A protocol for the purification of carp eosinophils on metrizamide gradients is presented. Density gradient centrifugation is a new approach to the study of eosinophils in fish. It provides enriched populations of viable eosinophils necessary for in vitro functional assays. Eosinophils purified from the peripheral blood of carp resemble mammalian eosinophils by light and electron microscopy. They are indistinguishable from carp neutrophils which have a different density and morphology.
In both primary and secondary infections using 3 different strains of mice (DBA/1, B10 X D2, and C3H/HeJ), 2 peaks of peripheral blood eosinophilia were noted. Depending upon the strain, the first of the biphasic peaks in primary infections occurred between days 5 and 9, and the second peak occurred between days 16 and 22. The relative heights of the peak varied with the strain. In secondary infections, the initial peak was lower and the larger second peak occurred earlier. Methyridine treatment to eliminate adults prior to release of newborn larvae altered neither the course of eosinophilia in primary infection nor the biphasic distribution of eosinophilia. These studies demonstrate that the adult worm is responsible for inducing eosinophilia in murine infections, that 2 peaks of eosinophilia exist in primary and secondary infections, and that eosinophilia in secondary infections is an anamnestic response.
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