title: Characterization of the fetal blood transcriptome and proteome in maternal anti-fetal rejection: evidence of a distinct and novel type of human fetal systemic inflammatory response.
abstract: BACKGROUND
  The human fetus is able to mount a systemic inflammatory response when exposed to microorganisms. This stereotypic response has been termed the 'fetal inflammatory response syndrome' (FIRS), defined as an elevation of fetal plasma interleukin-6 (IL-6). FIRS is frequently observed in patients whose preterm deliveries are associated with intra-amniotic infection, acute inflammatory lesions of the placenta, and a high rate of neonatal morbidity. Recently, a novel form of fetal systemic inflammation, characterized by an elevation of fetal plasma CXCL10, has been identified in patients with placental lesions consistent with 'maternal anti-fetal rejection'. These lesions include chronic chorioamnionitis, plasma cell deciduitis, and villitis of unknown etiology. In addition, positivity for human leukocyte antigen (HLA) panel-reactive antibodies (PRA) in maternal sera can also be used to increase the index of suspicion for maternal anti-fetal rejection. The purpose of this study was to determine (i) the frequency of pathologic lesions consistent with maternal anti-fetal rejection in term and spontaneous preterm births; (ii) the fetal serum concentration of CXCL10 in patients with and without evidence of maternal anti-fetal rejection; and (iii) the fetal blood transcriptome and proteome in cases with a fetal inflammatory response associated with maternal anti-fetal rejection.
  
  
  METHODS
  Maternal and fetal sera were obtained from normal term (n = 150) and spontaneous preterm births (n = 150). A fetal inflammatory response associated with maternal anti-fetal rejection was diagnosed when the patients met two or more of the following criteria: (i) presence of chronic placental inflammation; (ii) ≥80% of maternal HLA class I PRA positivity; and (iii) fetal serum CXCL10 concentration >75th percentile. Maternal HLA PRA was analyzed by flow cytometry. The concentrations of fetal CXCL10 and IL-6 were determined by ELISA. Transcriptome analysis was undertaken after the extraction of total RNA from white blood cells with a whole-genome DASL assay. Proteomic analysis of fetal serum was conducted by two-dimensional difference gel electrophoresis. Differential gene expression was considered significant when there was a P < 0.01 and a fold-change >1.5.
  
  
  RESULTS
  (i) The frequency of placental lesions consistent with maternal anti-fetal rejection was higher in patients with preterm deliveries than in those with term deliveries (56% versus 32%; P < 0.001); (ii) patients with spontaneous preterm births had a higher rate of maternal HLA PRA class I positivity than those who delivered at term (50% versus 32%; P = 0.002); (iii) fetuses born to mothers with positive maternal HLA PRA results had a higher median serum CXCL10 concentration than those with negative HLA PRA results (P < 0.001); (iv) the median serum CXCL10 concentration (but not IL-6) was higher in fetuses with placental lesions associated with maternal anti-fetal rejection than those without such lesions (P < 0.001); (v) a whole-genome DASL assay of fetal blood RNA demonstrated differential expression of 128 genes between fetuses with and without lesions associated with maternal anti-fetal rejection; and (vi) comparison of the fetal serum proteome demonstrated 20 proteins whose abundance differed between fetuses with and without lesions associated with maternal anti-fetal rejection.
  
  
  CONCLUSIONS
  We describe a systemic inflammatory response in human fetuses born to mothers with evidence of maternal anti-fetal rejection. The transcriptome and proteome of this novel type of fetal inflammatory response were different from that of FIRS type I (which is associated with acute infection/inflammation).
authors.affiliation: Perinatology Research Branch, NICHD/NIH/DHHS, Bethesda, MD, Detroit, MI, USA.
authors.forename: Joonho
authors.initials: J
authors.lastname: Lee
authors.forename: Roberto
authors.initials: R
authors.lastname: Romero
authors.forename: Tinnakorn
authors.initials: T
authors.lastname: Chaiworapongsa
authors.forename: Zhong
authors.initials: Z
authors.lastname: Dong
authors.forename: Adi L
authors.initials: AL
authors.lastname: Tarca
authors.forename: Yi
authors.initials: Y
authors.lastname: Xu
authors.forename: Po Jen
authors.initials: PJ
authors.lastname: Chiang
authors.forename: Juan Pedro
authors.initials: JP
authors.lastname: Kusanovic
authors.forename: Sonia S
authors.initials: SS
authors.lastname: Hassan
authors.forename: Lami
authors.initials: L
authors.lastname: Yeo
authors.forename: Bo Hyun
authors.initials: BH
authors.lastname: Yoon
authors.forename: Nandor Gabor
authors.initials: NG
authors.lastname: Than
authors.forename: Chong Jai
authors.initials: CJ
authors.lastname: Kim
mesh_terms: D000293:Adolescent; D000328:Adult; D054357:Chemokine CXCL10 / Q000097:blood; D002821:Chorioamnionitis / Q000175:diagnosis / Q000276:immunology*; D005260:Female; D005312:Fetal Blood / Q000276:immunology / Q000378:metabolism*; D005333:Fetus / Q000276:immunology; D020869:Gene Expression Profiling; D006680:HLA Antigens / Q000276:immunology; D054502:Histocompatibility, Maternal-Fetal / Q000235:genetics; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D007518:Isoantibodies / Q000097:blood; D010920:Placenta / Q000276:immunology*; D011247:Pregnancy / Q000276:immunology; D047928:Premature Birth / Q000175:diagnosis / Q000276:immunology*; D020543:Proteome; D055815:Young Adult
chemical_list: D054357:Chemokine CXCL10; D006680:HLA Antigens; D018836:Inflammation Mediators; D007518:Isoantibodies; D020543:Proteome
keywords: Apolipoprotein C-III; CD 34; CXCL10; chronic placental inflammation; pregnancy
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