Autoantibodies against p53 are not increased in human ascites and pleural effusions

Abstract

Mutated human p53 may give rise to the formation of autoantibodies and may be a marker for a worse prognosis. We speculated that ascites or pleural effusions may enhance the formation of such autoantibodies in cancer patients and, therefore, we measured the presence of autoantibodies in the ascites or pleural effusion of 40 patients with advanced malignancies. As controls, p53 autoantibodies were measured in 15 patients with effusions who did not have a malignancy. Using a specific enzyme-linked immunosorbent assay, p53 autoantibodies could only be detected in the effusions of 5/40 patients (12.5%) with known malignancies. The formation of autoantibodies did not correlate with the presence or absence of tumor cells in the effusion. The effusions of the patients without tumor were all negative for p53 autoantibodies. Our study shows that malignant or reactive effusions do not stimulate the local or systemic production of autoantibodies against p53.

Autoantibodies against tumour suppressor protein p53 in pleural effusions of patients with tuberculosis pleurisy

“…Membrane containing purified recombinant p53 proteins was immunoblotted with mouse anti-human p53 monoclonal antibody Pab240. M, molecular weight marker; lane 1, bacterial total lysate containing (his) 6 -p53 fusion proteins; lane 2, flow-through fraction of metal affinity chromatography; lanes 3, 4, 5 and 6, fractions of washing buffer containing 50 mM, 100 mM and two fractions of 200 mM imidazole, respectively Table 2. Seven of these p53 Abs-positive patients su¡ered from malignancy (one malignant oesophageal carcinoma, two hepatoma and four lung cancer).…”

“…The molecular mass of the purified protein was 53 kDa. M, molecular weight marker; lane 1, bacterial total lysate containing (his) 6 -p53 fusion proteins; lane 2, flowthrough fraction of the metal affinity chromatography; lanes 3, 4, 5 and 6, fraction of washing buffer containing 50 mM, 100 mM and two fractions of 200 mM imidazole, respectively as antigens for the detection of p53 Abs to monitor the immune response against p53 in pleural e¡usions.…”

“…Construction, expression and purification of full-length p53 fusion protein Full-length p53 cDNAwas in-frame subcloned into hexahistidine-tag (his) 6 -containing expression vector pET22b( þ ) to create (his) 6 -p53 fusion protein. (His) 6 -p53 fusion proteins were expressed in BL21(DE3) upon isopropyl-1-thio-b-D-galactopyranoside induction and puri¢ed by metal a⁄nity chromatography.…”

“…Recombinant full-length (his) 6 -p53 fusion proteins were puri¢ed by metal a⁄nity chromatography. Most of the fusion proteins were puri¢ed to at least 90% homogeneity analysed by SDS-PAGE (lanes 5 and 6, Figure 1).…”

“…(His) 6 -p53 fusion proteins were expressed in BL21(DE3) upon isopropyl-1-thio-b-D-galactopyranoside induction and puri¢ed by metal a⁄nity chromatography. Puri¢ed p53 fusion protein was further characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE).…”