a qualitative study

Objectives: It is known that transition, as a shift of care, marks a vulnerable phase in the adolescents’ lives with an increased risk for non- adherence and allograft failure. Still, the transition process of adolescents and young adults living with a kidney transplant in Germany is not well defined. The present research aims to assess transition-relevant structures for this group of young people. Special attention is paid to the timing of the process. Setting: In an observational study, we visited 21 departments of paediatric nephrology in Germany. Participants were doctors (n=19), nurses (n=14) and psychosocial staff (n=16) who were responsible for transition in the relevant centres. Structural elements were surveyed using a short questionnaire. The experiential viewpoint was collected by interviews which were transcribedverbatim before thematic analysis was performed. Results: This study highlights that professionals working within paediatric nephrology in Germany are well aware of the importance of successful transition. Key elements of transitional care are well understood and mutually agreed on. Nonetheless, implementation within daily routine seems challenging, and the absence of written, structured procedures may hamper successful transition. Conclusions: While professionals aim for an individual timing of transfer based on medical, social, emotional and structural aspects, rigid regulations on transfer age as given by the relevant health authorities add on to the challenge. Trial registration number: ISRCTN Registry no 22988897; results (phase I) and pre-results (phase II)

The TRANSNephro-study examining a new transition model for post-kidney transplant adolescents and an analysis of the present health care: study protocol for a randomized controlled trial

Kreuzer M, Prüfe J, Bethe D, et al. The TRANSNephro-study examining a new transition model for post-kidney transplant adolescents and an analysis of the present health care: study protocol for a randomized controlled trial. Trials. 2014;15(1): 505

CD49a Expression Identifies a Subset of Intrahepatic Macrophages in Humans

Macrophages play central roles in inflammatory reactions and initiation of immune responses during infections. More than 80% of total tissue macrophages are described to be located in the liver as liver-resident macrophages, also named Kupffer cells (KCs). While studies in mice have established a central role of liver-resident KCs in regulating liver inflammation, their phenotype and function are not well-characterized in humans. Comparing paired human liver and peripheral blood samples, we observed significant differences in the distribution of macrophage (Mφ) subsets, with lower frequencies of CD14hiCD16lo and higher frequencies of CD14int−hiCD16int Mφ in human livers. Intrahepatic Mφ consisted of diverse subsets with differential expression of CD49a, a liver-residency marker previously described for human and mice NK cells, and VSIG4 and/or MARCO, two recently described human tissue Mφ markers. Furthermore, intrahepatic CD49a+ Mφ expressed significantly higher levels of maturation and activation markers, exhibited higher baseline levels of TNF-α, IL-12, and IL-10 production, but responded less to additional in vitro TLR stimulation. In contrast, intrahepatic CD49a− Mφ were highly responsive to stimulation with TLR ligands, similar to what was observed for CD49a− monocytes (MOs) in peripheral blood. Taken together, these studies identified populations of CD49a+, VSIG4+, and/or MARCO+ Mφ in human livers, and demonstrated that intrahepatic CD49a+ Mφ differed in phenotype and function from intrahepatic CD49a− Mφ as well as from peripheral blood-derived monocytes

Unsupervised analysis of intrahepatic CD49a+ and CD49a- NK cells.

<p><b>(A)</b> Gated CD49a+ and <b>(B)</b> CD49a- NK cells from 19 donors were concatenated and represented in t-SNE maps for the expression of chemokine receptors, activation and residency markers. Color coding indicates the expression intensity of the surface marker, pink being higher expressed and green being lower expressed.</p

Proliferative capacity exhibited by human liver-resident CD49a+CD25+ NK cells

<div><p>The recruitment and retention of Natural Killer (NK) cells in the liver are thought to play an important role during hepatotropic infections and liver cirrhosis. The aims of this study were to determine differences between liver-derived and peripheral blood-derived NK cells in the context of liver inflammation and cirrhosis. We conducted a prospective dual-center cross-sectional study in patients undergoing liver transplantation or tumor-free liver resections, in which both liver tissue and peripheral blood samples were obtained from each consenting study participants. Intrahepatic lymphocytes and PBMCs were stained, fixed and analyzed by flow cytometry. Our results showed that, within cirrhotic liver samples, intrahepatic NK cells were particularly enriched for CD49a+ NK cells when compared to tumor-free liver resection samples. CD49a+ liver-derived NK cells included populations of cells expressing CD25, CD34 and CXCR3. Moreover, CD49a+CD25+ liver-derived NK cells exhibited high proliferative capacity <i>in vitro</i> in response to low doses of IL-2. Our study identified a specific subset of CD49a+CD25+ NK cells in cirrhotic livers bearing functional features of proliferation.</p></div

Immune phenotyping of combined peripheral and intrahepatic NK cells.

