Chickens

Coordinator: Pete Kaiser

Pete Kaiser is Head of the Avian Infectious Diseases Programme and Acting Head of Immunology at IAH and leads the avian component of the Toolbox. His contribution to the Toolbox focuses on the development of reagents to study avian immunology. This includes the identification of genes involved in immune regulation in chickens and turkeys, the cloning and expression of avian cytokines and CD molecules, the production of mAb for functional analyses and the development of downstream assays such as ELISA and ELIspot.

Key targets

chicken

  • IL-4 (mAbs, ELISA)
  • IL-13 (mAbs, ELISA)
  • IL-10 (mAbs, ELISA)

Summary of progress

Other targets

  • IL-6
  • IL-19
  • IL-22
  • CD14
  • CD25
  • CD34
  • FLT-3

Summary of progress

 

Progress on key targets for chickens
Updated 05 August 2009

The chicken genome encodes 85 cytokines that have been identified to date. The Toolbox has developed quantitative RT-PCR (Taqman) assays to over 70 of these, with antibodies also produced to chicken cytokines.

IL-4
Chicken IL-4 is among the family of cytokine genes comprising the first Th2 gene cluster to be characterised in a non-mammalian species along with GM-CSF, IL-3, IL-5 and IL-13 (see below).  Recombinant chicken IL-4 has been expressed and six mAb of different isotypes have been produced (see table).

IL-13  
In order to further understand the Th2 repertoire in chickens we have expressed recombinant chicken IL-13. Four mAb have been produced that are specific for chicken IL-13 (see table).

IL-10
Recombinant chicken IL-10 has been expressed and one mAb of IgM isotype has been produced and characterised (see table). 

Anti-avian cytokine mAb are being commercialised through SoftFlow (http://www.softflow.com) with whom we collaborate to develop cytometric bead arrays, ELISA and ELISPOT. 

 

Progress on other targets for chickens
Updated 05 August 2009

IFN-γ
Three mAbs to chicken IFN-γ have been generated, and a capture ELISA and ELISpot developed using two of these mAbs.

IL-6
Recombinant chicken IL-6 has been expressed and expressed in COS cells and assayed for biological function on neonatal chicken heterophils (see Ferro et al, 2005 in the publications list). Recently, three mAb have been produced against recombinant chicken IL-6 (see table, manuscript in preparation). A capture ELISA has been established.

IL-12
A capture ELISA for chicken IL-12 has recently been established using mAbs previously generated against IL-12β. The capture ELISA recognises both recombinant and native IL-12 p70.

IL-19
Chicken IL-19 has been cloned, expressed and a single antibody produced and characterised against it (see table). 

IL-22
Two anti-chicken IL-22 mAb have been raised (see table)

 CD14
Chicken CD14 has been cloned, expressed and antibodies produced which are being utilised in the purification of monocyte precursors from chicken bone marrow for in vitro growth of dendritic cells (see Wu et al, 2009 in publications list).  

 CD25
We have recently cloned chicken CD25 and expressed the recombinant protein. Immunisations have been carried out for mAb production.  

CD34
Chicken CD34 has been cloned, expressed and antibodies produced against the recombinant protein to be used in the purification of monocyte precursors from chicken bone marrow in conjunction with anti-CD14 antibodies (see above) for in vitro growth of chicken dendritic cells.

DC-LAMP
Chicken DC-LAMP has been cloned and expressed and immunisations have been carried out for mAb production.

Flt3
A full-length chicken Flt3 cDNA clone has recently been isolated.

Additional progress
We have cloned turkey IL-1β, IL-6, IL-10, IL-13, CXCLi2 and TGF-β4 and developed real-time qRT-PCR assays.

Five mAb have been developed and characterised against chicken IL-12b. Two mAb raised against chicken DEC205 and three mAb against chicken CD83 will facilitate studies of avian dendritic cells.

Turkey CD4, CD8 and CD28 have been cloned (see Powell et al, 2009 in publications list) and cross-reactivity of anti-chicken antibodies against turkey CD4 and CD8 has been demonstrated.

Antibodies to avian cell surface markers produced within the Toolbox are available from AbD Serotec (http://www.abdserotec.com)

new chicken reagents    

 

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