title: Semen lactoferrin promotes CCL20 production by epithelial cells: Involvement in HIV transmission.
abstract: OBJECTIVE
  To study the effect of seminal plasma on Chemokine (C-C motif) ligand 20 (CCL20) production by epithelial cells and its relationship with lactoferrin.
  
  
  METHODS
  HEC-1A cells, a cell line derived from a monostratified endocervical epithelium, were incubated with samples of seminal plasma (diluted 1:10 in culture medium) recovered from human immunodeficiency virus (HIV) seronegative (HIV-) or HIV seropositive (HIV+) subjects. Recombinant human interleukin 1 beta (IL-1β) was used as positive control, and culture medium only as negative control. The measurement of CCL20 production in the supernatants of HEC-1A cells and of lactoferrin in seminal plasma was determined by enzyme-linked immunosorbent assay techniques. A fractionation of seminal plasma proteins was performed by ion exchange chromatography on a pool of seminal plasma specimens from HIV- subjects. Each fraction was tested for its ability to stimulate the production of CCL20 by HEC-1A cells and for its lactoferrin concentration. The HIV viral load in seminal plasma samples from HIV+ patients was measured using the HIV-Monitor kit (Roche Diagnostic Systems, Branchburg, NJ, United States).
  
  
  RESULTS
  The positive control IL-1β was responsible for an increase of 11.36 ± 3.36 times in the production of CCL20. Stimulation of HEC-1A cells was performed in 34 seminal plasma samples (22 from HIV+ subjects and 12 from HIV- subjects). The mean production of CCL20 by HEC-1A in presence of seminal plasma from HIV- and HIV+ subjects was respectively 5.38 ± 0.91 and 7.57 ± 3.26 times higher than that obtained with the untreated cells (P < 0.05 between the two groups). Using the same 34 specimens of seminal plasma, no correlation was observed between the concentration of total proteins in seminal plasma and their ability to stimulate the secretion of CCL20 by HEC-1 cells. In contrast, the ability to produce CCL20 by HEC-1A cells correlated to the concentration of lactoferrin in the seminal plasma samples (r coefficient = 0.56; CI: 0.26-0.76; P < 0.001). After fractionation by ion exchange chromatography, the seminal plasma fractions exhibiting the highest concentrations of lactoferrin were responsible for the greatest stimulation of CCL20 production by HEC-1A cells (r coefficient = 0.89; CI: 0.78-0.95; P < 0.0001).
  
  
  CONCLUSIONS
  Lactoferrin present in seminal plasma correlated with an increased production of CCL20 by HEC-1A cells and therefore could facilitate HIV entry through the genital mucosa.
authors.affiliation: Alan Grupioni Lourenço, Alcyone Artioli Machado, Silvana Maria Quintana, Division of Infectious Diseases, Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14049-900 São Paulo, Brazil.
authors.forename: Alan Grupioni
authors.initials: AG
authors.lastname: Lourenço
authors.affiliation: Alan Grupioni Lourenço, Alcyone Artioli Machado, Silvana Maria Quintana, Division of Infectious Diseases, Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14049-900 São Paulo, Brazil.
authors.forename: Marilena Chinalli
authors.initials: MC
authors.lastname: Komesu
authors.affiliation: Alan Grupioni Lourenço, Alcyone Artioli Machado, Silvana Maria Quintana, Division of Infectious Diseases, Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14049-900 São Paulo, Brazil.
authors.forename: Alcyone Artioli
authors.initials: AA
authors.lastname: Machado
authors.affiliation: Alan Grupioni Lourenço, Alcyone Artioli Machado, Silvana Maria Quintana, Division of Infectious Diseases, Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14049-900 São Paulo, Brazil.
authors.forename: Silvana Maria
authors.initials: SM
authors.lastname: Quintana
authors.affiliation: Alan Grupioni Lourenço, Alcyone Artioli Machado, Silvana Maria Quintana, Division of Infectious Diseases, Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14049-900 São Paulo, Brazil.
authors.forename: Thomas
authors.initials: T
authors.lastname: Bourlet
authors.affiliation: Alan Grupioni Lourenço, Alcyone Artioli Machado, Silvana Maria Quintana, Division of Infectious Diseases, Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14049-900 São Paulo, Brazil.
authors.forename: Bruno
authors.initials: B
authors.lastname: Pozzetto
authors.affiliation: Alan Grupioni Lourenço, Alcyone Artioli Machado, Silvana Maria Quintana, Division of Infectious Diseases, Department of Internal Medicine, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, 14049-900 São Paulo, Brazil.
authors.forename: Olivier
authors.initials: O
authors.lastname: Delézay
keywords: CCL20; Endocervical epithelial cells; Human immunodeficiency virus/acquired immunodeficiency syndrome; Lactoferrin; Seminal plasma; Sexual transmission