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1.
Article in English | WPRIM | ID: wpr-906678

ABSTRACT

@#BACKGROUND: To investigate effects of Maxingloushi decoction on lung inflammation and programmed death markers (programmed death-1 [PD-1], programmed death-ligand 1 [PD-L1]) in the lung tissue, peripheral blood, and bronchoalveolar lavage fluid (BLF) in a mouse model of chronic obstructive pulmonary disease (COPD). METHODS: Thirty-six mature male BALB/C mice were randomly divided into normal group (group A, n=6), COPD model group (group B, n=10), Maxingloushi decoction + COPD group (group C, n=10), and PD-1 inhibitor + COPD group (group D, n=10). The COPD model was established by smoke inhalation combined with lipopolysaccharide (LPS). Levels of PD-1 and PD-L1 in plasma and BLF were measured by enzyme-linked immunosorbent assay (ELISA). Histopathological techniques were used to semi-quantitatively analyze the immuno-fluorescence optical density (IOD) value of the lung tissue. RESULTS: In plasma and BLF, the expression of PD-1 in the group B was higher than that in the group A, and the expression of PD-L1 was lower than that in the group A. The expression of PD-1 and PD-L1 in the lung tissue was normalized in the group C in comparison with the group B (P<0.05) and the group D (P<0.05), and inflammatory cell infiltration in the lung tissue was also improved. CONCLUSIONS: These findings reveal that COPD causes an immune imbalance in the peripheral blood and lung tissue, and that both Maxingloushi decoction and PD-1 inhibitor treatment can mitigate lung inflammation in COPD by reducing PD-1 expression and increasing PD-L1 expression. The treatment effect of Maxingloushi decoction may be superior to that of PD-1 inhibitor.

2.
Acta Pharmaceutica Sinica B ; (6): 1163-1185, 2022.
Article in English | WPRIM | ID: wpr-929376

ABSTRACT

Cancer immunotherapy has become a new generation of anti-tumor treatment, but its indications still focus on several types of tumors that are sensitive to the immune system. Therefore, effective strategies that can expand its indications and enhance its efficiency become the key element for the further development of cancer immunotherapy. Natural products are reported to have this effect on cancer immunotherapy, including cancer vaccines, immune-check points inhibitors, and adoptive immune-cells therapy. And the mechanism of that is mainly attributed to the remodeling of the tumor-immunosuppressive microenvironment, which is the key factor that assists tumor to avoid the recognition and attack from immune system and cancer immunotherapy. Therefore, this review summarizes and concludes the natural products that reportedly improve cancer immunotherapy and investigates the mechanism. And we found that saponins, polysaccharides, and flavonoids are mainly three categories of natural products, which reflected significant effects combined with cancer immunotherapy through reversing the tumor-immunosuppressive microenvironment. Besides, this review also collected the studies about nano-technology used to improve the disadvantages of natural products. All of these studies showed the great potential of natural products in cancer immunotherapy.

3.
Acta Pharmaceutica Sinica B ; (6): 1254-1270, 2022.
Article in English | WPRIM | ID: wpr-929346

ABSTRACT

Molecular targeted therapy has become an emerging promising strategy in cancer treatment, and screening the agents targeting at cancer cell specific targets is very desirable for cancer treatment. Our previous study firstly found that a secretory peroxidase of class III derived from foxtail millet bran (FMBP) exhibited excellent targeting anti-colorectal cancer (CRC) activity in vivo and in vitro, whereas its underlying target remains unclear. The highlight of present study focuses on the finding that cell surface glucose-regulated protein 78 (csGRP78) abnormally located on CRC is positively correlated with the anti-CRC effects of FMBP, indicating it serves as a potential target of FMBP against CRC. Further, we demonstrated that the combination of FMBP with the nucleotide binding domain (NBD) of csGRP78 interfered with the downstream activation of signal transducer and activator of transcription 3 (STAT3) in CRC cells, thus promoting the intracellular accumulation of reactive oxygen species (ROS) and cell grown inhibition. These phenomena were further confirmed in nude mice tumor model. Collectively, our study highlights csGRP78 acts as an underlying target of FMBP against CRC, uncovering the clinical potential of FMBP as a targeted agent for CRC in the future.

