Enzyme-Linked Immunosorbent Assay Specific for (1→6) Branched, (1→3)-β- d -Glucan Detection in Environmental Samples

Abstract

(133)-␤-D-Glucans are carbohydrate polymers found in the cell walls of most fungi, some bacteria, yeasts, and higher and lower plants. The molecular weight, degree of (136)-linked side branching, helical conformation, and solubility are the most important factors which determine the biological activities of (133)-␤-D-glucans. The biological activities of (133)-␤-D-glucans include host-mediated antitumor activity (11), adjuvant effects (21), and activation of neutrophils (34), eosinophils (23), macrophages (15, 27), and complement (26).There is increasing evidence that (133)-␤-D-glucans cause nonspecific inflammatory reactions (9, 10) and are responsible for bioaerosol-induced respiratory symptoms observed in both indoor (24,25,32) and occupational (4, 5) environments. Dose-response relationships have been found between levels of (133)-␤-D-glucans and eye and throat irritation, dry cough, and itching skin (25). Relationships have also been found among exposure to (133)-␤-D-glucans and an increased prevalence of atopy, a decrease in forced expiratory volume in 1 s in adults (30), and peak flow variability in children (6). Recent studies have reported significantly high levels of (133) (18,20).In the present study, we describe the use of an enzymelinked immunosorbent assay (ELISA), which was originally developed as a diagnostic assay for systemic fungemia (8), to assay (133)-␤-D-glucans in house dust samples, some mould cultures, and metal working fluid samples. The assay is based on the use of a high-affinity receptor (galactosyl ceramide, a sphingolipid) specific for (133)-␤-D-glucans as a capture reagent and a monoclonal antibody specific for complex fungal cell wall (133)-␤-D-glucans as a detector reagent. The assay is reported to be rapid, specific for fungal ␤-glucans, and insensitive to interference caused by agents which affect the Limulus assay. MATERIALS AND METHODSGalactosyl ceramide (Galactercerebroside type II; Sigma, St. Louis, Mo.) and 1% bovine serum albumin (BSA) (Sigma) in Tris-buffered saline (pH 8.0) (TBS) were used, respectively, for coating and blocking. Immulon 96-well polystyrene

Characterization of Airborne Molds, Endotoxins, and Glucans in Homes in New Orleans after Hurricanes Katrina and Rita

“…We found that glucan levels correlated significantly with airborne spore counts but not culturable fungal levels, possibly because measures of culturable fungi underestimate the airborne fungal load (10). In addition, different fungal species have different concentrations of (133,136)-␤-D-glucans (11).…”

Therapeutic Activity of an Engineered Synthetic Killer Antiidiotypic Antibody Fragment against Experimental Mucosal and Systemic Candidiasis

“…Finally, a major interest of our approach is the potential antimicrobial spectrum of KP, which may include most microorganisms possessing ␤-glucan in their cell wall, inclusive of yeasts, most filamentous fungi, and some bacteria (5,7,16,21). The observation that an engineered antibody fragment exerts an antimicrobial activity in vitro and in vivo comparable to that displayed by the original KT-scFv could open a new scenario in the biochemistry of antimicrobial peptides for the prevention and therapy of the numerous superficial and systemic diseases caused by the numerous KT-susceptible microorganisms.…”

β-(1,3)-Glucan Exposure Assessment by Passive Airborne Dust Sampling and New Sensitive Immunoassays

“…The assay developed by Sander et al (27) has been applied to only a limited number of samples from the work environment, and the EIA described by Blanc et al (2) and Rao et al (23) has been used only to analyze reservoir and airborne dust samples from heavily mold-contaminated houses in New Orleans after the hurricanes Katrina and Rita. A third sensitive EIA makes use of galactosyl ceramide, a receptor specific for ␤-(1,3)-glucans (41), as the capture reagent and of a monoclonal antibody specific for ␤-(1,3)-(1,6)-glucans as the detecting antibody (20). Application of this EIA in population studies has, however, not yet been reported.…”