Chest ACCP Member Benefits
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Guest Access | Sign In via User Name/Password
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF) Free
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Article Archive
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (28)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ashitani, J.-i.
Right arrow Articles by Matsukura, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ashitani, J.-i.
Right arrow Articles by Matsukura, S.
(Chest. 2001;119:1131-1137.)
© 2001 American College of Chest Physicians

Plasma and BAL Fluid Concentrations of Antimicrobial Peptides in Patients With Mycobacterium avium- intracellulare Infection*

Jun-ichi Ashitani, MD; Hiroshi Mukae, MD; Takeaki Hiratsuka, MD; Masamitsu Nakazato, MD; Kenshi Kumamoto, MD and Shigeru Matsukura, MD

* From the National Sanatorium Miyazakihigashi Hospital (Drs. Ashitani and Kumamoto), Miyazaki, Japan; and the Third Department of Internal Medicine (Drs. Mukae, Hiratsuka, Nakazato, and Matsukaura), Miyazaki Medical College, Miyazaki, Japan.

Correspondence to: Hiroshi Mukae, MD, Third Department of Internal Medicine, Miyazaki Medical College, Kihara 5200, Kiyotake, Miyazaki 889-1692, Japan; e-mail: hmukae{at}post.miyazaki-med.ac.jp


   Abstract
TOP
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Study objectives: To investigate the roles of human {alpha}-defensin (HAD), human ß-defensin (HBD)-1, and HBD-2, novel antimicrobial peptides, in patients with Mycobacterium avium-intracellulare infection (MAI).

Patients: The study included 25 patients (10 men) with MAI who visited our hospital between June 1998 and August 1999.

Measurements and results: In patients with pulmonary MAI, we measured HAD and HBD-1, and HBD-2 levels in plasma and in BAL fluid (BALF) by radioimmunoassay. Plasma concentrations of HAD and HBD-2 in those patients were higher than those in control subjects, whereas HBD-1 levels were similar to those in the control subjects. High levels of HAD and HBD-2, but not HBD-1, also were observed in the BALF of MAI patients. There was a positive correlation between HAD and interleukin (IL)-8 concentrations in the BALF of patients with MAI. BALF HBD-2 concentrations also correlated positively with those of plasma HBD-2 and BALF IL-1ß in MAI patients. Patients with cavity formation on the chest roentgenogram had higher HAD and HBD-2 levels in their BALF than those of patients without cavity formation. Treatment with clarithromycin combined with two or three other antibiotics, including ethambutol, rifampicin, ofloxacin, or ciprofloxacin, for at least 6 months resulted in a significant fall in plasma HBD-2 concentrations in responders, but not in nonresponders.

Conclusion: Our findings suggest that HAD and HBD-2 may participate in host defense and local remodeling of the respiratory tract in patients with MAI and that plasma HBD-2 levels may be a useful marker of disease activity in patients with pulmonary MAI.

Key Words: {alpha}-defensin • ß-defensin • Mycobacterium avium-intracellulare infection


   Introduction
TOP
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Mycobacterium avium-intracellulare is widely distributed in the environment, especially in water and soil.1 2 Humans are exposed to the pathogen through airborne acquisition, and, therefore, respiratory epithelial cells and alveolar macrophages are important components as the first defense system against Mycobacterium avium-intracellulare infection (MAI). A previous study3 has shown high concentrations of inflammatory cytokines such as interleukin (IL)-1ß and IL-8 in the BAL fluid (BALF) of MAI patients who were free of predisposing lung diseases, including AIDS. Such levels may induce the host defense system in respiratory epithelial cells through antimicrobial peptides. MAI without predisposing lung diseases commonly occurs in elderly women and nonsmokers, and some roentgenographic studies4 5 using chest CT scanning indicated that MAI might lead to bronchiectasis or cavity formation. However, the mechanisms of such pathologic changes are not yet fully understood.

