IN PIGS . Hermans L., Devriendt B., Favoreel H., Cox E . Haemolytic - - PowerPoint PPT Presentation

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IN PIGS . Hermans L., Devriendt B., Favoreel H., Cox E . Haemolytic - - PowerPoint PPT Presentation

Apr 01, 2023 122 likes •358 views

DEPARTMENT VIROLOGY, PARASITOLOGY AND IMMUNOLOGY RESEARCH GROUP IMMUNOLOGY THE EFFECTS OF -1,3-1,6-GLUCANS ON INNATE IMMUNE RESPONSES IN PIGS . Hermans L., Devriendt B., Favoreel H., Cox E . Haemolytic uremic syndrome STEC infection Renal

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THE EFFECTS OF β-1,3-1,6-GLUCANS ON INNATE IMMUNE RESPONSES IN PIGS.

Hermans L., Devriendt B., Favoreel H., Cox E.

DEPARTMENT VIROLOGY, PARASITOLOGY AND IMMUNOLOGY RESEARCH GROUP IMMUNOLOGY

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Haemolytic uremic syndrome

STEC infection

Renal failure Haemolytic anaemia Thrombocytopenia Public health Oedema Ataxia Death

C.L. Gyles (Ed.), Escherichia coli in Domestic Animals and Humans, CAB International, Wallingford, Oxon, UK (1994), p. 337 Wöchtl B et al. Comparison of clinical and immunological findings in gnotobiotic piglets infected with Escherichia coli O104 : H4 outbreak strain and EHEC O157 : H7. Gut Pathog. BioMed Central; 2017;1–18. Tseng M et al. Shiga toxin-producing Escherichia coli in swine: the public health perspective. Anim Health Res Rev. 2014;15(1):63–75.

β-glucans Economic losses Modulate innate immune response in pigs

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β-glucans stimulate PBMC ➔ cytokine production ➔ NK cell activation

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NK cells

APC features Memory-like PT CD3- CD172a- CD8a+ Cytotoxicity Shape immune responses

Rupert Handgretinger et al. Blood 2016;127:3341-3349 De Pelsmaeker et al. , J. Leukoc. Biol., 2018; 103(1); 129-140 Gerner et al., J Allergy Clin Immunol. 2013 September ; 132(3): 536–544

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STEC INFECTION IN GNOTOBIOTIC PIGLETS

Wöchtl B et al. Comparison of clinical and immunological findings in gnotobiotic piglets infected with Escherichia coli O104 : H4 outbreak strain and EHEC O157 : H7. Gut Pathog. BioMed Central; 2017;1–18

Potential role for NK cells during STEC infection

Increase of NKp46+ NK cells in PBMC: IFN-γ production

1. Control group 2.

  • E. Coli O104:H4 infection

3.

  • E. Coli O157:H7 infection
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Beta-glucans?!

Proven to modulate immunity MAMP PRR ▪ Dectin-1 ▪ CR3 Carbohydrate polymers

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FEED TRIAL WITH ETEC CHALLENGE

In-feed administered β-glucans can protect against ETEC infection

Stuyven et al., Vet Immunol Immunopathol, 2009; 128(1-3):60-6

Decrease in faecal ETEC excretion Lower antigen-specific IgM response

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Macrogard (Saccharomyces cerevisiae) Curdlan (Alcaligenes faecalis)

STRUCTURE?!

