Analytical assessment of predictive value of immune parameters in tumorigenesis of NSCLC
To explore the predictive role of immune cells in untreated NSCLC patients, a total of 487 Chinese adults (305 lung cancer patients and 132 healthy controls) were enrolled in this study. We did not analyze inflammatory cells due to lack these data for controls. It is verified that levels of lymphocyte subsets were significantly associated with gender and age in healthy controls and cancer patients, thus we carefully avoided age- and sex-related biases.
We compared levels of immune cells in all patients and controls based on t-test and Mann–Whitney test. In this study, low levels of T lymphocytes (p<0.001), NK cells (p<0.001), CD8+ T cells (p=0.008), naïve CD4+/CD4+ (p<0.001), and naïve CD4+ T cells (p<0.001) was observed in lung cancer patients compared to controls. However, levels of CD4+ T cells (p=0.042), memory CD4+/CD4+ (p<0.001), memory CD4+ T cells (p<0.001), CD4+CD28+/CD4+ (p<0.001), CD4+CD28+ T cells (p=0.002), CD8+CD28+/CD8+ (p=0.004), CD8+HLA-DR+/CD8+ (p<0.001), CD8+HLA-DR+ T cells (p=0.022), CD8+CD38+/CD8+ (p<0.001), CD8+CD38+ T cells (p=0.001) and CD4+/CD8+ (p<0.001) were higher in patients than those in controls. There is no significant difference for B cells and CD8+ CD28+ T cells counts between patients and controls (p>0.05). The result was shown in table 2.
Predictive value of immune parameters in NSCLC progression
To further analyze the role of immune cells in NSCLC progression, the 305 NSCLC patients were divided into 4 group by the stages. In this study, a trend of decrease in B cells counts (r=-0.193, p=0.001, Fig. 1A), CD4+ T cells counts (r=-0.135, p=0.020, Fig. 1C), naïve CD4+/CD4+ percentage (r=-0.122, p=0.037, Fig. 1D), naïve CD4+ T cells counts (r=-0.144, p=0.013, Fig. 1E), CD4+CD28+ T cells counts (r=-0.137, p=0.019, Fig. 1F), and CD8+CD28+ T cells counts (r=-0.186, p=0.001, Fig. 1G) was noted for patients with advancing stage. On the contrary, there were increasingly advancing stage related trend for NK cells counts (r=0.117, p=0.045, Fig. 1B), WBC counts (r=0.177, p=0.002, Fig. 1H), monocytes (r=0.186, p=0.001, Fig. 1I), neutrophils (r=0.158 p=0.007, Fig. 1J), eosinophils (r=0.171, p=0.003, Fig. 1K), basophils (r=0.203, p<0.001, Fig. 1L), MLR (r=0.206, p<0.001, Fig. 1M), NLR (r=0.165, p=0.005, Fig. 1N), ELR (r=0.188, p=0.001, Fig. 1O), BLR (r=0.230, p<0.001, Fig. 1P), PLR (r=0.121, p=0.038, Fig. 1Q).There were no significant correlation between other immune cells levels and advancing stages (Supplementary Table 1). Noteworthily, stage II patients had highest levels of T lymphocytes, NK cells, CD4+ T cells, CD8+ T cells, memory CD4+ T cells, CD4+CD28+ T cells, CD8+CD28+ T cells, CD8+HLA-DR+ T cells, lymphocytes and lowest counts of WBC, neutrophils than those in other stages.
Correlation of immune cells levels with clinicopathologic characteristics
To further demonstrate the relationship between immune cells levels and clinicopathologic characteristics based on t text, Mann–Whitney test for 2 group, and Spearman’s rank correlation test for more than 2 groups, which was summarized in Table 3-4 and figure 2-3. There are high B cells counts (p<0.001) and CD8+CD28+/CD8+ percentage (p=0.047) in female patients than those in male. On the contrary, we discovered low counts of WBC (p=0.005), monocytes (p<0.001), neutrophils (p=0.001), eosinophils (p=0.006), RBC (p<0.001), hemoglobins (p<0.001), and MLR (p<0.001), NLR (p=0.001), ELR (p=0.002), HLR (p=0.007) in female patients compared to those in male patients. Although there are significant difference for B cells counts (p=0.018), CD8+CD28+ T cells (p=0.026) and CD8+HLA-DR+/CD8+ percentage (p=0.033) in patients with allergic history than those in patients without allergic history, we only discovered significant difference for those data between patients with allergic history except antibiotic and patients without allergic. Low CD8+CD28+/CD8+ percentage (p=0.008), CD4+/CD8+ ratio (p=0.039), and high percentage of CD8+HLA-DR+ T cells (p=0.019), CD8+CD38+/CD8+ (p=0.016), CD8+CD38+ T cells (p=0.013), RBC (p= 0.001), hemoglobins (p<0.001) were discovered in patients with surgery than patients without surgery. Patients with history of diseases had higher CD4+CD28+/CD4+ percentage (p=0.043) and lower CD8+CD38+/CD8+ percentage (p=0.012), CD8+CD38+ T cells (p=0.022) than those in patients without diseases. There were significant difference for memory CD4+/CD4+ percentage (p=0.034), naïve CD4+/CD4+ percentage (p=0.034), CD8+CD28+ T cells (p=0.031), and monocytes (p=0.002) in various histology. Patients with distant metastases had lower levels of B cells (p=0.001), CD4+ T cells (p=0.040), naïve CD4+ T cells (p=0.049), CD4+CD28+/ CD4+ (p=0.040), CD4+CD28+ T cells (p=0.015), CD8+CD28+ T cells (p=0.001) and higher counts of NK cells (p=0.049), WBC (p=0.003), monocytes (p=0.011), neutrophils (p=0.004), eosinophils (p=0.003), basophils (p=0.001), RBC (p=0.034), platelets (p=0.013), and ratio of MLR (p=0.003), NLR (p=0.002), ELR (p<0.001),BLR (p<0.001), PLR (p=0.011) than those in patients without distant metastases.
