CN109402256B

CN109402256B - System for evaluating clinical prognosis of glioma by using expression quantity of multiple immune regulatory point molecules

Info

Publication number: CN109402256B
Application number: CN201811197997.9A
Authority: CN
Inventors: 吴安华; 程文; 邹存义; 朱晨
Original assignee: First Hospital of China Medical University
Current assignee: First Hospital of China Medical University
Priority date: 2018-10-15
Filing date: 2018-10-15
Publication date: 2021-07-23
Anticipated expiration: 2038-10-15
Also published as: CN109402256A

Abstract

The system comprises a computing device, an input device and an output device, wherein the input device is used for inputting the expression quantity of a plurality of immune regulatory point molecules of an individual glioma patient, and the output device is used for outputting the prognosis of the glioma; wherein the plurality of immunomodulating point molecules comprises CD48, SLAMF8 and PD-L1. The disclosure also provides an immune regulation and control point molecule for evaluating glioma prognosis, application of the immune regulation and control point molecule in preparing a kit for evaluating glioma prognosis of Chinese population, and application of a reagent for detecting expression quantity of the immune regulation and control point molecule in preparing the kit for evaluating glioma prognosis of Chinese population. The invention provides a biomarker with high specificity and high sensitivity and a prognosis prediction model for glioma prognosis of Chinese population.

Description

System for evaluating clinical prognosis of glioma by using expression quantity of multiple immune regulatory point molecules

Technical Field

The disclosure relates to the field of biological medicine and health, in particular to a system for evaluating clinical prognosis of glioma of Chinese population by using expression quantity of multiple immune regulation and control point molecules.

Background

Glioma is the most common primary tumor of the central nervous system, and has high malignancy, invasive growth, poor prognosis and easy relapse. Despite the progress of glioma treatment, the median survival of the most aggressive Glioblastoma (GBM) after receiving postoperative chemoradiotherapy is only 14.6 months. Despite the advent of new glioma immunotherapy strategies such as Dendritic Cell (DC) therapy and immune checkpoint blockade, the efficacy remains suboptimal and there is an urgent need to explore glioma immunity in depth to find more effective therapeutic approaches. Meanwhile, for glioma patients, effective glioma diagnosis and prognosis evaluation indexes are urgently needed clinically so as to achieve the purposes of early diagnosis and early treatment, and a corresponding reasonable treatment scheme is selected, so that the survival period of the patients is prolonged, and the prognosis is improved.

In gliomas, various immune cells and stroma make up the non-tumor components of the tumor parenchyma, such as T cells, tumor-associated macrophages (TAMs), NK cells, and the like. They transmit information through secretory cell factors or ligand-receptor interaction, and form a hotbed which is beneficial to malignant progression of glioma. Meanwhile, strong immunosuppression is generated among various cells in the glioma through a multiple method, and effective antitumor immune response is suppressed. It has been reported that TAMs can promote tumor progression and immunosuppression by interacting with glioma stem cells. In addition, the immune regulatory point plays an important role in inhibiting anti-tumor immunity, and CTLA-4 and PD-1/PD-L1 are key co-inhibitory molecules of tumor escape T cells and B cells mediated anti-tumor immunity. However, glioma immune checkpoint blockade therapy against CTLA-4, PD-1 is only effective in a fraction of glioma patients and is often accompanied by immunoinflammation-related side effects.

Signaling lymphocyte activation molecule family 2(SLAMF2, CD48) is an adhesion co-stimulatory molecule that is expressed on most hematopoietic cells, especially high expression in Antigen Presenting Cells (APCs). By binding to its receptor CD2, CD48 is involved in a variety of innate and adaptive immune responses, including regulation of granulocyte activity and inflammatory responses, T cell activation and autoimmune responses, and in regulation of CTL or NK cell function. The expression level of CD48 is increased in autoimmune diseases and allergic diseases, and the anti-CD 48 monoclonal antibody can effectively treat autoimmune encephalomyelitis. It has been found that the interaction of CD48 with its high affinity receptor 2B4(CD244) in liver cancer can lead to monocyte/macrophage induced NK cell dysfunction. The above suggests that CD48 plays an important role in immune activation or immunosuppression. However, few reports have been made on the prognostic evaluation and immunomodulatory effects of CD48 in gliomas. Moreover, the Chinese population still lacks a glioma clinical prognosis evaluation method suitable for the ethnic group.

