Induction of IL-22 protein and IL-22-producing cells in rainbow - - PowerPoint PPT Presentation

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Induction of IL-22 protein and IL-22-producing cells in rainbow - - PowerPoint PPT Presentation

Jul 15, 2023 343 likes •481 views

Induction of IL-22 protein and IL-22-producing cells in rainbow trout Oncorhynchus mykiss Yehfang Hu, Yamila Carpio, Callum Scott, Ayham Alnabulsi, Abdo Alnabulsi, Tingyu Wang, Fuguo Liu, Milena Monte, Tiehui Wang and Christopher J. Secombes

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SLIDE 1

Yehfang Hu, Yamila Carpio, Callum Scott, Ayham Alnabulsi, Abdo Alnabulsi, Tingyu Wang, Fuguo Liu, Milena Monte, Tiehui Wang and Christopher J. Secombes 18/06/2019 ISFSI

Induction of IL-22 protein and IL-22-producing cells in rainbow trout Oncorhynchus mykiss

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SLIDE 2

Interleukin (IL)-22

  • Belongs to IL-10 family

Anti-infection Anti- inflammatory IL-10 subfamily

IL-10 IL-19 IL-20 IL-22 IL-24 IL-26 Anti-inflammatory

  • Mainly produced by T cells, NK

cells and innate lymphoid cells (ILCs)

  • Mucosal defence and tissue

protection

  • Tissue regeneration
  • Cell proliferation
  • Antimicrobial molecule induction

Tissue Cell Antimicrobial peptide (AMP)

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SLIDE 3

Fish IL-22

  • Fish IL-22 has been identified
  • Similar functions to mammalian

IL-22 as mucosal barrier

  • High transcripts in mucosal tissues

(gill, intestine, fin, skin)

Mammalian Th17 immune network

(Tiehui Wang and Christopher J. Secombes. 2013)

Transcript IL-22 expression Protein IL-22 secretion

Monoclonal antibody

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SLIDE 4

Monoclonal antibody (mAb)

  • A. mAb production

Linear, accessible, hydrophilic, antigenic and low complexity region

Recognise induced native IL-22

(1) HK cells + PBS 72 h (2) HK cells + rIL21 72 h

  • B. mAb characterisation

Specific to peptide immunogen and rIL-22

(1) 100 μg rIL-22 (2) 100 μg rIL-2B

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SLIDE 5

IL-22 producing cells

  • A. Total PBLs gating
  • B. IL-22+ gating
  • Native IL-22 is

detectable by intracellular staining

  • Number of IL-22+ cells

increased by IL-22 inducing stimulants

  • Hypotonic PBL- rapid and high quantity (Hu et al., 2018)
  • IL-22 transcript inducing stimulants
  • C. Frequency of IL-22+
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SLIDE 6

Gill IL-22 post A. sal infection

% of IL-22+ cells

*

10 20 30 40 50 60 70

CTL AS

% of IL-22+ cells

*

  • B. Frequency of IL-22+
  • C. IL-22 transcript
  • D. IL-22 protein expression

(1) AS gill (2) Control gill (3) rIL-22

  • A. Total leucocytes/IL-22+ gating
  • Mucosal tissue-gill
  • Bacterial infection
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SLIDE 7

PBL IL-22 post A. sal infection

  • IL-22 transcript and protein are

increased post A. sal infection

  • IL-22 producing neutrophils?
  • C. IL-22 transcript
  • B. Frequency of IL-22+
  • A. Total leucocytes/IL-22+ gating
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SLIDE 8

Control SGL PGL SGL PGL: primary gill lamellae, SGL: secondary gill lamellae

IL-22+ cells in gill

  • A. sal 24 hpc

SGL PGL SGL Epithelial cells Undifferentiated basal cells

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SLIDE 9

IL-22+ cells in ILT

ILT

Control

  • A. sal 24 hpc

lu is

ILT

lu is

ILT: interbranchial lymphoid tissue, LU: lumen, IS: interbranchial septum ILT

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SLIDE 10

Conclusion

  • Two mAbs (L7 & L8) developed against rtIL-22
  • IL-22 protein expression is up-regulated in

respond to bacterial infection

  • Monitoring IL-22 to assess mucosal vaccines
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SLIDE 11

Acknowledgements

  • Prof. Chris Secombes
  • Dr. Tiehui Wang
  • Dr. Ayham Alnabusi
  • Dr. Dawn Shewring
  • Ms. Anna Harte

This work was funded by the Biotechnology and Biological Sciences Research Council (BBSRC, BB/N024052/1 and BB/R008442/1). This research was also funded by the European Commission under the 7th Framework Programme for Research and Technological Development (FP7)

  • f

the European Union (grant agreement No. 311993 TARGETFISH).

  • PGR award for traveling
  • ISFSI student bursary
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SLIDE 12