Lung rejection
Transcript Signatures of Lymphocytic Bronchitis in Lung Allograft Biopsy Specimens

https://doi.org/10.1016/j.healun.2004.06.016Get rights and content

Background

Rejection and obliterative bronchiolitis are barriers to sustained graft function in recipients of transplanted lungs. Early detection is hindered by inadequate tests and an incomplete understanding of the molecular events preceding or accompanying graft deterioration.

Methods

Hypothesizing that genes involved in immune responses and tissue remodeling produce biomarkers of rejection, we measured the expression of 192 selected genes in 72 sets of biopsy specimens from human lung allografts. Gene transcripts were quantified using a 2-step, multiplex, real-time polymerase chain reaction approach in endobronchial and transbronchial biopsy specimens from transplant recipients without acute infections undergoing routine surveillance bronchoscopy.

Results

Comparisons of histopathology in parallel biopsy specimens identified 6 genes correlating with rejection as manifested by lymphocytic bronchitis, a suspected harbinger of obliterative bronchiolitis. For example, β2-defensin and collagenase transcripts in inflamed bronchi increased 37-fold and 163-fold, respectively. By contrast, these transcripts did not correlate with acute rejection in transbronchial specimens. Further, no correspondence was noted between histopathologic bronchitis and parenchymal rejection when endobronchial and transbronchial samples were obtained from the same patient.

Conclusions

Our highly sensitive method permits quantitation of many gene transcripts simultaneously in small, bronchoscopically acquired biopsy specimens of allografts. Transcript signatures obtained by this approach suggest that airway and alveolar responses to rejection differ and that endobronchial biopsy specimens assess lymphocytic bronchitis and chronic rejection but are not proxies for transbronchial biopsy specimens. Further, they reveal changes in airway expression of the specific genes involved in host defense and remodeling and suggest that the measurement of transcripts correlating with lymphocytic bronchitis may be diagnostic adjuncts to histopathology.

Section snippets

Study Design, Biopsy Specimens, and Grading

The subjects in this prospective study were consecutively enrolled lung transplant recipients undergoing surveillance bronchoscopy according to the following schedule: every 2 weeks for the first 2 months after transplantation, then monthly for 6 months, quarterly for the next year, and every 6 months thereafter. EBB and TBB were performed at the same sitting. Pre-bronchoscopic monitoring on the same day included pulmonary function testing and computed tomography scans. Six to 10 TBB specimens

Patient Characteristics

We recruited 22 patients prospectively. Of these, 10 had 1 bronchoscopy during the recruitment interval and 12 patients had 2 or more. Forty bronchoscopies were performed, for 32 of which paired EBB and TBB specimens were obtained at the same sitting. In 7 participants, an EBB specimen was obtained without TBB. In 1 instance, a TBB specimen was obtained without EBB. Patient characteristics are summarized in Table 1. All participants received maintenance immunosuppressive medications, including

Discussion

Gene expression profiling is rapidly becoming an essential tool for research and drug discovery and may soon play a central role in clinical diagnostics. This study profiles a large cohort of genes in human lung allografts by applying a high-sensitivity PCR approach to bronchoscopically acquired biopsy specimens. We identify several genes whose transcripts correlate strongly with histopathologic airway rejection/lymphocytic bronchitis. The data also identify genes overexpressed in EBB or TBB

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    Supported by grant HL024136 from the National Institutes of Health and by the Diamond Family Foundation.

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    Copyright © 2005 International Society for Heart and Lung Transplantation. Published by Elsevier Inc. All rights reserved.