The Use of Technology to Support Precision Health in Nursing Science

PurposeThis article outlines how current nursing research can utilize technology to advance symptom and self‐management science for precision health and provides a roadmap for the development and use of technologies designed for this purpose.ApproachAt the 2018 annual conference of the National Institute of Nursing Research (NINR) Research Centers, nursing and interdisciplinary scientists discussed the use of technology to support precision health in nursing research projects and programs of study. Key themes derived from the presentations and discussion were summarized to create a proposed roadmap for advancement of technologies to support health and well‐being.ConclusionsTechnology to support precision health must be centered on the user and designed to be desirable, feasible, and viable. The proposed roadmap is composed of five iterative steps for the development, testing, and implementation of technology‐based/enhanced self‐management interventions. These steps are (a) contextual inquiry, focused on the relationships among humans, and the tools and equipment used in day‐to‐day life; (b) value specification, translating end‐user values into end‐user requirements; (c) design, verifying that the technology/device can be created and developing the prototype(s); (d) operationalization, testing the intervention in a real‐world setting; and (e) summative evaluation, collecting and analyzing viability metrics, including process data, to evaluate whether the technology and the intervention have the desired effect.Clinical RelevanceInterventions using technology are increasingly popular in precision health. Use of a standard multistep process for the development and testing of technology is essential.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/151985/1/jnu12518.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/151985/2/jnu12518_am.pd

Double Negative (CD3+4−8−) TCRαβ Splenic Cells from Young NOD Mice Provide Long-Lasting Protection against Type 1 Diabetes

-reactive T-cells. Herein, we analyzed the function and phenotype of DNCD3 splenic cells in young NOD mice predisposed to several autoimmune disorders among which, the human-like autoimmune diabetes. with a predominant Vβ13 gene usage.T-regulatory cells. DNCD3 splenic cells could be potentially manipulated towards the development of autologous cell therapies in autoimmune diabetes

Increased Membrane Cholesterol in Lymphocytes Diverts T-Cells toward an Inflammatory Response

Cell signaling for T-cell growth, differentiation, and apoptosis is initiated in the cholesterol-rich microdomains of the plasma membrane known as lipid rafts. Herein, we investigated whether enrichment of membrane cholesterol in lipid rafts affects antigen-specific CD4 T-helper cell functions. Enrichment of membrane cholesterol by 40–50% following squalene administration in mice was paralleled by an increased number of resting CD4 T helper cells in periphery. We also observed sensitization of the Th1 differentiation machinery through co-localization of IL-2Rα, IL-4Rα, and IL-12Rβ2 subunits with GM1 positive lipid rafts, and increased STAT-4 and STAT-5 phosphorylation following membrane cholesterol enrichment. Antigen stimulation or CD3/CD28 polyclonal stimulation of membrane cholesterol-enriched, resting CD4 T-cells followed a path of Th1 differentiation, which was more vigorous in the presence of increased IL-12 secretion by APCs enriched in membrane cholesterol. Enrichment of membrane cholesterol in antigen-specific, autoimmune Th1 cells fostered their organ-specific reactivity, as confirmed in an autoimmune mouse model for diabetes. However, membrane cholesterol enrichment in CD4+ Foxp3+ T-reg cells did not alter their suppressogenic function. These findings revealed a differential regulatory effect of membrane cholesterol on the function of CD4 T-cell subsets. This first suggests that membrane cholesterol could be a new therapeutic target to modulate the immune functions, and second that increased membrane cholesterol in various physiopathological conditions may bias the immune system toward an inflammatory Th1 type response

Genotype Prevalence and Risk Factors for Severe Clinical Adenovirus Infection, United States 2004-2006

Recently, epidemiological and clinical data have revealed important changes with regard to clinical adenovirus infection, including alterations in antigenic presentation, geographical distribution, and virulence of the virus

Tyrosine phosphorylation patterns of major signaling modules of T helper differentiation upon enrichment of membrane cholesterol.