Defensins are endogenous antibiotics that contribute to host defense by disrupting the cytoplasmic membrane of microorganisms.6 Human {alpha}-defensin (HAD) and two human ß-defensins (HBDs) have been isolated as a family of small (3.5 to 4.5 kd) cationic antimicrobial peptides. HAD, localized in azurophil granules in the neutrophil, plays an important role in the non-oxygen-dependent killing of phagocytized bacteria7 and has been demonstrated to have potent activity in killing M avium-intracellulare.8 However, HBDs are produced by epithelial cells9 10 and can be released on microbial invasion or up-regulated by stimulation with a lipopolysaccharide.11 12 13 Their structures are characterized by a conserved cysteine motif that forms three disulfide linkages, imposing a characteristic ß-sheet structure. This structure is associated with an amphiphilic charge distribution that enables the defensins to interact with and disrupt target cell membranes and to function in forming channels in the target membrane, leading to cell lysis and eventually to cell death.11 The HBD family is distinguished from HAD by differences in the peptide folding that is created by the linkage of six cysteine residues.9

In the present study, we investigated the inflammatory process of MAI by measuring HAD, HBD-1, and HBD-2 concentrations in the plasma and BALF of patients with MAI by using a radioimmunoassay (RIA). We also measured the concentrations of several cytokines in BALF because IL-1ß is known to induce the production of HBD-2 in respiratory tract epithelial cells,14 while IL-8 induces the release of HAD from neutrophils dose-dependently.15


   Materials and Methods
TOP
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
ample Preparation
We studied 25 consecutive patients with MAI who visited our hospital between June 1998 and August 1999. The group consisted of 15 women and 10 men with a mean (± SEM) age of 60.9 ± 3.5 years and included 3 smokers, 2 ex-smokers, and 20 nonsmokers. MAI was diagnosed and treated according to the guidelines set by the American Thoracic Society.16 The patients were deemed to be free of other lung diseases based on the following findings: (1) negative history; (2) normal findings on a chest roentgenogram and/or chest CT scan taken > 1 year after entry into the study; and (3) a positive culture of only MAI in the sputum on at least two separate occasions or a positive culture in samples obtained from the affected lesion using a sterile bronchoscope. Patients with any type of bacterial infections and AIDS, and a history of skin diseases, were excluded from the study. No patients had previously received treatment for MAI or any anti-inflammatory therapy including corticosteroids. All MAI patients were treated with clarithromycin (400 mg twice a day), combined with two or three other antibiotics, including ethambutol (25 mg/kg/d), rifampicin (450 mg/d), ofloxacin (400 mg twice daily), or ciprofloxacin (750 mg three times daily). Although the therapeutic regimen was modified during the treatment period in 9 of the 25 patients because of adverse effects, all patients continued to use clarithromycin throughout the treatment. MAI patients visited our hospital twice a month, and sputum samples were taken for cultures once a month. In this study, the treatment end point was set to coincide with the first negative sputum culture after at least 6 months of continued treatment. Responders, who were considered to have been successfully treated, were defined as those patients who showed negative results of sputum cultures for 6 months. Nonresponders were defined as patients in whom sputum culture results failed to remain negative for > 6 months. We selected healthy volunteers (n = 20) matched for age or sex as control subjects.

Blood Sampling
Blood samples (2 mL from each patient) were obtained before the commencement of antimicrobial therapy and at the treatment end point. The blood sample was anticoagulated with ethylenediaminetetraacetic acid-2Na, then was centrifuged to obtain plasma. The plasma (0.01 to 0.1 mL) was subjected to RIA for HAD, HBD-1, and HBD-2.

BAL
After informed consent was obtained, BAL was performed in 15 of the 25 patients before treatment, as previously described.17 The bronchoscope was wedged into one of the segments or segmental bronchi of the most heavily involved lobe, as determined by the chest CT scan. An aliquot of 50 mL sterile saline solution at body temperature was instilled through the bronchoscope. The fluid was retrieved immediately by gentle suction using a sterile syringe, and the procedure was repeated three times. BALF was passed through two sheets of gauze and then was centrifuged at 500g for 10 min at 4°C. After washing twice with phosphate-buffered saline solution that was free of calcium or magnesium (Life Technologies; Rockville, MD), the remaining cells were suspended in phosphate-buffered saline solution supplemented with 10% heat-inactivated fetal calf serum and were counted using a hemocytometer. An aliquot then was diluted to a concentration of 2 x 105 cells/mL, and a 0.2-mL cell suspension was spun down onto a glass slide at 1,100 revolutions per minute for 2 min using a cytocentrifuge (Cytospin 2; Shandon Instruments; Sewickley, PA). The remaining fluid was centrifuged at 500g for 5 min, and the supernatant was stored at -80°C until it was examined. The prepared slides were dried, fixed, and then stained using a May-Giemsa method. More than 200 cells were identified using a photomicroscope.

