PcpA of Streptococcus pneumoniae mediates adherence to nasopharyngeal and lung epithelial cells and elicits functional antibodies in humans

M. Nadeem Khan, Sharad K. Sharma, Laura M. Filkins, Michael E. Pichichero

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Streptococcus pneumoniae (pneumococci) adhere to human nasopharyngeal (NP) epithelial cells as a first step in colonization and adherence of pneumococci to lung epithelia may be required to establish pneumonia. We sought to determine if PcpA can serve as an adhesin to human NP (D562) and lung (A549) epithelial cells and whether PcpA mediated adherence can be inhibited by human anti-PcpA antibodies. A PcpA isogenic mutant was constructed in a pneumococcal TIGR4 background. When the mutant and wild type strains were compared for their adherence to D562 and A549 cell lines, a reduction in adherence by the mutant was observed (p = 0.0001 for both cell types). PcpA was ectopically expressed on the surface of minimally-adherent heterologous host Escherichia coli resulting in augmented adherence to D562 (p = 0.002) and A549 (p = 0.015) cells. Total IgG was purified from a pool of 6 human sera having high IgG titers of anti-pneumococcal proteins. The purified IgG reduced TIGR4 adherence to D562 cells but we determined that this effect was largely due to bacterial cell aggregation as determined by flow cytometry and confocal microscopy. Fab fragments were prepared from pooled IgG sera. Inhibition of TIGR4 adherence to D562 cells was observed using the Fab fragments without causing bacterial aggregation (p = 0.0001). Depletion of PcpA-specific Fab fragments resulted in an increase in adherence of TIGR4 to D562 cells (p = 0.028). We conclude that PcpA can mediate adherence of pneumococci to human NP and lung epithelial cells and PcpA mediated adherence can be inhibited by human anti-PcpA antibodies.

Original languageEnglish (US)
Pages (from-to)1102-1110
Number of pages9
JournalMicrobes and Infection
Volume14
Issue number12
DOIs
StatePublished - Oct 1 2012
Externally publishedYes

Fingerprint

Streptococcus pneumoniae
Epithelial Cells
Lung
Immunoglobulin Fab Fragments
Antibodies
Immunoglobulin G
Anti-Idiotypic Antibodies
Cell Aggregation
Serum
Confocal Microscopy
Pneumonia
Flow Cytometry
Epithelium
Escherichia coli
Proteins

Keywords

  • Adhesins
  • PcpA
  • Pneumococci

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Infectious Diseases

Cite this

PcpA of Streptococcus pneumoniae mediates adherence to nasopharyngeal and lung epithelial cells and elicits functional antibodies in humans. / Khan, M. Nadeem; Sharma, Sharad K.; Filkins, Laura M.; Pichichero, Michael E.

In: Microbes and Infection, Vol. 14, No. 12, 01.10.2012, p. 1102-1110.

Research output: Contribution to journalArticle

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