4.
Article in Chinese | WPRIM | ID: wpr-875866

ABSTRACT

@#[Abstract] Objective: To investigate the effects of antibiotics on the treatment efficacy of immune checkpoint inhibitors in NSCLC (non-small cell lung cancer) with Meta-analysis. Methods: Literatures regarding the effects of antibiotics on the treatment efficacy of immune checkpoint inhibitors in NSCLC were searched in Pubmed, Cochrane Library, Embase, EBSCO, Chinese Biomedical Literature Database(CBM) and Chinese Journal Full-text Database(CNKI). RevMan 5.3 software was used in this Meta-analysis. Results: Fourteen articles involving 2 505 NSCLC patients were included in this study. Meta-analysis showed that the application of antibiotics could significantly shorten the PFS (HR=1.14, 95%CI =1.04-1.26, P=0.005) and OS (HR=1.30, 95%CI =1.14-1.47, P<0.0001) of NSCLC patients treated with immune checkpoint inhibitors. Conclusion: Application of antibiotics before, concurrently or after immune checkpoint inhibitors in the treatment of NSCLC may significantly shorten PFS and OS, resulting in adverse effect on treatment efficacy.

5.
Article in Chinese | WPRIM | ID: wpr-907994

ABSTRACT

Objective:To investigate the role and clinical significance of T follicular helper (Tfh) cells in the pathogenesis of eosinophilic gastroenteritis (EG) in children.Methods:A total of 17 children diagnosed with EG in the Department of Gastroenterology, Children′s Hospital of Nanjing medical University from October 2018 to January 2020 were recruited as EG group.During the same period, 15 children diagnosed with colon polyps were included as control group.Flow cytometry was used to detect Tfh cells and their functional molecules, including inducible costimulatory molecules (ICOS) and programmed cell death protein 1 (PD-1) in peripheral blood of the 2 groups.Results:The median (interquartile range) of Tfh cell frequency in peripheral blood of children with EG group was 7.3 (2.6)%, which was significantly higher than that of controls 2.8 (1.4)% ( P<0.05). There were significant differences in the median (interquartile range) of ICOS [1.5 (1.3)% vs.0.1 (0.2)%] and PD-1 expressions [1.8 (3.2)% vs.0.7 (0.6)%] on Tfh cells between children with EG group and control group (all P<0.05). The frequency of Tfh cells in the peripheral blood of children with EG was positively correlated with the expressions of ICOS ( r=0.746, P<0.05) and PD-1 ( r=0.893, P<0.05), and immunoglobulin E (IgE) level ( r=0.587, P<0.05). Conclusions:The frequency of Tfh cells in peripheral blood of children with EG significantly increases, which are proliferative and overexpressed with ICOS and PD-1.Moreover, the frequency of Tfh cells of EG patients is positively correlated with the level of IgE.The abnormal expression of Tfh cells may play a promoting role in the mechanism of EG.

6.
Article in Chinese | WPRIM | ID: wpr-907923

ABSTRACT

Objective:To explore the expressions of programmed death-1 (PD-1) and programmed death-ligand 1 (PD-L1) and clinicopathological characteristics in post-transplant lymphoproliferative disorder (PTLD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children, with the aim of clarifying whether checkpoint inhibition of PD-1/PD-L1 inhibitors may serve as a therapy option.Methods:The clinical data of 13 cases of PTLD after allo-HSCT pathologically confirmed in Shenzhen Children′s Hospital from January 1, 2012 to December 30, 2019 were retrospectively analyzed.The detection was performed by immunohistochemical staining by MaxVision? method, Epstein-Barr virus(EBV) in situ hybridization and lymphoma gene rearrangement.The relationship between the expression of PD-1 and PD-L1 and the clinicopathological characteristics of PTLD were analyzed.Results:The expression of PD-1 was not correlated with gender, age, primary diseases, histopathological types, transplantation mode and the expression of EBV in situ hybridization (all P>0.05). The expression of PD-L1 was correlated with histopathological types ( P<0.05). Furthermore, the expression rate of PD-L1 on severe β-thalassemia was significantly higher than that of severe aplastic anemia [90.0%(9/10 cases) vs. 66.7%(2/3 cases)] and monomorphic PTLD was higher than that of polymorphic PTLD [100.0%(2/2 cases) vs. 83.3%(5/6 cases)]. Moreover, the positive PTLD in EBV was higher than the negative PTLD in EBV [90.9%(10/11 cases) vs. 50.0%(1/2 cases)]. The positive rates of PD-1 and PD-L1 in 13 cases with PTLD were 46.2%(6/13 cases) and 61.5%(8/13 cases) in tumor cells, 92.3% (12/13 cases) and 76.9% (10/13 cases) in microenvironmental cells, and 84.6%(11/13 cases) in EBV, respectively. Conclusions:PD-L1 has a higher positive rate in tumor cells with monomorphic PTLD; and routine staining for PD-1 and PD-L1 can be performed in all types of PTLD when standard immunotherapy and chemotherapy are ineffective.

