A monocyte-derived factor interferes with detection of reverse transcriptase in HIV-1 infection

D. P. Recker, H. Kulaga, D. Dorsett, T. Folks, T. J. Kindt

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Culture supernatants from the rabbit macrophage cell line 6083 infected with a retrovirus, human immunodeficiency virus type 1 (HIV-1), were negative for reverse transcriptase (RT) expression although the line was shown to be productively infected by all other criteria tested. Supernatants from uninfected cultures of 6083, the human monocyte line U937, and from freshly isolated peripheral human monocytes, were found to contain a monocyte-derived inhibitory factor (MDIF) which interferes with a standard assay for RT. MDIF is a heat-labile activity of approximately of 40 kD. Both substrates and products of the reverse transcriptase assay are degraded by MDIF which is not affected by reduction and alkylation of disulfide bonds. MDIF is inhibited by the addition of a particular thioated oligonucleotide (S-dG30) to the reaction mixture but this addition also inhibits RT. The optimum method to minimize MDIF interference in the RT assay is by addition of ethylene glycol bis-(β-aminoethyl ether)N,N,N',N'-tetraacetic acid (EGTA); MDIF requires divalent cations for activity and has a strong preference for calcium which is preferentially chelated by EGTA. The potential presence of this inhibitory activity should be considered when using RT levels as a measure of retroviral infection.

Original languageEnglish (US)
Pages (from-to)73-81
Number of pages9
JournalAIDS Research and Human Retroviruses
Volume7
Issue number1
StatePublished - 1991
Externally publishedYes

Fingerprint

Virus Diseases
HIV-1
Monocytes
RNA-Directed DNA Polymerase
Egtazic Acid
Ethylene Glycol
Human immunodeficiency virus 1 reverse transcriptase
Divalent Cations
Alkylation
Retroviridae
Oligonucleotides
Disulfides
Ether
Hot Temperature
Macrophages
Rabbits
Calcium
Cell Line
Acids
Infection

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Recker, D. P., Kulaga, H., Dorsett, D., Folks, T., & Kindt, T. J. (1991). A monocyte-derived factor interferes with detection of reverse transcriptase in HIV-1 infection. AIDS Research and Human Retroviruses, 7(1), 73-81.

A monocyte-derived factor interferes with detection of reverse transcriptase in HIV-1 infection. / Recker, D. P.; Kulaga, H.; Dorsett, D.; Folks, T.; Kindt, T. J.

In: AIDS Research and Human Retroviruses, Vol. 7, No. 1, 1991, p. 73-81.

Research output: Contribution to journalArticle

Recker, DP, Kulaga, H, Dorsett, D, Folks, T & Kindt, TJ 1991, 'A monocyte-derived factor interferes with detection of reverse transcriptase in HIV-1 infection', AIDS Research and Human Retroviruses, vol. 7, no. 1, pp. 73-81.
Recker, D. P. ; Kulaga, H. ; Dorsett, D. ; Folks, T. ; Kindt, T. J. / A monocyte-derived factor interferes with detection of reverse transcriptase in HIV-1 infection. In: AIDS Research and Human Retroviruses. 1991 ; Vol. 7, No. 1. pp. 73-81.
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