High-throughput analysis of amniotic fluid proteins associated with histological chorioamnionitis in preterm premature rupture of membranes using an antibody-based microarray

Seung Mi Lee, Kyo Hoon Park, Eunwook Joo, Da Eun Jeong, Ji Eun Lee, Kyong No Lee, Sue Shin

Research output: Contribution to journalArticlepeer-review

Abstract

Problem: To identify potential proteins in the amniotic fluid (AF) that may be associated with histologic chorioamnionitis (HCA) in patients with preterm premature rupture of membranes (PPROM) using antibody-based microarray analysis. Method of study: This was a retrospective cohort study involving 100 singleton pregnant women with PPROM at 24–34 weeks who underwent amniocentesis and delivered within 120 h of amniocentesis. First, the AF proteomes of 15 patients with PPROM and HCA were compared with those of 15 gestational age-matched patients without HCA using a protein microarray. Next, 12 candidate proteins associated with HCA were further validated in 100 consecutive patients with PPROM by ELISA. Results: Of 507 proteins assessed in the microarray analysis, 46 showed significant intergroup differences. Further quantification confirmed that the levels of EN-RAGE, IL-6, MMP-9, TNFR2, SPARC, TSP2, and uPA were higher in the AF of PPROM patients with HCA than in those without. Multivariate analyses also showed that elevated AF EN-RAGE, IL-6, MMP-9, and TNFR2 levels were independently associated with HCA when adjusted for baseline variables. The frequency of the highest quartile of the aforementioned proteins significantly increased as the total grade of HCA increased; the risk of HCA significantly increased with increasing AF levels of each protein (P for trend <.001). Conclusions: Using protein–antibody microarray technology, we discovered several potential AF proteins (EN-RAGE, IL-6, MMP-9, and TNFR2) independently associated with HCA in patients with PPROM. Furthermore, we demonstrated a direct correlation between the gradation of the intra-amniotic inflammatory response and HCA severity.

Original languageEnglish
Article numbere13595
JournalAmerican Journal of Reproductive Immunology
Volume88
Issue number3
DOIs
StatePublished - Sep 2022
Externally publishedYes

Keywords

  • amniotic fluid
  • antibody microarray
  • histologic chorioamnionitis
  • preterm premature rupture of membranes
  • proteins

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