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Establishment of an MT4 cell line persistently producing infective HIV-1 particles
1Microbiological Research Group, B. Johan National Institute of Hygiene, Budapest, Hungary.
Abstract:
Human T cell Lymphotropic Virus-I (HTLV-I) carrying human T cell line MT4 is highly sensitive to Human Immunodeficiency Virus-1 (HIV-1). After HIV-1 infection cell clusters characteristic of intact MT4 rapidly disintegrate, syncytia appear and the cells die. Surviving MT4 cells were subcultured following HIV-1 infection of high multiplicity. We succeeded to establish an MT4 cell line continuously producing infective HIV (MT4/HIV-1). The original and the HIV-1 infected MT4 cells were morphologically similar. The MT4/HIV-1 cells proved to be nearly 100% positive in indirect immunofluorescence assay using the serum of an HIV-1 antibody positive individual. OKT4 surface antigen could not be demonstrated on MT4/HIV-1 cells. On electron microscopic pictures typical and atypical virus particles could be seen near the surface of the cell membrane. The persistently produced virus particles were infective for H9 and MT4 cells. The antigenic structure of the virus produced by MT4 cells was similar to that produced by H9 cells.
Insights
Researchers developed a new cell line, MT4/HIV-1, that continuously produces infectious Human Immunodeficiency Virus-1 (HIV-1). This discovery aids in studying HIV-1 replication and its effects on Human T cell Lymphotropic Virus-I (HTLV-I) infected cells.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Human T cell Lymphotropic Virus-I (HTLV-I) infected MT4 cells are highly susceptible to Human Immunodeficiency Virus-1 (HIV-1).
- HIV-1 infection leads to MT4 cell disintegration, syncytia formation, and cell death.
Purpose of the Study:
- To establish a stable cell line of MT4 cells that continuously produce infectious HIV-1.
- To characterize the properties of this novel HIV-1 producing cell line.
Main Methods:
- Subculturing surviving MT4 cells after high multiplicity HIV-1 infection.
- Morphological analysis using light and electron microscopy.
- Indirect immunofluorescence assay for HIV-1 antibodies and OKT4 surface antigen detection.
Main Results:
- Successfully established the MT4/HIV-1 cell line, continuously producing infectious HIV-1.
- MT4/HIV-1 cells showed morphological similarity to original MT4 cells but lacked OKT4 surface antigen.
- Immunofluorescence confirmed nearly 100% positivity for HIV-1 antibodies.
- Electron microscopy revealed typical and atypical virus particles.
- The produced virus was infective for H9 and MT4 cells, with similar antigenic structure to HIV-1 from H9 cells.
Conclusions:
- The MT4/HIV-1 cell line is a valuable tool for studying HIV-1 replication and pathogenesis.
- The loss of OKT4 antigen suggests alterations in cell surface markers upon persistent HIV-1 infection.
- The study demonstrates the production of infective HIV-1 from an HTLV-I carrying cell line.