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Journal of Virology, November 2002, p. 10608-10616, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.10608-10616.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Prevalence and Quantitation of Species C Adenovirus DNA in Human Mucosal Lymphocytes

C. T. Garnett,1 D. Erdman,2 W. Xu,2,{dagger} and Linda R. Gooding1*

Department of Microbiology and Immunology, Emory University, Atlanta, Georgia 30322,1 Centers for Disease Control and Prevention, Atlanta, Georgia 303332

Received 7 May 2002/ Accepted 20 July 2002

The common species C adenoviruses (serotypes Ad1, Ad2, Ad5, and Ad6) infect more than 80% of the human population early in life. Following primary infection, the virus can establish an asymptomatic persistent infection in which infectious virions are shed in feces for several years. The probable source of persistent virus is mucosa-associated lymphoid tissue, although the molecular details of persistence or latency of adenovirus are currently unknown. In this study, a sensitive real-time PCR assay was developed to quantitate species C adenovirus DNA in human tissues removed for routine tonsillectomy or adenoidectomy. Using this assay, species C DNA was detected in Ficoll-purified lymphocytes from 33 of 42 tissue specimens tested (79%). The levels varied from fewer than 10 to greater than 2 x 106 copies of the adenovirus genome/107 cells, depending on the donor. DNA from serotypes Ad1, Ad2, and Ad5 was detected, while the rarer serotype Ad6 was not. When analyzed as a function of donor age, the highest levels of adenovirus genomes were found among the youngest donors. Antibody-coated magnetic beads were used to purify lymphocytes into subpopulations and determine whether viral DNA could be enriched within any purified subpopulations. Separation of T cells (CD4/8- expressing and/or CD3-expressing cells) enriched viral DNA in each of nine donors tested. In contrast, B-cell purification (CD19-expressing cells) invariably depleted or eliminated viral DNA. Despite the frequent finding of significant quantities of adenovirus DNA in tonsil and adenoid tissues, infectious virus was rarely present, as measured by coculture with permissive cells. These findings suggest that human mucosal T lymphocytes may harbor species C adenoviruses in a quiescent, perhaps latent form.

* Corresponding author. Mailing address: Department of Microbiology and Immunology Emory University, 1510 Clifton Rd., 3107 Rollins Research Center, Atlanta, GA 30322. Phone: (404) 727-5948. Fax: (404) 727-0293. E-mail: gooding{at}microbio.emory.edu.

{dagger} Present address: Department of Medical Microbiology, University of Manitoba, Winnipeg, Manitoba R3E 0W3, Canada.

Journal of Virology, November 2002, p. 10608-10616, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.10608-10616.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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