APOL1 risk alleles modulate T cell receptor signaling to promote allograft rejection
John Pell, EM Tanvir, Zeguo Sun, Irene Chernova, Anand Reghuvaran, Soichiro Nagata, Mateus T. Guerra, John Choi, Soltan Al Chaar, Hiroki Mizuno, Ke Dong, Xin Tian, Reika Ishibe, Barbara Franchin, Paolo Cravedi, Ashwani Kumar, Gabriel Barsotti, Hongmei Shi, Bony De Kumar, Shinobu Smithson, Wenzhi Song, John Cijiang He, Anita S. Chong, Jordan S. Pober, Stefan Somlo, Ian W Gibson, Waldemar Popik, Zhongyang Zhang, Joseph Craft, Jamil Azzi, Naoka Murakami, Shuta Ishibe, Peter S Heeger, Madhav C. Menon
John Pell, EM Tanvir, Zeguo Sun, Irene Chernova, Anand Reghuvaran, Soichiro Nagata, Mateus T. Guerra, John Choi, Soltan Al Chaar, Hiroki Mizuno, Ke Dong, Xin Tian, Reika Ishibe, Barbara Franchin, Paolo Cravedi, Ashwani Kumar, Gabriel Barsotti, Hongmei Shi, Bony De Kumar, Shinobu Smithson, Wenzhi Song, John Cijiang He, Anita S. Chong, Jordan S. Pober, Stefan Somlo, Ian W Gibson, Waldemar Popik, Zhongyang Zhang, Joseph Craft, Jamil Azzi, Naoka Murakami, Shuta Ishibe, Peter S Heeger, Madhav C. Menon
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Research Article Immunology Nephrology

APOL1 risk alleles modulate T cell receptor signaling to promote allograft rejection

Abstract

The exonic variants G1 and G2 in apolipoprotein-L1 (APOL1) are linked to an increased risk of kidney disease as well as kidney transplant rejection. Outside of the association of these prevalent variants with African ancestry, the underpinning causal mechanisms for rejection are unknown. We investigated T cell function using transgenic mice with physiologic expression of WT (G0), G1 APOL1 (G1), or G2 APOL1 (G2). Mice with the G1 or G2 variant showed greater CD8+ T cell activation with expansion of a central memory T cell (Tcm) subset. Stimulated G1 CD8+ T cells showed enhanced proliferation and cytokine production, which was reversed with APOL1 inhibition. In MHC-mismatched cardiac transplants, G1 mice demonstrated greater CD8+ T cell infiltration and worse survival. The bulk transcriptome of G1 CD8+ T cells and the single-cell transcriptome of graft-infiltrating Tcms showed enrichment of canonical T cell receptor (TCR) pathways including Ca2+ signaling. G1 CD8+ T cells demonstrated baseline ER Ca2+ depletion followed by sustained increases in cytosolic Ca2+ upon TCR stimulation. G1 CD8+ T cells were more sensitive to Ca2+ chelation, or store-operated Ca2+ entry inhibition, and were relatively resistant to calcineurin antagonism compared with G0 CD8+ T cells. Analogously, in a kidney transplant cohort, transplant recipients carrying an APOL1 risk variant (G1 or G2) who had elevated peripheral Tcms before transplantation developed rejection despite having significantly higher tacrolimus levels than recipients with the G0/G0 APOL1 genotype. In summary, we have unraveled an excitatory mechanism for APOL1 variants in T cells that causally links them to kidney rejection.

Authors

John Pell, EM Tanvir, Zeguo Sun, Irene Chernova, Anand Reghuvaran, Soichiro Nagata, Mateus T. Guerra, John Choi, Soltan Al Chaar, Hiroki Mizuno, Ke Dong, Xin Tian, Reika Ishibe, Barbara Franchin, Paolo Cravedi, Ashwani Kumar, Gabriel Barsotti, Hongmei Shi, Bony De Kumar, Shinobu Smithson, Wenzhi Song, John Cijiang He, Anita S. Chong, Jordan S. Pober, Stefan Somlo, Ian W Gibson, Waldemar Popik, Zhongyang Zhang, Joseph Craft, Jamil Azzi, Naoka Murakami, Shuta Ishibe, Peter S Heeger, Madhav C. Menon

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Figure 8

ER calcium depletion causes activation of variant APOL1 CD8+ T cells.

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ER calcium depletion causes activation of variant APOL1 CD8+ T cells. (A...
(A–C) (A) Representative images (dots = T cells), (B) mean calcium flux curves, and (C) corresponding average AUCs of fluorescence measured from live G0 and G1 CD8+ T cells in response to thapsigargin (n = 5 mice each). Original magnification, ×20 (A). (D–F) Summarized calcium flux curve (D), AUCs of early versus late fluorescence (<50 seconds or >50 seconds) (E), and total fluorescence from CD8+ T cells from G1 versus G0 after anti-CD3-stimulation (>500 cells, n >6 mice) (F). (G) Representative plot and (H) quantified undivided total CD8+ T cells from G1 versus G0 mice with YM-58483 at 50 nM. (I) Proliferation plots of G1CD8+ T cells treated with BAPTA-AM (0.3 μM) and DMSO and (J) quantification of proliferation of G0 or G1 CD8+ T cells treated with BAPTA-AM (0.3 μM) or DMSO (triplicates of 5 mice), respectively. (K) Representative plots of G1CD8+ T cells treated with TAC at 5 and 10 ng/mL and (L) and the corresponding quantification of proliferation of G0 and G1 CD8+ T cells with 0, 5, and 10 ng/mL TAC. Box-and-whisker plots show the median and range of triplicates pooled from more than 5 mice each. *P < 0.05, ***P < 0.001, and ****P < 0.0001, by 1-way ANOVA with Tukey’s post hoc test.