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 (Corresponding Author), Christopher J. Secombes (Corresponding Author)

Research output: Contribution to journalArticle

Abstract

IL-22 is a critical cytokine which is involved in modulating tissue responses during inflammation, and is produced mainly by T cells and innate leucocytes. In mammals, IL-22 is a key component in mucosal defences, tissue repair, epithelial cell survival and proliferation. In teleosts, IL-22 has been cloned and studied in several species, and the transcript is highly expressed in mucosal tissues and induced by pathogen associated molecular patterns (PAMPs), suggesting IL-22 also functions as an important component of the innate immune response in fish. To investigate these immune responses further, we have validated and characterised two monoclonal antibodies (mAbs) which were raised against two different peptide immunogens of salmonid IL-22. Our results show that both mAbs specifically react to their own peptide immunogens and recombinant IL-22, and are able to detect the induction of native protein expression after stimulation. In flow cytometry, an increase in IL-22 positive cells was detected after stimulation in vitro with cytokines and PAMPs and in vivo after bacterial challenge. The immunohistochemistry results showed that IL-22 is highly upregulated in the gills after challenge, both in cells within the gill filaments and in the interbranchial lymphoid tissue. Such results suggest IL-22 may have a role in triggering local antimicrobial defences in fish that may facilitate efficient microbial clearance. Hence monitoring IL-22 producing cells/protein secretion may provide an alternative mean to assess the effectiveness of mucosal vaccines.
Original languageEnglish
Article number103449
JournalDevelopmental and Comparative Immunology
Volume101
Early online date12 Jul 2019
DOIs
Publication statusE-pub ahead of print - 12 Jul 2019

Fingerprint

Oncorhynchus mykiss
Proteins
Fishes
Mucous Membrane
Monoclonal Antibodies
interleukin-22
Cytokines
Peptides
Lymphoid Tissue
Innate Immunity
Mammals
Cell Survival
Flow Cytometry
Leukocytes
Vaccines
Epithelial Cells
Immunohistochemistry
Cell Proliferation
Inflammation
T-Lymphocytes

Keywords

  • rainbow trout
  • interleukin-22
  • transcript expression
  • protein expression
  • monoclonal antibody

Cite this

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

In: Developmental and Comparative Immunology, Vol. 101, 103449, 12.2019.

Research output: Contribution to journalArticle

Hu, Yehfang ; Carpio, Yamila ; Scott, Callum ; Alnabulsi, Ayham ; Alnabulsi, Abdo ; Wang, Tingyu ; Liu, Fuguo ; Monte, Milena ; Wang, Tiehui ; Secombes, Christopher J. / Induction of IL-22 protein and IL-22-producing cells in rainbow trout Oncorhynchus mykiss. In: Developmental and Comparative Immunology. 2019 ; Vol. 101.
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abstract = "IL-22 is a critical cytokine which is involved in modulating tissue responses during inflammation, and is produced mainly by T cells and innate leucocytes. In mammals, IL-22 is a key component in mucosal defences, tissue repair, epithelial cell survival and proliferation. In teleosts, IL-22 has been cloned and studied in several species, and the transcript is highly expressed in mucosal tissues and induced by pathogen associated molecular patterns (PAMPs), suggesting IL-22 also functions as an important component of the innate immune response in fish. To investigate these immune responses further, we have validated and characterised two monoclonal antibodies (mAbs) which were raised against two different peptide immunogens of salmonid IL-22. Our results show that both mAbs specifically react to their own peptide immunogens and recombinant IL-22, and are able to detect the induction of native protein expression after stimulation. In flow cytometry, an increase in IL-22 positive cells was detected after stimulation in vitro with cytokines and PAMPs and in vivo after bacterial challenge. The immunohistochemistry results showed that IL-22 is highly upregulated in the gills after challenge, both in cells within the gill filaments and in the interbranchial lymphoid tissue. Such results suggest IL-22 may have a role in triggering local antimicrobial defences in fish that may facilitate efficient microbial clearance. Hence monitoring IL-22 producing cells/protein secretion may provide an alternative mean to assess the effectiveness of mucosal vaccines.",
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note = "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) of the European Union (grant agreement No. 311993 TARGETFISH). YH was supported by a PhD Studentship from the Ministry of Education, Republic of China (Taiwan). Tingyu Wang was supported by the Ministry of Science and Technology, Republic of China (Taiwan) (MOST 107-2917-I-564-019). Fuguo Liu was supported by a Newton International Fellowship funded by the Academy of Medical Sciences, UK (AMS, NIF004\1036). Thanks also go to Dr. Dawn Shewring for excellent technical assistance and to Dr. Alex Douglas and Ms. Anna Harte for statistical advice.",
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AU - Hu, Yehfang

