File Download
There are no files associated with this item.
Supplementary
-
Citations:
- Appears in Collections:
Conference Paper: Human cytomegalovirus latent infection modulates HIV restriction factors in CD34+ hematopoietic stem cells
Title | Human cytomegalovirus latent infection modulates HIV restriction factors in CD34+ hematopoietic stem cells |
---|---|
Authors | |
Issue Date | 2016 |
Citation | The 21st International AIDS Conference (AIDS 2016), Durban, South Africa, 18-22 July 2016. How to Cite? |
Abstract | BACKGROUND: Individuals pre-exposed to human cytomegalovirus (HCMV) are more prone to HIV/AIDS disease progression but the reasons remain elusive. HCMV is a ubiquitous DNA virus that establishes natural lifelong latent infection in CD34+ progenitor cells, where latency-specific viral genes modulate the host cell environment. Myeloid-lineage cells are known to resist replicative HIV-1 infection but recent studies provided evidence that latent reservoir may be established in CD34+ myeloid progenitors. Therefore, an intricate relationship between HCMV and HIV-1 may occur in these cells. METHODS: CD34+ cells isolated from healthy PBMCs were cultured in specialized media for ~30 days to allow expansion. Phenotype of CD34-subsets were assessed by flow cytometry before infection by HCMV. Establishment of HCMV latency and modulation of cellular responses were assessed using real-time PCR and plaque assay. X4/R5-HIV-1 was used to infect CD34+ cells harboring latent HCMV and successful HIV-1 infection was examined for proviral DNA by digital PCR. RESULTS: Peripheral blood-derived (PB-)CD34+ cells expanded up to 50-fold after 30 days in culture and retained early progenitor phenotype. The success of the establishment of HCMV latency was assessed by the detection of latency-specific transcripts UL111.5A, LUNA and UL138, lack of infectious virions, and the ability to reactivate upon co-culture with permissive cells. Importantly, latent HCMV infection downregulated the HIV-1 restriction factors SAMHD1, APOBEC3A/G, tetherin and Mx2 in PB-CD34+ cells, which led to higher HIV-1 transcriptional activity and proviral DNA. Interestingly, these cells retained HCMV-induced downmodulation of MHC class II molecules and did not show differentiation tendency. Furthermore, we demonstrated that the expression of CXCR4 and CCR5 on CD34+ cells were not altered by HCMV latent infection, but HIV-1 infection can be inhibited by antiretrovirals. Lastly, using a dual-reporter pseudovirus, infection of latent HCMV PB-CD34+ cells resulted in increased active infection compared to mock cells. CONCLUSIONS: This study describes an in vitro CD34+ cell expansion culture model to study the establishment of HCMV and HIV-1 infections. Furthermore, we demonstrated that interplay exists between latent HCMV and HIV-1 infection in CD34+ progenitor cells. These findings provide insights into the importance of pre-existing latent HCMV that affects HIV-1 infection outcome. |
Description | Poster Exhibition: - Viral mechanisms of HIV/SIV persistence and latency: no. WEPEA016 |
Persistent Identifier | http://hdl.handle.net/10722/232502 |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Cheung, KLA | - |
dc.date.accessioned | 2016-09-20T05:30:28Z | - |
dc.date.available | 2016-09-20T05:30:28Z | - |
dc.date.issued | 2016 | - |
dc.identifier.citation | The 21st International AIDS Conference (AIDS 2016), Durban, South Africa, 18-22 July 2016. | - |
dc.identifier.uri | http://hdl.handle.net/10722/232502 | - |
dc.description | Poster Exhibition: - Viral mechanisms of HIV/SIV persistence and latency: no. WEPEA016 | - |
dc.description.abstract | BACKGROUND: Individuals pre-exposed to human cytomegalovirus (HCMV) are more prone to HIV/AIDS disease progression but the reasons remain elusive. HCMV is a ubiquitous DNA virus that establishes natural lifelong latent infection in CD34+ progenitor cells, where latency-specific viral genes modulate the host cell environment. Myeloid-lineage cells are known to resist replicative HIV-1 infection but recent studies provided evidence that latent reservoir may be established in CD34+ myeloid progenitors. Therefore, an intricate relationship between HCMV and HIV-1 may occur in these cells. METHODS: CD34+ cells isolated from healthy PBMCs were cultured in specialized media for ~30 days to allow expansion. Phenotype of CD34-subsets were assessed by flow cytometry before infection by HCMV. Establishment of HCMV latency and modulation of cellular responses were assessed using real-time PCR and plaque assay. X4/R5-HIV-1 was used to infect CD34+ cells harboring latent HCMV and successful HIV-1 infection was examined for proviral DNA by digital PCR. RESULTS: Peripheral blood-derived (PB-)CD34+ cells expanded up to 50-fold after 30 days in culture and retained early progenitor phenotype. The success of the establishment of HCMV latency was assessed by the detection of latency-specific transcripts UL111.5A, LUNA and UL138, lack of infectious virions, and the ability to reactivate upon co-culture with permissive cells. Importantly, latent HCMV infection downregulated the HIV-1 restriction factors SAMHD1, APOBEC3A/G, tetherin and Mx2 in PB-CD34+ cells, which led to higher HIV-1 transcriptional activity and proviral DNA. Interestingly, these cells retained HCMV-induced downmodulation of MHC class II molecules and did not show differentiation tendency. Furthermore, we demonstrated that the expression of CXCR4 and CCR5 on CD34+ cells were not altered by HCMV latent infection, but HIV-1 infection can be inhibited by antiretrovirals. Lastly, using a dual-reporter pseudovirus, infection of latent HCMV PB-CD34+ cells resulted in increased active infection compared to mock cells. CONCLUSIONS: This study describes an in vitro CD34+ cell expansion culture model to study the establishment of HCMV and HIV-1 infections. Furthermore, we demonstrated that interplay exists between latent HCMV and HIV-1 infection in CD34+ progenitor cells. These findings provide insights into the importance of pre-existing latent HCMV that affects HIV-1 infection outcome. | - |
dc.language | eng | - |
dc.relation.ispartof | International AIDS Conference, AIDS 2016 | - |
dc.title | Human cytomegalovirus latent infection modulates HIV restriction factors in CD34+ hematopoietic stem cells | - |
dc.type | Conference_Paper | - |
dc.identifier.email | Cheung, KLA: allenc@hku.hk | - |
dc.identifier.authority | Chen, Z=rp00243 | - |
dc.identifier.hkuros | 264773 | - |