Computer device and computer readable storage medium for evaluating post-ART immune reconstruction state of HIV infected person

Abstract

The invention discloses a computer device and a computer readable storage medium for evaluating the immune reconstitution state of an HIV infected person after ART. Belongs to the field of computer information. The technical problem to be solved by the invention is how to evaluate the immune reconstruction state of an HIV infected person after ART. The invention provides a computer program stored in a memory in a computer device. The computer program comprises the following steps: S1, receiving sample data, namely the proportion of the number of cells expressing PD-1 in CD8 + T cells of peripheral blood mononuclear cells of an HIV infected subject after ART and the proportion of the number of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN and CD107A; s2, obtaining a predicted value Y from the sample data through a model formula, and outputting an immune reconstruction condition according to the predicted value Y and a pre-judgment condition; the method can accurately and efficiently determine whether the immune reconstruction state of the HIV infected person subjected to antiviral treatment is good or not.

Description

Technical Field

[0001] The present invention belongs to the field of computer information, and relates to a computer device and a computer-readable storage medium for evaluating the immune reconstruction status of HIV-infected persons after ART. Background Art

[0002] Antiretroviral therapy (ART) has effectively reduced the morbidity and mortality of acquired immunodeficiency syndrome (AIDS) caused by infection with human immunodeficiency virus (HIV). Despite significant progress, AIDS remains a major global health problem. + T cells play a crucial role in the immune response to HIV infection, recognizing and killing HIV-infected cells, thereby preventing further spread of HIV. PD-1 is an immune checkpoint molecule, and its expression in T cells is considered an important immunosuppressive signal. In HIV-infected people (People Living With HIV, PLWH), PD-1 is expressed in CD8 + The expression of cytokines in T cells was significantly upregulated, and its expression was similar to that of HIV-specific CD8 + Impaired T cell function is also a predictor of disease progression and may lead to CD8 + T cell function further depleted.

[0003] ART can effectively inhibit HIV replication, thereby preventing HIV from further damaging the immune system. With the promotion of ART, some PLWH can achieve immune reconstruction. However, some patients still face the problem of incomplete immune reconstruction after treatment. In these patients, PD-1 may continue to be highly expressed, and PD-1 cannot accurately determine whether such PLWH patients have incomplete immune reconstruction after treatment, thus affecting the evaluation of the effect of immune reconstruction.

[0004] Previous studies have shown that the incomplete reconstitution of the immune system in PLWH may be related to the inability of ART to fully restore CD8 + Therefore, it is necessary to further explore the method of judging the immune reconstitution effect of PLWH patients after ART treatment. Summary of the invention

[0005] The technical problem solved by the present invention is to evaluate the immune reconstitution effect of PLWH patients after ART treatment.

[0006] In order to solve the above technical problem, the first aspect of the present invention provides a computer device, including a memory, a processor and a computer program stored in the memory, wherein the processor executes the computer program to implement the following steps:

[0007] S1. Receive data: Receive sample data, which is CD8 + The percentage of T cells expressing PD-1 and the percentage of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN, and CD107A;

[0008] S2. Output result: obtain a predicted value Y by applying the sample data to the following corresponding model formula, and output the immune reconstruction status of the HIV-infected subject who has undergone antiviral treatment according to the predicted value Y and the pre-determined condition;

[0009] The model formula is any of the following:

[0010] PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2;

[0011] PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2;

[0012] PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2;

[0013] PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2;

[0014] PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2;

[0015] PD-1 model formula: Y = -9.7176 + 5.1257X1;

[0016] Among them, Y is the predicted value, X1 is CD8 + The number of cells expressing PD-1 in T cells accounts for CD8 + The percentage of T cells, X2 is CD8 + The number of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in T cells accounted for CD8 + The percentage of T cells.

[0017] The immune reconstitution status of the HIV-infected subjects who have received antiviral treatment is a good immune reconstitution status or a poor immune reconstitution status.

[0018] In the above, the antiviral treatment may be antiviral treatment for more than two years.

[0019] In the above, the antiviral treatment may specifically be 55-102 months.

