Farmer's lung: thermophilic actinomycetes as a source of "farmer's lung hay" antigen. 1963.

Mouldy hay was produced in the laboratory by sterilising good hay, inoculating with aqueous suspensions of microorganisms, and incubating at 40 degrees or 60 degrees C. Extracts were tested for presence of farmer's lung hay (F.L.H.) antigen by agar-gel double-diffusion and immunoelectrophoresis tests against sixteen to twenty sera from patients with farmer's lung. F.L.H. antigen developed in hay after: (1) inoculating with mixed microbial suspensions from antigenically active hay; (2) inoculation with mixed suspensions of pure cultures of thermophilic actinomycetes, after raising the pH of the hay to 70 either by prior inoculation with fungi or by infiltration with ammonia vapour; and (3) inoculation at pH 70 with pure cultures of Thermopolyspora polyspora or with Micromonospora vulgaris. F.L.H. antigen did not develop in hay after inoculation with fungi only, or with six other actinomycetes tested, or after prior heating (though some sera reacted to fungal antigens in all these extracts). T. polyspora is the richest source yet found of F.L.H. antigen, and inhalation of an extract by affected subjects produces some of the features of farmer's lung. Pure cultures can produce F.L.H. antigen on artificial media without hay. Spores and mycelium are rich in F.L.H. antigen, and inhalation of the spores may play a part in farmer's lung disease. Other antigens relevant to farmer's lung may be found in other actinomycetes, not yet cultured.

Specificity of an antibody to a subunit of high-molecular-weight storage protein from wheat seed and its reaction with other cereal storage proteins (prolamins).

An antiserum to subunit 2 from the high-molecular-weight (HMW) subunits of the glutenin fraction of Triticum aestivum cv. Highbury was shown to react with related subunits from other cultivars of wheat. The reaction was measured quantitatively by laser nephelometry in polyethylene glycol phosphate-buffered saline after dissolving the HMW fraction in 0.1 M acetic acid; urea used to dissolve the HMW prolamins inhibited the reaction, in some cases at the low concentration of 0.06 M. A study of the comparative reactions of other cereal prolamins was made. 'D' hordein, the homologous HMW protein of barley, showed less reaction, which was more inhibited by urea than the wheat subunits. Some ω-gliadins from the wheat cultivars Chinese Spring and Cheyenne reacted more strongly than the injected fraction and there was less inhibition by urea. A-, ß- and γ3 of wheat also reacted with the antiserum while a secalin of rye of Mr 40000 gave a weak reaction.

Immunochemical studies on barley seed storage proteins : The specificity of an antibody to 'C' hordein and its reaction with prolamins from other cereals.

The antigenic relationships between the prolamins of barley, rye and wheat have been studied by examining the specificity of an antibody to 'C' hordein in a quantitative study using a laser nephelometer. The antibody reacts weakly with 'B' hordein and strongly with 75-kdalton and 40-kdalton γ-secalins from rye and γ3 some ω-gliadins from wheat. Absorption experiments and immunodiffusion tests indicate that there are shared antigenic determinants for most of the prolamins. All the species with reacting prolamins belong to the sub-family Festucoideae of the Gramineae. The prolamins of maize, pearl millet and sorghum, species of the sub-family Panicoideae, do not react. The results confirm the known lack of homology between the prolamins of the two sub-families and also indicate the presence of relationships not yet established between 'C' hordein, the 75-kdalton and 40-kdalton γ-secalins and also γ3 gliadin.