HAD Assay
The concentration of HAD was measured by the RIA established in our laboratory.18 A diluted sample or standard peptide solution (100 µL) was incubated for 24 h with 100 µL antiserum diluent (final dilution, 1:21,000). 125I-labeled HAD solution (16,000 counts per minute in 100 µL of solution) then was added, and the mixture was incubated again for 24 h. In the next step, normal rabbit serum and antirabbit IgG goat serum were added, and the samples were stored for 16 h. Bound and free ligands were separated by centrifugation. All procedures were performed at 4°C, and duplicate assays were performed. The respective intra-assay and interassay coefficients of variation were 3.5% and 8%, respectively, at 50% binding.

HBD Assay
The concentrations of HBD-1 and HBD-2 were measured by the RIA established in our laboratory.19 20 HBDs were radioiodinated by the lactoperoxidase method, and the 125I-labeled peptide was purified by reversed phase high-performance liquid chromatography on a column (model TSK ODS 120A; Tosoh Co; Tokyo, Japan). The incubation buffer for the RIA was 50 mM sodium phosphate (pH, 7.4) containing 0.25% bovine serum albumin treated with N-ethylmaleimide, 80 mM NaCl, 25 mM ethylenediaminetetraacetic acid-2Na, 0.05% NaN3, 0.1% octoxynol-9 (Triton X-100), and 3.1% dextran T-40. The diluted sample or a standard peptide solution (100 µL) was incubated for 24 h with 100 µL diluted antiserum (final dilutions, 1:460,000 and 1:4,200,000, respectively). The tracer solution (16,000 to 18,000 counts per minute in 100 µL of solution) was added, and the mixture was incubated for 24 h, after which normal rabbit serum and antirabbit IgG goat serum were added and the whole preparation was stored for a further 16 h. Bound and free ligands were separated by centrifugation. All procedures were performed at 4°C, and the samples were assayed in duplicate.

IL-1ß and IL-8 Assays
The concentration of IL-1ß was measured using a commercially available enzyme-linked immunosorbent assay kit (R&D Systems; Minneapolis, MN). The concentration of IL-8 was measured by another commercially available kit (Toray Fuji Bionics; Tokyo, Japan).

Radiologic Findings
Before treatment, all patients underwent chest CT analysis in the supine position. CT scans were obtained with the patient supine during breath-hold at end-expiration, and their findings were evaluated by experienced chest radiologists.

tatistical Analysis
Data were expressed as the mean ± SEM. Differences between groups were examined using the Mann-Whitney U test. Correlations between two groups were determined using the Spearman rank correlation analysis. For the comparison of plasma defensin levels measured before and after treatment, we used a paired-samples Wilcoxon test to determine statistical significance. A p value of < 0.05 denoted the presence of a statistically significant difference.


   Results
TOP
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Patients Characteristics
The main symptoms were cough (15 patients), sputum production (10 patients), and hemoptysis (6 patients). The chest CT scans revealed small nodules in all patients, ectasia of peripheral bronchi and/or bronchioles in 11 patients, and cavity formation in 10 patients. Seventeen of 25 MAI patients successfully responded to therapy, while the remaining 8 patients were nonresponders.

Laboratory Data and BALF Findings
The Gaffky scale number in MAI patients ranged from 0 to 6 (mean, 3). The mean values and ranges for erythrocyte sedimentation rate, C-reactive protein, and leukocyte count in MAI patients before treatment were 36 mm/h (range, 10 to 128 mm/h), 0.7 mg/dL (range, 0 to 5.3 mg/dL), and 5,319 cells/µL (2,400 to 11,000 cells/µL), respectively. Differential counts of cells obtained from the BALF of patients with MAI showed a higher percentage of neutrophils than in the control subjects (Table 1 ). The numbers of both neutrophils and lymphocytes in the BALF of MAI patients were also higher than those of the control subjects. There was no significant difference in the differential counts in BALF between responders and nonresponders (data not shown).


