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Non‐opsonic phagocytosis of homologous non‐toxigenic and toxigenic Corynebacterium diphtheriae strains by human U‐937 macrophages
Author(s) -
Dos Santos Cíntia Silva,
Dos Santos Louisy Sanches,
De Souza Monica Cristina,
Dos Santos Dourado Fernanda,
De Souza de Oliveira Dias Alexandre Alves,
Sabbadini Priscila Soares,
Pereira Gabriela Andrade,
Cabral Maulori Curié,
Junior Raphael Hirata,
De MattosGuaraldi Ana Luíza
Publication year - 2010
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2009.00179.x
Subject(s) - biology , microbiology and biotechnology , phagocytosis , corynebacterium diphtheriae , opsonin , diphtheria toxin , macrophage , diphtheria , virology , toxin , in vitro , biochemistry , vaccination
As interactions between bacteria and macrophages dictate the outcome of most infectious diseases, analyses of molecular mechanisms of non‐opsonic phagocytosis should lead to new approaches for the prevention of diphtheria and systemic Corynebacterium diphtheriae infections. The present study aimed to evaluate human macrophage–bacteria interactions in the absence of opsonin antibodies and the influence of the tox gene on this process. Homologous C. diphtheriae tox + and tox – strains were evaluated for adhesion, entering and survival within U‐937 human macrophages at different incubation periods. Higher numbers of viable bacteria associated with and internalized by macrophages were demonstrated for the tox + strain. However, viable intracellular bacteria were detected at T‐24 hr only for the tox – strain. Cytoskeletal inhibitors, cytochalasin E, genistein and colchicine, inhibited intracellular viability of both strains at different levels. Bacterial replication was evidenced at T‐24 hr in supernatants of monolayers infected with the tox – strain. Host cell death and nuclear alterations were evidenced by the Trypan blue exclusion assay and DAPI fluorescence microscopy. ELISA of histone‐associated DNA fragments allowed detection of apoptosis and necrosis induced by tox + and tox – strains at T‐1 hr and T‐3 hr. In conclusion, human macrophages in the absence of opsonins may not be promptly effective at killing diphtheria bacilli. The presence of the tox gene influences the susceptibility of C. diphtheriae to human macrophages and the outcome of non‐opsonic phagocytosis. C. diphtheriae strains exhibit strategies to survive within macrophages and to exert apoptosis and necrosis in human phagocytic cells, independent of the tox gene.