Study On The Immunity Of Patients With Lung Cancer And The Effects Of Chemotherapy On The Immunity

OBJECTIVE: Lung cancer is the leading cause of cancer deaths in many countries and in china. Non-small-cell lung cancer accounts for approximately 75%～80% of lung cancer cases, with the majority of patients having inoperable locally advanced or metastatic disease at the time of diagnosis. The 5-year survival rate is very low for all stages. As a non-specific cytoxicity treatment, chemotherapy can kill the cancer cells. However the damage of the immunological function sometimes makes the disease worse. So we evaluated the state of immunity before and after chemotherapy in patients with lung cancer.METHOD: (1) The FCM of 2002 to April of 2006, 189 patients PBL with lung cancer and by the SAS6. 02 microsoft we analyzed the difference between the different TNM stages, KPS and metastasis, of the ratio of T lymphocyte subgroups,B lymphocyte and NK. The change of immunity was analyzed before and after chemotherapy between 102 patients on emphasis. And the correlation between the clinical response and the change of the T lymphocyte subgroup,B lymphocyte and NK is also analyzed. (2) The FCM of the 103 patients' PBL with lung cancer from 2005 June to 2006 April:the ratio of CD3+HLA-DR+,CD4+CD25+ and CD8+CD28+ lymphocyte; At the single cell level by the FCM the ratio of the Th subgroup is measuered . By the SAS6. 02 microsoft we analyzed the difference between the different TNM stages, KPS and different metastasis, of CD3+HLA-DR+,CD4+CD25+ and CD8+CD28+ lymphocyte and the Th subgroup. The change of immunity was analyzed before and after chemotherapy between 83 patients on emphasis. And the correlation between the clinical response and the change of the ratio of CD3+HLA-DR+,CD4+CD25+ and CD8+CD28+ lymphocyte and the Th subgroup is also analyzed. (3) Peripheral blood from 47 patients before and after chemotherapy were collected respectively. The mononuclear cells were harvested by Ficoll density gradient centrifuge. Their RNAs were extracted and the cDNAs were synthesized by RT. Both of the competitor with known concentration and cDNA(2μg) were added in the PCR mixture in which the amount of primers were limited for competitive quantitative PCR to measure the amount of mRNAs which reflecting the gene expression of FasL, Perforin, Granzyme B. By this competitive quantitative RT-PCR technique the correlation of the expressions of FasL, Perforin, Granzyme B and the clinical response before and after chemotherapy were then analyzed.RESULTS AND DISCUSSION:1, The ratio of T lymphocyte subgroups,B lymphocyte and NK of patients PBL with lung cancer and the effect of chemotherapy on the immunity. (1)After single chemotherapy, the ratio of CD3+CD4+ and CD4+/CD8+ decreased significantly, NK and B cell increased unsignificantly . (2)Between the circles of chemotherapy >4 and≤4, the ratio of CD3+,CD4+ and CD4+/CD8+ dereased significantly, but the ratio of CD8+ increased significantly. (3)They had no predictive function. (4) Between the III stage and contral, the ratio of NK cell and the CD4+/CD8+ decreased significantly ;the IV stage and contral, the ratio of CD3+, CD3+CD4+,NK cell and CD4+/CD8+ decreased significantely, CD3+CD8+ cell increased significant -ly, B lymphocyte had no change; Between the III and IV stage, there is no significantly change. (5)Between KPS<70 and KPS≥70, the ratio of NK cell and the CD4+/CD8+ decreased significantly. (6) Between multiple and single metastasis, CD4+/CD8+ decreased significantly and CD3+CD8+ cell increasing significantly. 2, The level of the activated T lymphocytes and the effect of chemotherapy on the immunity. (1) After single chemotherapy, the ratio of CD4+CD25+ increased significantely, the others had no significantely change. (2) Between the circles of chemotherapy >4 and≤4, the ratio of CD3+HLA-DR+,CD8+CD28+ and Th1 dereased significantly, but the ratio of CD4+CD25+ and Th2 increased significantly. (3)To predict clinical response:the ratio of CD4+CD25+Treg increasing predict bad clinical response, the others has no predictive function. (4) Between theIII stage and contral, the ratio of CD3+HLA-DR+ cell decreased significantly; between the IV stage and contral, the ratio of CD3+HLA-DR+ and CD8+CD28+ cell decreased significantely, CD4+CD25+ and Th2 cell increased significantly, Th0 and Th1 lymphocyte had no change. Between theIV and III stage, Th1 lymphocyte decreased significantely. (5) Between KPS <70 and KPS≥70, the ratio of CD8+CD28+ cell decreased significantly, but CD4+CD25+ and Th2 increased significantly. (6)Between multiple and single metastasis, the ratio of CD8+CD28+ cell decreased significantly, but CD4+CD25+ and Th2 increased significantly. 3, The effect of chemotherapy on the gene expression of FasL,Granzyme B and Perforin:(1) after single chemotherapy, the level of the gene expression of FasL,Granzyme B and Perforin were downregulated significantly. (2) Between the patients with the circles of chemotherapy>4and≤4, the the gene expression of FasL,Granzyme B and Perforin were downregulated significantly. (3)The upregulated expression of Granzyme B and FasL were related to good response, but perforin has no predictive function.CONCLUSION:1, The effects of chemotherapy on the immunity . (1) After single chemotherapy , the ratio of CD3+CD4+, CD4+/CD8+ decreased, but the ratio of CD4+CD25+ and gene expressions of Granzyme B, Perforin, or FasL increased, P≤0.05 respectively. The others has no significant change. The ratio of NK cell and the activated CD8+ cell have no significant chang but the quality of the killer activation decreased significantely. (2)Between the patients with the circles of chemotherapy＞4 and≤4, the ratio of CD3+,CD3+CD4+,CD3+HLA-DR+,CD8+CD28+,Th1 cell,CD4+/CD8+ and the expression level of Perforin,Granzyme B,FasL decreased(P≤0.05), the ratio of CD3+CD8+,CD4+CD25+ and Th2 cell increased(P≤0.05);the ratio of NK,B lymphocyte and Th0 cell has no significant change. (3)To predict clinical response:the ratio of CD4+CD25+ cell increasing predict bad clinical response, but the upregulated expression of Granzyme B and FasL was related to good response. the others has no predictive function. 2, The relationship between the immunity of people with lung cancer and the TNM stages,KPS and different metastasis numbers. (1) TNM stages: between the III stage and contral, the ratio of the CD3+HLA-DR+,NK cell and CD4+/CD8+ decreased significantly; between the IV stage and contral, the ratio of CD3+,CD3+CD4+,CD3+HLA-DR+,CD8+CD28+,NK cell and CD4+/CD8+ decreased significantely, CD3+CD8+,CD4+CD25+and Th2 cell increasing significantly, B lymphocyte and Th0 have no change; between the III and IV stage, Th1 cell decreased significantly. (2) Between KPS<70 group and KPS≥70 group, CD4+/CD8+ and the ratio of NK cell decreased significantly. (3) Between multiple and single metastasis, the ratio of CD8+CD28+ and NK cell and CD4+/CD8+ decreased, the CD3+CD8+,CD4+CD25+ and Th2 cell increased respectively significantly. But the ratio of B lymphocyte,Th1 and Th0 cell had no significantly difference.