View this table:
[in this window]
[in a new window]

  		Table 1. BALF Findings in MAI Patients*

 
HAD and HBD Levels in Plasma and BALF
HAD concentrations in plasma and BALF were higher in patients with MAI before treatment than those in control subjects (Table 2 ), but there was no significant correlation between HAD levels in the plasma and BALF of our patients (data not shown). Interestingly, the HBD-1 levels of plasma and BALF in patients with MAI before treatment were similar to those in the control subjects. In contrast, HBD-2 concentrations in both the plasma and the BALF were significantly higher in the patients than in the control subjects (Table 2) . Further analysis showed a positive correlation between the concentrations of HBD-2 in the BALF and plasma in MAI patients (r = 0.86; p < 0.0001; Fig 1 ), while HBD-1 levels in BALF did not correlate with plasma HBD-1 levels in patients with MAI (data not shown). Neither HBD-1 nor HBD-2 levels correlated with HAD levels in the BALF of patients with MAI (data not shown).


View this table:
[in this window]
[in a new window]

  		Table 2. Levels of Antimicrobial Peptides in Plasma and BALF in MAI Patients*

 


View larger version (25K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 1. Correlation between HBD-2 concentrations in the plasma and BALF of patients with MAI. Plasma HBD-2 levels correlated positively with those in BALF.

 
BALF IL-1ß and IL-8 Levels
The mean concentrations of IL-1ß and IL-8 in the BALF of MAI patients were 502 ± 142 pg/mL and 568 ± 57 pg/mL, respectively, which were significantly higher than those in the control subjects (14 ± 9 pg/mL and 4 ± 4 pg/mL, respectively; p < 0.001). There was a positive correlation between HAD and IL-8 levels in BALF (r = 0.67; p < 0.01; Fig 2 A) and between HBD-2 and IL-1ß levels in BALF (r = 0.96; p < 0.0001; Fig 2 , middle, B) in MAI patients. There was also a positive correlation between IL-1ß and IL-8 levels in the BALF of MAI patients (r = 0.71; p < 0.005; Fig 2 , bottom, C).



View larger version (19K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 2. Correlation between antimicrobial peptides and inflammatory cytokines in the BALF of MAI patients. Note the positive correlation between HAD and IL-8 levels (top, A) and also between HBD-2 and IL-1ß levels in BALF (middle, B). There was a positive correlation between IL-8 and IL-1ß levels in BALF (bottom, C).

 
Relationship Between Radiographic Findings and HAD and HBD Levels in BALF
Of the 15 patients in whom BAL was performed, 6 patients showed cavity formation and 8 showed ectasia of the bronchi and/or bronchioles on the chest CT scan. Both findings were found in five patients. Higher concentrations of HAD and HBD-2 were detected in the BALF of patients with cavity formation or in the BALF of patients with ectasia of the bronchi and/or bronchioles than in the BALF of patients without either cavity formation or ectasia (Table 3 ). In addition, the patients with cavity formation had higher plasma HBD-2 levels than those without cavity formation and/or ectasia (data not shown). There were no significant differences in HBD-1 levels in BALF between the groups with and without cavity formation or ectasia.


View this table:
[in this window]
[in a new window]

  		Table 3. Radiographic Findings and Antimicrobial Peptides in BALF*

 
Effects of Treatment on Plasma HAD and HBD Concentrations
Plasma HBD-1 and HAD concentrations did not change after treatment in both responders and nonresponders. However, plasma HBD-2 concentrations were significantly lower in responders after treatment than before treatment, whereas no change was seen in nonresponders (Table 4 ).


View this table:
[in this window]
[in a new window]

  		Table 4. Antimicrobial Peptide Levels in Plasma Before and After Treatment in MAI Patients*

 

   Discussion
TOP
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
The major finding of the present study was that MAI patients, especially those with cavity formation and/or bronchiectasis, showed high concentrations of HAD and HBD-2 both in plasma and BALF. HAD in neutrophils plays an important role in host defense against pathogens. However, once HAD is released from the neutrophils, it participates in lung injury, as observed in their cytotoxic effects in neutrophil-mediated pulmonary diseases such as diffuse panbronchiolitis,15 cystic fibrosis,21 and ARDS.22 Accumulated neutrophils in the lungs together with high concentrations of HAD in the BALF of the patients suggested that lung injury in the disorder is caused, at least partly, by neutrophils through the cytotoxic effect of the HAD released by these cells. This conclusion is consistent with the findings by Condos et al,23 who demonstrated that neutrophils were the predominant BALF cell type in patients with pulmonary tuberculosis with cavitary lesions. Mycobacterium tuberculosis and its cellular components are capable of stimulating both the biosynthesis and secretion of IL-8 from alveolar macrophages.24 The HAD released by activated neutrophils stimulates IL-8 synthesis by way of airway epithelial cells and may mediate the recruitment of additional neutrophils into the airways.25 IL-8 is a potent neutrophil attractant and can induce the release of HAD from neutrophils.16 In our study, high IL-8 levels were detected in the BALF of MAI patients, and a positive correlation was found between IL-8 and HAD levels in BALF. These findings suggest that IL-8 may be a key cytokine that contributes to the high levels of HAD in the BALF of patients with MAI.