Disclosure of Invention

The purpose of the disclosure is to provide a new molecular model of immune regulation and control points closely related to glioma prognosis of Chinese population, thereby enriching evaluation methods of glioma prognosis and improving the accuracy of glioma prognosis evaluation.

To achieve the above object, a first aspect of the present disclosure: the system is characterized by comprising a computing device, an input device and an output device, wherein the input device is used for inputting the expression quantity of a plurality of immune regulation point molecules of an individual glioma patient, and the output device is used for outputting the prognosis of the glioma; wherein the plurality of immunomodulating point molecules comprises CD48, SLAMF8 and PD-L1; the computing device comprises a memory having a computer program stored therein and a processor configured to execute the computer program stored in the memory to implement an algorithm of a discriminant function as shown in equation (1);

f is 0.2029A +0.2942B +0.4562C formula (1)

In the formula (1), F represents prognosis of glioma, F return value is not less than 0.4710, F return value represents poor prognosis of glioma, A, B and C represent expression levels of PD-L1, CD48 and SLAMF8 respectively.

Optionally, the system further comprises a device for detecting the expression level of the plurality of immunomodulating point molecules.

Optionally, the device for detecting the expression levels of the plurality of immunoregulation point molecules comprises an immunoregulation point molecule expression level detection chip and a chip signal reader, and the immunoregulation point molecule expression level detection chip comprises probes for detecting the expression levels of CD48, SLAMF8 and PD-L1 respectively.

Optionally, the device for detecting the expression levels of the plurality of immune regulatory site molecules comprises a real-time quantitative PCR instrument and real-time quantitative PCR primers of the molecules, and the real-time quantitative PCR primers of the molecules comprise real-time quantitative PCR primers for respectively detecting the expression levels of CD48, SLAMF8 and PD-L1.

In a second aspect of the present disclosure: there is provided use of a system according to the first aspect of the present disclosure in the manufacture of a medical device for assessing the prognosis of glioma in the population of china.

A third aspect of the disclosure: provides an immune regulatory point molecule for evaluating glioma prognosis of Chinese population, which comprises CD 48.

Optionally, the immune checkpoint molecule further comprises SLAMF8 and PD-L1.

A fourth aspect of the present disclosure: provides the application of the immune regulatory point molecule in the preparation of a kit for evaluating glioma prognosis of Chinese population, wherein the immune regulatory point molecule comprises CD 48.

Optionally, the immune checkpoint molecule further comprises SLAMF8 and PD-L1.

The fifth aspect of the present disclosure: the application of the reagent for detecting the expression quantity of the immune regulatory point molecules in preparing the kit for evaluating the glioma prognosis of Chinese population is provided, wherein the immune regulatory point molecules comprise CD48, SLAMF8 and PD-L1.

Glioma is a disease which is difficult to treat along with infiltration of various immune cells. Although immunotherapy with blockade of the site of immune regulation holds promise for tumor patients, treatment with blockade of the site of glioma is still not ideal. The inventor screens out high-reliability disease diagnosis and treatment immune regulation and control point molecules by analyzing RNA sequencing data of a patient, establishes a glioma prognosis judgment and prediction model by a calculation formula, and evaluates the performance of the glioma prognosis judgment and prediction model. The evaluation result shows that the technical scheme of the disclosure provides a high-specificity and high-sensitivity immune regulation point molecule and a prognosis prediction model for diagnosis and treatment of glioma of Chinese population, provides a new method for prognosis evaluation and immunotherapy of glioma, and provides a new idea for treatment of immune regulation point.

Additional features and advantages of the disclosure will be set forth in the detailed description which follows.

Drawings

The accompanying drawings, which are included to provide a further understanding of the disclosure and are incorporated in and constitute a part of this specification, illustrate embodiments of the disclosure and together with the description serve to explain the disclosure without limiting the disclosure. In the drawings:

