Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Nov 8;6(21):e153175.
doi: 10.1172/jci.insight.153175.

NK and CD8+ T cell phenotypes predict onset and control of CMV viremia after kidney transplant

Harry Pickering  1 , Subha Sen  1 , Janice Arakawa-Hoyt  2 , Kenichi Ishiyama  2 , Yumeng Sun  1 , Rajesh Parmar  1 , Richard S Ahn  3   4 , Gemalene Sunga  1 , Megan Llamas  1 , Alexander Hoffmann  4   5 , Mario Deng  5 , Suphamai Bunnapradist  6 , Joanna M Schaenman  5 , David W Gjertson  1   7 , Maura Rossetti  1 , Lewis L Lanier  2 , Elaine F Reed  1 ; CMV Systems Immunobiology Group
Affiliations

NK and CD8+ T cell phenotypes predict onset and control of CMV viremia after kidney transplant

Harry Pickering et al. JCI Insight. .

Abstract

CMV causes mostly asymptomatic but lifelong infection. Primary infection or reactivation in immunocompromised individuals can be life-threatening. CMV viremia often occurs in solid organ transplant recipients and associates with decreased graft survival and higher mortality. Furthering understanding of impaired immunity that allows CMV reactivation is critical to guiding antiviral therapy and examining the effect of CMV on solid organ transplant outcomes. This study characterized longitudinal immune responses to CMV in 31 kidney transplant recipients with CMV viremia and matched, nonviremic recipients. Recipients were sampled 3 and 12 months after transplant, with additional samples 1 week and 1 month after viremia. PBMCs were stained for NK and T cell markers. PBMC transcriptomes were characterized by RNA-Seq. Plasma proteins were quantified by Luminex. CD8+ T cell transcriptomes were characterized by single-cell RNA-Seq. Before viremia, patients had high levels of IL-15 with concurrent expansion of immature CD56bright NK cells. After viremia, mature CD56dim NK cells and CD28-CD8+ T cells upregulating inhibitory and NK-associated receptors were expanded. Memory NK cells and NK-like CD28-CD8+ T cells were associated with control of viremia. These findings suggest that signatures of innate activation may be prognostic for CMV reactivation after transplant, while CD8+ T cell functionality is critical for effective control of CMV.

Keywords: Immunology; NK cells; Organ transplantation; T cells; Transplantation.