<p>(<b>A</b>) SDS-PAGE silver stain of pooled, negatively-sorted CD4⁺ splenic T-cell lysates from F1 mice (n = 4/group) untreated (<i>left lane</i>) or squalene treated (single dose of 180 µg, <i>right lane</i>) 3 days post-injection shows no detectable quantitative alterations in the protein bands between the two groups of mice. (<b>B</b>) Tyrosine phosphorylation patterns of the same samples in panel <i>A</i> were blotted with anti-phosphorylated tyrosine Ab-HRP conjugate. Of note, the amount of 55–100 kDa phosphorylated protein bands is increased in mice treated with squalene. (<b>C</b>) Immunoprecipitation of pooled splenic CD4⁺ T-cell lysates from F1 mice treated or not with squalene (180 µg/mouse, n = 4/group) 3 days post-injection was carried out for IL-12Rβ2, IL-2Rα, IL-4Rα, CD28, or CD3 receptors using specific Abs, and probed with specific anti-phospho Abs for STAT4, STAT5, STAT6, PI3K, and ZAP-70 kinases. Only the phosphorylated STAT-4, STAT-5, and ZAP-70 in squalene treated mice were significantly enhanced. Shown is one of two representative experiments.</p

Alteration in cytokine receptors mRNA expression after squalene enrichment of membrane cholesterol in resting lymphocytes.

<p>(<b>A</b>) Quantitative real-time RT-PCR of IL-4Rα, IL-12Rβ2, and IL-2Rα mRNA extracted from peripheral blood lymphocytes of individual F1 mice (n = 5/group) analyzed before squalene treatment (dark bars) and 7 days after squalene injection (180 µg/mouse) (light bars). Y axis indicates the mean fold increase in mRNA expression level relative to the endogenous 18S rRNA expression level (control ± SD). Shown are two combined separate experiments (*<i>p</i> values<0.05). (<b>B</b>) Aliquots samples in panel A were stained with CD4-FITC conjugate, co-stained either with IL-4Rα-PE or IL-12Rβ2-PE or IL-2Rα-PE conjugates, and analyzed by FACS at the single-cell level for the surface IL-Rs expression level based on MFI measurements. Shown are the IL-Rs MFI values ± SD measured in individual mice before and after squalene treatment. Of note, no significant changes occurred in the IL-Rs expression on cell surface after squalene treatment (*<i>p</i> values>0.05).</p

Squalene administration leads to accumulation of membrane cholesterol in resting CD4 T-cells.

<p>(<b>A</b>) F1 hybrid mice (n = 5/group) were injected i.p. or not (purple line) with a single dose (black line) or 4 doses of squalene (red line) within a week interval (180 µg/dose/mouse). Seven days after the last injection, negatively-sorted splenic CD4 cells from individual mice were co-stained with CD3-PE, CD4-FITC and Filipin III. Shown is the amount of cholesterol in plasma cell membrane of gated CD3⁺CD4⁺ splenic cells as measured by MFI of Filipin III in FACS at single-cell level in one representative mouse from each group (<i>left panel</i>). <i>Right panel</i>, F1 hybrid mice (n = 7/group) were injected i.p. (black line) or not (red line) with a single dose of squalene (180 µg/mouse) and 7 days later negatively-sorted splenic CD4 cells from individual mice were co-stained with CD3-PE, CD4-FITC, and Filipin III. Shown is the percentage of gated CD4⁺ T-cells ± standard deviation (SD) and MFI values of Filipin III ± SD before and after squalene injection as collected among 700 cell events in gated population of CD3⁺4⁺ T-cells for one of three representative experiments. (<b>B</b>) Cholesterol accumulation in the spleen was identified by Oil Red O (ORO) staining of frozen spleen sections, counter-stained with hematoxylin from untreated or squalene treated mice (180 µg/mouse) given one or four doses, and analyzed 7 days post-injection (n = 3/group). <i>Left panel,</i> spleen section from untreated mouse. <i>Middle panels,</i> spleen section from squalene treated mice. <i>Right panel,</i> positive control for ORO lipid droplet staining in adipocytes. Shown is one representative ORO stained section in each group. Dark arrows indicate ORO stain. (<b>C</b>) Quantitative real-time RT-PCR of HMG-CoA reductase mRNA and Squalene epoxidase mRNA extracted from negatively-sorted CD4 splenocytes isolated from individual F1 hybrid mice (n = 5/group) that were treated (light bars) or not treated (dark bars) with a single dose of squalene (180 µg/mouse) and analyzed 7 days post-injection. Y axis indicates the mean fold increase in mRNA expression level relative to the endogenous 18S rRNA expression level (control ± standard deviation (SD). Shown are two combined separate experiments (*<i>p</i> value<0.05). (<b>D</b>) FACS measurements of CD3⁺4⁺<i>Foxp3</i>⁺ T-reg cells from negatively-sorted CD4⁺ splenic cells of the same F1 mouse groups analyzed in panel A that were co-stained with CD3-PE and Filipin III. Shown is the percentage of gated CD4⁺Foxp3⁺ T-reg cells ± SD and MFI values of Filipin III ± SD collected among 500 cell events in the gated population of GFP⁺-<i>Foxp3</i>/GFP⁺ cells from one mouse in each group from two representative experiments.</p

