The lower panel of Figure 2A shows the semiquantitative measurement of band www.selleckchem.com/products/azd9291.html densities. Figure 2 Detection of Ma2 autoantibodies in serum of healthy controls and primary SI-NET patients. Next we confirmed the specificity of Ma2-specific autoantibodies in serum samples from the patients by sequential immunoprecipitation using 35S-methionine-labeled human Ma2 protein generated by in vitro transcription-coupled translation as described in Material and Methods. Ma2-specific autoantibodies were detected in serum samples from primary SI-NET patients expressing high titer of Ma2 autoantibodies (Figure 2B, lanes 6, 7, and 8) but not in serum samples from healthy controls (lanes 3, 4, and 5). Lane 1 shows immune precipitation with a commercial goat anti-human PNMA2 (positive control) and lane 2 shows immunoprecipitation without antibody or serum (negative control).

The lower panel of Figure 2B shows the result of autoradiography of the blot. Recently, we showed that serum antibodies of patients detect Ma2 and faintly Ma1 by using commercial immuno-dot-blot (Figure S1). Progression free survival (PFS) and recurrence free survival (RFS) of primary SI-NET patients, after surgery with curative intent, depend on Ma2 autoantibody titer We have evaluated the clinical data of 36 patients followed up after radical operation of primary tumors with a curative intent. We evaluated Kaplan-Meyer survival curves, followed by a log-rank test to determine whether the curves were different. The hazard ratios were calculated, based on Cox regression function as described in Material and Methods, and found to be 4.

31 (p-value=0.011) for progression free survival (PFS) and 4.24 (p-value=0.012) for recurrence free survival (RFS). The analysis showed that 19 patients with Ma2 autoantibody titer below the cutoff had a longer PFS compared to 17 patients with Ma2 autoantibody titer higher than the cutoff, Figure 3A. The same was true for patients with RFS, Figure 3B. The significance of the analyses is clearly expressed by both p-values=0.006. The median survival time for each group of patients was estimated from the survival curves. It was clearly shorter for patients with Ma2 autoantibody titer higher than the cutoff with an estimated time of about 40 months compared to those with levels below the cutoff with an estimated survival time of about 125 months. The results are summarized in Table 2. Clinical information for patients with Ma2 autoantibodies concentration < cutoff are to the left in Table 2 and clinical information for patients with Ma2 autoantibodies concentration > cutoff are to the right. Brefeldin_A Figure 3 PFS and RFS of primary SI-NET patients after surgery with curative intent depend on Ma2 autoantibody titer.