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Investigation on Intranuclear Paracrystalline Inclusions Induced by Adenovirus 5 in KB Cells^*

P. A. Boulanger

G. Torpier

Unité de Recherches no. 16 de l'I.N.S.E.R.M. sur la Biochimie des Protéines, Place de Verdun, 59, Lille, France

G. Biserte

Laboratoire de Virologie de l'Institut Pasteur de Lille, France

Intranuclear paracrystalline bodies have been observed in cells infected with adenovirus type 5 (Leuchtenberger & Boyer, 1957; Morgan et al. 1957, 1960). The present study reports the attempts to isolate these paracrystalline proteins from KB cell nuclei and discusses their nature and possible role.

KB cells were grown as monolayers in 1 l. bottles (5 x 10⁷ to 6 x 10⁷ cells/bottle) in Eagle's basal medium supplemented with 10% calf serum. The cells were infected at a multiplicity of 100/infecting cell units/cell (Warocquier, Ménard & Samaille, 1966) and the virus was allowed to grow for 48 hr. The paracrystalline bodies were extracted directly from infected KB cells by homogenizing them in 1% (w/v) citric acid (Mirsky & Pollister, 1946) or 0.15% (v/v) Cemulsol (Zalta et al. 1962) using a Dounce homogenizer. The suspension was centrifuged at 1000 g for 10 min. and the pellet examined in the electron microscope, after fixation in glutaraldehyde + osmium tetroxide and embedding in Araldite (Richardson, Jarett & Finke, 1960).

^* This work was supported in part by a Convention de Recherche 67-00-537 from the Délégation Générale à la Recherche Scientifique et Technique (Action Concertée: Biologie Moléculaire).

Present address: Institute for Molecular Virology, 3681 Park Avenue, Saint Louis, Missouri 63110, U.S.A.

Recipient of a fellowship from the Institut National de la Santé et de la Recherche Médicale (Action Concertée: Cancer-Leucémie, DGRST, CR 66-209).

Received 26 August 1969; accepted 14 October 1969.