Tracking bacterial infection of macrophages using a novel red-emission pH sensor

Yuguang Jin, Yanqing Tian, Weiwen Zhang, Sei Hum Jang, Alex K Y Jen, Deirdre Meldrum

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The relationship between bacteria and host phagocytic cells is key to the induction of immunity. To visualize and monitor bacterial infection, we developed a novel bacterial membrane permeable pH sensor for the noninvasive monitoring of bacterial entry into murine macrophages. The pH sensor was constructed using 2-dicyanomethylene-3-cyano-4,5,5-trimethyl-2,5-dihydrofuran (TCF) as an electron-withdrawing group and aniline as an electron-donating group. A piperazine moiety was used as the pH-sensitive group. Because of the strong electron-donating and -withdrawing units conjugated in the sensing moiety M, the fluorophore emitted in the red spectral window, away from the autofluorescence regions of the bacteria. Following the engulfment of sensor-labeled bacteria by macrophages and their subsequent merger with host lysosomes, the resulting low-pH environment enhances the fluorescence intensity of the pH sensors inside the bacteria. Time-lapse analysis of the fluorescent intensity suggested significant heterogeneity of bacterial uptake among macrophages. In addition, qRT-PCR analysis of the bacterial 16 S rRNA gene expression within single macrophage cells suggested that the 16 S rRNA of the bacteria was still intact 120 min after they had been engulfed by macrophages. A toxicity assay showed that the pH sensor has no cytotoxicity towards either E. coli or murine macrophages. The sensor shows good repeatability, a long lifetime, and a fast response to pH changes, and can be used for a variety of bacteria.

Original languageEnglish (US)
Pages (from-to)1375-1384
Number of pages10
JournalAnalytical and Bioanalytical Chemistry
Volume398
Issue number3
DOIs
StatePublished - Oct 2010

Fingerprint

pH sensors
Macrophages
Bacterial Infections
Bacteria
Electrons
Fluorophores
Sensors
Cytotoxicity
Gene expression
Escherichia coli
Phagocytes
Toxicity
Lysosomes
rRNA Genes
Assays
Fluorescence
Immunity
Membranes
Monitoring
Gene Expression

Keywords

  • Bacterial infection
  • Mouse macrophage
  • pH sensor
  • Red emitter

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Tracking bacterial infection of macrophages using a novel red-emission pH sensor. / Jin, Yuguang; Tian, Yanqing; Zhang, Weiwen; Jang, Sei Hum; Jen, Alex K Y; Meldrum, Deirdre.

In: Analytical and Bioanalytical Chemistry, Vol. 398, No. 3, 10.2010, p. 1375-1384.

Research output: Contribution to journalArticle

Jin, Yuguang ; Tian, Yanqing ; Zhang, Weiwen ; Jang, Sei Hum ; Jen, Alex K Y ; Meldrum, Deirdre. / Tracking bacterial infection of macrophages using a novel red-emission pH sensor. In: Analytical and Bioanalytical Chemistry. 2010 ; Vol. 398, No. 3. pp. 1375-1384.
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