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Recombinant cytokines as immunological adjuvants
A D Nash1, S A Lofthouse, G J Barcham
1Centre for Animal Biotechnology, School of Veterinary Science, University of Melbourne, Parkville, Victoria, Australia.
Immunology and Cell Biology
|October 1, 1993
Summary
Researchers produced bioactive recombinant ovine interleukins and tumor necrosis factor. Recombinant ovine interleukin-1 alpha and beta demonstrated significant adjuvant activity, enhancing sheep immune responses to antigens but not protection against parasitic infection.
Area of Science:
- Immunology
- Veterinary Science
- Biotechnology
Background:
- Ovine cytokines like interleukin-2 (IL-2), interleukin-1 alpha (IL-1α), and tumor necrosis factor alpha (TNFα) play crucial roles in immune regulation.
- Understanding the function of these cytokines in sheep is vital for developing effective vaccines and immunotherapies.
- Bacterial expression systems offer a scalable method for producing recombinant proteins for research and therapeutic applications.
Purpose of the Study:
- To express and purify bioactive recombinant ovine interleukin-2 (rovIL-2), interleukin-1 alpha (rovIL-1α), and tumor necrosis factor alpha (TNFα).
- To evaluate the immunological adjuvant potential of recombinant ovine interleukin-1 alpha (rovIL-1α) and interleukin-1 beta (rovIL-1β) in sheep.
- To assess the impact of these cytokines on humoral immune responses and protection against parasitic infections.
Main Methods:
- Bacterial expression and purification of recombinant ovine cytokines (rovIL-2, rovIL-1α, TNFα).
- Bioassays to determine the specific activity of purified recombinant proteins.
- Immunization of sheep with model antigens (avidin, H. contortus antigen) combined with recombinant ovine IL-1α or IL-1β, with or without aluminum hydroxide adjuvant.
- Measurement of humoral immune responses (specific antibody levels and isotype profiles) and assessment of protection against H. contortus larval challenge.
Main Results:
- Purified recombinant ovine cytokines exhibited specific activities of 1 x 10^7 U/mg (rovIL-2, rovIL-1α) and 1 x 10^5 U/mg (TNFα).
- Recombinant ovine IL-1α significantly enhanced the secondary humoral response to avidin when administered intradermally or intramuscularly.
- Recombinant ovine IL-1β significantly boosted the humoral response to an H. contortus antigen but did not improve protection against larval infection or alter antibody isotype profiles.
Conclusions:
- Bacterial expression and purification of bioactive recombinant ovine cytokines are feasible.
- Recombinant ovine IL-1α and IL-1β possess significant adjuvant properties, enhancing humoral immune responses in sheep.
- While enhancing antibody production, rovIL-1β did not confer protection against H. contortus infection, highlighting the complexity of immune responses and vaccine development.