Abstract:
:In HIV+ patients, the presence of HIV-RNA in plasma and circulating cells has been reported to be a marker of progression but the percentage of transcriptionally active infected cells remains unclear. We have developed a reliable fluorescent in situ hybridization method for the detection of HIV specific RNA by flow cytometry. The procedure was applied to a panel of chronically infected cell lines and to an acutely infected cell line mimicking normal peripheral blood lymphocytes in susceptibility to HIV-1. The cells were fixed in suspension and hybridized by means of an HIV-1 genomic probe labeled with digoxigenin-11-dUTP. An FITC-labeled anti-digoxigenin antiserum was then applied and the resulting fluorescence signals were analyzed both by flow cytometry and confocal microscopy. Different procedures for double staining HIV-RNA together with virus induced proteins or surface markers were also developed. Flow cytometric detection of in situ hybridization offers the possibility of analyzing thousands of cells in a few seconds and of collecting multiparametric information at the single cell level, thus providing a potential tool for detecting the rare HIV-RNA expressing cells in peripheral blood samples.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Borzì RM,Piacentini A,Monaco MC,Lisignoli G,Degrassi A,Cattini L,Santi S,Facchini Adoi
10.1016/0022-1759(96)00070-1subject
Has Abstractpub_date
1996-06-21 00:00:00pages
167-76issue
2eissn
0022-1759issn
1872-7905pii
0022175996000701journal_volume
193pub_type
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