Performance of enhanced liver fibrosis plasma markers in asymptomatic individuals with ZZ α1-antitrypsin deficiency.

OBJECTIVES: Alpha1-antitrypsin deficiency (AATD) is a common genetic cause of chronic liver disease. According to retrospective studies, up to 25% of those with homozygous ZZ (Glu 342 to Lys) AATD suffer from liver cirrhosis and/or liver cancer in late adulthood. We hypothesized that the plasma markers for liver fibrosis, necrosis, and apoptosis may identify AATD individuals at higher risk for liver diseases. METHODS: The study cohort included 52 clinically healthy ZZ AATD individuals of 34 years of age, identified in the Swedish neonatal screening of 1972-1974, and 81 age-matched controls with normal MM AAT variant. We analyzed plasma levels of the enhanced liver fibrosis (ELF) panel, including plasma tissue inhibitor of metalloprotease-1, amino-terminal propeptide of type III collagen and hyaluronic acid (HA), and the M30 and M65 antigens, markers for apoptosis/necrosis. RESULTS: Higher levels of tissue inhibitor of metalloprotease-1 (52%, P<0.001), amino-terminal propeptide of type III collagen (12%, P<0.05), HA (17% not significant), and M65 (13.4%, P=0.043) were found in ZZ than in MM patients. In the ZZ group, plasma levels of AAT correlated with M65 (P<0.01) and with HA (P<0.05). On the basis of the ELF panel, M30 and M65, a logistic regression model enabled us to correctly classify 81.2% of the originally grouped ZZ and MM cases with a sensitivity of 73.1% and a specificity of 86.4%. CONCLUSION: The ELF markers are associated with ZZ AATD at early adulthood, and can be considered as a useful tool to identify ZZ cases at an increased risk of developing liver diseases later in life.

Chronic intestinal pseudo-obstruction due to buserelin-induced formation of anti-GnRH antibodies.

BACKGROUND & AIMS: A 30-year-old woman, treated with buserelin, an analogue of gonadotropin-releasing hormone (GnRH) (also called luteinizing hormone-releasing hormone, LH-RH), developed chronic intestinal pseudo-obstruction (CIPO). The sudden onset of this disease in a previously healthy woman perplexed us. CIPO refers to a gastrointestinal disorder that can have a variety of causes, such as drugs, among others. Thus, we wanted to examine whether in this patient the development of CIPO is related to the treatment with buserelin. METHODS: The patient was examined using esophagogastroduodenoscopy, esophageal, and antroduodenojejunal manometry, gastric emptying tests, and histologic analyses and immunohistochemistry on full-thickness biopsies including staining with anti-GnRH antibody. Plasma samples were examined by the standard serologic analyses and specifically for the occurrence of anti-GnRH antibodies by enzyme-linked immunosorbent assay methods. RESULTS: CIPO was diagnosed based on symptoms (abdominal pain, vomiting, and constipation), and the results of the clinical examinations, such as signs of esophageal aperistalsis, delayed gastric emptying, and small intestinal bursts. Histologic examination revealed a decreased number of myenteric neurons as well as increased neuronal degeneration and an abnormal immune profile. There was a loss of GnRH-containing neurons. The patient had high plasma titers of anti-GnRH antibodies, which occurred on the occasions of the treatment with buserelin. CONCLUSIONS: Our findings suggest that the patient has developed CIPO due to buserelin-induced formation of anti-GnRH antibodies destroying GnRH-producing neurons of the myenteric plexus.

Plasma levels of alpha1-antichymotrypsin and secretory leukocyte proteinase inhibitor in healthy and chronic obstructive pulmonary disease (COPD) subjects with and without severe alpha1-antitrypsin deficiency.

BACKGROUND: Individuals with severe Z alpha1-antitrypsin (AAT) deficiency have a considerably increased risk of developing chronic obstructive lung disease (COPD). It has been hypothesized that compensatory increases in levels of other protease inhibitors mitigate the effects of this AAT deficiency. We analysed plasma levels of AAT, alpha1-antichymotrypsin (ACT) and secretory leukocyte protease inhibitor (SLPI) in healthy (asymptomatic) and COPD subjects with and without AAT deficiency. METHODS: Studied groups included: 71 asymptomatic AAT-deficient subjects (ZZ, n = 48 and SZ, n = 23, age 31 +/- 0.5) identified during Swedish neonatal screening for AAT deficiency between 1972 and 1974; age-matched controls (MM, n = 57, age 30.7 +/- 0.6); older asymptomatic ZZ (n = 10); healthy MM (n = 20, age 53 +/- 9.6); and COPD patients (ZZ, n = 10, age 47.4 +/- 11 and MM, n = 10, age 59.4 +/- 6.7). Plasma levels of SLPI, AAT and ACT were analysed using ELISA and immunoelectrophoresis. RESULTS: No significant difference was found in plasma ACT and SLPI levels between the healthy MM and the ZZ or SZ subjects in the studied groups. Independent of the genetic variant, subjects with COPD (n = 19) had elevated plasma levels of SLPI and ACT relative to controls (n = 153) (49.5 +/- 7.2 vs 40.7 +/- 9.1 ng/ml, p < 0.001 and 0.52 +/- 0.19 vs 0.40 +/- 0.1 mg/ml, p < 0.05, respectively). CONCLUSION: Our findings show that plasma levels of ACT and SLPI are not elevated in subjects with genetic AAT deficiency compared MM controls and do not appear to compensate for the deficiency of plasma AAT.

