A novel in vitro system to generate and study latently HIV-infected long-lived normal CD4+ T-lymphocytes.
Studies of mechanisms of HIV-latency and its reactivation in long-lived resting CD4+ T-lymphocytes in patients have been limited due to the very low frequency of these cells ( approximately 1-10 cells per 10(6) CD4+ T-cells). To circumvent this obstacle, an in vitro culture system for post-activation long-term survival of normal CD4+ T-cells in a quiescent (non-cycling) state was developed and used to generate latently infected, long-lived quiescent CD4+ T-cells from HIV-infected, activated normal CD4+ T-lymphocytes. This yielded a frequency of approximately 5x10(4) latently infected cells per 10(6) cells in culture, which is approximately 10(3)- to 10(4)-fold higher than that available from patients. Moreover, 5-10% of long-term surviving non-cycling T-cells were found to make infectious HIV continuously at low levels, showing that HIV production from infected T-cells does not require full cellular activation. This model system should facilitate studies of long-lived, latently infected and persistently HIV-producing quiescent normal CD4+ T-lymphocytes.
Sahu
Gautam K
GK
Department of Microbiology and Immunology, The University of Texas Medical Branch, 301 University Blvd, BSB #3.132, Galveston, TX 77555, USA.
Lee
Kyeongeun
K
Ji
Jiaxiang
J
Braciale
Vivian
V
Baron
Samuel
S
Cloyd
Miles W
MW
D015703:Antigens, CD / Q000032:analysis; D000956:Antigens, Viral / Q000032:analysis; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology*; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D005434:Flow Cytometry; D006678:HIV / Q000502:physiology*; D006801:Humans; D007156:Immunologic Memory; D008213:Lymphocyte Activation; D016131:Lymphocyte Subsets; D011948:Receptors, Antigen, T-Cell / Q000032:analysis; D017735:Virus Latency; D014779:Virus Replication
D015703:Antigens, CD; D000956:Antigens, Viral; D011948:Receptors, Antigen, T-Cell