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. 2022 Apr 12;19(1):68.
doi: 10.1186/s12985-022-01776-4.

Increased expression of TIGIT and KLRG1 correlates with impaired CD56bright NK cell immunity in HPV16-related cervical intraepithelial neoplasia

You Nie #  1   2 , Dandan Liu #  1 , Wen Yang #  3 , Yazhuo Li  1 , Lihua Zhang  1 , Xia Cheng  1 , Ruyu Chen  1 , Bingbing Yuan  2 , Guangzheng Zhang  4 , Hongwei Wang  5
Affiliations

Increased expression of TIGIT and KLRG1 correlates with impaired CD56bright NK cell immunity in HPV16-related cervical intraepithelial neoplasia

You Nie et al. Virol J. .

Abstract

Background: The onset and progression of cervical intraepithelial neoplasia (CIN) are closely associated with the persistent infection of high-risk HPV (especially type16), which is mainly caused by immune escape. Natural killer (NK) cells play an important role against virally infected cells and tumor cells through a fine balance of signals from multiple surface receptors. Overexpression of non-MHC-I specific inhibitory receptors TIGIT, KLRG1, Siglec-7, LAIR-1, and CD300a on NK cells correlates with cellular exhaustion and immune evasion, but these receptors have not been investigated in CIN. The aim of the present study was to examine the potential role of NK cell non-MHC-I specific inhibitory receptors expression in immune escape from HPV16(+)CIN patients.

Methods: The subset distribution, IFN-γ and TNF-α expression levels and immunophenotype of TIGIT, KLRG1, Siglec-7, LAIR-1, and CD300a of NK cells were investigated in peripheral blood mononuclear cell samples by flow cytometry from 82 women who were HPV16(+) with CIN grades 0, I, II-III or HPV(-) CIN 0. Immunohistochemistry was applied to detect the expression of ligands for NK receptors in the cervical tissues. HPV types were identified by PCR assays.

Results: The HPV16(+) subjects with high-grade lesions had an increased number of circulating peripheral blood CD56bright NK cells with reduced functionality and IFN-γ secretion. The expression levels of the inhibitory molecules TIGIT and KLRG1 on CD56bright NK cells increased in parallel with increasing CIN grade. In addition, TIGIT and KLRG1 related ligands, Poliovirus receptor (PVR), N-Cadherin and E-Cadherin expression level was also elevated with increasing CIN grade.

Conclusions: Our results suggest that up-regulation of the inhibitory TIGIT, KLRG1 and their ligands may negatively regulate cervical CD56bright NK-mediated immunity to HPV16 and contribute to the progression of CIN. These results may facilitate the development of early-warning immune predictors and therapeutic strategies for HPV16(+) CIN based on the TIGIT and KLRG1 inhibitory pathways of NK cells.

Keywords: Cervical intraepithelial neoplasia; Cytokines; Human papillomavirus 16; KLRG1-Cadherin; Natural killer cell; TIGIT-PVR.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Increased number of circulating peripheral blood CD56bright NK cells counts in HPV16(+) CIN II–III. A(a) Representative flow cytometry dot plots (SSC-A vs FSC-A). A(b) gated from A(a) and stained to obtain NK cells using CD3− CD56+ as a marker. A(c) A representative dot plot in which two NK cell subsets were gated based on CD56 and CD16 expression: CD56dim and CD56bright NK cells. B(a–c) The percentage of NK cells and its subsets was analyzed from HPV16(+) women with different CIN grades or HPV(−) women using violin plots, in which each dot represents a donor. B(d–f) the absolute count of NK cells and their subsets was analyzed from HPV16(+) women with different CIN grades or HPV(−) women using violin plots, in which each dot represents a donor. The Kruskal–Wallis test was used to determine statistical significance
Fig. 2
Fig. 2
Circulating CD56bright NK cells from HPV16(+) subjects with CIN showed a reduced ability to secrete IFN-γ. A Statistical graph displays percentage of IFN-γ created by CD56dim and CD56bright NK cells in the HPV16(+) women with different CIN grades and HPV(−) women. B Statistical graph displays percentage of TNF-α created by CD56dim and CD56bright NK cells in the HPV16(+) women with different CIN grades and HPV(−) women. C Representative dot plots of IFN-γ and TNF-α created by CD56bright NK cells in the HPV(−) CIN 0 group and HPV16(+) CIN II–III groups
Fig. 3
Fig. 3
Increased expression of TIGIT and KLRG1 expression on CD56bright NK cells in HPV16(+)-related CIN. a–l The percentage and MFI of inhibitory receptors TIGIT, NKG2A, CD300a, KLRG1, LAIR1, and Siglec-7 on NK cells were analyzed from HPV16(+) women with different CIN grades or HPV(−) women.. The Kruskal–Wallis test was used to test for statistical significance
Fig. 4
Fig. 4
Increased expression of PVR, N-Cadherin and E-Cadherin in HPV16(+) CIN. A(a–c) Representative immunohistochemical sections of PVR, E-Cadherin and N-Cadherin expression in cervical tissues with HPV(−) CIN 0 and HPV16(+) CIN. B(a, b) Summary of the Allred scores in each group. Error bars indicate SD, original magnification: × 200
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