[0020] In the computer device described above,

[0021] The pre-determination conditions of each model formula are as follows:

[0022] If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good.

[0023] If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

[0024] In a second aspect, the present invention provides a computer program product, comprising a computer program, which, when executed by a processor, implements the steps in the computer device described in the first aspect.

[0025] In a third aspect, the present invention provides a computer-readable storage medium storing a computer program, wherein the computer program enables a computer to execute the steps in the computer device according to the first aspect.

[0026] In a fourth aspect, the present invention provides a device for evaluating immune reconstitution in HIV-infected subjects undergoing antiviral treatment, the device comprising:

[0027] S1, data receiving module: used to receive sample data, the sample data is CD8 + The percentage of T cells expressing PD-1 and the percentage of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN, and CD107A;

[0028] S2, result output module: obtain the predicted value Y by the sample data through the following corresponding model formula, and output the immune reconstruction status of the HIV-infected subject who has undergone antiviral treatment according to the predicted value Y and the pre-determined conditions;

[0029] The model formula is any of the following:

[0030] PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2;

[0031] PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2;

[0032] PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2;

[0033] PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2;

[0034] PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2;

[0035] PD-1 model formula: Y = -9.7176 + 5.1257X1;

[0036] Among them, Y is the predicted value, X1 is CD8 + The number of cells expressing PD-1 in T cells accounts for CD8 + The percentage of T cells, X2 is CD8 + The number of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in T cells accounted for CD8 + The percentage of T cells.

[0037] The immune reconstitution status of the HIV-infected subjects who have received antiviral treatment is a good immune reconstitution status or a poor immune reconstitution status.

[0038] In the device described above,

[0039] The pre-determination conditions of each model formula are as follows:

[0040] If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good.

[0041] If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

[0042] In a fifth aspect, the present invention provides a method for evaluating or assisting in evaluating the immune reconstitution of HIV-infected subjects undergoing antiviral treatment, comprising the following steps:

[0043] S1. Receive sample data, wherein the sample data is CD8 +The percentage of T cells expressing PD-1 and the percentage of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN, and CD107A;

[0044] S2. Obtain a predicted value Y by applying the sample data to the following corresponding model formula, and output the immune reconstruction status of the HIV-infected subject who has undergone antiviral treatment according to the predicted value Y and the pre-determined condition;

[0045] The model formula is any of the following:

[0046] PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2;

[0047] PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2;

[0048] PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2;

[0049] PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2;

[0050] PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2;

[0051] PD-1 model formula: Y = -9.7176 + 5.1257X1;

[0052] Among them, Y is the predicted value, X1 is CD8 + The number of cells expressing PD-1 in T cells accounts for CD8 + The percentage of T cells, X2 is CD8 + The number of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in T cells accounted for CD8 + The percentage of T cells.

[0053] The immune reconstitution status of the HIV-infected subjects who have received antiviral treatment is a good immune reconstitution status or a poor immune reconstitution status.

[0054] In the method described above,

[0055] The pre-determination conditions of each model formula are as follows:

[0056] If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good.

[0057] If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

[0058] The above threshold is the difference between the CD8 + The threshold is determined by the Y value calculated by the corresponding model formula, which is the percentage of cells expressing PD-1 in T cells and the percentage of cells expressing at least one of the five factors CD57, HLA-DR, CD38, PERFORIN and CD107A.

[0059] The experiment of the present invention proves that by detecting the CD8 + The proportion of PD-1 expressing cells in T cells and the proportion of cells expressing at least one of the following factors: the expression levels of CD38, HLA-DR, CD57, PERFORIN and CD107A, can be used to judge the immune reconstruction status of HIV-infected patients undergoing antiviral treatment, and can accurately and efficiently determine whether the immune reconstruction status of HIV-infected patients undergoing antiviral treatment is good. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 Flowchart for identifying immune reconstitution status in HIV-infected individuals undergoing antiviral therapy.

[0061] Figure 2 CD8 expressing PD-1 in INRs + T cells have a higher frequency. Note: (A) Distribution and number of each cell cluster; (B) CD3 + t-SNE plot of T cells; (C) Expression distribution of selected markers in each cluster (dark red, high expression; dark blue, low or no expression); (D) CD8 + PD-1 + Comparison of T cell counts; (E) CD8 + PD-1 + Comparison of T cell proportions.

