Increased levels of nerve growth factor in the airways of patients with sarcoidosis.

OBJECTIVES: Nerve growth factor (NGF) is a potent neuronal growth factor with inflammatory properties that recently has been proposed to be of importance in airway pathology. A role for NGF in the inflammatory granulomatous lung disease sarcoidosis is not well elucidated. The aims of this study were to investigate the secreted levels of NGF in bronchoalveolar lavage fluid (BALF) from sarcoidosis patients compared with patients with resolved disease, patients with another granulomatous disease--chronic beryllium disease (CBD)--and healthy subjects and also to investigate the relationship between NGF levels and markers of inflammation. METHODS AND RESULTS: NGF levels in BALF from 56 patients with active sarcoidosis (22 with Löfgren's syndrome), nine subjects with resolved sarcoidosis, six patients with CBD, and 31 healthy subjects were compared. A 10-fold elevation of NGF levels was found in patients with active sarcoidosis compared with subjects with clinically resolved sarcoidosis, patients with CBD and healthy subjects. In sarcoidosis patients, positive correlations between concentrations of NGF and lymphocytes, eosinophils and interferon-gamma, interleukin (IL)-4, IL-10, IL-12 were found. CONCLUSIONS: We demonstrate that secreted levels of NGF are markedly enhanced in the airways in active pulmonary sarcoidosis. Furthermore, a relationship between NGF and pulmonary inflammation in sarcoidosis is supported.

No serological evidence of Rickettsia helvetica infection in Scandinavian sarcoidosis patients.

Many agents have been suggested to elicit sarcoidosis, and, recently, an association was presented between this disease and the bacterium Rickettsia helvetica. The aim of this study was to investigate if serological support for such an association could be detected. Sera from 20 well-characterised sarcoidosis patients were investigated for anti-rickettsial immunoglobulin G antibodies with a micro-immunofluorescence technique. R. helvetica, R. conorii and R. typhi served as antigens. In conclusion, none of the investigated sera displayed detectable titres of anti-rickettsial immunoglobulin G antibodies. Thus, the current study does not support an association between rickettsia and sarcoidosis.

Markers of activity in clinically recovered human leukocyte antigen-DR17-positive sarcoidosis patients.

Scandinavian human leukocyte antigen-DR17-positive (DR17+) sarcoidosis patients are characterised by a good prognosis. They also reveal an accumulation in bronchoalveolar lavage fluid of T-lymphocytes expressing the T-cell receptor V gene segment AV2S3 at disease onset. The authors of this study wished to establish whether AV2S3 T-lymphocyte accumulation changes from disease onset to clinically resolved disease and how this relates to other activity parameters. Bronchoalveolar lavage fluid and serum from nine DR17+ sarcoidosis patients were examined at disease onset and after spontaneous resolution of clinical and radiographical signs of disease. Nine DR17+ patients with lung accumulated CD4+ AV2S3+ T-cells were investigated after clinically recovery. At re-examination the percentage of CD4+ AV2S3+ lymphocytes in bronchoalveolar lavage fluid was normalised (29 versus 5.4%). A significant reduction in lymphocyte percentage (14 versus 4.4%) and a decrease in cellular concentration (179x10(6) x L(-1) versus 111x10(6) x L(-1)) and CD4/CD8 ratio (5.2 versus 2.4) were also seen. In serum, the activity of angiotensin-converting enzyme (24.9 versus 14.0 U x mL(-1)) as well as the levels of neopterin (7.8 versus 5.3 nmol x L(-1)) decreased significantly after recovery. These results indicate that the locally accumulated AV2S3 positive T-lymphocytes in bronchoalveolar lavage are involved in the pathogenic process of sarcoidosis in this patient group.

Apoptosis resistant bronchoalveolar lavage (BAL) fluid lymphocytes in sarcoidosis.

