Toward a humanized mouse model of Pneumocystis pneumonia
Guixiang Dai, Alanna Wanek, Taylor Eddens, Paul Volden, Jay K. Kolls
Guixiang Dai, Alanna Wanek, Taylor Eddens, Paul Volden, Jay K. Kolls
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Research Article Infectious disease Pulmonology

Toward a humanized mouse model of Pneumocystis pneumonia

Abstract

Pneumocystis is an important opportunistic fungus that causes pneumonia in children and immunocompromised individuals. Recent genomic data show that divergence of major surface glycoproteins may confer speciation and host range selectivity. On the other hand, immune clearance between mice and humans is well correlated. Thus, we hypothesized that humanize mice may provide information about human immune responses involved in controlling Pneumocystis infection. CD34-engrafted huNOG-EXL mice controlled fungal burdens to a greater extent than nonengrafted mice. Moreover, engrafted mice generated fungal-specific IgM. Fungal control was associated with a transcriptional signature that was enriched for genes associated with nonopsonic recognition of trophs (CD209) and asci (CLEC7A). These same genes were downregulated in CD4-deficient mice as well as twins with bare lymphocyte syndrome with Pneumocystis pneumonia.

Authors

Guixiang Dai, Alanna Wanek, Taylor Eddens, Paul Volden, Jay K. Kolls

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Figure 3

Human lymphocytes generated Pneumocystis-specific human cytokines in lung cells.

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Human lymphocytes generated Pneumocystis-specific human cytokines in lun...
huNOG-EXL mice engrafted with human umbilical cord blood–derived CD34+ hematopoietic stem cells and unengrafted mice were infected with 100 μl (approximately 2 × 105 asci) P. murina inoculum by oral pharyngeal aspiration. Six weeks after infection, single lung cells were prepared and cultured with or without Pneumocystis antigen for cytokine production, as measured by flow cytometry. The NOG-EXL group of 4 mice did not have human cells and, thus, no cytokine-generating human cells were detected. In vitro antigen stimulation did not have any effect on cytokine-generating cells. Analysis using a 2-tailed, unpaired t test (A) indicated that there were significant differences in cytokine-generating cells between the huNOG-EXL and NOG-EXL groups (4 mice each group); P < 0.0001 for IFN-γ+CD4+, IL-17A+CD4+, and IFN-γ+CD8+; P < 0.05 for IL-5+CD4+; and P > 0.05 for IL-4+CD4+ cells. Individual values are shown with a horizontal bar indicating the mean of the group. Representative flow cytometry gating plots are shown for huNOG-EXL (B) and NOG-EXL (C) mice.