Circulating Antibodies to Linear Peptide Antigens Derived from ANXA1 and FOXP3 in Lung Cancer.

BACKGROUND: Our previous studies revealed that concentrations of circulating antibodies to annexin A1 (ANXA1) and forkhead-box P3 (FOXP3) increased significantly in patients with non-small cell lung cancer (NSCLC). This study was thus undertaken to replicate our initial findings with different sample sets. PATIENTS AND METHODS: Antibodies were tested in 108 patients with NSCLC and 216 controls, who were divided into the discovery (49 vs. 108) and validation (60 vs. 108) group based on the time of enrolment. RESULTS: Analysis of the discovery group showed a significant increase in circulating anti-ANXA1 IgG levels in the patient group compared with the control group (p=0.005) but the validation group simply exhibited a trend toward an increase in IgG levels in NSCLC (p=0.238), generating a combined p-value of 0.009. CONCLUSION: The findings of this study support the notion that circulating IgG antibodies to ANXA1 could be used as a biomarker for early diagnosis of NSCLC but failed to replicate such findings for FOXP3.

Further study of circulating IgG antibodies to CD25-derived peptide antigens in nonsmall cell lung cancer.

A recent study reported that circulating antibodies to CD25-derived peptide antigens were significantly higher in patients with nonsmall cell lung cancer (NSCLC) than control subjects. The present study was, thus, undertaken to replicate the initial finding with different sample sets. An in-house ELISA was applied to determine circulating IgG antibodies to linear peptide antigens derived from CD25. A total of 111 patients with NSCLC and 216 control subjects were recruited and divided into the discovery sample (51 vs 108) and the validation sample (60 vs 108) based on the time of sampling. Student's t test showed that circulating anti-CD25 IgG levels were significantly higher in the patient group than the control group (t = 2.23, P = 0.027) and the validation sample replicated this finding (t = 3.31, P = 0.0012), generating a combined P value of 0.0004 (χ(2) = 20.8, df = 4). Fisher's combining probability revealed that patients with stage IV NSCLC had a significant increase in anti-CD25 IgG levels compared with control subjects (χ(2) = 22.1, df = 4, P = 0.0002) but those with the other three stages did not. This study suggests that circulating anti-CD25 IgG antibodies may have prognostic rather than early diagnostic values for lung cancer.

Study of circulating IgG antibodies to BIRC5 and MYC in non-small cell lung cancer.

An in-house enzyme-linked immunosorbent assay (ELISA) was developed in this study to detect circulating IgG antibodies to peptide antigens derived from baculoviral IAP repeat-containing protein 5 isoform 2 (BIRC5) and myc proto-oncogene protein (MYC) in non-small cell lung cancer (NSCLC). Student's t-test revealed that circulating anti-MYC IgG levels were significantly increased in patients with NSCLC compared with control subjects in the discovery sample (t = 3.96, P = 0.0001) but not in the validation sample (t = 1.24, P = 0.217), generating a combined P-value of 0.0003. Neither the discovery sample nor the validation sample showed a significant change in anti-BIRC5 IgG levels in NSCLC. Further analysis was performed to investigate whether circulating IgG antibodies to these two tumor-associated antigens (TAAs) significantly changed with early (stages I + II) and late (stages III + IV) NSCLC stages. The results showed that neither anti-MYC IgG nor anti-BIRC5 IgG levels significantly changed in patients with early stage NSCLC, while patients with late stage NSCLC had higher levels of circulating anti-MYC IgG than control subjects in the discovery sample (t = 4.74, P < 0.0001) but not in the validation sample (t = 0.80, P = 0.423), generating a combined P-value of 0.00003 (X (2) = 26.13, df = 4). In conclusion, circulating IgG antibodies to MYC and BIRC5 do not appear to serve as biomarkers for early diagnosis of lung cancer but anti-MYC IgG might have a prognostic value.

Circulating antibodies to p16 protein-derived peptides in breast cancer.

Overexpression of the p16 protein has been reported in breast cancer and may trigger the secretion of antibodies against itself. Circulating anti-p16 antibodies that were detected with a recombinant protein have been reported in breast cancer. The present study was designed to determine whether the levels of circulating IgG antibody to p16 protein-derived linear antigens are altered in breast cancer. An enzyme-linked immunosorbent assay (ELISA) was developed in-house to determine circulating IgG against peptide antigens derived from the p16 protein in 152 female breast cancer patients and 160 healthy female subjects. The Student's T-test revealed that breast cancer patients exhibited significantly higher levels of anti-p16 IgG antibody compared to control subjects (T=2.02, P=0.045). In addition, ductal cancer appeared to be the main type contributing to the increased levels of circulating anti-p16 antibodies (T=2.08, P=0.038). Of all four stages of breast cancer, stage I was associated with the highest levels of IgG antibody (T=2.02, P=0.045) and receiver operating characteristic (ROC) analysis demonstrated that the area under the ROC curve was 0.74 (95% confidence interval: 0.65-083) and that the sensitivity against a specificity of 90% was 30.3%. Therefore, the levels of circulating IgG antibody to the p16 protein may be a potential biomarker for early diagnosis of breast cancer.

Investigation of circulating antibodies to ANXA1 in breast cancer.

Our recent work demonstrated that circulating levels of IgG antibody to linear peptide antigens derived from annexin A1 (ANXA1) were significantly increased in lung cancer. The present study was then undertaken to test whether circulating anti-ANXA1 antibodies were also altered in breast cancer. An enzyme-linked immunosorbent assay was developed in-house to determine circulating IgG against ANXA1-derived peptide antigens in 152 female patients with breast cancer and 160 female control subjects. Student's t test revealed that patients with breast cancer had significantly higher levels of anti-ANXA1 IgG than control subjects (t = 4.75, P < 0.0001). Receiver operating characteristic (ROC) analysis showed that the area under the ROC curve was 0.73 with 95% confidence interval (CI) 0.67-0.78, and the sensitivity of anti-ANXA1 IgG assay was 23.2% against the specificity of 90%. The levels of anti-ANXA1 IgG did not appear to be stage-dependent, and Pearson correlation analysis showed no correlation between the anti-ANXA1 IgG levels and the stages of breast cancer (r = -0.02, df = 149, P = 0.796). This work suggests that circulating IgG for ANXA1-derived peptide antigens may have both diagnostic and prognostic values for breast cancer although further screening is needed to identify more such peptide antigens derived from tumor-associated antigens.