stimulation

DCs may undergo a number of phenotypical changes, following maturation by the uptake and processing of an antigen. The process of DC maturation, in general, involves an increase in the surface expression of co-stimulatory molecules, like CD40, CD80, CD83, and CD86, which can be measured by flow cytometry. As a negative control unloaded DCs can be taken along, as well as a DC positively stimulated with a known maturation cocktail, like IL-1beta, IL-6, TNF and PGE-2 or TLR ligands.

Cytokine Production

Besides up-regulation of co-stimulatory molecules, DCs produce a large variety of cytokines that could be either pro/anti-inflammatory or skew the phenotype of T cells. Therefore, a quite simple procedure to test the activity of the DCs towards the food bioactives is the measurement of cytokine levels. A wide variety of cytokines may be expressed by mature DCs including IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-8, IL-10, IL-12, IL-15, IL-16, IL-17, IL-18, IFN-alpha, IFN-beta, IFN-gamma, and TNF. These

cytokines can be measured by ELISA, or multiplex assays.

Other DC Readouts

Co-stimulation and cytokine production are effect parameters used in the vast majority of studies on effects of food bioactives on DCs (see Table 17.3 for an overview). Studying the induction of specific T cells is complicated due to the low

Table 17.3 In vitro studies using food-associated antigens in in vitro DC models

DC source

Food bioactive group

Specific food bioactives

Additional DC activator

Expression costimulation

Cytokine production

Other

MLR

References

CD34+

derived DC

Probiotics

L. paracasei CNCM I-4034

B. breve CNCM I-4035

S. typhi CECT 725 or LPS

No

Yes: various

TLR expression

No

BermudezBrito et al. (2012, 2013)

pDC (PBMC)

Probiotics

L. lactis JCM5805

Yes: HLA-DR and CD86

Yes:

IFN-alpha

IFN and IRF expression

No

Sugimura

et al. (2013)

Lamina propria and PBMC

Probiotics

Probiotic preparation VSL#3

LPS

Yes: CD40 and CD80

Yes: IL-10

and IL-12

Effect on T cell responses

No

Hart et al. (2004)

Lamina propria and PBMC

Probiotics

L. plantarum

BMCM12

No

Yes: various

Yes

Bernardo

et al. (2012)

MoDC

Probiotics

Lactobacillus rhamnosus Lcr35

Yes: CD83, CD86,

CD209 and HLA-DR

Yes: various

Microarray gene expression profile

No

Evrard et al. (2011)

MoDC

Polyphenols

Protocatechuic acid

LPS

Yes: CD83, CD86

and HLA-DR

Yes: various

Migration

PPARg activation

No

Del Cornò

et al. (2014b)

THP-1-

derived

Polyphenols

Apple polyphenol extract

OVA

(endotoxin?)

Yes: CD86 and HLA-DR

Yes: IL-1b, IL-10, IL-12

and TNF-a

Antigen uptake

No

Katayama

et al. (2013)

MoDC

Isoflavones

Genistein and Daidzein

LPS, TNFa, CT

Yes: CD83, CD80,

CD86, and HLA-DR

Yes: IL-6,

IL-12, TNF-a

and IL-10

DC–NK interaction

Yes

Wei et al. (2012)

MoDC

Isoflavones

Genistein, Daidzein and Glycitein

CT

Yes: CD83, CD80

and CD86

Yes: IL-6

and IL-8

Th-cytokine induction (IL-5, IL-9, IL-13)

Yes

Masilamani et al. (2011)

MoDC

Curcuminoids

Curcumin

LPS or Poly I:C

Yes: CD86, CD83, CD54 and HLA-DR

Yes: various

Migration Endocytosis

Yes

Shirley et al. (2008)

MoDC

Curcuminoids

Curcumin

LPS

Yes: CD40, CD83, CD80, CD86 and HLA-DR

Yes: various

FOXP3 induction Negative regulatory molecules (PDL1/2)

Yes

Rogers et al. (2010)

MoDC

Vitamins

1alpha,25-(OH)2 D3

LPS or PGN

Yes: CD40, CD83, CD80, CD86 and HLA-DR

Yes: IL-6, IL-8,

IL-12p70

and IL-10

No

BrosbølRavnborg et al. (2013)

MoDC

Vitamins

1,25-(OH)2 D3

LPS or CD40L

Yes: CD40, CD83, CD80, CD86 and HLA-DR

Yes:

IL-12p70

Endocytosis

Yes

Piemonti

et al. (2000)

MoDC

Proteins

Peptic fragments of gliadin, pRQP and pDAC (decapeptides)

Yes: CD83, CD80, CD86, CCR7 and HLA-DR

Yes: IL-12,

TNF-a and IL-10

Yes

Giordani

et al. (2014)

MoDC

Proteins

Soluble peanut Ag (PNAg) and purified Ara h 1

LPS and CT (positive controls)

Yes: CD83, CD40,

CD86, and HLA-DR

No

Th-cytokine induction (IFN-g, IL4 and

IL-13)

Yes

Shreffler

et al. (2006)

MoDC

Proteins

Bet v 1, Mal d 1, Api g 1 and Dau c 1

Yes: CD40, CD83, CD80, CD86 and HLA-DR

No

Th-cytokine induction (IFN-g, IL4, IL-10

and IL-13)

No

Smole et al. (2010)

MoDC

Proteins

Peptic fragments of gliadin, soya protein, and OVA

Yes: CD83, CD80, CD86 and HLA-DR

Yes: various

Endocytosis and signal transduction

Yes

Palova-

Jelinkova

et al. (2005)

frequency of T cell precursors with the corresponding TCR and requires extensive culture conditions and broad read-out tools as the TCR specificity in these conditions is unknown. With regard to the T cell stimulatory potential of DCs the mixed lymphocyte reaction (MLR), in which DCs are used to stimulate alloreactive (naïve) T cells, can be considered. Proliferation in combination with cytokine production can be used as a read-out for T cell skewing.

 
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