Sampson J E, Theve R P, Blatman R N, Shipp T D, Bianchi D W, Ward B E, Jack R M
Department of Obstetrics and Gynecology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Am J Obstet Gynecol. 1997 Jan;176(1 Pt 1):77-81. doi: 10.1016/s0002-9378(97)80015-4.
Although polymorphonuclear leukocytes are the inflammatory cells most frequently recovered from the amniotic cavity in cases of suspected intrauterine infection, the source of these cells has not been definitively determined. We took advantage of the gender difference between the mother and her male fetus, and we report four cases in which amniotic fluid polymorphonuclear leukocytes were identified as fetal by fluorescence in situ hybridization with probes specific for X and Y chromosomes. Fetal membranes were intact at the time amniotic fluid was obtained in all cases.
Amniotic fluid was obtained from women with male fetuses in premature labor with clinical or laboratory evidence of infection. Cytospin preparations of amniotic fluid samples with polymorphonuclear leukocytes were prepared and sequentially stained with fluorescent reagents. To determine which cells were polymorphonuclear leukocytes, all replicate samples were stained with the fluorescent nuclear stain 4'-6-diamidino-2-phenyl-indole. This allowed definition of the characteristic multilobed polymorphonuclear leukocytes nuclear morphologic features. The sample was then probed with a rhodamine-labeled probe specific for the X chromosome and a fluorescein-labeled probe specific for the Y chromosome to assess whether the polymorphonuclear leukocytes were male or female.
Ninety percent to 99% of polymorphonuclear leukocytes identified by normal multiple lobed nuclear morphologic study on 4'-6-diamidino-2-phenyl-indole staining had an X and Y chromosome and were therefore fetal cells.
These data demonstrate a fetal response during intraamniotic infection. Further investigation of the roles for maternal and fetal polymorphonuclear leukocytes in chorioamnionitis may provide valuable information about the critical interaction of the two immune responses in this setting.
尽管在怀疑有宫内感染的病例中，多形核白细胞是最常从羊膜腔中发现的炎性细胞，但其来源尚未明确确定。我们利用母亲与其男性胎儿之间的性别差异，报告了4例通过使用X和Y染色体特异性探针进行荧光原位杂交，将羊水多形核白细胞鉴定为胎儿来源的病例。在所有病例中，获取羊水时胎膜均完整。
从有早产男性胎儿且有临床或实验室感染证据的女性中获取羊水。制备含有多形核白细胞的羊水样本的细胞涂片，并依次用荧光试剂染色。为了确定哪些细胞是多形核白细胞，所有重复样本均用荧光核染料4′-6-二脒基-2-苯基吲哚染色。这使得能够明确特征性的多叶核多形核白细胞的核形态特征。然后用罗丹明标记的X染色体特异性探针和荧光素标记的Y染色体特异性探针探测样本，以评估多形核白细胞是男性还是女性来源。
通过对4′-6-二脒基-2-苯基吲哚染色进行正常多叶核形态学研究鉴定出的多形核白细胞中，90%至99%具有X和Y染色体，因此是胎儿细胞。
这些数据证明了羊膜腔内感染期间的胎儿反应。进一步研究母体和胎儿多形核白细胞在绒毛膜羊膜炎中的作用，可能会提供有关这种情况下两种免疫反应关键相互作用的有价值信息。
