Multiplication of adenovirus type 5 studied by infectivity titrations and by the fluorescent antibody technique

Antibody to the Type 5 Adenovirus Hexon Polypeptide: Detection of Nascent Polypeptides in the Cytoplasm of Infected KB Cells

“…The data presented demonstrate that the nascent polypeptide chains are present in the cytoplasm but are not detectable in the nucleus, whereas intact hexon is predominantly found in the nucleus [2][3][4][19][20][21]. The evidence implies that either the hexon assembles in the nucleus without permitting accumulation of the polypeptide chains or the hexon assembles in the cytoplasm and is rapidly transported into ihe nucleus [ 10], The latter interpretation is consistent with the finding that the polypeptide chains of adenovirus capsid proteins synthesized in vitro in cytoplasmic extracts and ribosomal fractions can assemble into multimeric structures having the physical and immunological characteristics of adenovirus capsomers [25].…”

“…KB monolayers were also infected with type 5 adenovirus ts mutants, H5ts 116 or H5ts115, and incubated at either 32 or 39.5°. The adenovirus mutants employed are termed hexon-minus because they are defective in the synthesis of immunologically detectable hexon at the nonpermissive temperature, 39.5 [5], Cells infected with adenovirus type 5 or 2, incubated at 37c, and treated with anti-hexon sera had fluorescence mainly in the nucleus, as previously reported for cells infected with adenoviruses and stained with anti-virion or capsid sera [2][3][4][19][20][21]. In contrast, type 5 ( fig.4A) or 2 adenovirus-infected cells treated with anti-type 5 hexon polypeptide serum had mainly cytoplasmic fluorescence.…”

“…Adenovirus capsid proteins are synthesized on cytoplasmic polyribosomes of infected cells [I], but immunofiuorescent and ferritin-labeled antibody techniques primarily detect the viral proteins in nuclei [2][3][4], Thus, the nascent polypeptide chains are rapidly transported from their site of biosynthesis to the locus of viral assembly [I], This report presents data showing that antibody to the native hexon capsomer reacts solely with the multimeric proteins (trimers) and not with its component polypeptide chains (monomers) and that antibody prepared by immunization with the isolated hexon polypeptide chains reacts with polypeptide chains and not with intact hexons. It will also be demonstrated that, although intact hexons cannot be immunologically under a partial vacuum that was created by aspirating through a hole in one of the silicone sheets.…”

Evaluation of sampling technique and transport media for the diagnostics of adenoviral eye infections

“…More rapid assays have been developed, including enzyme immunoassay, direct immunofluorescence, DNA hybridisation and electron microscopy [4,16,17,19,22]. These techniques can be complex, laborious and often inappropriate for routine situations.…”

The kinetics of adenovirus infection and spread in cell cultures infected with low multiplicity

“…The number of infectious particles produced per cell has been reported to be l03.5 to 104.0 (6,2). The number of infectious particles produced per cell has been reported to be l03.5 to 104.0 (6,2).…”