Quantitation by Competitive PCR of HIV‐1 Proviral DNA in Epidermal Langerhans Cells of HIV‐Infected Patients
作者:
Andrea Cimarelli,
Giovanna Zambruno,
Alessandra Marconi,
Giampiero Girolomoni,
Umberto Bertazzoni,
Alberto Giannetti,
期刊:
Journal of Acquired Immune Deficiency Syndromes
(OVID Available online 1994)
卷期:
Volume 7,
issue 3
页码: 230-235
ISSN:0894-9255
年代: 1994
出版商: OVID
关键词: Langerhans cells;Human immunodeficiency virus Type 1 (HIV - 1) Proviral DNA;Competitive polymerase chain reaction (PCR).
数据来源: OVID
摘要:
SummaryLangerhans cells (LC) belong to the dendritic cell family and represent the principal antigen presenting cells populating squamous epithelia. We have reported the presence of human immunodeficiency virus Type 1 (HIV-1) proviral DNA and RNA in purified LC from the epidermis of seropositive patients. The aim of this study was to quantify HIV-1 proviral DNA in LC of infected patients using a competitive polymerase chain reaction (PCR) assay. Bulk epidermal cell (EC) suspensions were obtained from the skin of nine AIDS patients and six seronegative subjects. Purified LC and LC-depleted EC were prepared by immunomagnetic separation using an anti-CD la monoclonal antibody. LC preparations did not contain T cells, as assessed by reverse transcription PCR analysis of the T cell receptor β-chain gene (C region). In addition, no CD 14+cells could be detected in LC fractions by immunostaining of cytospin preparations. To quantify HIV-1 DNA, a new competitive PCR system was devised using SK145/150 as primers (gag) and a competitor plasmid DNA with a modified sequence (209 instead of 142 bp). The number of HIV-1 DNA copies found in the LC of AIDS patients ranged from 107 to 3,645/105LC. In contrast, LC-depleted EC from the same subjects were all negative. The results indicate that in AIDS patients the frequency of infected LC is comparable to that reported for peripheral blood CD4+T cells.
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