fig. 1 shows the expression and prognostic value of CD48 in gliomas, wherein a shows the expression levels of CD48 in different grades, histopathological classifications, molecular typing and TCGA subtype gliomas in the CGGA database, with the abscissa representing grade II, grade III, grade IV, oligodendroglioma, oligodendroastrocytoma, astrocytoma, glioblastoma, low-grade glioma with IDH mutation and 1p19q co-deletion, low-grade glioma with IDH mutation and 1p19q non-co-deletion, low-grade glioma with IDH wild type, glioblastoma with IDH mutation, glioblastoma with IDH wild type, neuronal glioma, pre-neuronal glioma, transcorneal glioma, interstitial glioma, and the ordinate represents the expression level of CD 48; b shows that CD48 is a good predictor of mesenchymal subtype in the CGGA database with sensitivity on the abscissa and 1-specificity on the ordinate; c and D show the effect of CD48 expression levels on glioma prognosis in the CGGA database, with overall survival on the abscissa, overall survival on the ordinate, Low for Low CD48 expression, High for High CD48 expression, and n for the number of included cases per group; e and F show the effect of CD48 expression levels on glioma prognosis in the TCGA database, with overall survival on the abscissa, overall survival on the ordinate, Low for Low CD48 expression, High for High CD48 expression, and n for the number of included cases per group.

Fig. 2 shows CD48 expression and predicted clinical prognosis, wherein a shows the expression levels of CD48 in various grades, histopathological classifications, molecular typing and TCGA subtypes in the TCGA database, with the abscissa representing grade II, grade III, grade IV, oligodendroglioma, oligodendroastrocytoma, astrocytoma, glioblastoma, low grade glioma with IDH mutation and 1p19q co-deletion, low grade glioma with IDH mutation and 1p19q non-co-deletion, low grade glioma with IDH wild type, glioblastoma with IDH mutation, glioblastoma with IDH wild type, neuronal glioma, pre-neuronal glioma, classical glioma, interstitial glioma, and the ordinate representing the expression of CD 48; b shows enriched high expression in SLAMF 8-specific interstitial subtypes in the TCGA database, with sensitivity on the abscissa and 1-specificity on the ordinate; C-E shows CD48 predicts 1, 2, and 3 year survival, with sensitivity on the abscissa and 1-specificity on the ordinate.

Fig. 3 shows the prognostic value of the predictive models used to determine glioma prognosis in the system of the present disclosure, wherein a shows the correlation of the predictive model (checkpoint Risk Score) with glioma grade, histopathological classification, molecular typing and TCGA subtype, with abscissa representing grade II, grade III, grade IV, oligodendroglioma, oligodendroastrocytoma, astrocytoma, glioblastoma, low grade glioma with IDH mutation and 1p19q co-deletion, low grade glioma with IDH mutation and 1p19q non-co-deletion, low grade glioma with IDH wild type, glioblastoma with IDH mutation, glioblastoma with IDH wild type, neuronal glioma, pre-neuronal glioma, trans-glioma, mesenchymal glioma, with ordinate representing F value (Risk Score); b shows that the prediction model in the CGGA database is a good prediction molecule of mesenchymal subtype, the abscissa represents sensitivity, and the ordinate represents 1-specificity; C-E shows that a High F-number in the CGGA database indicates poor clinical prognosis of glioma, F-H shows that a High F-number in the TCGA database indicates poor clinical prognosis of glioma, the abscissa represents overall survival, the ordinate represents overall survival, Low represents a lower F-number (Risk Score), and High represents a High F-number (Risk Score).

Fig. 4 shows the distribution of the predictive model for determining prognosis of glioma in the system of the present disclosure in the TCGA database, wherein a shows that the malignant phenotype of glioma in the TCGA database has a significantly higher F-value (regulatory point Risk Score), with the abscissa representing class II, class III, class IV, oligodendroglioma, oligodendro-astrocytoma, glioblastoma, low grade glioma with IDH mutation and 1p19q co-deletion, low grade glioma with IDH mutation and 1p19q non-co-deletion, low grade glioma with IDH wild-type, glioblastoma with IDH mutation, glioblastoma with IDH wild-type, neurogenic glioma, anterior neurogenic glioma, classical glioma, interstitial glioma, with the ordinate representing F-value (Risk Score); b shows that F-values (regulatory point risk scores) are good predictors of mesenchymal subtypes.

FIG. 5 shows the prognostic value of GBM of different subtypes for the predictive model for assessing glioma prognosis in the system of the present disclosure, wherein A-D shows that F-value (Risk Score) has predictive value in both male and female GBM, E-H shows that High-Low expression of F-value (Risk Score) has prognostic difference in patients over 40 years of age but not in patients under 40 years of age, and I-P shows that High expression of F-value (Risk Score) in IDH wild-type and MGMT promoter methylated patients suggests poor prognosis, but not significant prognostic difference in High-Low expression F-value (Risk Score) patients in IDH mutant and MGMT unmethylated state, wherein abscissa represents overall survival, ordinate represents overall survival, and Low represents lower F-value (Risk Score) and High F-value (Risk Score).