A trend of decreased CD8+CD28+/CD8+ percentage (r=-0.170, p=0.006, Fig. 2A), CD8+CD38+/CD8+ percentage (r=-0.264, p<0.001, Fig. 2B), and increased CD8+HLA-DR+/CD8+ percentage (r=0.179, p=0.002, Fig. 2C) with ages were found in our study. However, we did not find the similar trend in RBC and hemoglobins in spite of statistically significant difference (r=-0.047, p=0.416; r=0.004, p=0.943) for these data. There were increased WBC (r=0.227, p<0.001, Fig. 2D), monocytes (r=0.293, p<0.001, Fig. 2E), neutrophils (r=0.207, p<0.001, Fig. 2F), RBC (r=0.194, p=0.001, Fig. 2G), hemoglobins (r=0.277, p<0.001, Fig. 2H), and MLR (r=0.226, p<0.001, Fig. 2I), NLR (r=0.150, p=0.011, Fig. 2J) with the various status of smoking history. In addition, we also found patients with smoking cessation had low B cells counts (r=-0.082, p=0.166) compared to that in patients with smoking or without smoking. There were a decreased trend in B cells counts (r=-0.139, p=0.018, Fig. 2K) and increased trend in WBC (r=0.146, p=0.013, Fig. 2L), monocytes counts (r=210, p<0.001, Fig. 2M), hemoglobins counts (r=0.194, p=0.001, Fig. 2N) and MLR (r=0.200, p<0.001, Fig. 2O) with various status of drinking history. A trend of increase in WBC (r=0.198, p=0.001, Fig. 2P), neutrophils (r=0.174, p=0.003, Fig. 2Q), and platelets (r=0.140, p=0.017, Fig. 2R) was found with increased ECOG.
Tumor stages were negatively associated with counts of B cells (r=-0.216, p<0.001, Fig. 3A), CD4+ T cells (r=-0.117, p=0.047, Fig. 3B), CD8+CD28+ T cells (r=-0.142, p=0.016, Fig. 3C) and positively associated with counts of WBC (r=0.117, p=0.048, Fig. 3D), monocytes (r=0.165, p=0.005, Fig. 3E), neutrophils (r=0.120, p=0.043, Fig. 3F), eosnophils (r=0.165, p=0.005, Fig. 3G), basophils (r=0.175, p=0.003, Fig. 3H), platelets (r=0.125, p=0.035, Fig. 3I), and MLR (r=0.242, p<0.001, Fig. 3J), NLR (r=0.196, p=0.001, Fig. 3K), ELR (r=0.208, p<0.001, Fig. 3L), BLR (r=0.220, p<0.001, Fig. 3M). However, there are no significant correlation between lymphocytes counts (r=-0.110, p=0.063) and tumor stages. There were negative correlation for B cells counts (r=-0.174, p=0.003, Fig. 3N), CD8+CD28+ T cells counts (r=-0.167, p=0.005, Fig. 3O) with lymph nodes metastases. But an increases trend with lymph nodes metastases in WBC counts (r=0.181, p=0.002, Fig. 3P), neutrophils counts (r=0.137, p=0.021, Fig. 3Q), MLR (r=0.231, p<0.001, Fig. 3R), NLR (r=0.187, p=0.002, Fig. 3S), ELR (r=0.161, p=0.006, Fig. 3T), BLR (r=0.222, p<0.001, Fig. 3U), and PLR (r=0.152, p=0.010, Fig. 3V) was discovered in this study. We did not find the relationship between other cells and clinicopathologic characteristics.