The finding that HBD-2 levels were high in the plasma and BALF of MAI patients indicates that HBD-2, in addition to HAD, also might play an important pathophysiologic role in MAI. This conclusion is supported by the fall in plasma HBD-2 levels after treatment in those patients who responded to therapy. HBD-2 is produced after stimulation of the epithelial cells by contact with microorganisms or cytokines such as tumor necrosis factor-{alpha} or IL-1ß.26 Our finding of a positive correlation between HBD-2 and IL-1ß in the BALF of the patients suggests that HBD-2 also may be regulated by IL-1ß in vivo. In this regard, Singh et al14 showed that IL-1ß stimulated the expression of HBD-2 messenger RNA and peptide production but not those of HBD-1 in primary cultures of airway epithelial cells. The authors suggested that HBD-2 expression in the lung is induced by inflammation, whereas HBD-1 may serve as a defense in the absence of inflammation. Their speculation was consistent with our results showing that the levels of HBD-1 in the BALF or plasma of MAI patients did not increase.

In this study, we could not clarify how HBD-2 is involved in pulmonary MAI, because HBD-2 was not shown to be cytotoxic to human cells. However, we believe that HBD-2 may be a useful marker of disease activity in patients with pulmonary MAI because plasma levels of HBD-2 fall after treatment in responders. Because HBD-2 is localized only in the epithelial cells of the respiratory tract, except in the skin in humans, plasma HBD-2 levels may reflect the severity of epithelial injury. Plasma HAD levels in responders also showed a tendency to decrease after treatment, although this reduction was not statistically significant. In this regard, our RIA data on HAD were expressed as the sum of mature HAD and their precursors, and their precursors are released from the bone marrow into systemic circulation by stimuli.27 Therefore, HAD levels in plasma are the cumulative effects of local inflammation in the lung and systemic inflammation. Plasma levels of HBD-2 probably reflect more directly the degree of epithelial injury than those of HAD.

The optimal duration of drug therapy for MAI patients has not yet been established, and relapse is a serious problem. Patients with MAI have to be treated over a long period with multiple antimicrobial agents because there is no sensitive test to assess the response to such therapy. Wallace et al28 and Dautzenberg et al29 used 12 and 7 to 9 months, respectively, of negative culture results as the treatment end point to confirm the lack of relapse of pulmonary MAI. In our study, we used a 6-month period of negative sputum culture results as an index of the response to therapy, although such a period really may be short compared with previous long-term studies for the clarithromycin-containing regimen.28 29 In fact, plasma HBD-2 levels remained high in 3 of 17 responders in our study, even after treatment, but relapse of MAI occurred in 2 of these 3 responders within 6 months after the finish of treatment.

In conclusion, we have demonstrated the presence of high plasma and BALF levels of antimicrobial peptides in the respiratory tracts of patients with MAI. Antimicrobial peptides may participate not only in opposing actions such as host defense but also tissue injury. Further studies of the roles of antimicrobial peptides should enhance our understanding of the pathogenesis of MAI.


   Footnotes
 
Abbreviations: BALF = BAL fluid; HAD = human {alpha}-defensin; HBD = human ß-defensin; IL = interleukin; MAI = Mycobacterium avium-intracellulare infection; RIA = radioimmunoassay

Received for publication July 19, 2000. Accepted for publication November 16, 2000.