Detailed Description

The following detailed description of specific embodiments of the present disclosure is provided in connection with the accompanying drawings. It should be understood that the detailed description and specific examples, while indicating the present disclosure, are given by way of illustration and explanation only, not limitation.

The first aspect of the disclosure: the system comprises a computing device, an input device and an output device, wherein the input device is used for inputting the expression quantity of the plurality of immune regulatory point molecules of an individual glioma patient, and the output device is used for outputting the prognosis of the glioma; wherein the plurality of immunomodulating point molecules comprises CD48, SLAMF8 and PD-L1; the computing device comprises a memory having a computer program stored therein and a processor configured to execute the computer program stored in the memory to implement an algorithm of a discriminant function as shown in equation (1);

f is 0.2029A +0.2942B +0.4562C formula (1)

SLAMF8 is an immune co-stimulatory molecule of the same family of CD 48. The inventor of the application finds in research that SLAMF8 is highly expressed in the process of glioma malignancy and is an independent prognostic factor of glioma, and the high expression of SLAMF8 indicates that the prognosis of a patient is poor. Through functional analysis, the SLAMF8 is involved in glioma immune response, and the high-expression SLAMF8 glioma presents an immune activation state, but an effective anti-tumor immune response is inhibited, and an active inflammatory response state is presented. Further analysis shows that CD48 and SLAMF8 are in strong positive correlation with most of classical immune regulatory points, and are in weak correlation with PD-L1, so that the combination of CD48, SLAMF8 and PD-L1 is presumed to represent a classical immune regulatory molecule to evaluate the prognosis of glioma. Based on the above, the clinical prognosis evaluation system of glioma suitable for Chinese population is established based on CGGA database.

Wherein CD48 is a Gene with Gene ID 962 in NCBI database, and the expression level of CD48 may be the protein expression level of CD48, preferably the protein expression level on the cell membrane surface. SLAMF8 refers to the Gene whose Gene ID is 56833 in NCBI database, and the expression level of SLAMF8 may be the protein expression level of SLAMF8, preferably the protein expression level on the cell membrane surface. PD-L1 refers to Gene with Gene ID 29126 in NCBI database, and the expression level of PD-L1 may be PD-L1 protein expression level, preferably cell membrane surface protein expression level.

According to the present disclosure, the system may further comprise a means for detecting the expression level of the plurality of immunomodulating point molecules.

Further, the device for detecting the expression levels of the plurality of immunoregulatory site molecules may include an immunoregulatory site molecule expression level detection chip including probes for detecting the expression levels of CD48, SLAMF8, and PD-L1, respectively, and a chip signal reader. Or the detection device for the expression quantity of the multiple immune regulation point molecules comprises a real-time quantitative PCR instrument and real-time quantitative PCR primers of the molecules, and the real-time quantitative PCR primers of the molecules comprise real-time quantitative PCR primers for respectively detecting the expression quantity of CD48, SLAMF8 and PD-L1.

The inventors of the present application found in studies that the expression level of CD48 increases with the increase of glioma grade, for example, CD48 is highly expressed in gliomas having a malignant phenotype, such as glioblastoma, IDH wild-type glioma, and mesenchymal subtype glioma; furthermore, the total survival time (OS) of the patient with high expression of CD48 is obviously shorter than that of the patient with low expression, and CD48 has certain influence on glioma microenvironment and can promote immunosuppression and inflammatory reaction. Therefore, the CD48 is used as a good prediction molecule of the mesenchymal glioma to evaluate the clinical prognosis of the glioma, so that a reasonable clinical treatment strategy can be selected in time to improve the prognosis.

Further, the immunomodulating point molecules may also include SLAMF8 and PD-L1.

The reagent for detecting the expression level of the immune regulatory point molecule can be a probe and/or a primer.

Hereinafter, the present disclosure will be described in further detail by examples.

Examples

This example serves to illustrate the discovery of biomarkers and the establishment of predictive models of the present disclosure.

