Department of Biology, University of Padova, Via Ugo Bassi 58/B, 35131 Padova, Italy.
Fish Shellfish Immunol. 2010 Apr;28(4):534-41. doi: 10.1016/j.fsi.2009.12.003. Epub 2009 Dec 29.
For the first time, a functional study of haemocytes from the crab Carcinus aestuarii was performed in order to evaluate their involvement in immune responses. Total haemocyte count (THC), phagocytosis, haemolymph opsonisation properties, hydrolytic and oxidative enzyme activities, and production of intracellular superoxide anion were evaluated. A great variability in THC was recorded among individuals, and haemocyte mean number was 6.4 (x10(6)) cells/ml haemolymph. Although only hyalinocytes were able to phagocytose yeast cells or Zymosan, phagocytic index was low (3%) and did not increase significantly (4%) after pre-incubation of yeast and Zymosan in cell-free haemolymph, suggesting that haemolymph did not have opsonising properties. All haemocyte types produced superoxide anion, whereas only granulocytes were positive to the hydrolytic enzymes assayed. In addition, only granulocytes were positive to phenoloxidase activity. Both Petri dish and spectrophotometric assays revealed a very low lysozyme-like activity in cell-free haemolymph (CFH) and haemocyte lysate (HL), although enzyme activity was higher in CFH than in HL. Interestingly, normalisation of data as to total protein content in CFH and HL resulted in an opposite situation, lysozyme-like activity being higher in HL than in CFH. This demonstrated that haemolymph of C. aestuarii has a high quantity of total proteins, functional properties of which need to be better investigated in future studies. Overall, the results obtained in the present study indicated that C. aestuarii haemocytes are not very active phagocytic cells, but they are more active in terms of both hydrolytic and oxidative enzyme activities and superoxide anion production.
首次对蟹类 Carcinus aestuarii 的血细胞进行了功能研究，以评估其在免疫反应中的作用。评估了总血细胞计数 (THC)、吞噬作用、血淋巴调理特性、水解和氧化酶活性以及细胞内超氧阴离子的产生。个体之间 THC 的变化很大，血细胞的平均数量为 6.4(x10(6))个细胞/ml 血淋巴。尽管只有透明细胞能够吞噬酵母细胞或 Zymosan，但吞噬指数很低（3%），并且在用无细胞血淋巴孵育酵母和 Zymosan 后没有显著增加（4%），表明血淋巴没有调理特性。所有血细胞类型都产生超氧阴离子，而只有粒细胞对测定的水解酶呈阳性。此外，只有粒细胞对酚氧化酶活性呈阳性。在 Petri 盘和分光光度法测定中，无细胞血淋巴 (CFH) 和血细胞裂解液 (HL) 中的溶菌酶样活性都非常低，尽管 CFH 中的酶活性高于 HL。有趣的是，将 CFH 和 HL 中的总蛋白含量数据归一化后，情况正好相反，HL 中的溶菌酶样活性高于 CFH。这表明蟹类 C. aestuarii 的血淋巴含有大量总蛋白，其功能特性需要在未来的研究中进一步研究。总的来说，本研究的结果表明，C. aestuarii 的血细胞不是非常活跃的吞噬细胞，但它们在水解和氧化酶活性以及超氧阴离子产生方面更为活跃。
