Laboratory of Pathology and Immunology of Aquatic Animals, LMMEC, Ocean University of China, Qingdao 266003, PR China.
Fish Shellfish Immunol. 2010 Feb;28(2):333-43. doi: 10.1016/j.fsi.2009.11.015. Epub 2009 Nov 26.
The humoral and cellular immune responses of Japanese flounder, Paralichthys olivaceus, were investigated following intraperitioneal injection with outer membrane protein (OMP) of Edwardsiella tarda in Freund's incomplete adjuvant (FIA). The specific serum antibody titre against OMP of E. tarda were measured using ELISA for 14 weeks, and the total serum antibody concentrations were also determined according to the sandwich ELISA standard model constructed using purified IgM. Both of the specific and total antibodies had an increase and reached their peaks 4 weeks after immunization. Simultaneously, the percentages of sIg + lymphocytes in blood, spleen, pronephros and mesonephros were detected by flow cytometry. It was shown that the percentages of sIg + lymphocytes in all lymphoid organs reached their peak levels 4 weeks after immunization, and then decreased gradually. To investigate the protection against infection, three challenges were performed in the same way at day 14, 30 and 100 after immunization, fish challenged at day 30 showed a higher relative percentage survival (RPS) of 71 compared to the 14-day group (30) and 100-day group (53), which indicated a positive correlation between the survival and the levels of the antibody.
本研究旨在探讨牙鲆外周血、头肾、脾脏和中肾 sIg+淋巴细胞的百分比变化，探讨牙鲆外周血、头肾、脾脏和中肾 sIg+淋巴细胞的百分比变化与体液免疫和细胞免疫之间的关系。采用弗氏不完全佐剂（FIA）对牙鲆进行腹腔注射，研究爱德华氏菌外膜蛋白（OMP）对牙鲆体液免疫和细胞免疫的影响。采用 ELISA 法检测牙鲆血清中针对 E. tarda OMP 的特异性抗体效价，根据建立的纯化 IgM 夹心 ELISA 标准曲线，测定总抗体浓度。结果表明，免疫后 4 周，特异性和总抗体均呈上升趋势，并达到峰值。同时，采用流式细胞术检测牙鲆外周血、头肾、脾脏和中肾中 sIg+淋巴细胞的百分比。结果显示，免疫后 4 周，所有免疫器官中 sIg+淋巴细胞的百分比均达到峰值，随后逐渐下降。为了探讨免疫保护作用，在免疫后第 14、30 和 100 天进行了 3 次攻毒试验。结果表明，在第 30 天攻毒的牙鲆的相对百分比存活率（RPS）为 71%，显著高于第 14 天（30%）和第 100 天（53%）组，这表明存活率与抗体水平之间存在正相关关系。