Case source and sample size: 946 glioma-sequencing data with detailed clinical information were analyzed, 310 of which were derived from CGGA databases (http:// www.cgga.org.cn, a Chinese glioma genomic profile based on a sample of glioma from the Chinese population), and 636 of which were derived from TCGA databases (https:// TCGA-data. nc. nih. gov/TCGA Download. jsp, a tumor genomic profile based on a sample of glioma from a foreign population). The CGGA database was used as the discovery database and the TCGA database was used as the verification database. Overall Survival (OS) is the time from diagnosis to death or end of final follow-up.

Bioinformatics analysis: glioma stromal score, immune score, and glioma purity were calculated using the R language to assess the proportion of non-tumor cell components in the tumor. The R package software calculates the microenvironment cell subset to calculate the absolute enrichment degree of eight immune cells and two interstitial cells. The corresponding gene is screened by utilizing the Pearson correlation analysis for functional analysis. DAVID software was used for gene ontology functional analysis. GSEA software was used to analyze the different functional phenotypes between high and low expression groups of CD 48. Principal component analysis and gene set variation analysis are used to differentiate differences in genomic, immune function and inflammatory responses caused by differences in CD48 expression.

Statistical analysis: SPSS, Graphpad Prism 7 and R3.3.3 (https:///www.r-project. org) software for statistical analysis. the t-test was used to evaluate the differential expression between the different groups, and the Pearson correlation coefficient was used to calculate the correlation. The low expression group and the high expression group were grouped according to median expression level. The prognosis results were evaluated using Kaplan-Meier survival analysis, and the log-rank test was used to evaluate the difference in prognosis between the different groups. Cox single factor and multivariate regression analysis were used to determine whether it was an independent prognostic factor. Receiver Operating Curves (ROCs) were plotted using Medcalc software. Other statistical calculations and statistical maps are plotted using an R-package (ggplot2, corrplot, pheatmap, etc.). A two-tailed P value of < 0.05 was defined as statistically significant.

Analysis of RNA sequencing data in the CGGA database revealed that CD48 expression levels increased with increasing glioma grade, particularly high expression at grade IV. With reference to histopathological typing, CD48 is most highly expressed in Glioblastoma (GBM). CD48 expression was significantly higher in IDH wild-type gliomas, both in lower-grade gliomas (LGG) and GBM, among different molecular subtypes (fig. 1A). Similar results were obtained after analysis of the TCGA database (fig. 2A), indicating that CD48 expression levels are closely correlated with glioma malignant progression.

The interstitial subtype is the most malignant subtype among the glioma TCGA database subtypes. The receiver operating characteristic curve (ROC) was plotted and CD48 was found to be a good predictor of mesenchymal glioma in the CGGA database (AUC 85%) (fig. 1B) and the TCGA database (AUC 91.9%) (fig. 2B).

In the CGGA database, patients with high expression of CD48 had short patients with significantly lower expression of OS, as well as High Grade Glioma (HGG) (fig. 1C and 1D). And the results were further validated in the TCGA database (fig. 1E and 1F). Given the importance of CD48 in influencing glioma prognosis, we evaluated the prognostic predictive value of CD48 in CGGA, and the results showed that CD48 is an accurate predictor for assessing glioma survival for 1, 2 and 3 years (AUC 77.7%, 79.2% and 81.3%, respectively) (fig. 2C-E).

Single and multifactorial regression analyses combined with clinical factors revealed that CD48 is an independent prognostic factor for glioma in the CGGA database (HR 1.202, P0.006) (see table 1). CD48 also independently predicts the adverse prognosis of glioma (HR 1.144, P0.006) in the TCGA database (see table 2). These indicate that CD48 plays an important role in determining the prognosis of glioma.

TABLE 1

TABLE 2

Both CD48 and SLAMF8 have strong associations with classical immune regulatory points, but have little association with PD-L1. Therefore, CD48, SLAMF8 and PD-L1 are used to establish a model for judging and predicting glioma prognosis, which is shown in formula (1).

F is 0.2029A +0.2942B +0.4562C formula (1)

In the formula (1), A, B and C represent the expression levels of PD-L1, CD48 and SLAMF8, respectively, in this order.

It was detected and calculated that F values were significantly higher in the malignant glioma subtypes (about 2-10 in CGGA database and about 5-10 in TCGA database), such as GBM, IDH-wild type LGG and GBM, and mesenchymal subtypes (CGGA database is shown in FIG. 3A, TCGA database is shown in FIG. 4A). Moreover, the model has accurate prediction value on the mesenchymal glioma, and the area under the curve AUC is 94.2% in the CGGA database and 94.8% in the TCGA database (the CGGA database is shown in figure 3B, and the TCGA database is shown in figure 4B).

