| Objective: PD-1(Programmed death-1,programmed death molecule 1)is a marker of T lymphocytes in immunosuppressive state.The overexpression of PD-1 on T lym phocytes affects its proliferation and killing activity.To investigate the expression of PD-1 in peripheral blood mononuclear cells(PBMCs)of patients with nasopharyngeal carcinoma(NPC),and to analyze the relationship between the expression of PD-1 an d the clinical stage in patients with nasopharyngeal carcinoma(NPC)The effects of NKT lymphocytes,??T lymphocytes and NK lymphocytes on the killing of nasopharyn geal carcinoma cell line CNE-1 were studied by immunizing the threekinds lymphocy tes of peripheral blood of patients with nasopharyngeal carcinoma.The effect of pd-1on the lymphocyte type of PD-1 monoclonal antibody was studied to influence the kill ing effect of nasopharyngeal carcinoma cell line.Methods: The expression of PD-1 on PBMC surface of 53 patients with nasopharyng eal carcinoma and 58 healthy people was detected by flow cytometry.The expression of PD-1 and CD3 + CD8 + cytotoxic T lymphocytes and CD3 + CD4 + T cell correlation s and clinical staging were used to evaluate the cellular immune status of patients with nasopharyngeal carcinoma.At the same time,PBMC cells were isolated from periphe ral blood of patients with nasopharyngeal carcinoma,and the cytokines such as GM-C SF and IL-4 were used ??T lymphocytes,NK cells and NKT lymphocytes were cultur ed in vitro by ?-Galcer,anti-CD3 m Ab,IFN-?,IL-2 and other cytokines.Cells,?? T ly mphocytes and NKT lymphocytes and nasopharyngeal carcinoma tumor antigen pre-e xcitation DC co-incubation,and ultimately the development of antigen pre-stimulated T cells,using no marker tracking method to compare the three lymphocytes to kill nas opharyngeal carcinoma The expression of PD-1 on the surface of T lymphocytes was blocked by the addition of anti-PD-1 monoclonal antibody to detect the proliferation o f T lymphocytes after blocking And PD-1 expression.Finally,the specific killing acti vity of nasopharyngeal carcinoma cell line CNE-1 was detected by marker-free metho d before and after blocking T lymphocytes with the strongest killing ability.Results:(1)The expression rate of PD-1 in PBMC of patients with nasopharyngeal ca rcinoma was significantly higher than that of healthy people(P <0.0001),but there was no significant difference with clinical stage of nasopharyngeal carcinoma.(2)The e xpression of PD-1 was negatively correlated with the contents of CD3 + CD8 + CTL(c ytotoxic lymphocyte)and CD45RO+CD62L TEM(effector memory T cell)(P <0.0001).(3)Unlabeled tracing experiments showed that NKT lymphocytes were the most capable of killing CNE-1 cells in three kinds of lymphocytes,followed by NK lympho cytes and ?? T lymphocytes(P <0.05(P <0.05),but the expression of PD-1 on NKT c ells was significantly decreased(P <0.0001).(4)The anti-PD-1 monoclonal antibody had no effect on the proliferation of NKT lymphocytes(P> 0.05).(5)The killing activ ity of NKT lymphocytes in PD-1 block group was higher than that in control group(P<0.05),and the cytotoxicity of CNE-1 was significantly higher than that of control gr oup(P <0.05).Conclusion:(1)The expression rate of PD-1 in PBMC of patients with nasopharynge al carcinoma is higher than that in healthy PBMC.The high expression of PD-1 is in a way that reflects the inhibition of T lymphocyte function in some way.The expressio n of PD-1 in PBMC of patients with nasopharyngeal carcinoma by flow cytometry ca n be used as an evaluation index for the clinical application of anti-PD-1 monoclonal antibody.(2)The expression of PD-1 in PBMC was not correlated with the staging of nasopharyngeal carcinoma patients,and it could not be used as a prognostic indicator of nasopharyngeal carcinoma.(3)The high expression of PD-1 on the surface of adop tive NKT lymphocytes in vitro can effectively improve its ability to kill CNE-1 in vitr o,and the combination of anti-PD-1 monoclonal antibody and adoptive NKT cells bec ome a new way of immunotherapy for nasopharyngeal carcinoma. |
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