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A macaque adherent cell line that expresses human CD4 is susceptible to SIV: utility for assessing neutralizing

S Goldstein1, B Hague, D Montefiori

  • 1Immunodeficiency Viruses Section, LID, NIAID, NIH Twinbrook Facility, Rockville, MD 20852, USA.

Journal of Virological Methods
|May 1, 1995
PubMed

Insights

Researchers developed a new cell line, CMMT/CD4, for studying simian immunodeficiency virus (SIV). This cell line enables a sensitive microassay to detect neutralizing antibodies in animal plasma, aiding SIV research.

Area of Science:

  • Virology
  • Immunology
  • Cell Biology

Background:

  • Simian immunodeficiency virus (SIV) poses a significant challenge in non-human primate research.
  • Developing robust cell lines susceptible to SIV infection is crucial for studying viral pathogenesis and immune responses.
  • Existing methods for detecting neutralizing antibodies against SIV can be labor-intensive or lack sensitivity.

Purpose of the Study:

  • To generate a stable macaque CD4+ cell line for SIV infection studies.
  • To develop a sensitive microassay for quantifying SIV-specific neutralizing antibodies in plasma.
  • To compare the sensitivity of the new microassay with established methods.

Main Methods:

  • Stable expression of the human CD4 gene in a rhesus macaque mammary tumor cell line (CMMT) to create CMMT/CD4.
  • Infection of CMMT/CD4 cells with various SIV isolates to assess susceptibility.
  • Development of an immunoperoxidase-based microassay to detect single infected cells.
  • Quantification of neutralizing activity by measuring the reduction in infected cells in response to plasma dilutions.

Main Results:

  • The CMMT/CD4 cell line expressed surface CD4 and was susceptible to a broad range of SIV isolates but not HIV-1.
  • The developed microassay could detect single infected cells in CMMT/CD4 monolayers.
  • A 90% reduction in positive cells indicated neutralizing activity in two-fold plasma dilutions.
  • The microassay demonstrated comparable sensitivity to methods measuring reverse transcriptase activity, viral antigen reduction, or cytopathic effect inhibition.

Conclusions:

  • The CMMT/CD4 cell line provides a valuable tool for SIV research, particularly for studying viral entry and infectivity.
  • The developed microassay offers a sensitive and efficient method for measuring SIV-neutralizing antibodies in plasma.
  • This assay facilitates the evaluation of vaccine efficacy and the characterization of immune responses in SIV-infected or immunized animals.

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