Abstract
Background:Identifying biomarkers for cells harboring replication-competent HIV is a major research priority. Recently, there have been mixed reports addressing the possibility that CD32-expressing T cells are enriched for HIV. There is growing evidence that CD32 expression increases with cellular activation that may be related to, but not necessarily specific for, infection with HIV. However, the relationship of CD32 expression to HIV-infection in subtypes of tissue-resident leukocytes is unclear.Methods:First, we used duplex chromogenic in situ hybridization to identify cells actively transcribing RNA for both CD32 and HIV on human gut tissues. Then we performed multiplexed immunofluorescence and in situ hybridization (mIFISH) on sections from the same tissues to determine the phenotype of individual cells co-expressing HIV-RNA and CD32-RNA.Results:HIV-RNA+ cells were more abundant in tissues from viremic individuals than in those receiving suppressive anti-retroviral therapy (ART). However, staining by both methods indicated that a higher proportion of HIV-RNA+ cells co-expressed CD32-RNA in ART-suppressed individuals than in those with viremia. The majority of HIV-RNA+ cells were CD3+.Conclusions:Our data suggest that the transcription of CD32-RNA is correlated with HIV transcriptional activity in CD3+ cells found within human gut tissue. Whether or not up-regulation of CD32-RNA is a direct result of HIV transcription or more global T-cell activation remains unclear.
Highlights
Whereas cell populations harboring HIV in the peripheral blood have been studied extensively [1], tissue reservoirs are less well described and are thought to exist within tissue resident lymphocyte subsets that may not be found in circulating blood [2, 3]
Recent observations suggest that the expression of CD32 on T cells may be associated with activation that is not specific to HIV infection [6]
To explore these issues further we sought to assess the transcriptional activity of CD32-RNA and HIV-RNA in gut tissue from viremic and antiretroviral therapy (ART)-suppressed HIV-infected individuals using RNAscope in situ hybridization (ISH) or multiplexed immunofluorescence with ISH [13]
Summary
Whereas cell populations harboring HIV in the peripheral blood have been studied extensively [1], tissue reservoirs are less well described and are thought to exist within tissue resident lymphocyte subsets that may not be found in circulating blood [2, 3]. Recent observations suggest that the expression of CD32 on T cells may be associated with activation that is not specific to HIV infection [6]. Since CD32 is most frequently associated with myeloid cells, the questions remain regarding the phenotype of CD32+ HIV-infected cells and the extent to which CD32 co-localizes with HIV-infected cells in other tissues. To explore these issues further we sought to assess the transcriptional activity of CD32-RNA and HIV-RNA in gut tissue from viremic and ART-suppressed HIV-infected individuals using RNAscope in situ hybridization (ISH) or multiplexed immunofluorescence with ISH (mIFISH) [13]. The relationship of CD32 expression to HIV-infection in subtypes of tissue-resident leukocytes is unclear
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