Quantitation of simian cytokine and beta-chemokine mRNAs, using real-time reverse transcriptase-polymerase chain reaction: variations in expression during chronic primate lentivirus infection

AIDS Res Hum Retroviruses. 2002 Jun 10;18(9):627-39. doi: 10.1089/088922202760019329.

Abstract

Cytokines and beta-chemokines are important mediators of the immune system and are expressed in many infectious diseases. To study cytokine and beta-chemokine profiles during pathogenesis of lentiviral infection and progression to AIDS in rhesus macaques, we established new quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assays based on TaqMan chemistry. Using synthetic RNA standards, we quantified mRNAs of IL-2, IL-4, IL-6, IL-10, IL-12 p40, interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), RANTES, macrophage inflammatory protein 1 alpha (MIP-1 alpha), and MIP-1 beta in unstimulated peripheral blood mononuclear cells (PBMCs) and lymph nodes from macaques chronically infected with SIV or SHIV. Viremic monkeys with decreased CD4(+) T cell counts (<500 cells/microl) had significantly higher IL-10 mRNA expression than uninfected controls, which parallels the findings in HIV-1-infected humans. In addition, MIP-1 alpha, MIP-1 beta, and RANTES mRNA expression increased in viremic monkeys with decreased CD4(+) T cell counts; gene expression was inversely correlated with CD4(+) T cell counts, but not viral load. The newly established quantitative real-time RT-PCR assays will allow the determination of cytokine and beta-chemokine patterns in rhesus macaques in studies of microbial pathogenesis or vaccine development.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • CD4 Lymphocyte Count
  • Chemokine CCL3
  • Chemokine CCL4
  • Chemokines, CC / analysis*
  • Chemokines, CC / genetics
  • Cytokines / analysis*
  • Cytokines / genetics
  • Disease Progression
  • Gene Expression
  • Interleukin-10 / analysis
  • Interleukin-10 / genetics
  • Lentiviruses, Primate*
  • Leukocytes, Mononuclear / metabolism
  • Lymph Nodes / cytology
  • Lymph Nodes / metabolism
  • Macaca mulatta
  • Macrophage Inflammatory Proteins / genetics
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Simian Acquired Immunodeficiency Syndrome / immunology
  • Simian Acquired Immunodeficiency Syndrome / metabolism*
  • Viral Load

Substances

  • Chemokine CCL3
  • Chemokine CCL4
  • Chemokines, CC
  • Cytokines
  • Macrophage Inflammatory Proteins
  • RNA, Messenger
  • Interleukin-10