Secreted leukocyte protease inhibitor is present in aqueous humours from cataracts and other eye pathologies.

Previous studies identified serine, cysteine and metalloproteases in normal aqueous humours (AH) and suggested that a balance between proteases and their inhibitors may play a role in the modulation of the AH outflow. We aimed to determine whether secretory leukocyte protease inhibitor (SLPI), a serine protease inhibitor, is present in AH of patients with cataract and other eye pathologies. AH was collected from 117 cataract patients of which 55 were diagnosed with more when one eye disease: cataract only (n=62), pseudoexfoliation (PEX) (n=26), glaucoma (n=6), diabetes retinopathy (n=4), iritis-uveitis (n=4) and macular degeneration (n=28). The total protein in AH was determined by a Bradford assay and SLPI was analyzed by Western blot and ELISA methods. The average concentration of total protein and SLPI in AH samples was 160+/-15 microg/ml (n=117, +/-SEM) and 500+/-94 pg/ml (n=105), respectively. The cataract patients with additional eye disease(s) showed higher protein levels (201+/-35 microg/ml) than cataract (controls) (128+/-31 microg/ml), P<0.01. It is noteworthy that no correlation was found between SLPI and the total protein concentrations in AH, but SLPI was positively correlated with age (r=0.2, P<0.05). No statistical difference in SLPI levels was found between controls (cataract) and other pathologies, while patients with iritis/uveitis had higher SLPI levels compared to those with diabetes (P<0.05). We show here for the first time that SLPI is present in AH and may play a role as well as serve as a marker in pathological states.

Increased plasma levels of serine proteinase inhibitors in lung cancer patients.

BACKGROUND: Tumor growth and invasiveness occur through infiltration of tumor cells into the host cells and by angiogenesis, which is modulated by proteinases and antiproteinases released from tumor cells that carry out tissue remodelling. A number of studies have revealed variations in the plasma levels of serine proteases and their inhibitors among tumor types. PATIENTS AND METHODS: By immunological methods we analysed the levels of serine protease inhibitors AAT, ACT and SLPI in newly diagnosed lung cancer patients (n=14) compared to non-smoker and smoker, age- and gender-matched control groups (n=16), and also in an expanded group of lung cancer patients with local tumors (n=14) and with metastasis (n=18). RESULTS: Our data show that plasma levels of AAT, ACT and SLPI were elevated in lung cancer patients by 1.43-fold, p<0.01, 2.57-fold, p<0.01 and 1.6-fold, p<0.001, respectively when compared to controls. In addition, we found that levels of AAT and ACT were higher by 1.47-fold, p<0.001 and 2.27-fold, p<0.001, respectively in lung cancer cases with metastasis compared to localized tumor. CONCLUSION: These inhibitor levels may provide measures of cancer progression in individual patients and possibly offer useful information for an understanding of the mechanisms of metastasis.

Analysis of systemic biomarkers in COPD patients.

The finding that alphal-antitrypsin (AAT) deficiency, PiZZ, a well-established genetic risk factor for COPD, is related to high levels of circulating AAT polymers, prompted us to measure serum levels of such polymers and selected markers of inflammation in age- and gender-matched patients with stable COPD and control subjects with and without severe AAT deficiency, and to assess their relationship with each other and with the genetic AAT-variant. We found that COPD individuals (n= 20), independent of AAT-variant, had significantly higher serum levels of AAT and its polymers, MMP-9, sICAM-1, VEGF and sE-selectin than controls (n=30). Subjects with PiZZ COPD (n= 10) showed significantly elevated serum levels of AAT-polymers, sE-selectin and sICAM-1, while patients with PiMM COPD (n= 10) showed higher levels of MMP-9, VEGF, IL-8 and MCP-1 than controls. By using factor analysis we were able to split the analysed biomarkers into two independent components: the first containing MMP-9, MCP-1, IL-8 and VEGF and the second-AAT and its polymers and sE-selectin. The result from the binomial logistic regression showed that 95.2 percent of the control individuals and 94.7 percent of the COPD patients can be correctly classified on the basis of the measured serum biomarkers. These observations highlight the importance of the finding sets of biomolecules, which could offer new strategies for the diagnosis of COPD and may have value for monitoring progression of COPD.