A blood atlas of COVID-19 defines hallmarks of disease severity and specificity.

Treatment of severe COVID-19 is currently limited by clinical heterogeneity and incomplete description of specific immune biomarkers. We present here a comprehensive multi-omic blood atlas for patients with varying COVID-19 severity in an integrated comparison with influenza and sepsis patients versus healthy volunteers. We identify immune signatures and correlates of host response. Hallmarks of disease severity involved cells, their inflammatory mediators and networks, including progenitor cells and specific myeloid and lymphocyte subsets, features of the immune repertoire, acute phase response, metabolism, and coagulation. Persisting immune activation involving AP-1/p38MAPK was a specific feature of COVID-19. The plasma proteome enabled sub-phenotyping into patient clusters, predictive of severity and outcome. Systems-based integrative analyses including tensor and matrix decomposition of all modalities revealed feature groupings linked with severity and specificity compared to influenza and sepsis. Our approach and blood atlas will support future drug development, clinical trial design, and personalized medicine approaches for COVID-19

The Future of RIP2/RICK/CARDIAK as a Biomarker of the Inflammatory Response to Infection

Biological markers of disease have become increasingly important for the clinician to diagnose, predict and monitor progression, and assess the therapeutic effect of interventions on underlying pathogenic mechanisms. Robust and specific biomarkers would be very useful in inflammation, where they may facilitate early identification of tissue injury, predict disease progression and help to modify disease outcomes. However, at present, there are no robust biomarkers to predict the course of inflammation. Here, we discuss emerging data indicating that RIP2, a putative serine/threonine protein kinase, may serve as a biomarker for the resolution of peritoneal dialysis-associated peritonitis and, more generally, of the acute inflammatory response to infection

A Retrospective Comparison of the Safety and Efficacy of 3 months vs. 6 months Valganciclovir for Cytomegalovirus Prophylaxis in Renal Transplant Recipients

Pharmacy residents have the opportunity to complete a research project during their residency training, which provides them with skills on how to conduct and manage a research project. Projects often represent an area of interest and need that has been recognized by the host institution’s pharmacy department. Projects are presented as a poster at an annual CSHP Ontario Branch Residency Research Night, and many eventually go on to be published in a peer-reviewed journal.Abstract Background: Antiviral prophylaxis has been shown to be effective in reducing the risk of CMV disease in renal transplant recipients and reducing all-cause mortality in solid organ transplant recipients. Extending valganciclovir prophylaxis from 100 to 200 days was associated with a further reduction of CMV disease post-renal transplant. Longer valganciclovir prophylaxis can induce leukopenia, increase risk of other infections, and lead to alteration in immunosuppression, which may lead to rejection. It is currently unknown how the extension from 3 to 6-month prophylaxis with valganciclovir has impacted outcomes in our institution. Methods: A retrospective chart review was conducted from January 1st 2010 to May 31st 2014 (followed until May 31st 2015). Patients were included if they had received a renal transplant and were prescribed 3 months (group 1; from January 2010 to December 2011) or 6 months (group 2; from January 2012 to May 2014) of valganciclovir and were at least 18 years of age at time of transplant. Results: Both groups experienced high rates of leukopenia; 78% in 3-month prophylaxis group compared to 85% in 6-month prophylaxis group (P = 0.284). There is a statistically insignificant increase in patients who developed CMV viremia in 6-month prophylaxis group (19.8%) compared to the 3-month group (14.3%). There was one patient in 3-month prophylaxis group (2.0%) and three patients in 6-month prophylaxis group (3.5%) (P=0.633) who experienced acute rejection. Conclusion: The change of TOH Renal Transplant Protocol to extend duration of CMV prophylaxis from 3 to 6 months for high-risk recipients did not result in statistically significant change in incidence of leukopenia; CMV viremia; or rates of graft rejection

Mouse Nkrp1-Clr gene cluster sequence and expression analyses reveal conservation of tissue-specific MHC-independent immunosurveillance.

The Nkrp1 (Klrb1)-Clr (Clec2) genes encode a receptor-ligand system utilized by NK cells as an MHC-independent immunosurveillance strategy for innate immune responses. The related Ly49 family of MHC-I receptors displays extreme allelic polymorphism and haplotype plasticity. In contrast, previous BAC-mapping and aCGH studies in the mouse suggest the neighboring and related Nkrp1-Clr cluster is evolutionarily stable. To definitively compare the relative evolutionary rate of Nkrp1-Clr vs. Ly49 gene clusters, the Nkrp1-Clr gene clusters from two Ly49 haplotype-disparate inbred mouse strains, BALB/c and 129S6, were sequenced. Both Nkrp1-Clr gene cluster sequences are highly similar to the C57BL/6 reference sequence, displaying the same gene numbers and order, complete pseudogenes, and gene fragments. The Nkrp1-Clr clusters contain a strikingly dissimilar proportion of repetitive elements compared to the Ly49 clusters, suggesting that certain elements may be partly responsible for the highly disparate Ly49 vs. Nkrp1 evolutionary rate. Focused allelic polymorphisms were found within the Nkrp1b/d (Klrb1b), Nkrp1c (Klrb1c), and Clr-c (Clec2f) genes, suggestive of possible immune selection. Cell-type specific transcription of Nkrp1-Clr genes in a large panel of tissues/organs was determined. Clr-b (Clec2d) and Clr-g (Clec2i) showed wide expression, while other Clr genes showed more tissue-specific expression patterns. In situ hybridization revealed specific expression of various members of the Clr family in leukocytes/hematopoietic cells of immune organs, various tissue-restricted epithelial cells (including intestinal, kidney tubular, lung, and corneal progenitor epithelial cells), as well as myocytes. In summary, the Nkrp1-Clr gene cluster appears to evolve more slowly relative to the related Ly49 cluster, and likely regulates innate immunosurveillance in a tissue-specific manner

Wide-spread expression of <i>Clr-b</i> in lymphoid and non-lymphoid tissues and cell types.

<p>In situ hybridization was performed on tissue sections from a PBS-perfused B6 mouse. Hybridization of the indicated (A) primary and secondary lymphoid organs and (B) non-lymphoid organs was performed with a DIG-labeled anti-sense <i>Clr-b</i> RNA probe and revealed with an alkaline phosphatase-conjugated anti-DIG secondary mAb. Control in situ hybridization with a DIG-labeled <i>Clr-b</i> sense RNA probe is shown. Anti-sense <i>Clr-g</i> staining of hind leg muscle tissue is provided as a hybridization specificity control. H&E staining is provided to identify cell types and reveal organ structure. The scale bars indicate 200 µm in the lymph node and spleen, 500 µm in the liver and thymus, and 50 µm in the kidney, lung, heart, and muscle.</p

Comparison of repeats in the <i>Nkrp1-Clr</i> and <i>Ly49</i> gene clusters from B6 mice.^a

a<p>As detected by RepeatMasker.</p

Tissue and organ expression of <i>Nkrp1</i> genes as determined by RT-PCR.

a<p>-, no expression; −/+, very weak expression; +, weak to moderate expression; ++, strong expression; +++, very strong expression.</p>b<p>Microarray data were obtained from the Mouse MOE430 Gene Atlas database at BioGPS portal server in order to compare with results of RT-PCR.</p>c<p>ND, not determined.</p

Primers used to amplify <i>Nkrp1- Clr</i> genes from tissue cDNA and plasmids.

<p>Primers used to amplify <i>Nkrp1- Clr</i> genes from tissue cDNA and plasmids.</p