[0062] Figure 3 PD-1 for INRs + CD8 +Cells tend to express higher levels of activation and senescence. (A) Comparison of CD57 expression in C9 cell clusters between INRs and IRs; (B) Comparison of HLA-DR expression in C9 cell clusters between INRs and IRs; (C) Comparison of CD57 expression in C14 cell clusters between INRs and IRs; (D) Comparison of HLA-DR expression in C14 cell clusters between INRs and IRs.

[0063] Figure 4 PD-1 in INRs + CD8 + Cells express higher levels of senescence, activation, and cytotoxic functions.

[0064] Figure 5 This is the ROC curve of the PD-1 prediction model.

[0065] Figure 6 This is the ROC curve of the PD-1 and CD57 prediction model.

[0066] Figure 7 This is the ROC curve of the PD-1 and HLA-DR prediction model.

[0067] Figure 8 This is the ROC curve of the PD-1 and CD107A prediction model.

[0068] Fig. 9 This is the ROC curve of the PD-1 and PERFORIN prediction model.

[0069] Fig.10 This is the ROC curve of the PD-1 and CD38 prediction model. DETAILED DESCRIPTION

[0070] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0071] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0072] Unless otherwise specified, the quantitative tests in the following examples were performed three times and the results were averaged.

[0073] The research object information in the following embodiments is as follows:

[0074] From April 1, 2022, to September 30, 2022, a total of 69 PLWH (Table 1) were recruited from the Infection Center of Beijing You'an Hospital Affiliated to Capital Medical University, all of whom were Chinese. Patients with opportunistic infections, autoimmune diseases, epilepsy, brain trauma, medication use, suppressive treatment, or other serious diseases were excluded from the study. All patients receiving ART had undetectable HIV RNA load. Clinical characteristics of the study subjects (including demographic characteristics, CD4 + T cell count, HIV RNA load, HIV infection date, HIV infection duration, ART initiation date, and ART duration) were obtained from the medical records of the Beijing You'an Hospital AIDS Research Cohort.

[0075] Table 1 shows the demographic and clinical characteristics

[0076]

[0077]

[0078] Note: IRs: Immunological Responders; INRs: Immunological Non-Responders; IRs: immunological responders; BMI: Body Mass Index; ART: Antiretroviral Therapy; IQR: Interquartile Range; SD, Standard Deviation. The Shapiro-Wilk test was used to assess whether the data conformed to normal distribution; the t-test was used to analyze the differences between groups for data sets that conformed to normal distribution, and the U-test was used to analyze the differences between groups for data sets that did not conform to normal distribution; a two-tailed P value less than 0.05 was used as the standard for statistical significance.

[0079] The CD4 count after antiviral treatment and the CD8 count after antiviral treatment in the above table are the test results after the antiviral treatment duration in Table 1.

[0080] The above-mentioned initial refers to the CD4 information, CD8 information or HIV RNA load first retained when HIV is diagnosed.

[0081] According to the data in Table 1 above, for patients after antiviral treatment (undetectable HIV RNA load 2 years after starting ART), CD4 + Total T cells less than 350 / μL or CD4 +Individuals with a T cell count increase of less than 30% from baseline were classified as immunological non-responders (INRs); and those with CD4 + Total T cells greater than 500 / μL or CD4 + Individuals whose total T cell count increased by 30% or more compared with baseline were classified as immunological responders (IRs).

[0082] Plasma and peripheral blood mononuclear cells (PBMCs) samples of each subject were stored at -80 °C and in liquid nitrogen tanks, respectively. Approved by the Ethics Committee of Beijing You'an Hospital Affiliated to Capital Medical University (2023 / 057), all participants provided written informed consent.

