Carnation Etched Ring Virus Inclusions: Serology and Ultrastructure of Alkaline-Treated Inclusions

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Abstract
Carnation etched ring virus (CERV) cytoplasmic inclusion bodies induced the formation of antibodies in immunized rabbits to intact virions (S-Ag) and to 2 rapid-diffusing antigens. One of the rapid-diffusing antigens (R-AG) may be matrix protein. The 2nd rapid-diffusing antigen was absorbed by sap from healthy Saponaria vaccaria. The effects of extraction medium, cations and pH on the stability of CERV inclusions were determined by serological tests and EM. Agar gel double diffusion tests and EM showed that R-Ag was dispersed more readily from inclusions extracted in distilled water than from those extracted in Tris buffer. Increased amounts of R-Ag were released from inclusions treated in distilled water at increasingly alkaline pH. Inclusions in distilled water showed a progressive increase in the loss of dense-staining matrix protein in ultrathin section with increasing pH. In Tris buffer at high pH the inclusion matrix remained more electron-dense than in distilled water. Weak precipitin lines were formed to R-Ag in Tris. S-Ag was detected with prolonged incubation in both distilled water and Tris at neutral and alkaline pH. The addition of cations stabilized the matrix protein. R-Ag was detected when protoinclusions were first observed in infected S. vaccaria cells.