Characterisation of CD4 T cells in healthy and diseased koalas (Phascolarctos cinereus) using cell-type-specific monoclonal antibodies

Chandan Mangar; Charles W Armitage; Peter Timms; Lynn M Corcoran; Kenneth W Beagley

doi:10.1016/j.dci.2016.02.018

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Characterisation of CD4 T cells in healthy and diseased koalas (Phascolarctos cinereus) using cell-type-specific monoclonal antibodies

Journal article

Peer reviewed

Characterisation of CD4 T cells in healthy and diseased koalas (Phascolarctos cinereus) using cell-type-specific monoclonal antibodies

Chandan Mangar, Charles W Armitage, Peter Timms, Lynn M Corcoran and Kenneth W Beagley

Development and Comparative Immunology, Vol.60, pp.80-90

2016

DOI: https://doi.org/10.1016/j.dci.2016.02.018

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Abstract

koala

chlamydia

CD4 T cell

monoclonal antibody

The koala (Phascolarctos cinereus) is an arboreal herbivorous marsupial that is an Australian icon. Koalas in many parts of Australia are under multiple threats including habitat destruction, dog attacks, vehicular accidents, and infectious diseases such as Chlamydia spp. and the koala retrovirus (KoRV), which may contribute to the incidence of lymphoma and leukaemia in this species. Due to a lack of koala-specific immune reagents and assays there is currently no way to adequately analyse the immune response in healthy, diseased or vaccinated animals. This paper reports the production and characterisation of the first anti-koala CD4 monoclonal antibody (mAb). The koala CD4 gene was identified and used to develop recombinant proteins for mAb production. Fluorochrome-conjugated anti-CD4 mAb was used to measure the levels of CD4+ lymphocytes collected from koala spleens (41.1%, range 20-45.1%) lymph nodes (36.3%, range 19-55.9%) and peripheral blood (23.8%, range 17.3-35%) by flow cytometry. Biotin-conjugated anti-CD4 mAb was used for western blot to determine an approximate size of 52 kDa for the koala CD4 molecule and used in immunohistochemistry to identify CD4+ cells in the paracortical region and germinal centres of spleen and lymph nodes. Using the anti-CD4 mab we showed that CD4 cells from vaccinated, but not control, koalas proliferated following in vitro stimulation with UV-inactivated Chlamydia pecorum and recombinant chlamydial antigens. Since CD4+ T cells have been shown to play a pivotal role in clearing chlamydial infection in both human and mouse infections, using this novel antibody will help determine the role CD4+ T cells play in protection against chlamydial infection in koalas and also enhance our knowledge of how KoRV affects the koala immune system.

Details

Title: Characterisation of CD4 T cells in healthy and diseased koalas (Phascolarctos cinereus) using cell-type-specific monoclonal antibodies
Authors: Chandan Mangar (Author) - Queensland University of Technology
Charles W Armitage (Author) - Queensland University of Technology
Peter Timms (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Lynn M Corcoran (Author) - University of Melbourne
Kenneth W Beagley (Author) - Queensland University of Technology
Publication details: Development and Comparative Immunology, Vol.60, pp.80-90
Publisher: Pergamon
Date published: 2016
DOI: 10.1016/j.dci.2016.02.018
ISSN: 0145-305X
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99449175002621
Output Type: Journal article

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Collaboration types: Domestic collaboration
Web Of Science research areas: Fisheries; Immunology; Veterinary Sciences; Zoology

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