Analysis of cell cycle and replication of mouse macrophages after in vivo and in vitro Cryptococcus neoformans infection using laser scanning cytometry

Carolina Coelho, Lydia Tesfa, Jinghang Zhang, Johanna Rivera, Teresa Gonçalves, Arturo Casadevall

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

We investigated the outcome of the interaction of Cryptococcus neoformans with murine macrophages using laser scanning cytometry (LSC). Previous results in our lab had shown that phagocytosis of C. neoformans promoted cell cycle progression. LSC allowed us to simultaneously measure the phagocytic index, macrophage DNA content, and 5-ethynyl-2'-deoxyuridine (EdU) incorporation such that it was possible to study host cell division as a function of phagocytosis. LSC proved to be a robust, reliable, and high-throughput method for quantifying phagocytosis. Phagocytosis of C. neoformans promoted cell cycle progression, but infected macrophages were significantly less likely to complete mitosis. Hence, we report a new cytotoxic effect associated with intracellular C. neoformans residence that manifested itself in impaired cell cycle completion as a consequence of a block in the G 2/M stage of the mitotic cell cycle. Cell cycle arrest was not due to increased cell membrane permeability or DNA damage. We investigated alveolar macrophage replication in vivo and demonstrated that these cells are capable of low levels of cell division in the presence or absence of C. neoformans infection. In summary, we simultaneously studied phagocytosis, the cell cycle state of the host cell and pathogen-mediated cytotoxicity, and our results demonstrate a new cytotoxic effect of C. neoformans infection on murine macrophages: fungus-induced cell cycle arrest. Finally, we provide evidence for alveolar macrophage proliferation in vivo.

Original languageEnglish (US)
Pages (from-to)1467-1478
Number of pages12
JournalInfection and Immunity
Volume80
Issue number4
DOIs
StatePublished - Apr 2012

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Laser Scanning Cytometry
Cryptococcus neoformans
Cell Cycle
Phagocytosis
Macrophages
Infection
Alveolar Macrophages
Cell Cycle Checkpoints
Cell Division
Cell Membrane Permeability
Mitosis
DNA Damage
In Vitro Techniques
Fungi
DNA

ASJC Scopus subject areas

  • Immunology
  • Microbiology
  • Parasitology
  • Infectious Diseases

Cite this

Analysis of cell cycle and replication of mouse macrophages after in vivo and in vitro Cryptococcus neoformans infection using laser scanning cytometry. / Coelho, Carolina; Tesfa, Lydia; Zhang, Jinghang; Rivera, Johanna; Gonçalves, Teresa; Casadevall, Arturo.

In: Infection and Immunity, Vol. 80, No. 4, 04.2012, p. 1467-1478.

Research output: Contribution to journalArticle

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