7.
Article in Chinese | WPRIM | ID: wpr-907557

ABSTRACT

With the development of immunotherapy in clinical application, immunotherapy also takes advantage in esophageal squamous cell carcinoma (ESCC). Immune checkpoint inhibitors such as programmed death-1 (PD-1) and its ligand PD-L1 and cytotoxic T lymphocyte antigen-4 show significant antitumor activity and safety in immunotherapy for patients with advanced ESCC.

8.
Journal of Leukemia & Lymphoma ; (12): 508-512, 2021.
Article in Chinese | WPRIM | ID: wpr-907206

ABSTRACT

When chimeric antigen receptor T cells (CAR-T) failed to treat relapsed and refractory B-cell malignancies, some researchers try to improve the efficacy by enhancing the function of CAR-T. It is a new hotspot of drug development and clinical research, and significant achievements have been made in this area recently. Multi-targeted CAR-T, lenalidomide, decitabine, programmed death 1 inhibitor and ibrutinib can all enhance the function of CAR-T. This article reviews the recent progress of enhancing the function of CAR-T in relapsed and refractory B-cell malignancies.

9.
Journal of Leukemia & Lymphoma ; (12): 449-454, 2021.
Article in Chinese | WPRIM | ID: wpr-907198

ABSTRACT

The programmed death-1 (PD-1) inhibitor has a unique mechanism and better efficacy in classic Hodgkin lymphoma (cHL). Multiple PD-1 inhibitors have been approved for the salvage treatment of patients with cHL in advanced stage. The combination of PD-1 with other small molecule targeted drugs, immunoactive drugs or cytotoxic drugs can further improve the efficacy and have a better safety compared with the traditional treatment, meanwhile, the regimens have also been explored in treatment of recurrent and refractory patients and frontline therapy. In addition, 9P24.1 amplification, PD-1 ligand expression and circulating tumor DNA level may be potential biomarkers for predicting the efficacy of PD-1 inhibitors.

10.
Cancer Research and Clinic ; (6): 641-644, 2021.
Article in Chinese | WPRIM | ID: wpr-912939

ABSTRACT

With the breakthrough of immune checkpoint blocking therapy in the clinical treatment of a variety of malignant solid tumors, tumor immunotherapy has opened a new era. However, clinical practice has proved that the response rate of this therapy is low. Seeking for the key factors limiting its response rate has become a research hotspot in this field. Programmed death ligand 2 (PD-L2) is the second important ligand binding to programmed death 1 (PD-1) after programmed death ligand 1 (PD-L1). The encoded protein can bind to PD-1 and then inhibit immune cells. This paper reviews the biological characteristics of PD-L2, its mechanism in immune regulation and its research progress in solid tumor immunotherapy to provide more theoretical support for solid tumor immunotherapy.

11.
Article in Chinese | WPRIM | ID: wpr-912065

ABSTRACT

Programmed death-1 (PD-1) is an immunosuppressive molecule on the surface of several cells, and functions as a negative regulatory factor in cellular and humoral immune responses via interaction with its ligand PD-L1 or PD-L2. The expression of PD-1 on the peripheral blood lymphocytes of human immunodeficiency virus (HIV) infected people is commonly up-regulated, which not only affects the susceptibility of the virus to target cells and the host′s antiviral immune response, but also affects the effect of antiretroviral therapy. In vitro blocking PD-1/PD-L pathway showed improved HIV-specific immune response of host cells, and PD-1 blockade and/or PD-L1 blockade have been used as an auxiliary means to enhance the efficacy of antiviral therapy and HIV vaccines. This article reviews the progress of the studies on PD-1 in HIV infection, prevention and treatment.

