利用实时定量PCR扩增MHC多态性分析恒河猴母系微嵌合现象。

Chimerism Pub Date : 2014-01-01 Epub Date: 2014-01-17 DOI:10.4161/chim.27778
Sonia Bakkour, Chris A R Baker, Alice F Tarantal, Li Wen, Michael P Busch, Tzong-Hae Lee, Joseph M McCune
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引用次数: 14

摘要

虽然已知人类存在妊娠相关的微嵌合现象,但其临床意义尚不清楚。胎儿微嵌合现象在恒河猴中已被证实,但母体细胞在猴胎儿和婴儿中的运输和持续存在尚未得到充分探讨。为了研究母系微嵌合现象在恒河猴(Macaca mulatta)中发生的频率,我们建立了一种实时聚合酶链反应(PCR)策略,以多态主要组织相容性复合体(MHC)基因序列为目标,并进行了验证。对25只母坝及其后代中的19只进行了信息丰富的PCR检测。对一组动物产前和产后收集的组织(胸腺、肝脏、脾脏、淋巴结和骨髓)和外周血单个核细胞(PBMCs)进行了分析。19只猴子中有7只在至少一个室中检测到母体微嵌合(嵌合细胞范围:0.001-1.9%)。在组织中,7个胎儿中有2个和12个幼体(1-1.5岁)中有3个存在母体微嵌合,大多数阳性动物在多个组织中存在微嵌合细胞。在所有(4 / 4)个胎儿的PBMCs中检测到母体微嵌合。这些观察结果表明,母体微嵌合发生在恒河猴胎儿中,并且可以在出生后的一部分后代的组织中检测到。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Analysis of maternal microchimerism in rhesus monkeys (Macaca mulatta) using real-time quantitative PCR amplification of MHC polymorphisms.

Analysis of maternal microchimerism in rhesus monkeys (Macaca mulatta) using real-time quantitative PCR amplification of MHC polymorphisms.

Analysis of maternal microchimerism in rhesus monkeys (Macaca mulatta) using real-time quantitative PCR amplification of MHC polymorphisms.

Although pregnancy-associated microchimerism is known to exist in humans, its clinical significance remains unclear. Fetal microchimerism has been documented in rhesus monkeys, but the trafficking and persistence of maternal cells in the monkey fetus and infant have not been fully explored. To investigate the frequency of maternal microchimerism in the rhesus monkey (Macaca mulatta), a real-time polymerase chain reaction (PCR) strategy was developed and validated to target polymorphic major histocompatibility complex (MHC) gene sequences. Informative PCR assays were identified for 19 of 25 dams and their respective offspring. Analyses were performed on tissues (thymus, liver, spleen, lymph nodes, and bone marrow) and peripheral blood mononuclear cells (PBMCs) collected prenatally and postnatally in a subset of animals. Seven of 19 monkeys had detectable maternal microchimerism in at least one compartment (range: 0.001-1.9% chimeric cells). In tissues, maternal microchimerism was found in 2 of 7 fetuses and 3 of 12 juveniles (1-1.5 years of age), and most of the animals that were positive had microchimeric cells in more than one tissue. Maternal microchimerism was detected in PBMCs from all (4 of 4) fetuses. These observations suggest that maternal microchimerism occurs in the rhesus monkey fetus and can be detected in tissues in a subset of offspring after birth.

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