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AU - Alnabulsi, Ayham

AU - Alnabulsi, Abdo

AU - Wang, Tingyu

AU - Liu, Fuguo

AU - Monte, Milena

AU - Wang, Tiehui

AU - Secombes, Christopher J.

N1 - 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) of the European Union (grant agreement No. 311993 TARGETFISH). YH was supported by a PhD Studentship from the Ministry of Education, Republic of China (Taiwan). Tingyu Wang was supported by the Ministry of Science and Technology, Republic of China (Taiwan) (MOST 107-2917-I-564-019). Fuguo Liu was supported by a Newton International Fellowship funded by the Academy of Medical Sciences, UK (AMS, NIF004\1036). Thanks also go to Dr. Dawn Shewring for excellent technical assistance and to Dr. Alex Douglas and Ms. Anna Harte for statistical advice.

PY - 2019/7/12

Y1 - 2019/7/12

N2 - IL-22 is a critical cytokine which is involved in modulating tissue responses during inflammation, and is produced mainly by T cells and innate leucocytes. In mammals, IL-22 is a key component in mucosal defences, tissue repair, epithelial cell survival and proliferation. In teleosts, IL-22 has been cloned and studied in several species, and the transcript is highly expressed in mucosal tissues and induced by pathogen associated molecular patterns (PAMPs), suggesting IL-22 also functions as an important component of the innate immune response in fish. To investigate these immune responses further, we have validated and characterised two monoclonal antibodies (mAbs) which were raised against two different peptide immunogens of salmonid IL-22. Our results show that both mAbs specifically react to their own peptide immunogens and recombinant IL-22, and are able to detect the induction of native protein expression after stimulation. In flow cytometry, an increase in IL-22 positive cells was detected after stimulation in vitro with cytokines and PAMPs and in vivo after bacterial challenge. The immunohistochemistry results showed that IL-22 is highly upregulated in the gills after challenge, both in cells within the gill filaments and in the interbranchial lymphoid tissue. Such results suggest IL-22 may have a role in triggering local antimicrobial defences in fish that may facilitate efficient microbial clearance. Hence monitoring IL-22 producing cells/protein secretion may provide an alternative mean to assess the effectiveness of mucosal vaccines.

AB - IL-22 is a critical cytokine which is involved in modulating tissue responses during inflammation, and is produced mainly by T cells and innate leucocytes. In mammals, IL-22 is a key component in mucosal defences, tissue repair, epithelial cell survival and proliferation. In teleosts, IL-22 has been cloned and studied in several species, and the transcript is highly expressed in mucosal tissues and induced by pathogen associated molecular patterns (PAMPs), suggesting IL-22 also functions as an important component of the innate immune response in fish. To investigate these immune responses further, we have validated and characterised two monoclonal antibodies (mAbs) which were raised against two different peptide immunogens of salmonid IL-22. Our results show that both mAbs specifically react to their own peptide immunogens and recombinant IL-22, and are able to detect the induction of native protein expression after stimulation. In flow cytometry, an increase in IL-22 positive cells was detected after stimulation in vitro with cytokines and PAMPs and in vivo after bacterial challenge. The immunohistochemistry results showed that IL-22 is highly upregulated in the gills after challenge, both in cells within the gill filaments and in the interbranchial lymphoid tissue. Such results suggest IL-22 may have a role in triggering local antimicrobial defences in fish that may facilitate efficient microbial clearance. Hence monitoring IL-22 producing cells/protein secretion may provide an alternative mean to assess the effectiveness of mucosal vaccines.

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