Assessing Lipid Derived Innate Immune Factors in Vernix Caseosa and Amniotic Fluid Protecting Against Intrauterine Infections

Innate immunity provides first line of defense against infectious diseases. Host derived lipids, including cholesteryl esters, have emerged as key effector molecules of innate immunity. Mother-to-child transmission of HIV and other infectious disease agents, such as Listeria monocytogenes (LM), pose high risks to unborn babies. Two protective newborn substances: Amniotic Fluid (AF), clear liquid in the amniotic sac that protects the fetus during pregnancy, and Vernix Caseosa (VC), lipid-rich wax-like substance coating a newborn, contains antibody-independent, innate anti-HIV factors. However, the molecular source of their antimicrobial properties is unknown. We hypothesize that AF and VC-derived lipids have anti-HIV and anti-LM activity. To test this hypothesis, we aim to subject liposomal preparation from AF and VC lipid extracts to an antiviral assay with a pseudotyped virus and LM antibacterial tests. Here we report on the methodology establishment for liposome preparation from AF and VC lipid extracts. Unilamellar liposomes were formulated via heat and sonication with extrusion through a filter under pressure. Light microscopy revealed successful generation of liposomal structures and thin layer chromatography with visualization in iodine vapor demonstrated up to 40% recovery of lipids. The lipid profiles were comparable to the lipid distribution in the original material. We are now conducting our first colony forming unit assays employing these liposomes and LM. Our efforts may lead to the identification of important novel factors preventing against HIV transmission and may have clinical applications in fetus protection in utero.