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Pigeon breeder's disease (PBD) is an interstitial lung disease induced by exposure to pigeon antigens. Search of antipigeon antigen antibodies (APSA) in serum or bronchoalveolar lavage is generally used for auxiliary diagnostic purposes. However, APSA can be present in a number of exposed but asymptomatic individuals as well as in patients with other interstitial lung diseases who live in areas where keeping pigeons is a common domestic habit. In this study, saliva was evaluated as an alternative means to serum for APSA detection by ELISA using pooled pigeon sera as antigen. Serum and saliva samples obtained from 17 patients with PBD, 14 with idiopathic pulmonary fibrosis (IPF), 19 asymptomatic relatives (AR) exposed to pigeon antigens, and 27 clinical healthy voluntary subjects (CHVS) were tested for IgG and IgA APSA. Our results showed that both fluids obtained from PBD patients exhibited a significantly higher specific IgG antibody activity compared to the other groups. Serum optical density (O.D.) values for PBD were 1.187 +/- 0.738 vs. 0.024 +/- 0.033, 0.255 +/- 0.471, and 0.204 +/- 0.346 for CHVS, AR and IPF, respectively (P < 0.05). Salivary O.D. for PBD were 0.801 +/- 0.447 vs 0.010 +/- 0.011, 0.104 +/- 0.151, and 0.22 +/- 0.447 (P < 0.05). In contrast, serum specific IgA did not discriminate between PBD and IPF patients. In addition, although the PBD group exhibited the highest values of IgA salivary APSA, high levels were also observed in saliva specimens from CHVS, a group of normal individuals who deny pigeon exposure. These findings suggest that measurement of IgG salivary APSA can play a role in the evaluation process of patients with pigeon breeder's disease.

Anticardiolipin antibodies (ACA) were evaluated using enzyme-linked immunosorbent assay (ELISA) with and without heat inactivation of bovine serum used for plastic surface blockade. Untreated sera samples from primary antiphospholipid syndrome patients (PAS) and healthy blood donors (HBD) were tested. A significant decrease of ACA titers of PAS sera occurred with inactivated bovine serum blockade ELISA when compared with basal ELISA. In HBD sera there was no significant change. Probably, as happens with normal human serum, heating for normal bovine serum produces an increase in ACA titers. This bovine ACA may react with cardiolipin, and when human samples are added, they find antigen sites occupied, resulting in a spurious decrease of ACA titers.

The standard enzyme linked immunosorbent assay (ELISA) currently in use for detection of anticardiolipin antibodies (ACA) was used to evaluate the influence of heat inactivation and sheep erythrocyte adsorption on individual optical density (OD) of sera from healthy blood donors or patients with primary antiphospholipid syndrome. Each sample was tested after single or combined maneuvers as follows: adsorbed, adsorbed and inactivated, only inactivated, and compared to basal readings. A significant increase of ACA titers did occur after inactivation of normal sera, but adsorption had no effect. In contrast, neither inactivation nor adsorption changed ACA titer in primary antiphospholipid syndrome sera as a group, although in certain sera there were changes. This observation may suggest the presence in normal serum of a thermolabile factor which modulates ACA binding to its antigen and the reactivity of the anticardiolipin antibodies of the primary antiphospholipid syndrome with sheep erythrocyte membrane phospholipids.

The study reported here evaluated the usefulness of the enzyme-linked immunosorbent assay (ELISA) in the detection of antibodies against pigeon antigens in the serum and bronchoalveolar lavage (BAL) of patients with clinical, radiological, and functional evidence of interstitial lung disease (ILD) with and without pigeon breeder's disease (PBD). The results were compared with those obtained by the simultaneous use of counterimmunoelectrophoresis (CIE) in the same patients. In PBD, ELISA detected antibodies against pigeon's sera in both serum and BAL in 100% of patients, while CIE failed to detect the antibodies in the serum of one patient and in most of the samples of BAL. In addition, we used enzyme immunotransfer blotting to determine the number of epitopes in pigeon serum recognized by antibodies present in serum and BAL. There was a heterogeneous response in both fluids, but the reaction pattern demonstrated that patient's sera recognize to-25 different pigeon epitopes. We conclude that ELISA is a highly sensitive and specific method for the detection of antibodies against pigeon antigens in the serum and BAL of patients with PBD and that the host response involves a great number of avian antigens.