Experiments are described in which the conditions for the production, assay and isolation of immunomodulatory factors from the excretory-secretory (ES) products of Heligmosomoides polygyrus have been standardized. For the inhibition of an in vitro antibody response to keyhole limpet haemocyanin, immunosuppressive activity was most reproducibly produced by 10-20-day-old adult worms maintained in culture for 24 h. This activity was relatively stable at room temperature, at 50-degrees-C and pH 2, but was destroyed by boiling. Immunosuppressive activity was eluted from Sephadex G-100 in fractions preceding those containing the bulk of ES proteins, and resolved on sodium dodecyl sulphate-polyacrylamide gel electrophoresis with molecular masses of 67, 54 and 20 kDa. The relative purity of these factors was confirmed by iso-electric focusing, where immunosuppressive activity was associated with proteins of pI values of approximately 4.2 and 4.35.