Pathobiology 1992:60:195-205
Molecular Virology Laboratory St. Luke's/Roosevelt Hospital Center and College of Physicians & Surgeons, Columbia University, New York, N.Y., USA
Regulated Expression of Human Immunodeficiency Virus Type 1 in Human Glial Cells: Induction of Dormant Virus
Key Words
Abstract
HIV-1 Dormant virus Virus expression Neural cells Human
Human neural cells are susceptible to infection with human immunodefi ciency virus type 1 (HIV-1) in vitro; however, virus replication in these cells is strongly restricted. To understand the mechanism of this restriction, we exam ined the regulation of HIV-1 expression in glial cell cultures expressing high levels of HIV-1 after transfection of infectious viral DNA and selection. In all cases, high HIV-1 expression declined to low basal levels within 4-8 weeks of cultivation. The decrease in HIV-1 protein production was paralleled by the decline in the relative levels of the 9.2-, 4.3- and 1.8-kilobase HIV-1 trans cripts, but not by significant loss of HIV-1 DNA. Analysis of one long-term cell culture revealed 5 full-length unrearranged HIV-1 DNA copies per cell, but no viral transcripts on Northern blots, and minimal production of infec tious virus. HIV-1 replication in these cells was markedly augmented by treat ment with sodium butyrate (Na But) and to a lesser extent by 5-azacytidine, dibutyryl AMP and human herpes virus type 6. The virus induced by Na But was infectious. Transient expression assays revealed that Na But was more effective than phorbol myristate acetate in increasing the HIV-1 promoter activity in glial cells. Thus, one phase where glial cells can limit HIV infection is the expression of viral RNA from stable HIV provirus. However, such pro virus remains responsive to inductive signals and may be activated to produce infectious HIV.
Introduction
er-inducer subset, monocyte/macrophages, B lympho cytes. fibroblasts, glial and neuronal cells, and bone mar row myeloid progenitor cells. Undoubtedly, the complex pathogenic outcomes of HIV-1 infection correlate with the ability of the virus to infect, injure and persist in the variety of cells and tissues [reviewed in ref. 1 and 2],
Human immunodeficiency virus type 1 (HIV-1) is the etiologic agent of acquired immune deficiency syndrome (AIDS). The virus has a remarkably wide host cell range in vitro and in vivo, including T lymphocytes of the help
Received: June 15. 1991 Accepted: July 29. 1991
Dr. David J. Volsky, Molecular Virology Laboratory St. Luke's/Roosevelt Hospital Center anc College of Physicians & Surgeons. Columbia University 432 West 58th Street. New York. NY 10019 (USA)
Downloaded by: King's College London 137.73.144.138 - 1/12/2019 9:33:05 PM
Muhammad Shahabuddin Barbara Volsky Helen Kim Koji Sakai David J. Volsky
Table 1. HIV-I expression in persistently infected glial cell lines
Cell line
251-MG/Ea, 251-MG/Ea2 251-MG/Eaj 251-MG/Aa 251-MG/Ab 251/Ea 148/Aa 148/Ab 148/Ea 148/Eh U l.l ACH-2 CR10/N1T
Cell origin
Reference3
Virus/DNAb
Characteristics of HIV-1 expression
glia' glia' glia' glia' glia' glia' neuroglia neuroglia neuroglia neuroglia monocyte T cell T cell
10,2 10.2 10.2 2, TW 2, TW 2. TW 2. TW 2. TW 2. TW 2, TW 4 3 23
pNIT-E + pSV2m> pNIT-E + pSV2neo pNIT-E + pSV2wo pNIT-Aww pNIT-Art«? pN 1T-E neo pN IT-A w o pNIT-A/i«> pNIT-E««) pNIT-En«> HIV-1/III b HIV-1/III b HIV-1/N1T
HIV-1 p24core % of viral protein in culture antigen positive cells1-' supernatant, pg/mld 30 5-10 < 1.0 5 < 0.1 < 0.1 60 < 0.1 40 < 0.1 < 1.0 < 1.0 90
4.005 ±455 465 ±79 54 ±15 344 ±34 < 10 < 10 2.6X 104±0.5X 10J < 10 9,626 ±1.200
