Jesse K. Mackey

Thirty-one human adenovirus serotypes (Ad1-Ad31) form five groups (A-E) based upon DNA genome homologies.

Abstract The DNA homology relationships among 31 human adenovirus serotypes (Ad1–Ad31) were investigated by liquid-phase molecular hybridization, using in vitro labeled viral DNA as probe. Hybridizations were carried to 40 times the C 0 t 1 2 and were assayed by batchwise chromatography on hydroxylapatite (HAP), and in some experiments by use of the more stringent S1 nuclease procedure. Five distinct DNA homology groups, A to E, were identified. DNAs of group A Ads (Ad12, 18, and 31) hybridized 48 to 69% with each other and 8 to 20% with DNAs of other serotypes (HAP). DNAs of group B Ads (Ad3, 7, 11, 14, 16, and 21) hybridized 89 to 94% (HAP; 81 to 89% by S1 nuclease) with each other and 9 to 20% (HAP; 8 to 15% by S1 nuclease) with DNAs of other types. DNAs of group C Ads (Ad1, 2, 5, and 6) hybridized 99 to 100% with each other and 10 to 16% with DNAs of other types (HAP). DNAs of group D Ads (Ad8–10, 13, 15, 17, 19, 20, and 22–30) hybridized 95 to 99% (HAP; 88 to 98% by S1 nuclease) with each other and 4 to 17% with DNAs of other types. Ad4 DNA hybridized to 4 to 23% (HAP; 3 to 22% by S1 nuclease) with DNAs of other types, and thus Ad4 is the only member of group E. Members within all groups except group A were closely related. Members within group A showed considerable heterology, and six isolates, classified as Ad12 by neutralization tests, were much more related to Ad31 than to Ad12 prototype Huie strain. These DNA homology groupings are consistent in the main with the properties of other “groupings” of human Ads, e.g., oncogenic groups (tumorigenicity in newborn hamsters), T-antigen groups, G + C content of viral DNA, hemagglutination groups, molecular characteristics of subviral particles and virion proteins (e.g., length of fiber), and human epidemiology and pathogenicity.

Analysis of human cancers, normal tissues, and verruce plantares for DNA sequences of human papillomavirus types 1 and 2

Abstract Comparatively little is known about human papillomaviruses (HPV) because they cannot be grown in tissue culture. We have in vitro labeled DNAs from two HPVs, HPV-1 which was isolated from plantar warts, and HPV-2 which was isolated from common hand warts, and used these DNAs to examine the homology between HPV-1 and HPV-2, to examine the state of the HPV genome in papillomavirus lesions, and to assay human cancer DNAs for HPV. The specific activities of the DNAs were 5.0 × 10 7 to 1.1 × 10 8 cpm/μg. The C 0 t 1 2 of the HPV-1 and HPV-2 DNAs were 5 and 7 × 10 −4 , respectively, consistent with a genome molecular weight of about 5.2 × 10 6 . Cross-hybridization of HPV-1 and HPV-2 DNAs revealed only 5–7% homology, confirming that these are distinct viruses. HPV-1 DNA was detected by Southern blot analysis in 9 of 10 plantar warts examined. No clear evidence was found for integrated viral sequences in DNAs from eight of the nine warts analyzed. Using these HPV-1 and HPV-2 probes, we have performed the first extensive and definitive molecular hybridization analysis of human cancer DNAs for HPV sequences. Human tumor DNAs were analyzed for HPV sequences by saturation hybridization using nick-translated HPV-1 and HPV-2 DNA probes. Reconstruction experiments with added HPV-1 or HPV-2 DNAs indicated that the probes could detect 0.1 copy of the viral genome per diploid equivalent of cellular DNA. No HPV-1 sequences were detected in DNAs from 156 human cancers (14 melanoma, 3 Ca skin, 5 Ca pharynx, 1 Ca esophagus, 4 Ca stomach, 5 Ca small intestine, 22 Ca colon, 14 Ca rectum, 25 squamous cell Ca lung, 3 adenocarcinoma lung, 4 oat cell Ca lung, 21 Ca kidney, 7 Ca bladder, 3 Ca ovary, 3 Ca cervix, 4 Ca prostate, 10 non-Hodgkin lymphoma, 2 reticulum cell sarcoma [spleen]), or 27 normal human tissues (1 skin, 10 tonsil, 8 colon, 8 kidney). No HPV-2 sequences were detected in DNAs from 145 human cancers (13 melanoma, 4 Ca skin, 2 Ca pharynx, 3 Ca mouth, 7 Ca esophagus, 4 Ca stomach, 3 Ca small intestine, 29 Ca colon, 15 Ca rectum, 25 Ca kidney, 15 Ca bladder, 2 Ca ovary, 6 Ca cervix, 4 Ca prostate, 2 Ca seminoma testes, 11 non-Hodgkin lymphoma) or 1 normal human ovary. These data are strong evidence that none of the cancer specimens assayed were induced by HPV-1 or HPV-2. However, additional work is required to fully evaluate whether HPVs are possible agents of human cancers, because the cancer types assayed in this study represent only about 50% of the cancer incidence in the United States, and because our probes would not detect sequences of other recognized HPV types (HPV-3, HPV-4, and HPV-5).

Evidence for post-transcriptional selection of viral mRNA in cells transformed by human adenovirus 12

CONTROL of gene expression occurs mainly at the transcriptional level in prokaryotic cells1. In eukaryotic cells, the sites of transcription and translation are physically separated by the nuclear membrane, giving rise to the possibility of regulation by post-transcriptional mechanisms. Control could be exercised at several stages of mRNA biogenesis: methylation, polyadenylation, cleavage, or transport across the nuclear membrane2,3.

Chronic Inflammatory Bowel Disease: Absence of Adenovirus DNA as Established by Molecular Hybridization

Because previous evidence indicated that adenovirus is tropic for gut wall cells (particularly terminal ileum) and can become chronically established in mammalian cells as a latent infection, we examined, through the use of sensitive molecular hybridization techniques capable of detecting minute amounts of adenovirus DNA, the hypothesis being that adenovirus is specificallly associated with both Crohns disease and ulcerative colitis. DNA of 21 coded specimens from 17 operated patients (6 with Crohns disease, 4 with ulcerative colitis, 7 controls) was hybridized with each of five [ 32 P]DNA probes, prepared by nick translation of adenovirus DNA with Escherichia coli polymerase I and representative of each major adenovirus subgroup (A through E). While quantities as small as 0.2 copies of purified adenovirus DNA per human cell were detectable in standardization experiments, none of the test specimens was positive, either in Crohns disease or ulcerative colitis, whether histologically normal or abnormal. Potential sources of false-negative results were analyzed and judged to be unlikely. Future investigation in this area may await the development of more sensitive assays for viral DNA or for evidence of new candidate virus systems.