12.
Article in Chinese | WPRIM | ID: wpr-910623

ABSTRACT

Biliary tract cancer is characterized by high heterogeneous, rare and refractory malignancy. Since patients are often diagnosed at late stage, the radical resection rate is low, and the effect of traditional adjuvant therapy is limited, therefore the prognosis of patients is poor. Recently Immunotherapy has opened up a new field for tumor therapy. Porgrammed death-1 (PD-1) antibody therapy has a great clinical application prospects. The efficacy of related therapies in biliary tract tumors is being evaluated under a number of clinical trials. One of the main challenges is to identify the biomarkers that can predict the response and prognosis of PD-1 antibody therapy. This article aims to summarize the research progress of PD-1 antibody therapy and related biomarkers such as PD-L1, tumor mutational burden, DNA damage repair in biliary tract cancers, and to prospect the future research direction.

13.
Acta Pharmaceutica Sinica B ; (6): 750-762, 2021.
Article in English | WPRIM | ID: wpr-881167

ABSTRACT

The protein tyrosine phosphatase Src homology phosphotyrosyl phosphatase 2 (SHP2) is implicated in various cancers, and targeting SHP2 has become a promising therapeutic approach. We herein described a robust cross-validation high-throughput screening protocol that combined the fluorescence-based enzyme assay and the conformation-dependent thermal shift assay for the discovery of SHP2 inhibitors. The established method can effectively exclude the false positive SHP2 inhibitors with fluorescence interference and was also successfully employed to identify new protein tyrosine phosphatase domain of SHP2 (SHP2-PTP) and allosteric inhibitors. Of note, this protocol showed potential for identifying SHP2 inhibitors against cancer-associated SHP2 mutation SHP2-E76A. After initial screening of our in-house compound library (∼2300 compounds), we identified 4 new SHP2-PTP inhibitors (0.17% hit rate) and 28 novel allosteric SHP2 inhibitors (1.22% hit rate), of which SYK-85 and WS-635 effectively inhibited SHP2-PTP (SYK-85: IC

14.
Acta Pharmaceutica Sinica B ; (6): 2983-2994, 2021.
Article in English | WPRIM | ID: wpr-922779

ABSTRACT

Genomic instability remains an enabling feature of cancer and promotes malignant transformation. Alterations of DNA damage response (DDR) pathways allow genomic instability, generate neoantigens, upregulate the expression of programmed death ligand 1 (PD-L1) and interact with signaling such as cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling. Here, we review the basic knowledge of DDR pathways, mechanisms of genomic instability induced by DDR alterations, impacts of DDR alterations on immune system, and the potential applications of DDR alterations as biomarkers and therapeutic targets in cancer immunotherapy.

15.
Cancer Research and Clinic ; (6): 32-35, 2020.
Article in Chinese | WPRIM | ID: wpr-799300

ABSTRACT

Objective@#To analyze the expressions of coordinated stimulating molecular programmed death 1(PD-1) and programmed death ligand 1 (PD-L1) in human glioma and their clinical significances.@*Methods@#A total of 70 postoperative paraffin specimens of brain glioma and 35 normal brain tissues in Heji Hospital Affiliated to Changzhi Medical College from January 2013 to December 2017 were collected. The expressions of PD-1 and PD-L1 in 70 glioma tissues and 35 normal brain tissues were detected by immunohistochemical SP method. The relationship between the expressions of PD-1 and PD-L1 and their correlation with the clinicopathological features were analyzed.@*Results@#The positive expression rates of PD-1 and PD-L1 in glioma tissues were 69% (48/70) and 62% (43/70), respectively, which were higher than those in normal brain tissues [29% (10/35), 31% (11/35)], the differences were statistically significant (χ2 values were 15.099 and 8.407, both P < 0.05). The positive expression rates of PD-1 and PD-L 1 in high-grade glioma were 81% (30/37) and 73% (27/37), respectively, which were higher than those in low-grade glioma [55% (18/33) and 49% (16/33)], the differences were statistically significant (χ 2 values were 5.699 and 4.415, both P < 0.05). There was no significant difference in the positive expression rates of PD-1 and PD-L1 among patients with different sex, age and maximum tumor diameter (all P > 0.05). There was a positive correlation between the expressions of PD-1 and PD-L1 proteins in glioma tissues (r= 0.372, P= 0.002).@*Conclusions@#The PD-1 and PD-L1 may become new biological indicators for evaluating the occurrence and development of glioma.