Molecular Virology Laboratory St. Luke's/Roosevelt Hospital Center and College of Physicians & Surgeons, Columbia University, New York, N.Y., USA
Regulated Expression of Human Immunodeficiency Virus Type 1 in Human Glial Cells: Induction of Dormant Virus
Key Words
Abstract
HIV-1 Dormant virus Virus expression Neural cells Human
Human neural cells are susceptible to infection with human immunodefi ciency virus type 1 (HIV-1) in vitro; however, virus replication in these cells is strongly restricted. To understand the mechanism of this restriction, we exam ined the regulation of HIV-1 expression in glial cell cultures expressing high levels of HIV-1 after transfection of infectious viral DNA and selection. In all cases, high HIV-1 expression declined to low basal levels within 4-8 weeks of cultivation. The decrease in HIV-1 protein production was paralleled by the decline in the relative levels of the 9.2-, 4.3- and 1.8-kilobase HIV-1 trans cripts, but not by significant loss of HIV-1 DNA. Analysis of one long-term cell culture revealed 5 full-length unrearranged HIV-1 DNA copies per cell, but no viral transcripts on Northern blots, and minimal production of infec tious virus. HIV-1 replication in these cells was markedly augmented by treat ment with sodium butyrate (Na But) and to a lesser extent by 5-azacytidine, dibutyryl AMP and human herpes virus type 6. The virus induced by Na But was infectious. Transient expression assays revealed that Na But was more effective than phorbol myristate acetate in increasing the HIV-1 promoter activity in glial cells. Thus, one phase where glial cells can limit HIV infection is the expression of viral RNA from stable HIV provirus. However, such pro virus remains responsive to inductive signals and may be activated to produce infectious HIV.
Introduction
er-inducer subset, monocyte/macrophages, B lympho cytes. fibroblasts, glial and neuronal cells, and bone mar row myeloid progenitor cells. Undoubtedly, the complex pathogenic outcomes of HIV-1 infection correlate with the ability of the virus to infect, injure and persist in the variety of cells and tissues [reviewed in ref. 1 and 2],
Human immunodeficiency virus type 1 (HIV-1) is the etiologic agent of acquired immune deficiency syndrome (AIDS). The virus has a remarkably wide host cell range in vitro and in vivo, including T lymphocytes of the help
Received: June 15. 1991 Accepted: July 29. 1991
Dr. David J. Volsky, Molecular Virology Laboratory St. Luke's/Roosevelt Hospital Center anc College of Physicians & Surgeons. Columbia University 432 West 58th Street. New York. NY 10019 (USA)
Downloaded by: King's College London 137.73.144.138 - 1/12/2019 9:33:05 PM
Muhammad Shahabuddin Barbara Volsky Helen Kim Koji Sakai David J. Volsky
Table 1. HIV-I expression in persistently infected glial cell lines
Cell line
251-MG/Ea, 251-MG/Ea2 251-MG/Eaj 251-MG/Aa 251-MG/Ab 251/Ea 148/Aa 148/Ab 148/Ea 148/Eh U l.l ACH-2 CR10/N1T
Cell origin
Reference3
Virus/DNAb
Characteristics of HIV-1 expression
glia' glia' glia' glia' glia' glia' neuroglia neuroglia neuroglia neuroglia monocyte T cell T cell
10,2 10.2 10.2 2, TW 2, TW 2. TW 2. TW 2. TW 2. TW 2, TW 4 3 23
pNIT-E + pSV2m> pNIT-E + pSV2neo pNIT-E + pSV2wo pNIT-Aww pNIT-Art«? pN 1T-E neo pN IT-A w o pNIT-A/i«> pNIT-E««) pNIT-En«> HIV-1/III b HIV-1/III b HIV-1/N1T
HIV-1 p24core % of viral protein in culture antigen positive cells1-' supernatant, pg/mld 30 5-10 < 1.0 5 < 0.1 < 0.1 60 < 0.1 40 < 0.1 < 1.0 < 1.0 90
4.005 ±455 465 ±79 54 ±15 344 ±34 < 10 < 10 2.6X 104±0.5X 10J < 10 9,626 ±1.200
