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Ex Vivo Infection of Human Lymphoid Tissue and Female Genital Mucosa with Human Immunodeficiency Virus 1 and Histoculture
Published on: October 12, 2018
Immunological landscape of human lymphoid explants during measles virus infection
Joshua A Acklin1,2, Aum R Patel1,2, Andrew P Kurland1
1Department of Microbiology.
Abstract:
In humans, lymph nodes are the primary site of measles virus (MeV) replication. To understand the immunological events that occur at this site, we infected human lymphoid tissue explants using a pathogenic strain of MeV that expresses GFP. We found that MeV infected 5%-15% of cells across donors. Using single-cell RNA-Seq and flow cytometry, we found that while most of the 29 cell populations identified in the lymphoid culture were susceptible to MeV, there was a broad preferential infection of B cells and reduced infection of T cells. Further subsetting of T cells revealed that this reduction may be driven by the decreased infection of naive T cells. Transcriptional changes in infected B cells were dominated by an interferon-stimulated gene (ISG) signature. To determine which of these ISGs were most substantial, we evaluated the proteome of MeV-infected Raji cells by mass spectrometry. We found that IFIT1, IFIT2, IFIT3, ISG15, CXCL10, MX2, and XAF1 proteins were the most highly induced and positively correlated with their expression in the transcriptome. These data provide insight into the immunological events that occur in lymph nodes during infection and may lead to the development of therapeutic interventions.
Insights
Measles virus (MeV) primarily replicates in human lymph nodes, preferentially infecting B cells over T cells. Infected B cells show a strong interferon-stimulated gene (ISG) response, highlighting key immune events during MeV infection.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Lymph nodes are the primary replication sites for measles virus (MeV) in humans.
- Understanding MeV's interaction with lymphoid cells is crucial for developing interventions.
Purpose of the Study:
- To investigate the immunological events during MeV infection in human lymphoid tissue.
- To identify specific cell types and molecular responses following MeV infection.
Main Methods:
- Infection of human lymphoid tissue explants with a GFP-expressing pathogenic MeV strain.
- Single-cell RNA-Seq and flow cytometry to analyze cell susceptibility and transcriptional changes.
- Mass spectrometry to evaluate protein induction in MeV-infected cells.
Main Results:
- MeV infected 5%-15% of cells, with preferential infection of B cells and reduced infection of T cells, particularly naive T cells.
- Transcriptional analysis of infected B cells revealed a significant interferon-stimulated gene (ISG) signature.
- Proteomic analysis confirmed high induction of IFIT1, IFIT2, IFIT3, ISG15, CXCL10, MX2, and XAF1 proteins.
Conclusions:
- Measles virus exhibits cell-type-specific tropism within human lymph nodes, favoring B cells.
- The interferon-stimulated gene response is a prominent feature of MeV-infected B cells.
- These findings offer insights into lymph node immunology during MeV infection and potential therapeutic targets.
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