恒河猴（猕猴）白细胞介素-9和白细胞介素-33的克隆及功能测试
Cloning and functional testing of rhesus macaque (Macaca mulatta) IL-9 and IL-33.
作者信息
Sarkar Sanghita, Hessell Ann J, Haigwood Nancy L, Kobie James J
机构信息
Infectious Diseases Division, University of Alabama at Birmingham, Birmingham, AL, USA.
Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, OR, USA.
出版信息
J Med Primatol. 2020 Jun;49(3):144-152. doi: 10.1111/jmp.12464. Epub 2020 Feb 4.
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.
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.
背景
白细胞介素-9（IL-9）和白细胞介素-33（IL-33）可深刻影响免疫反应。作为确定它们在恒河猴模型中作用的必要第一步，我们确认了它们的内源性表达和序列同一性，并构建了用于重组表达恒河猴IL-9和IL-33的表达载体。
方法
采用逆转录聚合酶链反应（RT-PCR）和桑格测序法来确定恒河猴IL-9和IL-33的表达及序列。通过酶联免疫吸附测定（ELISA）和增殖试验对所得重组细胞因子进行检测。
结果
恒河猴全长IL-9与成熟形式的恒河猴IL-33分别与人的核苷酸相似度为78%和73%。两种细胞因子均在淋巴细胞中表达，IL-9在CD4 + T细胞中的表达也很明显。重组表达的恒河猴IL-9和IL-33在体外均具有生物活性，包括增强恒河猴B细胞系的增殖。
结论
重组恒河猴IL-9和IL-33构建体产生的生物活性细胞因子可作用于恒河猴B细胞。
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