探明无脊椎动物血细胞的来源和发生机制是深入研究其免疫系统、免疫机制的基础，也是免疫基础研究的焦点问题。现有研究多集中于节肢动物，对其他门类的研究很少。海胆隶属棘皮动物门进化地位特殊，除作为特种水产养殖动物，还是一种重要的模式生物。体腔细胞是海胆免疫防御功能的主要执行者，并行使一定的生理功能。由于研究手段的限制，关于海胆体腔细胞发生的认识非常贫乏。本项目拟利用免疫学检测技术从分子水平上研究海胆体腔细胞的发生机制。通过制备可识别不同类型海胆体腔细胞的单克隆抗体，利用其与细胞增殖标记（BrdU）抗体、有丝分裂标记抗体等的免疫标记法，结合组织学、体外培养、细胞周期检测等研究手段，从造血组织和自我增殖等可能途径入手，检测分析体腔细胞的增殖情况，弄清体腔细胞的组织来源与增殖方式，明确其发生机制，为了解海胆免疫系统的结构组成、功能，以及比较免疫学研究提供重要资料。

Invertebrate blood cells (hemocytes) are mainly comparable to cells of the myeloid lineage in vertebrates, because their main function is to participate in innate immunity and blood clotting. Source of hemocytes remains unknown in most of the invertebrete. Relevent researches mainly focused on Arthropods. As member of phylum Echinodermata, Sea urchins occupied a key position in evolution. Furthermore, sea urchins have been recognized as important model organisms in biological research. Meanwhile, sea urchins have been explored as a target of aquaculture. Coelomocyte of sea urchin play key roles both in immunology and physiological fuctions. The study of coelomocyte origin, make great sense in the research of immunology system. Two theories have been proposed to address this issue: one involving specific organs or tissues as the source of celomocytes while the other points at the celomocytes themselves as self-replicating cells. However,the evidence to ascertain the origin of celomocytes is far from definitive, and would not be acceptable by modern scientific standards. Thus, it is necessary to use modern methodologies to verify the celomocyte origins. The present program propose to utilize monoclonal antibodies against coelomocyte and the methods combined of BrdU and H3 labeling, immunology, histology and primary culture to investigate the orgin of coelomocyte. It is supposed to clarify the cytopoietic organ and self-replicating capability of celomocytes. The results will provide vital informations for the study of composition, function and development of sea urchin immune system and comparative immunology.

体腔细胞是海胆免疫防御功能的主要执行者并行使一定的生理功能，然而有关其来源和发生机制的认识却非常贫乏。本项目即以探明体腔细胞的组织来源与增殖方式为目的。通过实施，主要取得以下重要结果：（1）构建中间球海胆体腔细胞单克隆抗体库，其可作为检测探针，用于海胆体腔细胞的分布、发生、来源以及功能等分析研究；（2）探明海胆在缺失本分体腔液后可迅速生产体腔细胞，并在机体正反馈与负反馈的共同调节作用下逐渐恢复。具体表现为6h即可达到甚至超过原有细胞密度，12h后体腔细胞密度基本与原水平持平，18h-24h细胞增殖又出现增强；（3）利用制备的体腔细胞单克隆抗体及细胞增殖标记确定海胆中轴器中存在体腔细胞及有丝分裂。在海胆体腔细胞修复期间，中轴器有丝分裂信号随修复时间延长呈现一定的变化，同时，修复期中轴器出现组织松散、空腔化，外壁上皮细胞层破解、脱落等情况，表明海胆中轴器具有体腔细胞造血功能；（4）经细胞增殖信号检测发现个别细胞呈现绿色的荧光信号，其中发现处于分裂末期的细胞，表明海胆的游离体腔细胞也具有增殖复制能力；（5）对正常及体腔细胞恢复期的海胆中轴器与体腔细胞进行转录组测序分析，显示中轴器与体腔细胞转录组数据库相似度较高，中轴器具有大量与细胞生长、分化相关的基因，其在体腔修复旺盛期表达上调。体腔细胞亦存在细胞生长相关基因，但在修复期变化较小。推测在急性缺血状态下，中轴器造血起到更为关键的作用。项目研究结果表明，海胆的体腔细胞可由中轴器造血产生，同时体腔细胞自身也具有自我增殖能力，研究结果填补了相关认识的空白，也为了解海胆免疫系统的结构组成、功能，以及比较免疫学研究提供了重要资料。本项目资助已发表SCI收录论文3篇，另Fish and shellfish immunology期刊在投论文1篇（小修中）。