机构地区:[1]Division of Infectious Diseases,Department of Internal Medicine,School of Medicine of Ribeir?o Preto,University of S?o Paulo,Ribeir?o Preto,14049-900 S?o Paulo,Brazil [2]Department of Morphology,Stomatology and Physiology,School of Dentistry of Ribeir?o Preto,University of S?o Paulo,Ribeir?o Preto,14049-900 S?o Paulo,Brazil [3]EA3064,Groupe Immunité des Muqueuses et Agents Pathogènes(GIMAP),Faculty of Medicine Jacques Lisfranc,University of Lyon,42023 Saint-Etienne,France
出 处:《World Journal of Virology》2014年第2期11-17,共7页世界病毒学杂志
基 金:Supported by The Brazilian Federal Agency CAPES(Coordenao de Aperfeioamento de Pessoal de nível Superior);evaluation of graduate education
摘 要:AIM: 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 iAIM: 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 i
关 键 词:Human IMMUNODEFICIENCY virus/acquired IMMUNODEFICIENCY syndrome Sexual transmission Seminal plasma CCL20 LACTOFERRIN ENDOCERVICAL epithelial cells
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