16.
Article in Chinese | WPRIM | ID: wpr-872829

ABSTRACT

Objective:To clarify the effect of Fangfeng Tongshengtang on early-stage serum endotoxin (ET) and programmed death-1/programmed ligand-1(PD-1/PD-L1) in patients with hepatitis B virus-related acute-on-chronic liver failure(HBV-ACLF)(early-stage), and exploring the mechanism of Fangfeng Tongshengtang in the treatment of early stage HBV-ACLF. Method:The 69 patients with early stage HBV-ACLF were enrolled in the study and all of them received antiviral drugs, liver protection and jaundice relieving drugs as well as supporting therapy. According to the random number table, 35 patients were randomly assigned to observation group (to take Fangfeng Tongshengtang, and 34 patients were assigned to control group to take placebo. The observation period was 3 weeks in both groups. Before treatment and 1, 2, and 3 weeks after treatment, theserum ET, expression of PD-1/PD-L1 in serum CD4+ and CD8+ T cells, liver function [alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), total bilirubin (TBIL), and direct bilirubin (DBIL)], coagulation function [prothrombin time (PT), and prothrombin activity (PTA)] were detected to verify the effect of Fangfeng Tongshengtang on HBV-ACLF (early-stage). Result:After 3 weeks of treatment, ET, expression of serum CD4+PD-1+, CD4+PD-L1+, CD8+PD-1+, CD8+PD-L1+, ALT, AST, TBIL, DBIL, and PT decreased significantly (P<0.01), while Alb and PTA increased significantly(P<0.01)in both groups. As compared with the control group, the ET in observation group was lower at 1st, 2nd and 3rd week after treatment (P<0.01), the CD4+PD-1+, CD4+PD-L1+, CD8+PD-1+ and CD8+PD-L1+ in observation group were lower at 2nd week and 3rd week(P<0.05, P<0.01), the ALT, AST, TBIL and DBIL in observation group were lower at 1st, 2nd and 3rd week(P<0.05, P<0.01), the PT in observation group was lower at 2nd and 3rd week(P<0.05), and the PTA in observation group was higher at the 2nd and 3rd week(P<0.01). Conclusion:Fangfeng Tongshengtang can achieve the therapeutic effect for HBV-ACLF (early-stage) probably by reducing the serum ET and the expression of PD-1 / PD-L1 in serum CD4 +, CD8 + T cells.

17.
Article in Chinese | WPRIM | ID: wpr-861609

ABSTRACT

Head and neck squamous cell carcinoma (HNSCC) is a kind of malignant tumor characterized by metastasis and local invasion. Its recurrence rate is high after surgery and radiotherapy, and the prognosis and quality of life are poor. In recent years, programmed death-1 (PD-1) inhibitors have been recommended in National Comprehensive Cancer Network (NCCN) guidelines for the treatment of recurrent, unresectable, and metastatic HNSCC, and their efficacy has been remarkable. PD-1 inhibitors constitute a new treatment for the patients with advanced HNSCC who are refractory to platinum-based chemotherapy and can increase the probability of surgical resection, reduce the risk of postoperative dysfunction, and improve the survival and quality of life. This article reviews the structure and mechanism of the PD-1/PD-L1 immunocheckpoint, as well as research progress on its inhibitors in the treatment of HNSCC.

18.
Article in Chinese | WPRIM | ID: wpr-843275

ABSTRACT

Programmed death-1 (PD-1) receptor and programmed death-ligand 1 (PD-L1) as a pair of T cell immune response co-stimulatory molecules play a negative role in adoptive immunity by inhibiting T lymphocyte function. Blocking the PD-1/PD-L1 signaling pathway has been a hot spot of research and treatment of cancer. Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer, and there is no approved targeted therapy. Immunotherapy represented by PD-1/PD-L1 blockers, is getting widely studied due to the medical potential of therapy for TNBC. Thereinto, the clinical researches of the monoclonal antibodies against PD-1 or PD-L1 have brought a promising future for the treatment of metastatic TNBC. Currently, there are several trials with anti-PD-1 or anti-PD-L1 monoclonal antibodies and their combination with other therapies in salvage, neoadjuvant, and adjuvant settings on going. In this article, the authors review the current studies about potential value of PD-1/PD-L1 in prognosis, prediction, and treatment in TNBC, intending to cast insight on future basic and clinical studies.

