1. Volume 88, Issue 6, Pages 1374-1382 (December 2015)
Peripheral natural killer cell and allo-stimulated T-cell function in kidney transplant recipients associate with cancer risk and immunosuppression-related complications 
Christopher M. Hope, Alexander Troelnikov, William Hanf, Shilpanjali Jesudason, Patrick T. Coates, Peter S. Heeger, Robert P. Carroll 
Kidney International 
Volume 88, Issue 6, Pages (December 2015)
DOI: /ki
Copyright © 2015 International Society of Nephrology Terms and Conditions

2. Figure 1
Dichotomized natural killer (NK) cell function in kidney transplant recipients (KTRs). The depicted NK cell function is dichotomized on the median amount of lactate dehydrogenase (LDH) released when KTR peripheral blood mononuclear cells (PBMCs) are co-cultured with K562 at a ratio of 20:1, high NK function (black)/low NK function (gray). (a) Bar graph of 62 KTRs. The 22 KTRs with current cancer have a greater proportion of KTRs with low NK cell function (68%) compared with the 19 KTRs with past cancer (26%) and 21 KTRs with no cancer (33%) with a χ2-test P-value of (b) A Kaplan–Meier survival curve depicting the dichotomized data of the 41 KTRs with current and past cancer, developing metastatic cancer, or dying of immunosuppressive-related diseases (i.e., cancer or sepsis) over 500 days. KTRs with low NK cell function (gray line) have lower survival proportions at 365 days (32% vs. 71%) and lower overall survival with a hazard ratio (95% confidence interval)=2.1 (0.97–5.00), log-rank P=0.063, when compared with KTRs with high NK cell function (black line).
Kidney International  , DOI: ( /ki )
Copyright © 2015 International Society of Nephrology Terms and Conditions

3. Figure 2
Allo-stimulated interferon-γ (IFN-γ) release from kidney transplant recipients (KTRs). Allo-response was measured by co-culturing 1 × 105 allogeneic B cells with 3 × 105 peripheral blood mononuclear cells (PBMCs) from KTRs and quantitating IFN-γ released from cells (spots) via enzyme-linked immunospot (ELISPOT). The intra-assay coefficient of variation (C.V.) was determined to be 31%. (a) Box plot representing median (range) numbers of spots from 77 KTRs. The KTRs with current (n=29) and past (n=24) cancer, both have decreased IFN-γ allo-responses when compared with KTRs with no cancer (n=24, P<0.001) using a Kruskal–Wallis non-parametric test. (b) A receiver operator characteristic (ROC) curve for the 53 KTRs who had current or past cancer, who went on to developed metastatic cancer or died of immunosuppressive drug-related diseases (i.e., cancer or sepsis) over the median (range) of days followed=303 (8–1712). The ROC has an accuracy of 69% (P=0.025) with trade-off of 280 spots per 3 × 105 PBMC (spots) leading to a sensitivity of 53% and specificity of 79%. The 280 spots trade-off was identified using a single value that had both the highest likelihood ratio and Youden’s index. (c) A Kaplan–Meier survival curve for metastatic cancer or immunosuppressive-related death (cancer or sepsis)-free days dichotomized on the trade-off value of 280 spots, over 500 days. KTRs with <280 spots (gray line) have lower survival rates (23% vs. 61%, at 500 days), log-rank test P=0.050, when compared to KTRs with >280 spots (black line).
Kidney International  , DOI: ( /ki )
Copyright © 2015 International Society of Nephrology Terms and Conditions

4. Figure 3
Immunosuppressive drug regimen and cancer associations to natural killer (NK) cell function in kidney transplant recipients (KTRs). A panel of bar plots depicting NK cell function dichotomized on the median NK cell function of 62 KTRs, with various drug regimens and with either current cancer (Cancer) or past/no cancer (No Cancer). (a) KTRs on triple therapy (three drugs) do not differ from their one to two drug counterparts. However, KTRs with cancer have a greater proportion of KTRs with low NK cell function than the KTR with no cancer when on 1-2 drugs (63 vs. 19%, P=0.029) but does not reach statistical significance when on three drugs (83% vs. 35%, P=0.073). (b) KTRs on mycophenolate (MMF)-based regimens do not differ from their non-MMF-based counterparts. However, there was a greater proportion of KTRs with cancer that have low NK function compared with KTRs with no cancer, which is only statistically significant when KTRs were not on MMF-based regimens (67% vs. 18%, P=0.021). (c) KTRs on calcineurin inhibitor (CNI)-based regimens do not differ from their non-CNI counterparts. Although KTR with cancer on CNI and not on CNI have a greater proportion of KTR with low NK function (70% and 67%, respectively), these do not statistically differ from their non-cancer counterparts (30% and 36%, respectively) with (P=0.056 and P=0.116, respectively). All statistical analysis performed using Fisher’s exact tests.
Kidney International  , DOI: ( /ki )
Copyright © 2015 International Society of Nephrology Terms and Conditions

5. Figure 4
Immunosuppressive drug regimen and cancer associations to allo-stimulated interferon-γ (IFN-γ) release in kidney transplant recipients (KTRs). A panel of box plots depicting median (range) of allo-stimulated IFN-γ release from 77 KTRs with various drug regimens and with either current cancer (Cancer) or past/no cancer (No Cancer). (a) IFN-γ release from KTRs on triple therapy (three drugs) does not differ from their one to two drug counterparts. However, KTRs with cancer had less IFN-γ release than the KTRs with no cancer for those on three drugs (P=0.001). (b) KTRs on mycophenolate (MMF)-based regimens do not differ from their non-MMF-based counterparts. However, KTRs with cancer have lower IFN-γ release than KTRs with no cancer when on MMF-based regimens (P=0.014). (c) KTRs on calcineurin inhibitor (CNI)-based regimens do not differ from their non-CNI counterparts. However, KTRs with cancer have less IFN-γ release than the KTR with no cancer for those on CNI-based regimens (P=0.014). All statistical analysis performed using Mann–Whitney tests.
Kidney International  , DOI: ( /ki )
Copyright © 2015 International Society of Nephrology Terms and Conditions

6. Figure 5
Associations of kidney graft function and proteinuria (>500mg/day) to natural killer (NK) cell function and allo-stimulated interferon-γ (IFN-γ) release in kidney transplant recipients (KTRs). (a and b) Bar graphs depicting proportions of KTRs with high (black) and low (gray) NK cell function dichotomized on median NK cell function. (a) Bar graph of 62 KTRs. Estimated glomerular filtration rate (eGFR) does not associate to dichotomized NK cell function (P=0.276), using χ2-analysis. (b) Bar graph depicting 57 KTRs. There is a larger proportion of KTRs with high NK function (85%), with proteinuria (>500mg/day) compared with KTR with lower or no detectable proteinuria (48%, P=0.026) using a Fisher’s exact test. (c and d) Box plots depicting the median (range) of allo-stimulated IFN-γ (IFN-γ) release (Spots) from 3 × 105 peripheral blood mononuclear cells (PBMCs) measured by enzyme-linked immunospot (ELISPOT) in KTRs. (c) For 77 KTRs tested, there are no differences between the different groups of eGFR and amount of IFN-γ release (P=0.860), using a Kruskal–Wallis test. (d) Out of 71 KTRs, there is no difference between those KTRs with proteinuria (n=18) and those without (n=53, P=0.473), using a Mann–Whitney test.
Kidney International  , DOI: ( /ki )
Copyright © 2015 International Society of Nephrology Terms and Conditions