<p>Gated NK cells from 19 donors were concatenated and represented in t-SNE maps for the expression of chemokine receptors, activation and residency markers. <b>(A)</b> peripheral and <b>(B)</b> intrahepatic NK cells are shown. Color coding indicates the expression intensity of the surface marker, pink being higher expressed and green being lower expressed. <b>(C)</b> Proportion of NK cells derived from the liver (ihNK) and the peripheral blood (pNK) on the liver transplantation cohort expressing CD49a (pNK median (IQR): 0.9 (0.3–3.9); ihNK median (IQR): 34.4 (27.6–40.5); p<0.0001), CD34 (pNK median (IQR): 2.2 (1–4.7); ihNK median (IQR): 12 (6.8–20.9); p<0.0001), CXCR4 (pNK median (IQR): 9.8 (4.9–22.2); ihNK median (IQR): 3.4 (1.3–7.7); p = 0.0024), CD57 (pNK median (IQR): 19 (22–38.5); ihNK median (IQR): 13.7 (9.4–23.3); p<0.0001) and DNAM-1 (pNK median (IQR): 79.6 (51.5–85.6); ihNK median (IQR): 26.5 (8.5–32.1); p<0.0001) (n = 19). <b>(D)</b> Proportion of NK cells from the tumor-free liver resections expressing CD49a, CD34, CD57, DNAM-1, CXCR3 and CXCR4 within the IHLs NK cells and pNK cells (n = 5). <b>(E)</b> Frequency of CD49a+ NK cell population within the IHLs NK cells in tumor-free liver resection cohort (HLR) and the liver retransplant cohort (cirrhotic livers, CL). Data is depicted as scatter plot, with each dot corresponding to a participant. Bars indicate median and IQR. Wilcoxon signed rank tests with adjustment of p-values by false discovery rate.</p

Unsupervised analysis of intrahepatic CD49a+ and CD49a- NK cells.

<p><b>(A)</b> Gated CD49a+ and <b>(B)</b> CD49a- NK cells from 19 donors were concatenated and represented in t-SNE maps for the expression of chemokine receptors, activation and residency markers. Color coding indicates the expression intensity of the surface marker, pink being higher expressed and green being lower expressed.</p

Demographics and clinical characteristics from explanted liver tissue samples.

<p>Demographics and clinical characteristics from explanted liver tissue samples.</p

Effects of cytokine stimulation on CD98 expression: Samples were compared using Wilcoxon matched-pairs signed rank tests and multiplicity was controlled for by FDR testing.

<p>Bars indicate the median, significance was defined as p≤0.05 (*). A. Representative histograms of CD98 expression on unstimulated and stimulated CD56^bright CXCR6⁺ (I, grey: unstimulated; purple: stimulated), CD56^dim CXCR6⁺ (II, grey: unstimulated; teal: stimulated), CD56^bright CXCR6^- (III, grey: unstimulated; purple: stimulated) and CD56^dim CXCR6^- (IV, grey: unstimulated; teal: stimulated) NK cells from blood (left), liver (middle) and spleen (right) samples. B. Expression (MdFI) of CD98 on unstimulated (grey) and CD56^brightCD16^- (purple) and CD56^dimCD16⁺ (teal) tissue-resident (TR), tissue-derived (TD) and peripheral blood (PB) NK cells from paired liver-blood (left diagram, n = 12) and spleen-blood (right diagram, n = 11) samples.</p

NK cell phenotype and baseline nutrient receptor expression: Samples were compared using Wilcoxon matched-pairs signed rank tests and multiplicity was controlled for by FDR testing.

<p>Bars indicate the median, significance was defined as p≤0.05 (*). A. viSNE representation of peripheral blood- (PBMC, top row), liver- (middle row) and spleen (bottom row) derived NK cells and their expression of CD56, CD16, CXCR6, CD57 and CD127. B. Expression (Median fluorescence intensity, MdFI) of Glut1 on CD56^brightCD16^- (purple) and CD56^dimCD16⁺ (teal) tissue-resident (TR), tissue-derived (TD) and peripheral blood (PB) NK cells from paired liver-blood (left diagram, n = 12) and spleen-blood (right diagram, n = 11) samples. C. Expression (MdFI) of CD98 on CD56^brightCD16^- (purple) and CD56^dimCD16⁺ (teal) tissue-resident (TR), tissue-derived (TD) and peripheral blood (PB) NK cells from paired liver-blood (left diagram, n = 12) and spleen-blood (right diagram, n = 11) samples. D. Expression (MdFI) of CD71 on CD56^brightCD16^- (purple) and CD56^dimCD16⁺ (teal) tissue-resident (TR), tissue-derived (TD) and peripheral blood (PB) NK cells from paired liver-blood (left diagram, n = 12) and spleen-blood (right diagram, n = 11) samples.</p