Cloning and functional testing of rhesus macaque (Macaca mulatta) IL-9 and IL-33

恒河猴(Macaca mulatta)IL-9 和 IL-33 的克隆和功能测试

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作者:Sanghita Sarkar, Ann J Hessell, Nancy L Haigwood, James J Kobie

Background

IL-9 and IL-33 can profoundly influence immune responses. As a necessary first step toward defining their impact in the rhesus macaque model, we confirmed their endogenous expression and sequence identity and generated expression vectors for the recombinant expression of rhesus IL-9 and IL-33.

Conclusions

The recombinant rhesus IL-9 and IL-33 constructs produce biologically active cytokines that can act upon rhesus B cells.

Methods

RT-PCR and Sanger sequencing was used to define the expression and sequences for rhesus IL-9 and IL-33. The resulting recombinant cytokines were tested by ELISA and proliferation assays.

Results

Full-length rhesus IL-9 and the mature form of rhesus IL-33 share 78% and 73% nucleotide similarity, respectively, with humans. Both cytokines are expressed in lymphocytes, with IL-9 expression also evident in CD4+ T cells. Recombinantly expressed rhesus IL-9 and IL-33 were each biologically active in vitro, including enhancing the proliferation of a rhesus B cell line. Conclusions: The recombinant rhesus IL-9 and IL-33 constructs produce biologically active cytokines that can act upon rhesus B cells.

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