In gliomas of the CGGA database, higher F values indicated shorter overall survival, and higher F values suggested poor prognosis in Lower Grade Gliomas (LGG) and GBM (fig. 3C-E). The same results were verified in the TCGA database (FIGS. 3F-H). The above model was found to be a glioma independent prognostic factor after single and multifactorial regression analysis (CGGA database is shown in Table 3 and TCGA database is shown in Table 4).

TABLE 3

TABLE 4

The CGGA and TCGA database analysis proves that the F value of an immune regulatory point model consisting of CD48, SLAMF8 and PD-L1 is an independent prognostic influence factor of glioma, and prognosis prediction of glioma can be carried out. The GBM median survival time is 14.6 months, the total survival time of glioma patients is defined to be poor prognosis when the total survival time is less than 14.6 months, and further, a ROC curve is utilized in a Chinese glioma database CGGA to determine that the F value is 0.4710, which is the best dividing point for poor clinical prognosis, namely, in Chinese population glioma patients, when the F return value is more than or equal to 0.4710, the clinical prognosis of glioma is represented.

GBM is glioma with high malignancy and poor prognosis, and immune regulatory point research and immune regulatory point blocking treatment have become hot spots for GBM treatment. The above model has predictive value in both the female and male groups (fig. 5A-D). The prognosis for patients with high F-numbers was significantly different in patients over 40 years of age (FIGS. 5E-H), i.e., the model evaluation prognosis was influenced by age factors.

In IDH wild-type GBM, patients with high F had a significantly poorer prognosis than those with low F, but no prognostic difference was observed between the two groups in IDH mutated GBM (FIGS. 5I-L). In GBM patients with MGMT promoter methylation, high F values suggested poor prognosis, but not in GBM with MGMT promoter unmethylated (fig. 5M-P). The method is used for prompting that the accuracy of the model evaluation is improved by carrying out prognosis evaluation on different molecular subtypes.

The preferred embodiments of the present disclosure are described in detail with reference to the accompanying drawings, however, the present disclosure is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present disclosure within the technical idea of the present disclosure, and these simple modifications all belong to the protection scope of the present disclosure.

It should be noted that, in the foregoing embodiments, various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various combinations that are possible in the present disclosure are not described again.

In addition, any combination of various embodiments of the present disclosure may be made, and the same should be considered as the disclosure of the present disclosure, as long as it does not depart from the spirit of the present disclosure.

Claims

1. A system for evaluating clinical prognosis of glioma in Chinese population by using expression quantity of molecules of a plurality of immune control points is characterized by comprising a computing device, an input device and an output device, wherein the input device is used for inputting the expression quantity of the molecules of the plurality of immune control points of individual glioma patients, and the output device is used for outputting prognosis of the glioma; wherein the plurality of immunomodulating point molecules comprises CD48, SLAMF8 and PD-L1; the computing device comprises a memory having a computer program stored therein and a processor configured to execute the computer program stored in the memory to implement an algorithm of a discriminant function as shown in equation (1);

f is 0.2029A +0.2942B +0.4562C formula (1)

2. The system of claim 1, further comprising a means for detecting the amount of expression of a plurality of immunomodulating point molecules.

3. The system of claim 2, wherein the means for detecting the expression levels of the plurality of immunomodulating site molecules comprises an immunomodulating site molecule expression level detection chip comprising probes for detecting the expression levels of CD48, SLAMF8 and PD-L1, respectively, and a chip signal reader.

4. The system of claim 2, wherein the means for detecting the expression levels of the plurality of immunomodulating point molecules comprises a real-time quantitative PCR instrument and real-time quantitative PCR primers for molecules, including real-time quantitative PCR primers for detecting the expression levels of CD48, SLAMF8 and PD-L1, respectively.

5. Use of the system of any one of claims 1 to 4 in the manufacture of a medical device for assessing the prognosis of glioma in the population of china.

6. The application of the reagent for detecting the expression quantity of the immune regulatory point molecules in the preparation of the kit for evaluating the glioma prognosis of Chinese population, wherein the immune regulatory point molecules comprise CD48, SLAMF8 and PD-L1.