Intracellular survival of Brucella spp. in human monocytes involves conventional uptake but special phagosomes.
Brucella spp. are facultative intracellular parasites of various mammals, including humans, typically infecting lymphoid as well as reproductive organs. We have investigated how B. suis and B. melitensis enter human monocytes and in which compartment they survive. Peripheral blood monocytes readily internalized nonopsonized brucellae and killed most of them within 12 to 18 h. The presence of Brucella-specific antibodies (but not complement) increased the uptake of bacteria without increasing their intracellular survival, whereas adherence of the monocytes or incubation in Ca(2+)- and Mg(2+)-free medium reduced the uptake. Engulfment of all Brucella organisms (regardless of bacterial viability or virulence) initially resulted in phagosomes with tightly apposed walls (TP). Most TP were fully fusiogenic and matured to spacious phagolysosomes containing degraded bacteria, whereas some TP (more in monocyte-derived macrophages, HeLa cells, and CHO cells than in monocytes) remained tightly apposed to intact bacteria. Immediate treatment of infected host cells with the lysosomotropic base ammonium chloride caused a swelling of all phagosomes and a rise in the intraphagosomal pH, abolishing the intracellular survival of Brucella. These results indicate that (i) human monocytes readily internalize Brucella in a conventional way using various phagocytosis-promoting receptors, (ii) the maturation of some Brucella phagosomes is passively arrested between the steps of acidification and phagosome-lysosome fusion, (iii) brucellae are killed in maturing but not in arrested phagosomes, and (iv) survival of internalized Brucella depends on an acidic intraphagosomal pH and/or close contact with the phagosomal wall.
Rittig
M G
MG
INSERM U-431, Université de Montpellier II, Montpellier, France. michael.rittig@nottingham.ac.uk
Alvarez-Martinez
M T
MT
Porte
F
F
Liautard
J P
JP
Rouot
B
B
D000818:Animals; D002002:Brucella / Q000254:growth & development* / Q000648:ultrastructure; D017347:Brucella melitensis / Q000254:growth & development / Q000648:ultrastructure; D002006:Brucellosis / Q000382:microbiology; D016466:CHO Cells / Q000276:immunology / Q000648:ultrastructure; D006224:Cricetinae; D006367:HeLa Cells / Q000276:immunology / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000276:immunology* / Q000382:microbiology* / Q000648:ultrastructure; D010587:Phagocytosis / Q000276:immunology*; D010588:Phagosomes / Q000382:microbiology*