[0083] The mass spectrometry flow cytometry data and data analysis in the following examples are as follows:

[0084] A total of 23 antibodies were used to analyze each immune cell. These antibodies were obtained from Fluidigm (South San Francisco, USA) in a pre-bound state. Among them, the product numbers of anti-PD-1 antibody, anti-CD38 antibody, anti-HLA-DR antibody, anti-CD57 antibody, anti-PERFORIN antibody, and anti-CD107A antibody are: anti-PD-1 antibody (3155009B); anti-CD38 antibody (3172007B); anti-HLA-DR antibody (3170013B); anti-CD57 antibody (3142007B); anti-PERFORIN antibody (3175004B); anti-CD107A antibody (3151002B). The gene names of the detected phenotypes and the corresponding GenBank numbers and first submission dates are: PD-1: gene name: PDCD1, GenBank number: U64863, first submission date: July 1996; CD38: gene name: CD38, GenBank number: M83652, first submission date: January 1993; HLA-DR: gene name: HLA-DRA, GenBank number: M11157, first submission date: January 1984; CD57: gene name: B3GAT1, GenBank number: NM_003783, first submission date: August 1998; PERFORIN: gene name: PRF1, GenBank number: M28393, first submission date: January 1990; CD107A: gene name: LAMP1, GenBank number: M16038, first submission date: January 1989.

[0085] The isolated PBMCs were washed and treated with cisplatin-195Pt (Fluidigm, 201064) to remove dead cells. Human TruStain FcX was used for Fc receptor blocking before antibody staining. All antibodies were used according to the manufacturer's instructions. Subsequently, the cell samples were washed and stained with cell surface antibodies at low temperature for 30 minutes. The antibody-labeled samples were then washed, incubated in 125nM Cell-ID Intercalator-Ir (Fluidigm, USA), and diluted in phosphate-buffered saline (SigmaAldrich, USA) and stored at 4°C. The samples were 5.5×10 5 The cells were resuspended in double distilled water containing 10% EQ Beads (Fluidigm, South San Francisco, USA) at a concentration of 10 cells / mL. The samples were then analyzed using a CyTOF2 mass spectrometry system (Fluidigm, South San Francisco, USA).

[0086] The raw data of each sample was de-barcoded by using a unique mass marker barcode, using a two-body filtering protocol. The .fcs files from different batches were normalized using a bead normalization method. Gating was performed using FlowJo (version 10.8.1) software to eliminate debris and dead cells. Subsequently, CD45 + CD3 + CD8 + T cells were gated for further analysis in R. PhenoGragh clustering algorithm was used to divide cells into different clusters according to their surface marker expression levels. t-Distributed Stochastic Neighbor Embedding (t-SNE) was applied to reduce the dimensionality and visualize the high-dimensional data. Cell cluster distribution, marker expression, and differential analysis between groups were performed in R software (version 3.6.0).

[0087] The statistical analysis in the following examples is as follows:

[0088] Statistical analysis was performed using SPSS (version 27.0) and R (version 3.6.0). For categorical variables, χ 2 INRs and IRs were compared using the t-test or Fisher's exact test; for continuous variables, the t-test or Mann-Whitney U test was used. Spearman correlation analysis was used to investigate the correlation between PD-1 expression and immune function. The results were analyzed using GraphPadPrism (version 9.5.1) and R (version 3.6.0). P < 0.05 was considered statistically significant.

[0089] Example 1. Selection of factors in a computer device for identifying immune reconstitution in ART-treated PLWH

[0090] A total of 16 patients in the INRs group and 53 patients in the IRs group were included. INRs and IRs were matched in age, time of first ART, duration of HIV infection, and duration of ART. + T cell count, initial CD8 + T cell count, CD4 after antiviral treatment + T cell counts and CD8 after antiviral therapy + T cell counts were significantly lower than INRs, while initial HIV RNA loads were higher than INRs.

[0091] 1. CD8 expressing PD-1 + Difference in T Quantity Between INRs and IRs

[0092] Peripheral blood mononuclear cells (PBMCs) were collected from 16 subjects in the INRs group and 53 subjects in the IRs group for mass spectrometry flow cytometry to investigate the CD8 + Immune characteristics of T cells.