  • Solubility
  • Size
  • Branching frequency
  • Length of the branches
  • Molecular conformation
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NK cells Beta-glucans

Activation Effects on innate immune responses

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EFFECTS OF β–GLUCANS ON PORCINE NK CELLS

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Direct effects of β-glucans on pNK cells

̶ NK isolation ̶ IL-2 Priming (16-18h) ̶ Beta-glucan (50 µg/ml) (2h) ̶ CFSE-labelled K562 (4h) IL-2 Lysis of target cells

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Direct effects of β-glucans on pNK cells

̶ NK isolation ̶ IL-2 Priming (16-18h) ̶ Beta-glucan (50 µg/ml) (2h) ̶ CFSE-labelled K562 (4h)

W ith o u t g lu c a n M a c ro g a rd C u rd la n 0 .0 0 .5 1 .0 1 .5

R e la tiv e N K -m e d ia te d ly s is

  • f K 5 6 2 c e lls a fte r 1 8 h IL -2 p rim in g

No direct effect of β-glucans on pNK cells

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Indirect effects of β-glucans on pNK cells

̶ PBMC isolation ̶ β-glucan priming (10 µg/ml)/ IL-2 priming (50ng/ml) (18h) ̶ Medium (2h) ̶ CFSE-labelled K562 (4h) M / IL-2 / Lysis of target cells

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Indirect effects of β-glucans on pNK cells

̶ PBMC isolation ̶ β-glucan priming (10 µg/ml)/ IL-2 priming (50ng/ml) (18h) ̶ Medium (2h) ̶ CFSE-labelled K562 (4h)

M e d iu m IL -2 M G C L 2 0 4 0 6 0 8 0

A b s o lu te P B M C -m e d ia te d ly s is

* * * * MG: Macrogard CL: Curdlan

p<0,05: * ; p<0,001: ***

β-glucans do indirectly activate pNK cells

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Are these effects NK-mediated?

R e la tiv e ly s is

P B M C P B M C  N K P B M C P B M C  N K P B M C P B M C  N K P B M C P B M C  N K 5 1 0 1 5

M e d iu m IL -2 M G (1 0 µ g /m l) C L (1 0 µ g /m l)

Yes

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HOW DOES THIS WORK?

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Effect of supernatant on pNK cells

̶ PBMC isolation ̶ β-glucan priming (10 µg/ml)/ IL-2 priming (50ng/ml) (18h) ̶ Collecting culture SN ̶ FACS purified NK cells (18h)

̶ CFSE-labelled K562 cells (4h)

M / IL-2 / SN

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SLIDE 18

Effect of supernatant on pNK cells

̶ PBMC isolation ̶ β-glucan priming (10 µg/ml)/ IL-2 priming (50ng/ml) (18h) ̶ Collecting culture SN ̶ FACS purified NK cells (18h)

̶ CFSE-labelled K562 cells (4h)

A b s o lu te N K -m e d ia te d ly s is

M I L

  • 2

M G C L 2 0 4 0 6 0 8 0 1 0 0

* * * *

p<0,05: * ; p<0,001: ***

PBMC produce NK cell stimulating factors upon β-glucan priming

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Effect of monocytes/DC

P B M C -m e d ia te d ly s is o f K 5 6 2 c e lls

% ly s e

F u l l  C D 1 7 2 a

+

F u l l  C D 1 7 2 a

+

F u l l  C D 1 7 2 a

+

1 0 2 0 3 0 4 0 5 0 M e d ium IL -2 M G (1 0 µ g /m l)

*** **

p<0,01: ** ; p<0,001: ***

Role for CD172a+ cells: monocytes and/or DC

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CONCLUSIONS

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Beta-glucans Monocytes / DC Cytokines Cytotoxicity Cytokines ??? Activation of monocytes Activation of Th1 Maturation of DC

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Beta-glucans Monocytes / DC Cytokines Cytotoxicity Cytokines ??? Activation of monocytes Activation of Th1 Maturation of DC Shape innate immune response → Stronger resistance to infection

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Leen Hermans

DEPARTMENT VIROLOGY, PARASITOLOGY AND IMMUNOLOGY

Promotors: Prof. Eric Cox, Prof. Herman Favoreel Co-promotor: Dr. Bert Devriendt

ICONS AND FIGURES Post Pigeon by Alina Oleynik from the Noun Project Mail Password by iconsmind.com from the Noun Project Human by Rose Alice Design from the Noun Project BioRender

Acknowledgements