Squalene induced accumulation of membrane cholesterol in resting CD4 T cells favors Th1 polarization under antigen-specific or non antigen specific stimulation.

<p>(A) Isolated adherent splenocytes (APCs, 5×10⁵) from individual F1 mice treated or not with 180 µg of squalene, were pulsed (+) or not pulsed (-) with HA_110–120 synthetic peptide (40 µg/mL/10⁶ cells) <i>in vitro</i> 7 days after squalene injection, and co-cultured with negatively-sorted CD4 splenic T-cells (10⁶ cells) from the same groups of mice, treated or not with squalene (Sq) (n = 4/group). Various cell co-culture combinations are shown on the X-axis, where (+) indicates presence and (–) indicates absence from the culture. Cell-culture supernatants were collected 24–48 h later, and secretion of IL-2, IL-4, and IFN-γ (Y-axis) was measured in pg/ml by Luminex. Bars represent average ± SD. Differences among groups were highly significant (<i>p</i><0.001) for cytokines with the following exceptions denoted as §: For IL-4, co-culutre in lane 6 differed significantly from lane 5 (<i>p</i> = 0.047) but not from lane 3 (<i>p</i> = 0.097). No significant difference was observed between co-culture in lane 3 and 5 for any of the three cytokines. (<b>B</b>) Intracellular cytokine staining for IFN-γ (<i>left panels</i>) and IL-4 (<i>right panels</i>) in splenic cells and CD4-gated splenic cells from individual untreated (<i>top panels</i>) and squalene treated F1 mice (<i>bottom panels</i>) (n = 3/group) were stimulated for 48 h with anti-CD3/CD28 Abs (2.5 µg each/10⁶ cells). Shown are the overlapped FACS histograms of gated CD4⁺ T cells synthesizing IL-4 or IFN-γ (red cell events), and total splenic cells (dark cell events) from squalene treated or untreated F1 mice, 7 days after squalene treatment. R1 gate indicates the low-proliferating cell population whereas the R2 gate indicates the high proliferating cell population in each experiment. Dead cells are shown in the un-gated cell population below the R1 gate. Shown is one of two representative experiments.</p

Co-localization of cytokine receptors with lipid rafts of resting CD4 T-cells before and after squalene treatment.

<p>Resting CD4 T-cells from individual F1 mice (n = 5/group) before and 7 days after squalene treatment (180 µg/mouse) were analyzed for interleukin receptor expression and distribution at the single-cell level by CLSM. Cells were stained with IL-4Rα-, IL-12Rβ2-, or IL-2Rα-PE conjugates, and co-stained for GM1 ganglioside by CTB- FITC conjugate and for nuclei with DAPI. First column indicates single-channel color for DAPI staining (blue), second column indicates GM1 staining (green), third column indicates interleukin receptor (IL-Rs) staining (red), and last column indicates merged channels at X63 magnification. <i>Top-two rows</i>, indicate cells from untreated (<i>upper row</i>) and squalene treated mice (<i>lower row</i>) stained for IL-4Rα. <i>Middle-two rows</i>, indicate cells from untreated (<i>upper row</i>) and squalene treated mice (<i>lower row</i>) stained for IL-12Rβ2. <i>Bottom-two rows</i>, indicate cells from untreated (<i>upper row</i>) and squalene treated mice (<i>lower row</i>) stained for IL-2Rα. Arrows indicate presence of IL-Receptor co-expression with the GM1 resident of lipid rafts. Enlargements of the merged channels are depicted to the <i>right</i> along with two different angles of the membrane for each IL-Receptor at X220 magnification. Shown are representative images in one of three experiments.</p