[0093] The results are as follows: CD8 + The four subsets of T cells were divided into 18 cell clusters according to the expression levels of markers such as PD-1, CD57, HLA-DR, CD38, CD107A, and PERFORIN in R software. The cell cluster numbers, distribution in IRs and INRs, and expression of marker PD-1 are shown in Figure 2 A-2C. In 18 CD8 + Among the T cell clusters, C7, C8, and C15 represent CD8+ naive T cells ( T, TN; CD3 + CD4 - CD8 + CD45RA + CCR7 + ) cells; C6, C10, C11, C16 represent CD8 + CD45RA + Effector memory T (CD45RA + Effector Memory T,TEM;CD3 + CD4 - CD8 + CD45RA + CCR7 - ) cells; C1, C3, C12, C14, C18 represent CD8+ CD45RA - Effector Memory T (TEM; CD3 + CD4 - CD8 + CD45RA - CCR7 - ) cells; C2, C4, C5, C9, C13, C17 represent CD8 + Central Memory T (TCM; CD3 + CD4 - CD8 + CD45RA - CCR7 + )cell.

[0094] We found that compared with IRs, the PD-1 expressing cell cluster C9 (PD-1 + TCM) and C14 (PD-1 + TEM) has a tendency to be higher, but it is not significant ( Figure 2 D).

[0095] Further, PD-1 was circled by manual gating in FlowJo. + CD8 + T cells, found in INRs, PD-1 + CD8 + The proportion of T was significantly higher than that of IRs ( Figure 2 E) These cell clusters represent CD8 + The frequencies of T cell subsets in INRs were significantly higher than those in IRs, indicating that there were more CD8 + T cells expressing PD-1, i.e., CD8 T cells expressing PD-1 in INRs + T cells are larger than IRs.

[0096] 2. CD57, HLA-DR, CD38, PERORIN, CD107A, CD8 expressing PD-1 in INRs and IRs + Differences between T cells

[0097] Then, the expression levels of each cell phenotype and cytotoxic function (CD57, HLA-DR, CD38, PERORIN, CD107A) in the above 18 cell clusters were obtained by analysis using R software.

[0098] The expression of PD-1 in CD8 +Differences in senescence (CD57), activation (HLA-DR, CD38), and cytotoxic function (PERFORIN, CD107A) of T cell clusters between the IRs and INRs groups.

[0099] The results are as follows Figure 3 As shown, in C14 (PD-1 + In TEM), the levels of CD57 and HLA-DR were higher in INRs compared with IRs.

[0100] These results indicate that terminally differentiated PD-1 + CD8 + T cells [C9 (PD-1 + TCM) and C14 (PD-1 + TEM)] showed higher levels of activation and senescence.

[0101] Other factors such as CD38, CD107A, and PERFORIN also showed the same trend.

[0102] Therefore, the following experiment was conducted to use markers such as CD57, HLA-DR, CD38, CD107A, PERFORIN and PD-1 to predict the immune reconstruction effect after 2 years of ART for HIV.

[0103] 3. PD-1 in INRs + CD8 + Cells express higher levels of senescence, activation, and cytotoxic functions

[0104] A separate analysis compared the PD-1 + CD8 + T cells and PD-1 - CD8 + Differences in T cell activation, senescence, and cytotoxic function.

[0105] The results are as follows Figure 4 Shown: In TCM cells, PD-1 - CD8 + Compared with T, PD-1 + CD8 + T cells had significantly higher levels of CD38, HLA-DR, CD57, PEFORIN, and CD107A. It was also observed that in INRs, PD-1 + CD8 + T cells expressed significantly higher levels of HLA-DR and CD38. This result suggests that in INRs, compared with PD-1 - CD8 + For T cells, PD-1+ CD8 + While expressing programmed cell death, T cells also exhibited high levels of activation, senescence, and cytotoxic functions, indicating that although the CD8+ T cells in INRs were numerous, they were in a state of cellular dysfunction and continuous exhaustion and were of poor quality.

[0106] Example 2: Preparation of a computer device for identifying immune reconstitution of PLWH treated with ART using PD-1 and a combination of five indicators

[0107] Figure 1 Flowchart for identifying immune reconstitution status in HIV-infected individuals undergoing antiviral therapy.

