Lin Yi, Zeng Yaoying, Zhao Jingxian, Zeng Shan, Huang Jintao, Feng Zheng, Di Jingfang, Zhan Meiyi
Institute of Tissue Transplantation and Immunology, Jinan University, Guangzhou 510632, China.
J Reprod Immunol. 2004 Dec;64(1-2):133-43. doi: 10.1016/j.jri.2004.08.002.
The present study aims to address whether the analysis of CD45+CD86+ cells isolated from para-aortic lymph nodes (pLNs) is valuable in assessment of the status of local immunity at the murine feto-maternal interface. CBA/J x DBA/2 mice, virgin CBA/J mice, and CBA/J x BALB/c mice were used as an abortion-prone model (group A), nonpregnant controls (group N), and fertile controls (group F), respectively. The percentage of CD45+CD86+ cells in the CD45+ cell group (CD45+CD86+ percentage for short) and the absolute number of these cells were determined by means of flow cytometry (FCM), using mononuclear cells isolated from pLNs collected 5.5, 9.5, and 13.5 days post-coitum (dpc), respectively, and mononuclear cells isolated from placentas 13.5 dpc. To clarify the identity of these CD86+ cells, FCM was also performed with CD3, CD19, and DX5 as specific markers for murine T-cells, B-cells, and NK cells, respectively. Both resorption rate and absolute number of resorptions were significantly higher in group A (29.3%, 1.8+/-1.0) than in group F (4.8%, 0.3+/-0.5, P<0.001, respectively). Similarly, both cell percentage and absolute number of CD45+CD86+ cells in pLNs collected 13.5 dpc were significantly higher in group A than in group F (27.5+/-14.0% versus 12.3+/-7.1%, and 1362+/-687 versus 615+/-353, P=0.001, respectively). The CD45+CD86+ percentage was around 7.5% in nonpregnant CBA/J mice, similar to the 10.6% in CBA/JxDBA/2 mice 5.5 dpc, but had increased dramatically, to 23.9%, by 9.5 dpc (P<0.001 versus nonpregnant mice and P=0.002 versus CBA/JxDBA/2 mice 5.5 dpc), and remained at a higher level (27.5%) until 13.5 dpc. However, this trend was not observed in group F during pregnancy. The increased CD45+CD86+ percentage at day 9.5 of gestation, when resorption begins, may support the assumption that CD45+CD86+ cells play a role in the course of embryo resorption. Lymphocyte phenotypic analysis in the lymph nodes that drain the pregnant uterus may be helpful to assess the status of local immunity at the feto-maternal interface.
本研究旨在探讨分析从腹主动脉旁淋巴结（pLNs）分离出的CD45+CD86+细胞是否有助于评估小鼠胎儿-母体界面的局部免疫状态。CBA/J×DBA/2小鼠、未孕CBA/J小鼠和CBA/J×BALB/c小鼠分别用作易流产模型（A组）、非孕对照（N组）和可育对照（F组）。采用流式细胞术（FCM）测定CD45+细胞群中CD45+CD86+细胞的百分比（以下简称CD45+CD86+百分比）及其绝对数量，分别使用从交配后5.5、9.5和13.5天（dpc）收集的pLNs中分离的单核细胞，以及从13.5 dpc的胎盘中分离的单核细胞。为了明确这些CD86+细胞的身份，还分别以CD3、CD19和DX5作为小鼠T细胞、B细胞和NK细胞的特异性标志物进行了FCM检测。A组的吸收率和吸收绝对数（29.3%，1.8±1.0）均显著高于F组（4.8%，0.3±0.5，P均<0.001）。同样，A组13.5 dpc收集的pLNs中CD45+CD86+细胞的百分比和绝对数均显著高于F组（分别为27.5±14.0%对12.3±7.1%，以及1362±687对615±353，P = 0.001）。未孕CBA/J小鼠的CD45+CD86+百分比约为7.5%，与CBA/J×DBA/2小鼠5.5 dpc时的10.6%相似，但到9.5 dpc时显著增加至23.9%（与未孕小鼠相比P<0.001，与CBA/J×DBA/2小鼠5.5 dpc时相比P = 0.002），并在13.5 dpc时保持在较高水平（27.5%）。然而，F组在孕期未观察到这种趋势。妊娠第9.5天吸收开始时CD45+CD86+百分比增加，这可能支持CD45+CD86+细胞在胚胎吸收过程中起作用的假设。对妊娠子宫引流淋巴结进行淋巴细胞表型分析可能有助于评估胎儿-母体界面的局部免疫状态。