   References
TOP
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. du Moulin, GC, Stottmeier, KD, Pelletier, PA, et al (1988) Concentration of Mycobacterium avium by hospital hot water systems JAMA 260,1599-1601[Abstract]
  2. Kennedy, TP, Weber, DJ (1994) Nontuberculous mycobacteria: an underappreciated cause of geriatric lung disease. Am J Respir Crit Care Med 149,1654-1658[Abstract]
  3. Yamazaki, Y, Kubo, K, Sekiguchi, M, et al (1998) Analysis of BAL fluid in M. avium-intracellulare infection in individuals without predisposing lung disease. Eur Respir J 11,1227-1231[Abstract]
  4. Elizabeth, H, Moore, EH (1993) Atypical mycobacterial infection in the lung: CT appearance. Radiology 187,777-782[Abstract/Free Full Text]
  5. Swensen, SJ, Hartman, TE, Williams, DE (1994) CT diagnosis of Mycobacterium avium-intracellulare complex in patients with bronchiectasis. Chest 105,49-52[Abstract/Free Full Text]
  6. Lehrer, RI, Barton, A, Daher, KA, et al (1989) Interaction of human defensins with Escherichia coli: mechanism of bactericidal activity. J Clin Invest 84,553-561
  7. Lehrer, RI, Lichtenstein, AK, Ganz, T (1993) Defensins: antimicrobial and cytotoxic peptides of mammalian cells. Annu Rev Immunol 11,105-128[CrossRef][ISI][Medline]
  8. Ogata, K, Linzer, BA, Zuberi, RI, et al (1992) Activity of defensins from human neutrophilic granulocytes against Mycobacterium avium-Mycobacterium intracellulare. Infect Immun 60,4720-4725[Abstract/Free Full Text]
  9. McCray, PB, Jr, Bentley, L (1997) Human airway epithelia express a ß-defensin. Am J Respir Cell Mol Biol 16,343-349[Abstract]
  10. Zhao, C, Wang, I, Lehrer, RI (1996) Widespread expression of ß-defensin hBD-1 in human secretory glands and epithelial cells. FEBS Lett 396,319-322[CrossRef][ISI][Medline]
  11. Stolzenberg, ED, Anderson, GM, Ackermann, MR, et al (1997) Epithelial antibiotic induced in states of disease. Proc Natl Acad Sci U S A 94,8686-8690[Abstract/Free Full Text]
  12. Diamond, G, Russell, JP, Bevins, CL (1996) Inducible expression of an antibiotic peptide gene in lipopolysaccharide-challenged tracheal epithelial cells. Proc Natl Acad Sci U S A 93,5156-5160[Abstract/Free Full Text]
  13. Harder, J, Bartels, J, Christophers, E, et al (1997) A peptide antibiotic from human skin [letter]. Nature 387,861[CrossRef][Medline]
  14. Singh, PK, Jia, HP, Wiles, K, et al (1998) Production of ß-defensins by human airway epithelia. Proc Natl Acad Sci U S A 95,14961-14966[Abstract/Free Full Text]
  15. Ashitani, J, Mukae, H, Nakazato, M, et al (1998) Elevated concentrations of defensins in bronchoalveolar lavage fluid in diffuse panbronchiolitis. Eur Respir J 11,104-111[Abstract/Free Full Text]
  16. . American Thoracic Society. (1997) Diagnosis and treatment of disease caused by nontuberculous mycobacteria. Am J Respir Crit Care Med 156,S1-S25
  17. Mukae, H, Kadota, J, Kohno, S, et al (1995) Increase in activated CD8+ cells in bronchoalveolar lavage fluid in patients with diffuse panbronchiolitis. Am J Respir Crit Care Med 156,613-618
  18. Shiomi, K, Nakazato, M, Ihi, T, et al (1993) Establishment of radioimmunoassay for human neutrophil peptides and their increases in plasma and neutrophil in infection. Biochem Biophys Res Commun 195,1336-1344[CrossRef][ISI][Medline]
  19. Hiratsuka, T, Nakazato, M, Matsukura, S, et al (2000) Structural analysis of human ß-defensin 1 and its significance in urinary tract infection. Nephron 85,34-40[CrossRef][ISI][Medline]
  20. Hiratsuka, T, Nakazato, M, Date, Y, et al (1998) Identification of human ß-defensin-2 in respiratory tract and plasma and its increase in bacterial pneumonia. Biochem Biophys Res Commun 249,943-947[CrossRef][ISI][Medline]
  21. Soong, LB, Ganz, T, Ellison, A, et al (1997) Purification and characterization of defensins from cystic fibrosis sputum. Inflamm Res 46,98-102[CrossRef][ISI][Medline]
  22. Ashitani, J, Mukae, H, Ihiboshi, H, et al (1996) Defensins in plasma and bronchoalveolar lavage fluid from patients with acute respiratory distress syndrome Nippon Kyobu Shikkan Gakkai Zasshi (in Japanese) 12,1349-1353
  23. Condos, R, Rom, WN, Ming, LYM, et al (1998) Local immune responses correlate with presentation and outcome in tuberculosis. Am J Respir Crit Care Med 157,729-735[Abstract/Free Full Text]
  24. Zhang, Y, Broser, M, Cohen, H, et al (1995) Enhanced interleukin-8 release and gene expression in macrophages after exposure to Mycobacterium tuberculosis and its components. J Clin Invest 95,586-592
  25. van Wetering, S, Mannesse-Lazeroms, SP, van Sterkenburg, MA, et al (1997) Effect of defensins on interleukin-8 synthesis in airway epithelial cells. Am J Physiol 272,L888-L896[Abstract/Free Full Text]
  26. Schroder, JM, Harder, J (1999) Human ß-defensin-2. Int J Biochem Cell Biol 31,645-652[CrossRef][ISI][Medline]
  27. Ashitani, J, Nakazato, M, Mukae, H, et al (2000) Recombinant granulocyte colony-stimulating factor induces production of human neutrophil peptides in lung cancer patients with neutropenia. Regul Pept 95,87-92[CrossRef][ISI][Medline]
  28. Wallace, RJ, Jr, Brown, BA, Griffith, DE, et al (1996) Clarithromycin regimens for pulmonary mycobacterium avium complex: the first 50 patients. Am J Respir Crit Care Med 153,1766-1772[Abstract]
  29. Dautzenberg, B, Piperno, D, Diot, P, et al (1995) Clarithromycin in the treatment of Mycobacterium avium lung infections in patients without AIDS: Clarithromycin Study Group of France. Chest 107,1035-1040[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 Eur Respir JHome page
H. Ishimoto, H. Mukae, Y. Date, T. Shimbara, M. S. Mondal, J. Ashitani, T. Hiratsuka, S. Kubo, S. Kohno, and M. Nakazato
Identification of hBD-3 in respiratory tract and serum: the increase in pneumonia
Eur. Respir. J., February 1, 2006; 27(2): 253 - 260.
[Abstract] [Full Text] [PDF]

