ELISA Development for Serum Hemeoxygenase-1 and Its Application to Patients with Acute Respiratory Distress Syndrome

Abstract

Background Hemeoxygenase-1 (HO-1) is an essential enzyme in heme catabolism and has been proposed as a biomarker of lung disease prognosis. We modified a commercial HO-1 enzyme-linked immunosorbent assay (ELISA) kit to achieve higher sensitivity and evaluated if serum HO-1 could be a biomarker to predict the prognosis of acute respiratory distress syndrome (ARDS) patients. Methods Serum samples were collected from 15 healthy volunteers to validate the modified ELISA. In the 22 patients with ARDS who were enrolled, serum HO-1 was measured upon diagnosis (D0) and at 7 days after diagnosis (D7). Results The serum HO-1 concentration could be measured in all healthy volunteers. The intra- and interassay tests and the percentage recovery test were acceptable. Compared with normal control subjects, patients with ARDS had significantly higher D0 HO-1 concentrations (75.4 ng/mL versus 31.7 ng/mL, P < 0.001). The 28-day survival was significantly better in patients with low D0 HO-1 (<75.8 ng/mL) than in those with high D0 HO-1 (≥75.8 ng/mL) (mortality rate: 18% versus 73%, P=0.016). Nonsurvivors had significantly higher D0 and D7 HO-1 concentrations than survivors (P < 0.05). Conclusion Serum HO-1 may be a useful biomarker to predict the prognosis of patients with ARDS.

Serum Heme Oxygenase-1 Measurement is Useful for Evaluating Disease Activity and Outcomes in Patients With Acute Respiratory Distress Syndrome and Acute Exacerbation of Interstitial Lung Disease

“…Mumby et al reported that HO-1 protein concentrations are signi cantly elevated in lung tissue and bronchoalveolar lavage uid taken from ARDS patients compared with controls, and HO-1 expression contributes to changes in iron mobilization, signalling, and regulation seen in this condition (11). We have also demonstrated the usefulness of measuring serum HO-1 in the diagnosis and prognosis of patients with ARDS and AE-ILDs (12,13).…”

“…The assay validation was performed reproducibility of ELISA standard curve for serum HO-1, the intra-and inter-assay tests, and the percentage recovery test. We con rmed all of these results were acceptable (12). Control subjects for serum HO-1 levels included 28 healthy, non-smoking adults who had been admitted to the hospital for a medical checkup.…”

“…HO-1, a rate-limiting enzyme in heme catabolism, has antioxidative activities in patients with diffuse parenchymal lung disease (17)(18)(19). We previously investigated whether evaluating the degree of oxidative stress by measuring serum HO-1 using the sandwich ELISA method is useful for assessing disease activities and predicting prognosis in patients with ARDS and AE-ILDs (12,13). The present study was an integrated analysis of these.…”

“…Serum HO-1 enzyme-linked immunosorbent assay (ELISA) measurement Serum HO-1 levels were measured at the time of ARDS or AE-ILD diagnosis (D0) and 7 (D7) and 14 (D14) days from the diagnosis using the IMMUNOSET® HO-1 (human) ELISA development set (Enzo, Farmingdale, NY, USA), according to the manufacturer's instructions. The details of this ELISA method have been described previously (12). The assay validation was performed reproducibility of ELISA standard curve for serum HO-1, the intra-and inter-assay tests, and the percentage recovery test.…”

Serum heme oxygenase-1 measurement is useful for evaluating disease activity and outcomes in patients with acute respiratory distress syndrome and acute exacerbation of interstitial lung disease

“…Mumby et al reported that HO-1 protein concentrations are signi cantly elevated in lung tissue and bronchoalveolar lavage uid taken from ARDS patients compared with controls, and HO-1 expression contributes to changes in iron mobilization, signalling, and regulation seen in this condition (18). Also, we have demonstrated the usefulness of measuring serum HO-1 in the diagnosis and prognosis of patients with ARDS and AE-ILDs (19,20). Therefore, we speculate that HO-1 characterized as stress response protein with rapid response induced by various stimuli and with various physiological activities caused by HO-1 metabolites could re ect pulmonary cellular damage induced by ROS and RNS more closely and directly.…”