[0108] 1. Construction of a model to identify immune reconstitution in ART-treated PLWH

[0109] In order to clarify whether PD-1 can predict the immune reconstitution effect with other cell phenotype and cytotoxic function markers CD38, HLA-DR, CD57, PEFORIN, and CD107A, the ROC curve was then drawn, as follows:

[0110] 1) Considering clinical applicability, according to the 16 subjects in the INRs group and the 53 subjects in the IRs group in Example 1, a total of 69 PLWH were analyzed by mass spectrometry flow cytometry of peripheral blood mononuclear cells (PBMCs) of each subject, and the CD8 + T was further manually gated to analyze the CD8 + T cells expressing PD-1, CD57, HLA-DR, CD38, CD107A or PERFORIN accounted for all CD8 + T cell percentage.

[0111] 2) CD8 + The proportion of all CD8 T cells expressing PD-1 + The percentage of T cells was taken as the independent variable X1, and the cells expressing CD38, HLA-DR, CD57, PERFORIN or CD107A accounted for all CD8 + T percentage was used as the independent variable X2, and logistic regression was performed separately.

[0112] 3) Through logistic regression analysis, the following 6 model formulas are obtained:

[0113] PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2;

[0114] PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2;

[0115] PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2;

[0116] PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2;

[0117] PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2;

[0118] PD-1 model formula: Y = -9.7176 + 5.1257X1;

[0119] Y is the predicted value, X1 is the percentage of cells expressing PD-1 in all CD8 + T cell percentage, X2 is the percentage of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in all CD8 + T cell percentage;

[0120] 4) Determine the threshold

[0121] Six ROC curves were drawn based on the Y values ​​of the 69 subjects calculated by each model, and each ROC curve was calculated based on the Youden index (e.g. Figures 5 to 10 ) is the threshold of the corresponding model formula.

[0122] In the PD-1 model formula, when the factor is PD-1, the optimal threshold is 0.352 (AUC: 0.929, sensitivity: 0.8125, specificity: 0.9245,)

[0123] In the PD-1+CD57 model formula, when the factor is PD-1+CD57, the optimal threshold is 0.194 (AUC: 0.927, sensitivity: 0.875, specificity: 0.868)

[0124] In the PD-1+HLA_DR model formula, when the factor is PD-1+HLA_DR, the optimal threshold is 0.544 (AUC: 0.958, sensitivity: 0.8125, specificity: 0.981)

[0125] In the PD-1+CD107A model formula, when the factor is PD-1+CD107A, the optimal threshold is 0.471 (AUC: 0.929, sensitivity: 0.8125, specificity: 0.943)

[0126] In the PD-1+PERFORIN model formula, when the factor is PD-1+PERFORIN, the optimal threshold is 0.206 (AUC: 0.927, sensitivity: 0.875, specificity: 0.887)

[0127] In the PD-1+CD38 model formula, when the factor is PD-1+CD38, the optimal threshold is 0.362 (AUC: 0.929, sensitivity: 0.8125, specificity: 0.925)

[0128] The predicted value Y obtained by the corresponding model formula of each subject is compared with the corresponding threshold value, and the following is obtained:

[0129] If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good.

[0130] If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

[0131] The above-mentioned predicted value Y is the Y value calculated by the corresponding model formula, and the threshold is the threshold of the corresponding model formula.

[0132] The present invention can accurately and efficiently determine the immune reconstitution status of HIV-infected persons who have undergone antiviral treatment.

[0133] The statistical results of the Y and X values ​​of each model formula for 69 subjects are shown in Tables 2 to 7 below:

