Developing the first six-plus multi-colour flow cytometry assays for measuring immune responses in sheep following nematode infection. — Australasian Cytometry Society

Developing the first six-plus multi-colour flow cytometry assays for measuring immune responses in sheep following nematode infection. (24136)

Joanna Roberts 1 , Saleh Umair 2 , Charlotte Bouchet 2 , Qing Deng 2 , Anton Pernthaner
  1. flowjoanna, RD5 Palmerston North, New Zealand
  2. Animal Health , AgResearch Ltd, Palmerston North, Manawatu, New Zealand

The immune system of the sheep has many similarities to humans and can offer strategies for modelling the human immune system that smaller rodent models cannot. These include ease of regular abundant blood sampling and the ability to collect afferent lymph draining from tissues to lymph nodes (using surgical canulation). There are also differences, in particular the abundance of γδ T cells which are relatively scarce in humans. The selection of monoclonal antibodies available for investigations of the ovine immune response is limited in comparison to the mouse and human reagent repertoires. Those that are available have not commonly been stocked in an extensive range of directly conjugated fluorphores. This has traditionally rendered ovine flow cytometry assays limited in their ability to conduct multi-parameter enquiries about the nature of the sheep immune response with the majority of published studies limited to two or four colours. With the advent in recent years of conjugation kits for the straight forward production of direct antibody-fluor conjugates such as the LYNX kit from BioRad and the Biotium kits from Biotium, we are developing multi-colour panels (six or more colours) to track immune responses in the abomasal (stomach) lymph node and blood of sheep using proprietary and in house monoclonal antibodies. We show data from these panels measuring the γδ T cell, CD4 T cell and antigen presenting cell response following vaccination against, and experimental parasitism of sheep.

 

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