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Production and characterization of monoclonal antibodies raised against recombinant human granzymes A and B and showing cross reactions with the natural proteins

机译：产生和表征针对重组人颗粒酶A和B的单克隆抗体，并显示与天然蛋白的交叉反应

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The human serine proteases granzymes A and B are expressed in cytotoplasmic granules of activated cytotoxic T lymphocytes and natural killer cells. Recombinant granzyme A and granzyme B proteins were produced in bacteria, purified and then used to raise specific mouse monoclonal antibodies. Seven monoclonal antibodies (mAb) were raised against granzyme A, which all recognized the same or overlapping epitopes. They reacted specifically in an immunoblot of interleukin-2 (IL-2) stimulated PBMNC with a disulfide-linked homodimer of 43 kDa consisting of 28 kDa subunits. Seven mAb against granzyme B were obtained, which could be divided into two groups, each recognizing a different epitope. On an immunoblot, all mAb reacted with a monomer of 33 kDa protein. By immunohistochemistry, these mAb could be used to detect granzymes A and B expression in activated CTL and NK cells. The availability of these mAb may facilitate studies on the role of human cytotoxic cells in various immune reactions and may contribute to a better understanding of the role of granzmes A and B in the cytotoxic response in vivo.
机译：人丝氨酸蛋白酶颗粒酶A和B在活化的细胞毒性T淋巴细胞和天然杀伤细胞的细胞质颗粒中表达。重组粒酶A和粒酶B蛋白在细菌中产生，纯化，然后用于产生特异性小鼠单克隆抗体。产生了七种针对颗粒酶A的单克隆抗体（mAb），它们均识别相同或重叠的表位。他们在白介素2（IL-2）刺激的PBMNC的免疫印迹中与由28 kDa亚基组成的43 kDa二硫键连接的同型二聚体反应。获得了针对粒酶B的七个mAb，可以将其分为两组，每个组识别不同的表位。在免疫印迹上，所有mAb与33 kDa蛋白的单体反应。通过免疫组织化学，这些mAb可用于检测活化的CTL和NK细胞中颗粒酶A和B的表达。这些mAb的可用性可能有助于研究人类细胞毒性细胞在各种免疫反应中的作用，并且可能有助于更好地了解颗粒酶A和B在体内细胞毒性反应中的作用。

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