Kaplan Meier survival curve showing death censored graft survival for the presence or absence of HLA-C2 allele in the recipient.

<p>The presence of an HLA-C2 allele in the recipient was associated with a significant improvement in death censored graft survival (10-year survival: 74.2% versus 54.3%).</p

Cox regression multivariable analysis for all factors that may influence outcomes after kidney transplantation.

<p>Statistically significant associations are indicated in bold.</p

Demographics and distribution of parameters that are known to affect graft and patient survival after kidney transplantation.

<p>As the analysis involved the comparison between absence of HLA-C2 in the recipient versus presence of HLA-C2 in the recipient groups were divided accordingly. Absence of HLA-C2 represents HLA-C1 homozygous whereas presence of HLA-C2 combines both heterozygous and HLA-C2 homozygous groups. CI is 95% confidence interval.</p><p>*indicates significance by ANOVA test,</p>+<p>indicates significance by Kendall's tau-b test.</p

Causes of graft loss following kidney transplantation.

<p>Causes of graft loss following kidney transplantation.</p

Kaplan Meier survival curve showing patient survival for the presence or absence of HLA-C2 allele in the recipient.

<p>The presence of an HLA-C2 allele in the recipient was associated with a significant improvement in patient survival (10-year survival: 88.5% versus 80.4%).</p

Comparisons are made for ΔMFI of CD86, HLA-DR and CCR7 expression between DCs with either HLA-C1 or HLA-C2 homozygous allele.

<p>Data shown are ΔMFI of CD86, HLA-DR and CCR7 expressed by DC in NK-DC co-culture in the presence of 1 ng/ml IL-15 at cell ratios of either 1∶1 or 1∶5. ΔMFI are calculated as the difference of MFI for DC in co-culture versus DC in isolation i.e. spontaneous expression. In NK-DC co-culture, in the presence of IL-15, DC with HLA-C1 homozygous allele express more co-stimulation molecules, and MHC class II molecules than DC with HLA-C2 homozygous alleles. Furthermore expression of trafficking chemokine CCR7 is virtually exclusive to HLA-C1 homozygotes indicating their predominant role in T-cell immune priming in secondary lymphoid tissues. Data shown for 4 independent experiments performed in each group and * indicates statistical significance with p<0.05 by Mann Whitney U test.</p

Flow cytometer data illustrating the impact of IL-15 (1 ng/ml) treated NK-DC co-culture on the expression of DC maturation (HLA-DR FITC) and chemokine (CCR7 PE) markers, comparing responses for DCs with either HLA-C1 (<b>figures 6b, d, f</b>) or HLA-C2 (<b>figures 6c, e, g</b>) homozygous alleles.

<p>(a) shows DC stained with isotype control for HLA-DR & CCR7, (b) & (c) shows background DC staining where DCs are in isolation in the presence of IL-15, (d) & (e) shows HLA-DR & CCR7 expression by DC in NK-DC co-culture at ratios of 1∶1 in the presence of IL-15, (f) & (g) shows DC markers in NK-DC co-culture at ratios 1∶5 in the presence of IL-15. This data clearly demonstrates that in NK-DC co-culture at ratios of 1∶1 in the presence of IL-15, HLA-C1 homozygous DCs undergo significantly greater maturation than HLA-C2 homozygous DCs and attain CCR7 chemokine expression required for trafficking to secondary lymphoid tissues. Results are representative of 4 experiments with 5,000 DC gated events captured.</p

Immunohistochemistry slide demonstrating donor derived CD56 positive cells (anti-CD56 antibody staining brown in colour: arrow as indicator) in pre-transplant kidney biopsy tissue.

<p>In brief, the method for development of this slide included dewaxing and antigen retrieval obtained by W-cap system (Bio-Optica) and staining with mouse monoclonal anti-CD56 antibody (IgG2b Novocastra) used at a dilution of 1∶50 and visualised with the EnVision detection system (DAKO). This image is representative of the observation made for biopsies taken from five different kidney transplants studied. Cell counts (degree of infiltration) were performed using light microscopy and counting 10 randomly selected high power fields at a magnification of 400× (area = 0.17 mm²). Mean (±SEM) of 3±2 CD56 positive cells were identified per high power field.</p

Supernatants were collected from NK-DC co-culture experiments at cell ratios of 1∶5 treated with IL-15 and tested for cytokine synthesis using 25 cytokine multiplex bead immunoassay.

<p>Data shown are for all the cytokines that were expressed in culture supernatants. Of these, comparisons were made between cytokine synthesis for DCs with either HLA-C1 homozygous allele or DCs with HLA-C2 homozygous alleles. Data shown excludes background synthesis by cells in isolation. These data indicates that DC with HLA-C1 alleles express significantly greater pro-inflammatory cytokines in NK-DC co-culture favouring immune maturation whereas DC with HLA-C2 alleles predominantly express anti-inflammatory cytokines that are inhibiting to DC maturation. Means ± SEM for 4 independent experiments in each group are shown and * indicates statistical significance with p<0.05 by Mann-Whitney U test.</p

Kaplan Meier survival curve showing death non-censored graft survival for the presence or absence of HLA-C2 allele in the recipient.

<p>The presence of an HLA-C2 allele in the recipient was associated with a significant improvement in death non-censored graft survival (10-year survival: 65.7% versus 43.8%).</p