NKG2D blockade impairs tissue-resident memory T cell accumulation and reduces chronic lung allograft dysfunction
Kaveh Moghbeli, Madeline A. Lipp, Marta Bueno, Andrew Craig, Michelle Rojas, Minahal Abbas, Zachary I. Lakkis, Byron Chuan, John Sembrat, Kentaro Noda, Daniel J. Kass, Kong Chen, Li Fan, Tim Oury, Zihe Zhou, Xingan Wang, John F. McDyer, Oliver Eickelberg, Mark E. Snyder
Kaveh Moghbeli, Madeline A. Lipp, Marta Bueno, Andrew Craig, Michelle Rojas, Minahal Abbas, Zachary I. Lakkis, Byron Chuan, John Sembrat, Kentaro Noda, Daniel J. Kass, Kong Chen, Li Fan, Tim Oury, Zihe Zhou, Xingan Wang, John F. McDyer, Oliver Eickelberg, Mark E. Snyder
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Research Article Immunology Transplantation

NKG2D blockade impairs tissue-resident memory T cell accumulation and reduces chronic lung allograft dysfunction

Abstract

Chronic lung allograft dysfunction (CLAD) substantially limits long-term survival following lung transplantation. To identify potential targets for CLAD prevention, T cells from explanted CLAD lungs and lung-draining lymph nodes, as well as diseased and nondiseased controls were isolated and single-cell RNA sequencing and TCR sequencing were performed. TCR sequencing revealed a clonally expanded population of CD8+ tissue-resident memory T cells (TRMs) with high cytotoxic potential, including upregulation of KLRK1, encoding the co-receptor NKG2D. These cytotoxic CD8+ TRMs accumulated around the CLAD airways and had a 100-fold increase in clonal overlap with lung-draining lymph nodes when compared with non-CLAD lungs. Using a murine model of orthotopic lung transplantation, we confirmed that cytotoxic CD8+ TRM accumulation was due to chronic rejection and not transplantation alone. Furthermore, blocking NKG2D in vivo attenuated the airway remodeling following transplantation and diminished airway accumulation of CD8+ T cells. Our findings support NKG2D as a potential therapeutic target for CLAD, affecting cytotoxic CD8+ TRM accumulation.

Authors

Kaveh Moghbeli, Madeline A. Lipp, Marta Bueno, Andrew Craig, Michelle Rojas, Minahal Abbas, Zachary I. Lakkis, Byron Chuan, John Sembrat, Kentaro Noda, Daniel J. Kass, Kong Chen, Li Fan, Tim Oury, Zihe Zhou, Xingan Wang, John F. McDyer, Oliver Eickelberg, Mark E. Snyder

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Figure 9

NKG2D blockade attenuates pathophysiologic changes in murine model of CLAD.

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NKG2D blockade attenuates pathophysiologic changes in murine model of CL...
(A) Experimental approach. (B) Respiratory system elastance and inspiratory capacity after syngeneic single lung transplant (n = 3), allogeneic single lung transplant (n = 5), and allogeneic single lung transplant with NKG2D blockade (n = 6). (C and D) Trichrome staining of connective tissue deposition in mouse allogeneic lung transplants with and without NKG2D blockade at (C) ×4 and (D) ×10 magnification. (E) Cumulative data quantifying trichrome staining. (F) Representative flow cytometry plot showing gating strategy for NKG2D positivity (left) and UMAP of concatenated CD8+ T cells obtained from murine allografts showing major T cell phenotypes, including TEM (CD44+CD62L–), TCM (CD44+CD62L+), and naive cells (CD44–CD62L–) (middle), and UMAP highlighting the NKG2D+ cells in the protected (orange) and labeled (blue) compartments (includes cells from n = 7 allografts, 3 from PBS and 4 from NKG2D blocker, downsampled to have equal numbers of cells from each lung). (G) Cumulative data comparing percentage of all CD8+ T cells that are NKG2D+ within the protected versus labeled allografts; square symbols are those from PBS-treated and circles are those from NKG2D blocker–treated allografts. (H) Immunofluorescence imaging for CD8 (purple) and NK1.1 (yellow) in mouse allogeneic lung transplants with and without NKG2D blockade (first column at ×4, remaining at ×40). (I) CD8+ T cell concentration in mouse allogeneic lung transplants with (Treated) and without (Untreated) NKG2D blockade. *P < 0.05; **P < 0.01 by unpaired t test.