Tucková L, Bilej M
Department of Immunology, Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague.
Immunol Lett. 1994 Jul;41(2-3):273-7. doi: 10.1016/0165-2478(94)90145-7.
The administration of protein antigens into earthworms Lumbricus terrestris and Eisenia foetida induces the formation of antigen-binding protein (ABP) with the maximum response occurring between days 4 and 8. High proteolytic activities observed both in coelomocytes and in coelomic fluids cause rapid antigen degradation; the majority of antigen is digested during the first 24 h. To analyze the role of proteolytic processing of antigen in ABP response in vitro, the intact antigen (ARS-HSA) as well as its proteolytic fragments were added to tissue explant cultures and ABP formation followed by indirect ELISA using mAb to ABP. The response to small fragments was comparable to that induced by intact ARS-HSA. Furthermore, the response to intact antigen was almost completely blocked by the non-toxic serine protease inhibitor Pefabloc, while the response to small fragments was only slightly reduced. The kinetics of response to intact antigen significantly differed from that induced by small (< 3 kDa) fragments. We suggest that proteolytic processing is involved in the stimulation of ABP formation.
将蛋白质抗原注射到蚯蚓(陆正蚓和赤子爱胜蚓)体内会诱导抗原结合蛋白(ABP)的形成,在第4天至第8天之间出现最大反应。在体腔细胞和体腔液中均观察到高蛋白水解活性,这会导致抗原迅速降解;大部分抗原在最初的24小时内被消化。为了在体外分析抗原的蛋白水解加工在ABP反应中的作用,将完整抗原(ARS-HSA)及其蛋白水解片段添加到组织外植体培养物中,并使用抗ABP单克隆抗体通过间接ELISA跟踪ABP的形成。对小片段的反应与完整ARS-HSA诱导的反应相当。此外,对完整抗原的反应几乎完全被无毒丝氨酸蛋白酶抑制剂苯甲磺酰氟阻断,而对小片段的反应仅略有降低。对完整抗原的反应动力学与小(<3 kDa)片段诱导的反应动力学显著不同。我们认为蛋白水解加工参与了ABP形成的刺激过程。