PubMed Disclaimer

Figures

Figure 1
Figure 1. Schematic of study design, systems immunology approach, and integrated, multiomic analysis.
Figure 2
Figure 2. CD56 expression on NK cells delineates stages of CMV viremia.
(A) Median number of CD56bright (blue) and CD56dim (red) NK cells at 3 months after transplant (post-Tx), before viremia, and longitudinally after detection of viremia; 2 standard deviations (dotted lines) and numbers per patient (nonbold lines) are indicated. (B) Ratio of CD56dim/CD56bright NK cells in CMV PCR– patients 3 months (dark blue, n = 17) and 12 months after transplant (light blue, n = 24) and CMV PCR+ patients 3 months after transplant (purple, n = 14), 1 week after viremia (1WK PV, red, n = 19), 1 month after viremia (1M PV, orange, n = 9), and 12 months after transplant (yellow, n = 22). P values comparing CMV PCR– and CMV PCR+ at 3 and 12 months after transplant, determined by binomial logistic regression, and change over time after detection of viremia in CMV PCR+ patients, determined by linear regression, including patient ID as a random effect, are shown. (C) Principal component analysis of individual marker expression values on CD56bright (blue) and CD56dim (red) NK cells. Ellipses represent 50% of patient variance per group, with increasing width of lines indicating increasing time after transplant for CMV PCR– patients and time after transplant and after detection of viremia for CMV PCR+ patients. Direction and strength of variance explained by each marker is indicated by annotated green arrows. Increasing evidence of CD56bright NK cell maturation (CD56b_maturation) and CD56dim NK cell memory-like (CD56d_memory) phenotypes are highlighted.
Figure 3
Figure 3. CD8+ T cells expressing inhibitory and NK-like receptors increase after CMV viremia.
(A) CD8+ T cells as a percentage of lymphocytes for CMV PCR– patients 3 months (dark blue, n = 17) and 12 months after transplant (light blue, n = 24) and CMV PCR+ patients 3 months after transplant (purple, n = 14), 1 week after viremia (1WK PV, red, n = 19), 1 month after viremia (1M PV, orange, n = 9),and 12 months after transplant (yellow, n = 22). P values comparing CMV PCR– and CMV PCR+ at 3 and 12 months after transplant, determined by binomial logistic regression, and change over time after detection of viremia in CMV PCR+ patients, determined by linear regression, including patient ID as a random effect, are shown. (B) Ratio of CD28–/CD28+ CD8+ T cells as described above. (C) Mean percentage of CD28– (blue) and CD28+ (red) T cells expressing each marker. (D) Principal component analysis of individual marker expression values on CD28– (blue) and CD28+ (red) T cells. Ellipses represent 50% of patient variance per group, with increasing width of lines indicating increasing time after transplant for CMV PCR– patients and time after transplant and after detection of viremia for CMV PCR+ patients; full and dashed lines represent CD28– and CD28+ cells, respectively. Direction and strength of variance explained by each marker is indicated by annotated green arrows. Increasing evidence of transitional (CD28p_transitional), inhibitory (CD28n_inhibitory), and NK-like (CD28n_NK-like) phenotypes are highlighted.
Figure 4
Figure 4. Cytotoxic, inhibitory, and NK-like phenotype of CD28–CD8+ T cells highlighted by single-cell transcriptomics.
(A) Single-cell transcriptomes of CD8+ T cells, purified by negative selection, were reduced to 2 dimensions by t-SNE for visualization. Cells were classified as early, transitional, or late by expression of CD28 and known markers of CD8+ differentiation. (B) Differentially expressed transcripts per classification. Proportion of cells expressing each transcript within each class (blue) and proportion expressing each transcript outside each class (gray) are shown. (C) Normalized expression of transcripts delineating early, transitional, and late differentiated CD8+ T cells, from low (gray) to high (dark blue).
Figure 5
Figure 5. Partial least squares regression defines CMV PCR+ patients before and long-term after viremia and highlights acute and longitudinal immunological changes after detection of CMV viremia.
Partial least squares (PLS) regression of NK and T cell phenotypes, plasma analytes, and whole-blood transcriptome modules was used to identify (A) variables important in differentiating CMV PCR– and PCR+ patients longitudinally after transplant and (B) those important in defining immunological phenotypes prior to and longitudinally after detection of CMV viremia. Top 10 most informative variables per component are indicated by arrows and linked text. CMV PCR– 3 months (dark blue, n = 17) and 12 months after transplant (light blue, n = 24), and PCR+ 3 months after transplant, before viremia, (purple, n = 14), 1 week after detection of viremia (1WK PV, red, n = 19), 1 month after detection of viremia (1M PV, orange, n = 9), and 12 months after transplant after viremia (yellow, n = 22) samples are indicated; ellipses capture 50% of the variance per group. Bold arrow highlights longitudinal progression immune profiles of CMV PCR+ patients. CD56b_maturation, CD56d_memory, CD28p_transitional, CD28n_inhibitory, and CD28n_NK-like were determined from protein surface expression profiles on NK and CD8+ T cells, as detailed in Figures 2 and 3.
Figure 6
Figure 6. Cellular activity and NK and CD8+ T cell phenotype identify effective controllers of CMV viremia.
(A) Duration of initial viremia in days was used to define patients as effective controllers (gray; duration ≤ 16 days, n = 12) and ineffective controllers (black; duration >16 days, n = 19). PLS regression of NK and T cell phenotypes, plasma analytes, and whole-blood transcriptome modules was used to identify variables predictive of control of CMV viremia (B) 3 months after transplant, before viremia, and (C) 1 month after detection of viremia. Top 10 most informative variables per component are indicated by arrows and linked text. Differential control of viremia is indicated by color of points. CD56b_maturation, CD56d_memory, CD28p_transitional, CD28n_inhibitory, and CD28n_NK-like were determined from protein surface expression profiles on NK and CD8+ T cells, as detailed in Figures 2 and 3.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Griffiths P, et al. The pathogenesis of human cytomegalovirus. J Pathol. 2015;235(2):288–297. doi: 10.1002/path.4437. - DOI - PubMed
    1. Dupont L, Reeves MB. Cytomegalovirus latency and reactivation: recent insights into an age old problem. Rev Med Virol. 2016;26(2):75–89. doi: 10.1002/rmv.1862. - DOI - PMC - PubMed
    1. Schultz DA, Chandler S. Cytomegalovirus testing: antibody determinations and virus cultures with recommendations for use. J Clin Lab Anal. 1991;5(1):69–73. doi: 10.1002/jcla.1860050113. - DOI - PubMed
    1. Lopez-Vergès S, et al. Expansion of a unique CD57+NKG2C hi natural killer cell subset during acute human cytomegalovirus infection. Proc Natl Acad Sci U S A. 2011;108(36):14725–14732. doi: 10.1073/pnas.1110900108. - DOI - PMC - PubMed
    1. Sester M, et al. Dominance of virus-specific CD8 T cells in human primary cytomegalovirus infection. J Am Soc Nephrol. 2002;13(10):2577–2584. doi: 10.1097/01.ASN.0000030141.41726.52. - DOI - PubMed

Publication types