Vibrio cholerae Proteome-Wide Screen for Immunostimulatory Proteins Identifies Phosphatidylserine Decarboxylase as a Novel Toll-Like Receptor 4 Agonist

Recognition of conserved bacterial components provides immediate and efficient immune responses and plays a critical role in triggering antigen-specific adaptive immunity. To date, most microbial components that are detected by host innate immune system are non-proteinaceous structural components. In order to identify novel bacterial immunostimulatory proteins, we developed a new high-throughput approach called “EPSIA”, Expressed Protein Screen for Immune Activators. Out of 3,882 Vibrio cholerae proteins, we identified phosphatidylserine decarboxylase (PSD) as a conserved bacterial protein capable of activating host innate immunity. PSD in concentrations as low as 100 ng/ml stimulated RAW264.7 murine macrophage cells and primary peritoneal macrophage cells to secrete TNFα and IL-6, respectively. PSD-induced proinflammatory response was dependent on the presence of MyD88, a known adaptor molecule for innate immune response. An enzymatically inactive PSD mutant and heat-inactivated PSD induced ∼40% and ∼15% of IL-6 production compared to that by native PSD, respectively. This suggests that PSD induces the production of IL-6, in part, via its enzymatic activity. Subsequent receptor screening determined TLR4 as a receptor mediating the PSD-induced proinflammatory response. Moreover, no detectable IL-6 was produced in TLR4-deficient mouse macrophages by PSD. PSD also exhibited a strong adjuvant activity against a co-administered antigen, BSA. Anti-BSA response was decreased in TLR4-deficient mice immunized with BSA in combination with PSD, further proving the role of TLR4 in PSD signaling in vivo. Taken together, these results provide evidence for the identification of V. cholerae PSD as a novel TLR4 agonist and further demonstrate the potential application of PSD as a vaccine adjuvant

Beyond Refugia: New insights on Quaternary climate variation and the evolution of biotic diversity in tropical South America

Haffer’s (Science 165: 131–137, 1969) Pleistocene refuge theory has provided motivation for 50 years of investigation into the connections between climate, biome dynamics, and neotropical speciation, although aspects of the orig- inal theory are not supported by subsequent studies. Recent advances in paleocli- matology suggest the need for reevaluating the role of Quaternary climate on evolutionary history in tropical South America. In addition to the many repeated large-amplitude climate changes associated with Pleistocene glacial-interglacial stages (~40 kyr and 100 kyr cyclicity), we highlight two aspects of Quaternary climate change in tropical South America: (1) an east-west precipitation dipole, induced by solar radiation changes associated with Earth’s precessional variations (~20 kyr cyclicity); and (2) periods of anomalously high precipitation that persisted for centuries-to-millennia (return frequencies ~1500 years) congruent with cold “Heinrich events” and cold Dansgaard-Oeschger “stadials” of the North Atlantic region. The spatial footprint of precipitation increase due to this North Atlantic forcing extended across almost all of tropical South America south of the equator. Combined, these three climate modes present a picture of climate change with different spatial and temporal patterns than envisioned in the original Pleistocene refuge theory. Responding to these climate changes, biomes expanded and contracted and became respectively connected and disjunct. Biome change undoubtedly influenced biotic diversification, but the nature of diversification likely was more complex than envisioned by the original Pleistocene refuge theory. In the lowlands, intermittent forest expansion and contraction led to species dispersal and subsequent isolation, promoting lineage diversification. These pulses of climate-driven biotic interchange profoundly altered the composition of regional species pools and triggered new evolutionary radiations. In the special case of the tropical Andean forests adjacent to the Amazon lowlands, new phylogenetic data provide abundant evidence for rapid biotic diversification during the Pleistocene. During warm interglacials and intersta- dials, lowland taxa dispersed upslope. Isolation in these disjunct climate refugia led to extinction for some taxa and speciation for others.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/155561/1/Baker2020.pdfDescription of Baker2020.pdf : Main articl