[0134] Table 2 shows the Y and X values ​​calculated by the PD-1 model formula

[0135] serial number X1 Y serial number X1 Y INR1 2.84 4.84 IRs20 1.45 -2.29 INR2 1.77 -0.65 IRs21 0.78 -5.72 INR3 1.32 -2.95 IRs22 1.56 -1.72 INR4 2.49 3.05 IRs23 0.5 -7.15 INR5 2.33 2.23 IRs24 0.56 -6.85 INR6 2.05 0.79 IRs25 1.01 -4.54 INR7 3.23 6.84 IRs26 1.15 -3.82 INR8 2.54 3.3 IRs27 0.65 -6.39 INR9 2.65 3.87 IRs28 1.28 -3.16 INRs10 1.33 -2.9 IRs29 1.14 -3.87 INRs11 1.91 0.07 IRs30 1.34 -2.85 INRs12 2.35 2.33 IRs31 1.54 -1.82 INRs13 1.6 -1.52 IRs32 0.5 -7.15 INRs14 2.57 3.46 IRs33 1.59 -1.57 INRs15 2.46 2.89 IRs34 0.66 -6.33 INRs16 1.95 0.28 IRs35 0.68 -6.23 IRs1 1.47 -2.18 IRs36 0.93 -4.95 IRs2 1.16 -3.77 IRs37 0.77 -5.77 IRs3 0.58 -6.74 IRs38 1.48 -2.13 IRs4 1.59 -1.57 IRs39 0.37 -7.82 IRs5 1.62 -1.41 IRs40 0.64 -6.44 IRs6 1.43 -2.39 IRs41 1.31 -3 IRs7 1.48 -2.13 IRs42 1.11 -4.03 IRs8 1.79 -0.54 IRs43 1.9 0.02 IRs9 1.55 -1.77 IRs44 1.22 -3.46 IRs10 1.63 -1.36 IRs45 1.42 -2.44 IRs11 1.34 -2.85 IRs46 1.13 -3.93 IRs12 1.61 -1.47 IRs47 2.04 0.74 IRs13 1.61 -1.47 IRs48 0.57 -6.8 IRs14 1.9 0.02 IRs49 1.09 -4.13 IRs15 1.4 -2.54 IRs50 1.24 -3.36 IRs16 0.39 -7.72 IRs51 0.58 -6.74 IRs17 1.4 -2.54 IRs52 1.22 -3.46 IRs18 0.87 -5.26 IRs53 0.76 -5.82 IRs19 0.53 -7 / / /

[0136] Table 3 shows the Y and X values ​​calculated by the PD1+CD57 model formula

[0137]

[0138]

[0139] Table 4 shows the Y and X values ​​calculated by the PD-1+HLA-DR model formula

[0140]

[0141]

[0142] Table 5 shows the Y and X values ​​calculated by the PD-1+CD38 model formula

[0143]

[0144]

[0145] Table 6 shows the Y and X values ​​calculated by the PD-1+PEFORIN model formula

[0146]

[0147]

[0148] Table 7 shows the Y and X values ​​calculated by the PD-1+CD107A model formula

[0149]

[0150]

[0151] II. Computational device to characterize immune reconstitution in PLWH receiving ART

[0152] The computer device includes a memory, a processor, and a computer program stored in the memory, and the processor executes the computer program to implement the following steps:

[0153] S1. Receive data: Receive sample data, which is the CD8 + The percentage of T cells expressing PD-1 and the percentage of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN, and CD107A;

[0154] S2. Output result: obtain a predicted value Y by applying the sample data to the following corresponding model formula, and output the immune reconstruction status of the HIV-infected subject who has undergone antiviral treatment according to the predicted value Y and the pre-determined condition;

[0155] The model formula is any of the following:

[0156] PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2;

[0157] PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2;

[0158] PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2;

[0159] PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2;

[0160] PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2;

[0161] PD-1 model formula: Y = -9.7176 + 5.1257X1;

[0162] Among them, Y is the predicted value, X1 is CD8 + The number of cells expressing PD-1 in T cells accounts for CD8 + The percentage of T cells, X2 is CD8 + The number of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in T cells accounted for CD8 + The percentage of T cell number;

[0163] The immune reconstitution status of the HIV-infected subjects who have received antiviral treatment is a good immune reconstitution status or a poor immune reconstitution status.

[0164] The pre-determination conditions corresponding to each model are as follows:

[0165] If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good.

[0166] If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

[0167] Results The immune reconstitution status after ART predicted by the six model formulas of 69 PLWH had no significant difference from the known INRs and IRs, indicating that the computer device can achieve the prediction purpose.