Home page
 Infect. Immun.Home page
B. Rivas-Santiago, S. K. Schwander, C. Sarabia, G. Diamond, M. E. Klein-Patel, R. Hernandez-Pando, J. J. Ellner, and E. Sada
Human {beta}-Defensin 2 Is Expressed and Associated with Mycobacterium tuberculosis during Infection of Human Alveolar Epithelial Cells
Infect. Immun., August 1, 2005; 73(8): 4505 - 4511.
[Abstract] [Full Text] [PDF]

Home page
 Eur Respir JHome page
R. Bals and P.S. Hiemstra
Innate immunity in the lung: how epithelial cells fight against respiratory pathogens
Eur. Respir. J., February 1, 2004; 23(2): 327 - 333.
[Abstract] [Full Text] [PDF]

Home page
 ThoraxHome page
T Hiratsuka, H Mukae, H Iiboshi, J Ashitani, K Nabeshima, T Minematsu, N Chino, T Ihi, S Kohno, and M Nakazato
Increased concentrations of human {beta}-defensins in plasma and bronchoalveolar lavage fluid of patients with diffuse panbronchiolitis
Thorax, May 1, 2003; 58(5): 425 - 430.
[Abstract] [Full Text] [PDF]

Home page
 ThoraxHome page
H Mukae, H Iiboshi, M Nakazato, T Hiratsuka, M Tokojima, K Abe, J Ashitani, J Kadota, S Matsukura, and S Kohno
Raised plasma concentrations of {alpha}-defensins in patients with idiopathic pulmonary fibrosis
Thorax, July 1, 2002; 57(7): 623 - 628.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF) Free
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Article Archive
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (28)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ashitani, J.-i.
Right arrow Articles by Matsukura, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ashitani, J.-i.
Right arrow Articles by Matsukura, S.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS