Nocardia Rubra Cell Wall Skeleton Up-Regulates T Cell Subsets and Inhibits PD-1/PD-L1 Pathway to Promote Local Immune Status of Patients With High-Risk Human Papillomavirus Infection and Cervical Intraepithelial Neoplasia

Abstract

The Nocardia rubra cell wall skeleton (Nr-CWS) for external use is an immune enhancer, which has been widely used in human cervix diseases such as cervical erosion, but the mechanism of Nr-CWS enhancing immunity is still unclear. The purpose of this study was to explore the effect and mechanism of Nr-CWS on the local immune status of cervical tissue in patients with high-risk human papillomavirus (HR-HPV) infection and cervical precancerous lesion, cervical intraepithelial neoplasia (CIN). The recruited patients with HR-HPV infection and CIN were treated with Nr-CWS. The specimens were taken from these patients before and after local application of Nr-CWS respectively. The normal control specimens were tested simultaneously. Serial section analysis of immunohistochemistry and co-expression analysis were performed to characterize populations of T cells and the expressions of programmed cell death-1 (PD-1) and programmed cell death-ligand 1 (PD-L1). The levels of cytokines in local cervical tissue were also detected. Nr-CWS significantly increased T cells including CD4+, CD8+ T cells, and reduced the expression of PD-L1 in the patients’ local cervical tissues. Co-expression analyses showed that the proportions of PD-1+CD4+ cells in CD4+ T cells and PD-1+CD8+ cells in CD8+ T cells decreased after Nr-CWS application. Furthermore, the increase in the number of immune cells was accompanied by increased pro-inflammatory cytokines interleukin-12 (IL-12), interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and decreased suppressive cytokine IL-10. The results indicate that Nr-CWS, as an immunotherapeutic agent for HR-HPV infection and CIN, plays an immune promoting role related to the upregulation of T cell subsets and the inhibition of PD-1/PD-L1 pathway.

Nocardia rubra cell‐wall skeleton influences the development of cervical carcinoma by promoting the antitumor effect of macrophages and dendritic cells

“…The secretion of these four cytokines was increased by the addition of both low and high doses of Nr‐CWS. Among them, the increasing trends of IL‐12 and TNF‐ɑ were consistent with the study results of Wei et al in high‐risk HPV infection and cervical intraepithelial neoplasia 13 . The trends in IL‐6 and IL‐1β levels in patients with bladder cancer were similar to the results of the study by Elizabeth et al However, none of their studies indicated the origin of the increased cytokines 28 .…”

“…Among them, the increasing trends of IL‐12 and TNF‐ɑ were consistent with the study results of Wei et al in high‐risk HPV infection and cervical intraepithelial neoplasia. 13 The trends in IL‐6 and IL‐1β levels in patients with bladder cancer were similar to the results of the study by Elizabeth et al However, none of their studies indicated the origin of the increased cytokines. 28 Our experiments suggest that these increased cytokines may come from the increase in the activity of dendritic cells and macrophages.…”

“…The immunostimulant of Nocardia rubra cell‐wall skeleton (Nr‐CWS), consisting of nocardomycolic acid, arabinogalactan, and mucopeptide, enhances the ability of the host to clear infection. Nr‐CWS significantly increased the number of CD4+ and CD8+T cells in the cervical tissues of patients with HR‐HPV infection and CIN, and decreased the expression of PD‐L1 13 . Nr‐CWS can significantly promote the activity of Cytokine‐Induced Killer cells, DCs and Natural Killer cells, which exert strong antitumor activity especially by inducing apoptosis 14 .…”

“…Nr‐CWS significantly increased the number of CD4+ and CD8+T cells in the cervical tissues of patients with HR‐HPV infection and CIN, and decreased the expression of PD‐L1. 13 Nr‐CWS can significantly promote the activity of Cytokine‐Induced Killer cells, DCs and Natural Killer cells, which exert strong antitumor activity especially by inducing apoptosis. 14 In addition, Nr‐CWS increases the response of CD8+T cell and activation of CD4+T cell, and promotes the differentiation of CD4+T cells into Th1 cells.…”

Enhancing Natural Killer Cell-Mediated Cancer Immunotherapy by the Biological Macromolecule Nocardia rubra Cell-Wall Skeleton

“…In addition, Nr-CWS can improve the therapeutic efficacy of PD-1/PD-L1 immunosuppressive pathway in patients with high-risk human papillomavirus (HR-HPV) infection and cervical precancerous lesion, cervical intraepithelial neoplasia (CIN), and mouse CT26. WT cancer cell [ 6 , 9 ]. Our previous study showed that Nr-CWS can promote CD4 + T cells activation, drive Th1 immune response, and enhance CD8 + T cells function [ 4 , 5 ].…”