Abstract
BackgroundThe female reproductive tract is amongst the main routes for Human Immunodeficiency Virus (HIV) transmission. Cervical mucus however is known to protect the female reproductive tract from bacterial invasion and fluid loss and regulates and facilitates sperm transport to the upper reproductive tract. The purpose of this study was to purify and characterize pregnancy plug mucins and determine their anti-HIV-1 activity in an HIV inhibition assay.MethodsPregnancy plug mucins were purified by caesium chloride density-gradient ultra-centrifugation and characterized by Western blotting analysis. The anti-HIV-1 activities of the crude pregnancy plug mucus and purified pregnancy plug mucins was determined by incubating them with HIV-1 prior to infection of the human T lymphoblastoid cell line (CEM SS cells).ResultsThe pregnancy plug mucus had MUC1, MUC2, MUC5AC and MUC5B. The HIV inhibition assay revealed that while the purified pregnancy plug mucins inhibit HIV-1 activity by approximately 97.5%, the crude pregnancy plug mucus failed to inhibit HIV-1 activity.ConclusionAlthough it is not clear why the crude sample did not inhibit HIV-1 activity, it may be that the amount of mucins in the crude pregnancy plug mucus (which contains water, mucins, lipids, nucleic acids, lactoferrin, lysozyme, immunoglobulins and ions), is insufficient to cause viral inhibition or aggregation.
Highlights
The female reproductive tract is amongst the main routes for Human Immunodeficiency Virus (HIV) transmission
Mucin purification Pregnancy plug mucins were purified by density gradient centrifugation, twice in caesium chloride/4 M GuHCl with a buoyant density between 1.39 and 1.40 g/ml to remove proteins and nucleic acids
In the case of breast milk we showed that its MUC1 component inhibited the Human Immunodeficiency Virus type 1 (HIV-1) from infecting CEM SS cells in an in vitro assay only after it was dissociated from the milk fat globules and isolated and purified by caesium chloride density gradient ultracentrifugation
Summary
The female reproductive tract is amongst the main routes for Human Immunodeficiency Virus (HIV) transmission. Cervical mucus is known to protect the female reproductive tract from bacterial invasion and fluid loss and regulates and facilitates sperm transport to the upper reproductive tract. Cervical mucus is reported to regulate sperm penetration and transport to the upper reproductive tract [1,2]. CFiageusiruem1chloride density gradient purification of the pregnancy plug mucins Caesium chloride density gradient purification of the pregnancy plug mucins. Samples in 4 M GuHCl were adjusted to a density of 1.39 to 1.40 g ml-1 with solid caesium chloride. Mucin positive fractions (u) at a density (s) between 1.37–1.42 and still associated with some protein (n) (a) were pooled and prepared for the second step centrifugation (b). Fractions (fraction number 3, 4 and 5) were pooled, dialysed against three changes of distilled water and freeze-dried
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