BACKGROUND: Sarcoidosis is a chronic granulomatous lung disease of unknown origin. The accumulation of activated T cells at sites of inflammation represents an early stage in granuloma formation. Since mechanisms governing the normal resolution of inflammatory processes are poorly understood, this study aimed to investigate the apoptotic phenotype of peripheral blood and lung T lymphocytes from patients with sarcoidosis. METHODS: Bronchoalveolar lavage (BAL) was performed in 10 patients with active sarcoidosis and five healthy controls. RESULTS: Virtually no lymphocyte apoptosis, as determined by annexin V or Hoechst staining, was seen in either patients or controls. Sustained caspase-3 activity in non-apoptotic BAL fluid lymphocytes of the patients was detected, however, in agreement with in vitro studies demonstrating caspase activation after T cell receptor (TCR) triggering as a physiological response required for efficient T cell activation. Only 11.0% (range 7.7-17.6) of the BAL lymphocytes from sarcoidosis patients were annexin V positive after exposure to the apoptotic stimulus tributyltin compared with 55.0% (range 42.0-62.0) of BAL lymphocytes from healthy controls (p<0.001). After anti-Fas treatment only 8.5% (range 6-10) of BAL fluid lymphocytes from patients but 45.5% (range 38-62) from healthy controls were apoptotic. CONCLUSION: BAL fluid lymphocytes from patients with sarcoidosis display a non-apoptotic morphology associated with endogenous caspase-3 activity. They seem to be resistant to apoptosis, which might contribute to the accumulation of inflammatory cells in the lungs, persistence of inflammation, and the development and maintenance of granuloma.

Angiotensin-converting enzyme gene polymorphism in relation to HLA-DR in sarcoidosis.

OBJECTIVES: To investigate if an insertion/deletion (I/D) polymorphism in the angiotensin-converting enzyme (ACE) gene associates with HLA-DR alleles previously found to be of prognostic interest in Scandinavian sarcoidosis patients. This may contribute to characteristics associated with these HLA-DR alleles, such as a good (DR17) or poor (DR14 or 15) prognosis. DESIGN, SETTINGS AND SUBJECTS: Polymerase chain reaction (PCR) was used for analysing an I/D polymorphism in the gene coding for ACE in 138 subjects; 65 controls and 73 sarcoidosis patients, and for HLA-DR genotyping 67 patients. Serum ACE level (S-ACE) was measured in all controls and 72 patients. Sixty-one patients were classified as chronic or nonchronic after 2 years follow-up. All patients were recruited and followed at our outpatient clinic. RESULTS: No significant differences in ACE alleles or genotypes were seen between controls and patients or between patients positive and negative for DR17 or DR14/15. The ACE genotype did not differ between nonchronic and chronic patients. The ACE genotype tended to influence the S-ACE in patients, whilst in controls S-ACE significantly differed between the ACE genotypes. CONCLUSION: This study does not support an association between ACE genotype and sarcoidosis or disease outcome. However, because significantly (P < 0.001) more DR17 positive (17 of 19) than DR14/15 positive (seven of 26) patients were classified as nonchronic, these results instead strengthen the prognostic importance of HLA-DR alleles in Scandinavian sarcoidosis patients.

Inflammatory BAL-fluid and serum parameters in HLA DR17 positive vs. DR17 negative patients with pulmonary sarcoidosis.

BACKGROUND AND AIM OF THE WORK: We have previously shown that HLA DR17 is associated with a favourable prognosis in Scandinavian sarcoidosis patients. By studying inflammatory parameters in these patients we wished to increase the knowledge of the involved mechanisms that may underlie good prognosis. METHODS: BAL was performed in 118 sarcoidosis patients, 67 HLA DR17 positive and 51 DR17 negative. BAL cell profile was analysed. The CD4 and CD8 lymphocyte phenotype was determined by flow cytometry. BALF-albumin, BALF-fibronectin (FN) and BALF-procollagen III aminoterminal peptide (PIIINP) were analysed by nephelometric, ELISA and RIA methods respectively. Neopterin and soluble interleukin-2 receptor (sIL2R) in serum were also determined by ELISA. Angiotensin-converting enzyme (ACE) in serum was analysed by spectrophotometry. RESULTS: In DR17+ patients, BAL lymphocytes and eosinophils were significantly decreased (median 30.2 and 0.19 x10(6)/L) compared to DR17- (median 50.2 and 0.64 x 10(6)/L respectively). However the BAL CD4/CD8 ratio was increased in the DR17+ group compared to DR17- (6.2 vs. 4.3). BALF-albumin, FN and PIIINP did not differ between the groups. Serum parameters were decreased in DR17+ patients compared to DR17- (ACE median 28.7 vs. 35.1 U/L, neopterin 9.1 vs. 12.7 nmol/L and sIL2R 296 vs. 605 U/L). CONCLUSIONS: The study revealed that an increased BAL CD4/CD8 ratio, in contrast to BAL lymphocytosis, was seen in a subgroup of sarcoidosis patients earlier associated with a favourable prognosis. Our results support that an increased BAL CD4/CD8 ratio is a favourable parameter in sarcoidosis. The lower ACE activity in the DR 17+ group indicates a reduced granuloma burden in this patient subgroup.