Immunological Studies of Insect Metamorphosis

1. The cell-free blood of the Cecropia silkworm produces a maximum of nine bands of antigen-antibody precipitate when reacted in antiserum-agar tests with antisera prepared by injecting Cecropia extracts into rabbits. The blood antigens producing these bands of precipitate have the properties of proteins in that they are non-dialyzable, labile at 75 degrees C., and salted out by 75 per cent saturated ammonium sulfate. One antigen was identified as a carotenoid protein. 2. Six bands of precipitate were selected for further study. Absorption tests revealed that the blood, at all stages of metamorphosis, is capable of precipitating the antibodies which produce five of these bands. This result indicates that five of the six antigens are present in the blood throughout metamorphosis. The sixth antigen is undetectable in blood from fourth instar larvae, appears in the blood late in the fifth instar, persists during the pupal stage, and disappears again during adult development. 3. When blood samples from various stages of metamorphosis were tested in antiserum-agar tubes, the rates of advance of the six bands of precipitate underwent systematic change in close correlation with the morphological stage of the silkworm's metamorphosis. These changes are interpreted in terms of concentration changes of the corresponding blood antigens. The validity of this interpretation was tested in several ways, with the conclusion that the interpretation was generally acceptable for the system under consideration. 4. All six antigens appear to increase in concentration during the last larval instar and to decrease in concentration during the period of adult development. However, each antigen has its own characteristic pattern of concentration change which differs from those of the other five. In order to explain this diversity, we conclude that the physiological mechanisms which regulate the synthesis and utilization of the blood antigens control each antigen on an individual basis.