RESUMO
The blood-brain barrier (BBB) is one of several barriers between the brain and the peripheral blood system to maintain homeostasis. Understanding the interactions between infectious agents such as human immunodeficiency virus type 1 (HIV-1), which are capable of traversing the BBB and causing neuroinflammation requires modeling an authentic BBB in vitro. Such an in vitro BBB model also helps develop means of targeting viruses that reside in the brain via natural immune effectors such as antibodies. The BBB consists of human brain microvascular endothelial cells (HBMECs), astrocytes, and pericytes. Here we report in vitro methods to establish a dual-cell BBB model consisting of primary HBMECs and primary astrocytes to measure the integrity of the BBB and antibody penetration of the BBB, as well as a method to establish a single cell BBB model to study the impact of HIV-1 infected medium on the integrity of such a BBB.
Assuntos
Astrócitos , Barreira Hematoencefálica , Células Endoteliais , Infecções por HIV , HIV-1 , Barreira Hematoencefálica/virologia , Barreira Hematoencefálica/metabolismo , Humanos , Astrócitos/virologia , Astrócitos/metabolismo , Astrócitos/imunologia , Células Endoteliais/virologia , Células Endoteliais/metabolismo , Células Endoteliais/imunologia , HIV-1/imunologia , HIV-1/fisiologia , Infecções por HIV/virologia , Infecções por HIV/imunologia , Pericitos/virologia , Pericitos/metabolismo , Pericitos/imunologia , Doenças Neuroinflamatórias/virologia , Doenças Neuroinflamatórias/imunologia , Técnicas de Cocultura/métodos , Células Cultivadas , Encéfalo/virologia , Encéfalo/imunologia , Encéfalo/metabolismoRESUMO
Although vaccination efforts have expanded, there are still gaps in our understanding surrounding the immune response to SARS-CoV-2. Measuring IgG Fc glycosylation provides insight into an infected individual's inflammatory state, among other functions. We set out to interrogate bulk IgG glycosylation changes from SARS-CoV-2 infection and vaccination, using plasma from mild or hospitalized COVID-19 patients, and from vaccinated individuals. Inflammatory glycans are elevated in hospitalized COVID-19 patients and increase over time, while mild patients have anti-inflammatory glycans that increase over time, including increased sialic acid correlating with RBD antibody levels. Vaccinated individuals with low RBD antibody levels and low neutralization have the same IgG glycan traits as hospitalized COVID-19 patients. In addition, a small vaccinated cohort reveals a decrease in inflammatory glycans associated with peak IgG concentrations and neutralization. This report characterizes the bulk IgG glycome associated with COVID-19 severity and vaccine responsiveness and can help guide future studies into SARS-CoV-2 protective immunity.