Microscopy-based assays for high-throughput screening of host factors involved in Brucella infection of hela cells

Journal of Visualized Experiments

Volumn 2016, Issue 114, 2016, Pages

  Casanova, Alain ^a   Low, Shyan H ^a   Emmenlauer, Mario ^a,d   Conde Alvarez, Raquel ^a,c   Salcedo, Suzana P ^b   Gorvel, Jean Pierre ^b   Dehio, Christoph ^a  

^a UNIVERSITY OF BASEL   (Switzerland)

^b CENTRE D IMMUNOLOGIE DE MARSEILLE LUMINY   (France)

^c UNIVERSITY OF NAVARRA   (Spain)

^d BioDataAnalysis GmbH   (Switzerland)

Author keywords

Automated image analysis; Brucella abortus; Cell biology; Cell invasion; Fluorescence microscopy; HeLa cells; High content high throughput screening assay; Infection; Infection biology; Intracellular pathogen; Issue 114; RNAi (RNA interference); SiRNA (small interfering RNA)

Indexed keywords

BRUCELLA; BRUCELLOSIS; HELA CELL LINE; HIGH THROUGHPUT SCREENING; HOST PARASITE INTERACTION; HUMAN; METABOLISM; MICROBIOLOGY; MICROSCOPY; PATHOGENICITY; PHAGOCYTE; PROCEDURES;

BRUCELLA; BRUCELLOSIS; HELA CELLS; HIGH-THROUGHPUT SCREENING ASSAYS; HOST-PARASITE INTERACTIONS; HUMANS; MICROSCOPY; PHAGOCYTES;

EID: 84983735396     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/54263     Document Type: Article

References (15)

1. Pappas, G., Papadimitriou, P., Akritidis, N., Christou, L., & Tsianos, E. V. The new global map of human brucellosis. Lancet Infect Dis. 6, 91-99 (2006).
2. Atluri, V. L., Xavier, M. N., de Jong, M. F., den Hartigh, A. B., & Tsolis, R. E. Interactions of the human pathogenic Brucella species with their hosts. Annu Rev Microbiol. 65, 523-541 (2011).
3. Guzman-Verri, C. et al. GTPases of the Rho subfamily are required for Brucella abortus internalization in nonprofessional phagocytes: direct activation of Cdc42. J Biol Chem. 276, 44435-44443 (2001).
4. Starr, T., Ng, T. W., Wehrly, T. D., Knodler, L. A., & Celli, J. Brucella intracellular replication requires trafficking through the late endosomal/ lysosomal compartment. Traffic. 9, 678-694 (2008).
5. Boschiroli, M. L. et al. The Brucella suis virB operon is induced intracellularly in macrophages. Proc Natl Acad Sci U S A. 99, 1544-1549 (2002).
6. Celli, J. et al. Brucella evades macrophage killing via VirB-dependent sustained interactions with the endoplasmic reticulum. J Exp Med. 198, 545-556 (2003).
7. Porte, F., Liautard, J. P., & Kohler, S. Early acidification of phagosomes containing Brucella suis is essential for intracellular survival in murine macrophages. Infect Immun. 67, 4041-4047 (1999).
8. Anderson, T. D., & Cheville, N. F. Ultrastructural morphometric analysis of Brucella abortus-infected trophoblasts in experimental placentitis. Bacterial replication occurs in rough endoplasmic reticulum. Am J Pathol. 124, 226-237 (1986).
9. Qin, Q. M. et al. RNAi screen of endoplasmic reticulum-associated host factors reveals a role for IRE1alpha in supporting Brucella replication. PLoS Pathog. 4, e1000110 (2008).
10. Ramo, P. et al. Simultaneous analysis of large-scale RNAi screens for pathogen entry. BMC genomics. 15, 1162 (2014).
11. Kuhbacher, A., Gouin, E., Cossart, P., & Pizarro-Cerda, J. Imaging InlC secretion to investigate cellular infection by the bacterial pathogen Listeria monocytogenes. J Vis Exp. e51043 (2013).
12. Kentner, D. et al. Shigella reroutes host cell central metabolism to obtain high-flux nutrient supply for vigorous intracellular growth. Proc Natl Acad Sci U S A. 111, 9929-9934 (2014).
13. Celli, J., Salcedo, S. P., & Gorvel, J. P. Brucella coopts the small GTPase Sar1 for intracellular replication. Proc Natl Acad Sci U S A. 102, 1673-1678 (2005).
14. von Bargen, K., Gorvel, J. P., & Salcedo, S. P. Internal affairs: investigating the Brucella intracellular lifestyle. FEMS microbiology reviews. 36, 533-562 (2012).
15. Kamentsky, L. et al. Improved structure, function and compatibility for CellProfiler: modular high-throughput image analysis software. Bioinformatics. 27, 1179-1180 (2011).