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Human immunodeficiency virus type-1 can be detected in monocytes by polymerase chain reaction
E Quirós1, F García, M C Maroto
1Department of Microbiology, Faculty of Medicine, University of Granada, Spain.
Insights
Detecting human immunodeficiency virus type-1 (HIV-1) DNA in monocytes and lymphocytes revealed distinct amplification patterns. Monocytes showed higher env gene detection, suggesting potential infection by a unique viral genetic variant.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Human immunodeficiency virus type-1 (HIV-1) infection impacts immune cells, including lymphocytes and monocytes.
- Understanding viral DNA distribution in different cell types is crucial for disease staging and management.
Purpose of the Study:
- To investigate the presence and amplification patterns of HIV-1 DNA in lymphocytes and monocytes from patients across different disease stages.
- To explore potential differences in viral genetic material within these distinct immune cell populations.
Main Methods:
- Polymerase chain reaction (PCR) was used to amplify HIV-1 DNA from lysed lymphocytes and monocytes using primer pairs targeting gag, env, and LTR regions.
- Amplified DNA was detected using solution hybridization with radiolabeled probes and PAGE-autoradiography.
Main Results:
- HIV-1 DNA was detected in both lymphocytes and monocytes across asymptomatic, AIDS-related complex (ARC), and acquired immunodeficiency syndrome (AIDS) patient groups.
- Monocytes showed consistent amplification of the env gene (100% in asymptomatic and AIDS patients), while LTR amplification failed in this cell type.
- Lymphocytes demonstrated high amplification rates for gag and env genes across all patient groups, with varying LTR amplification.
Conclusions:
- The presence of HIV-1 DNA was confirmed in monocytes, with the env gene being the most effectively amplified target in this cell subset.
- The distinct amplification pattern observed in monocytes, particularly the failure of LTR amplification, suggests these cells might harbor a genetically distinct variant of HIV-1.
Abstract:
Lymphocytes and monocytes from 25 patients infected with human immunodeficiency virus type-1 (HIV-1)--13 asymptomatic, seven with the AIDS-related complex (ARC) and five with the acquired immunodeficiency syndrome (AIDS)--were lysed and subjected to PCR with three primer pairs: SK38/SK39 (gag), SK68/SK69 (env) and SK29/SK30 (LTR). Amplified DNA was solution-hybridised with 32P-labelled probes (SK19, SK70 and SK31, respectively) and detected by PAGE-autoradiography. HIV-1 DNA was detected as follows. Asymptomatic patients: monocytes--gag 61.5%, env 100%, LTR 0%; lymphocytes--gag 100%, env 92.3%, LTR 53.84%. ARC patients: monocytes--gag 71.4%, env 57.1%, LTR 0%; lymphocytes--gag 100%, env 71.4%, LTR 71.4%. AIDS patients: monocytes--gag 80.0%, env 100%, LTR 0%; lymphocytes--gag 100%, env 60%, LTR 60%. The presence of HIV-1 DNA was confirmed in the monocyte fraction. In this cell subset, the env gene-directed primers were the most effective for amplification, whereas the LTR gene-directed primers failed to amplify HIV-1 DNA. The different pattern of amplification found in monocytes may suggest that these cells could be infected by a genetic variant of the virus.