Rickettsia rickettsii infection of cultured human endothelial cells induces NF-κB activation

Lee Ann Sporn, Sanjeev Sahni, Norma B. Lerner, Victor J. Marder, David J. Silverman, Loel C. Turpin, Amy L. Schwab

Research output: Contribution to journalArticle

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Abstract

Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever, is an obligate intracellular bacterial organism that infects primarily the vascular endothelial cells (EC). A component of the EC response to infection is transcriptional activation, which may contribute to the thrombotic and inflammatory consequences of disease. In this study, we explore R. rickettsii-induced activation of the nuclear factor-κB/Rel (NF- κB) family of transcription factors involved in early transcriptional responses to injurious stimuli. Two NF-κB species were activated by infection and reacted with a double-stranded oligonucleotide probe corresponding to the κB binding domain of the murine kappa light-chain gene enhancer. Gel supershift analysis demonstrated the reactivity of these complexes with antibodies against p65 and p50, and the induced species were tentatively identified as p50-p50 homodimers and p50-p65 heterodimers. Semiquantitative reverse transcription-PCR analysis revealed dramatic increases in the steady-state levels of mRNA coding for the inhibitory subunit of NF-κB (IκBα), transcription of which is enhanced by the binding of NF-κB within the IκBα promoter region. NF-κB activation was first detected 1.5 h following infection and was biphasic, with an early peak of activation at approximately 3 h, a return to baseline levels at 14 h, and even higher levels of activation at 24 h. It is likely that NF-κB activation requires cellular uptake of R. rickettsii, since treatment of EC with cytochalasin B during infection to block entry inhibited activation by only 70% at 3 h. R. rickettsii-induced activation of NF-κB may be an important controlling factor in the transcriptional responses of EC to infection with this obligate intracellular organism.

Original languageEnglish (US)
Pages (from-to)2786-2791
Number of pages6
JournalInfection and Immunity
Volume65
Issue number7
StatePublished - 1997
Externally publishedYes

Fingerprint

Rickettsia Infections
Rickettsia rickettsii
Endothelial Cells
Infection
Rocky Mountain Spotted Fever
Cytochalasin B
Oligonucleotide Probes
Genetic Promoter Regions
Transcriptional Activation
Reverse Transcription
Transcription Factors
Gels
Light
Polymerase Chain Reaction
Messenger RNA
Antibodies
Genes

ASJC Scopus subject areas

  • Immunology

Cite this

Sporn, L. A., Sahni, S., Lerner, N. B., Marder, V. J., Silverman, D. J., Turpin, L. C., & Schwab, A. L. (1997). Rickettsia rickettsii infection of cultured human endothelial cells induces NF-κB activation. Infection and Immunity, 65(7), 2786-2791.

Rickettsia rickettsii infection of cultured human endothelial cells induces NF-κB activation. / Sporn, Lee Ann; Sahni, Sanjeev; Lerner, Norma B.; Marder, Victor J.; Silverman, David J.; Turpin, Loel C.; Schwab, Amy L.

In: Infection and Immunity, Vol. 65, No. 7, 1997, p. 2786-2791.

Research output: Contribution to journalArticle

Sporn, LA, Sahni, S, Lerner, NB, Marder, VJ, Silverman, DJ, Turpin, LC & Schwab, AL 1997, 'Rickettsia rickettsii infection of cultured human endothelial cells induces NF-κB activation', Infection and Immunity, vol. 65, no. 7, pp. 2786-2791.
Sporn LA, Sahni S, Lerner NB, Marder VJ, Silverman DJ, Turpin LC et al. Rickettsia rickettsii infection of cultured human endothelial cells induces NF-κB activation. Infection and Immunity. 1997;65(7):2786-2791.
Sporn, Lee Ann ; Sahni, Sanjeev ; Lerner, Norma B. ; Marder, Victor J. ; Silverman, David J. ; Turpin, Loel C. ; Schwab, Amy L. / Rickettsia rickettsii infection of cultured human endothelial cells induces NF-κB activation. In: Infection and Immunity. 1997 ; Vol. 65, No. 7. pp. 2786-2791.
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