19.
Article in Chinese | WPRIM | ID: wpr-843260

ABSTRACT

Objective:To establish a lung cancer mouse model with humanized peripheral blood mononuclear cells (PBMC) expressing programmed death-ligand 1 (PD-L1), and study the role of the model in evaluating the efficacy of programmed death-1 (PD-1) inhibitors. Methods:Fresh biopsy tissue samples or tumor cells in malignant pleural effusion from the patients with advanced non-small cell lung cancer were inoculated subcutaneously in CB17-SCID mice to establish patient-derived xenograft (PDX) models. The expression of PD-L1 in PDX models was detected by immunohistochemistry. The mature human PBMC and PDX model tumor cells were mixed and then inoculated into NCG mice to establish a PDX model of lung cancer with humanized immunity, on which the efficacy of PD-1 inhibitor was verified. Results:Among the PDX models established by 16 clinical samples, 2 were strongly positive for PD-L1, 4 were positive, and the rest were negative. In the PDX model with strongly positive PD-L1, the tumor growth inhibition rate of cindilimab, an inhibitor of PD-1, was 82.6%, 21 days after the initial administration; in the PDX model with negative PD-L1, the inhibitor of PD-1 showed no antitumor activity. Conclusion:A PD-L1-expressing lung cancer mouse model with humanized immunity is successfully established and the efficacy of PD-1 inhibitor can be evaluated on the model.

20.
Article in Chinese | WPRIM | ID: wpr-828890

ABSTRACT

OBJECTIVE@#To investigate whether interleukin-12 (IL-12) over-expression in malignant melanoma B16 cells affects the expression level of programmed death-1 (PD-1) on T cells in mice during immune microenvironment reconstruction.@*METHODS@#B16 cells were transfected with an IL-12 expression lentiviral vector, and IL-12 over-expression in the cells was verified qPCR and ELISA. Plate cloning assay was used to compare the cell proliferation activity between B16 cells and B16/IL-12 cells. The expression of IL-12 protein in B16/IL-12 cells-derived tumor tissue were detected by ELISA. C57BL/6 mice were inoculated with B16 cells or B16/IL-12 cells, and 14 days later the proportion of T cells with high expression of PD-1 in the tumor-draining lymph nodes was detected by flow cytometry. Mouse models of immune reconstitution established by 650 cGy X-ray radiation were inoculated with B16 (B16+RT group) or B16/IL-12 (B16/IL-12+RT group) cells, with the mice without X-ray radiation prior to B16 cell inoculation as controls. Tumor growth in the mice was recorded at different time points, and on day 14, flow cytometry was performed to detect the proportion of T cells with high PD-1 expression in the tumor-draining lymph nodes and in the tumor tissue.@*RESULTS@#B16 cells infected with the IL-12-overexpressing lentiviral vector showed significantly increased mRNA and protein levels of IL-12 ( < 0.001) without obvious changes in cell viability (>0.05). B16/IL-12 cells expressed higher levels of IL-12 than B16 cells ( < 0.01). In the tumor-bearing mouse models, the proportion of CD4 PD-1 T cells was significantly lower in B16/IL-12 group than in B16 group ( < 0.01). In the mice with X-ray radiation-induced immune reconstitution, PD-1 expressions on CD4 T cells ( < 0.05) and CD8+ T cells ( < 0.01) were significantly higher in B16+ RT group than in the control mice and in B16/IL-12+RT group ( < 0.01 or 0.001); the tumors grew more slowly in B16/IL-12+RT group than in B16 + RT group ( < 0.001).@*CONCLUSIONS@#During immune microenvironment reconstruction, overexpression IL-12 in the tumor microenvironment can reduce the percentage of PD-1 T cells and suppress the growth of malignant melanoma in mice.


Subject(s)
Animals , CD8-Positive T-Lymphocytes , Cell Line, Tumor , Immune Reconstitution , Interleukin-12 , Melanoma, Experimental , Mice , Mice, Inbred C57BL , Tumor Microenvironment
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