Germline IgM predicts T cell immunity to Pneumocystis.

Pneumocystis is the most common fungal pulmonary infection in children under the age of 5 years. In children with primary immunodeficiency, Pneumocystis often presents at 3-6 months of age, a time period that coincides with the nadir of maternal IgG and when IgM is the dominant Ig isotype. Because B cells are the dominant antigen-presenting cells for Pneumocystis, we hypothesized the presence of fungal-specific IgMs in humans and mice and that these IgM specificities would predict T cell antigens. We detected fungal-specific IgMs in human and mouse sera and utilized immunoprecipitation to determine whether any antigens were similar across donors. We then assessed T cell responses to these antigens and found anti-Pneumocystis IgM in WT mice, Aicda-/- mice, and in human cord blood. Immunoprecipitation of Pneumocystis murina with human cord blood identified shared antigens among these donors. Using class II MHC binding prediction, we designed peptides with these antigens and identified robust peptide-specific lung T cell responses after P. murina infection. After mice were immunized with 2 of the antigens, adoptive transfer of vaccine-elicited CD4+ T cells showed effector activity, suggesting that these antigens contain protective Pneumocystis epitopes. These data support the notion that germline-encoded IgM B cell receptors are critical in antigen presentation and T cell priming in early Pneumocystis infection.

A Dual Omics Approach to Evaluate Transcriptional and Metabolic Responses During Lipid Deprivation in an Oyster Parasite, Perkinsus marinus.

Perkinsus marinus, a protozoan and the causative agent of perkinsosis (dermo disease) is a prevalent parasite found within the eastern oyster (Crassostrea virginica). In this study, we explore metabolic processes of P. marinus cells under lipid-depleted medium conditions to elucidate the interchanging flux of lipid and carbohydrate metabolism. Although P. marinus can synthesize their own lipids from available nutrients, they display a slower growth in medium not supplemented with lipids as opposed to medium with lipids. Under these conditions, using transcriptomics, we surprisingly observed evidence of stimulated lipid degradation through increased transcription of two core ß-oxidation pathway enzymes. Simultaneously, phospholipid biosynthetic pathways were downregulated. Metabolomic analysis supported the transcriptomic results. Most fatty acids were decreased in lipid-deplete medium as opposed to lipid-replete medium, and available glucose was fermented to lactate. A significant increase in the cholesterol derivative zymosterol further supported a downregulation of membrane synthesis under the experimental conditions. A robust tricarboxylic acid (TCA) cycle was apparent by enhanced citrate synthase transcription, and a simultaneous reduction in branched chain amino acids. It is concluded that although P. marinus has the capacity for synthesizing its own lipids, it can respond to lipid deprivation in medium by oxidizing readily available stores, and likely transitioning into a resting stage.

Effect of Subcutaneous Anti-CD20 Antibody-Mediated B Cell Depletion on Susceptibility to Pneumocystis Infection in Mice.

Prior work has shown that parenterally administered anti-CD20 (5D2) inhibits CD4+ T cell priming in response to challenge with Pneumocystis murina and predisposes to pneumonia. In this study, we investigated the effect of subcutaneous anti-CD20 antibody and Pneumocystis infection. In mice with primary infection, anti-CD20 antibody treatment depleted both CD19+ and CD27+ CD19+ cells but not T cells in the lung at days 14 and 28 after Pneumocystis inoculation. Although anti-CD20 antibody treatment impaired fungal clearance at day 14 postinfection, fungal burden in the lungs was substantially reduced at day 28 in both depleted and control mice in the low-dose group. Subcutaneous anti-CD20 antibody treatment did not alter antigen-specific serum immunoglobulin levels in mice compared with control mice, and there were no significant differences in the numbers of lung gamma interferon-positive (IFN-γ+) CD4+, interleukin 4-positive (IL-4+) CD4+, IL-5+ CD4+, and IL-17A+ CD4+ cells between depleted and control mice after infection. In mice with secondary infection, the lung fungal burden was comparable between depleted and control mice 14 days after reinfection. Low-dose subcutaneous anti-CD20 antibody treatment may delay fungal clearance, but it did not impair the ability of the host to clear Pneumocystis infection, irrespective of primary or secondary infection.IMPORTANCE Anti-CD20 antibody therapy is used for both cancer and autoimmune disease but has been shown to be associated with Pneumocystis pneumonia in humans. This study shows that low-dose subcutaneous anti-CD20 can modulate B cell populations without grossly perturbing fungal immunity against Pneumocystis lung infection.

Further Defining the Human Virome using NGS: Identification of Redondoviridae.

In this issue of Cell Host & Microbe, Abbas et al. (2019) uncover a previously undefined family of single-stranded DNA viruses, Redondoviridae, in human ororespiratory sites. The presence of Redondoviridae associates with critical illness such as respiratory failure and periodontitis, illustrating the power of metagenomics to define the human virome.