[0168] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

Claims

1. A computer device comprising a memory, a processor and a computer program stored in the memory, characterized in that: The processor executes the computer program to implement the following steps: S1. Receive data: Receive sample data, which is CD8 + The percentage of T cells expressing PD-1 and the percentage of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN, and CD107A; S2. Output result: obtain a predicted value Y by applying the sample data to the following corresponding model formula, and output the immune reconstruction status of the HIV-infected subject who has undergone antiviral treatment according to the predicted value Y and the pre-determined condition; The model formula is any of the following: PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2; PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2; PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2; PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2; PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2; PD-1 model formula: Y = -9.7176 + 5.1257X1; Among them, Y is the predicted value, X1 is CD8 + The number of cells expressing PD-1 in T cells accounts for CD8 + The percentage of T cells, X2 is CD8 + The number of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in T cells accounted for CD8 + The percentage of T cells.

2. The computer device according to claim 1, characterized in that: The pre-determination conditions of each model formula are as follows: If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good. If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

3. A computer program product, comprising a computer program, characterized in that: When the computer program is executed by a processor, the steps in the computer device according to claim 1 or 2 are implemented.

4. A computer-readable storage medium, characterized in that: The computer-readable storage medium stores a computer program, and the computer program causes a computer to execute the steps in the computer device according to claim 1 or 2.

5. A device for evaluating immune reconstitution in HIV-infected subjects undergoing antiviral therapy, the device comprising: S1, data receiving module: used to receive sample data, the sample data is CD8 + The percentage of T cells expressing PD-1 and the percentage of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN, and CD107A; S2, result output module: obtain the predicted value Y by the sample data through the following corresponding model formula, and output the immune reconstruction status of the HIV-infected subject who has undergone antiviral treatment according to the predicted value Y and the pre-determined conditions; The model formula is any of the following: PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2; PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2; PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2; PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2; PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2; PD-1 model formula: Y = -9.7176 + 5.1257X1; Among them, Y is the predicted value, X1 is CD8 + The number of cells expressing PD-1 in T cells accounts for CD8 + The percentage of T cells, X2 is CD8 + The number of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in T cells accounted for CD8 + The percentage of T cells.

6. The device according to claim 5, characterized in that: The pre-determination conditions of each model formula are as follows: If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good. If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

7. A method for evaluating or assisting in evaluating the immune reconstitution of HIV-infected subjects undergoing antiviral therapy, characterized in that: The method comprises the following steps: S1. Receive sample data, wherein the sample data is CD8 + The percentage of T cells expressing PD-1 and the percentage of cells expressing at least one of the following five factors: CD38, HLA-DR, CD57, PERFORIN, and CD107A; S2. Obtain a predicted value Y by applying the sample data to the following corresponding model formula, and output the immune reconstruction status of the HIV-infected subject who has undergone antiviral treatment according to the predicted value Y and the pre-determined condition; The model formula is any of the following: PD-1+CD57 model formula: Y=-9.7008+5.0551X1+0.0042X2; PD-1+HLA_DR model formula: Y=-12.2851+5.2778X1+0.1609X2; PD-1+CD107A model formula: Y=-11.0575+5.1849X1+0.0207X2; PD-1+PERFORIN model formula: Y=-10.2805+5.1479X1+0.0229X2; PD-1+CD38 model formula: Y=-9.7603+5.0304X1+0.0483X2; PD-1 model formula: Y = -9.7176 + 5.1257X1; Among them, Y is the predicted value, X1 is CD8 + The number of cells expressing PD-1 in T cells accounts for CD8 + The percentage of T cells, X2 is CD8 + The number of cells expressing any one of CD38, HLA-DR, CD57, PERFORIN and CD107A in T cells accounted for CD8 + The percentage of T cells.

8. The method according to claim 7, characterized in that: The pre-determination conditions of each model formula are as follows: If the predicted value Y of the subject is less than or equal to the threshold value, the immune reconstruction of the subject after antiviral treatment is good. If the predicted value Y of the subject is greater than the threshold, the immune reconstitution of the subject after antiviral treatment is poor.

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