An amyloid beta vaccine that safely drives immunity to a key pathological species in Alzheimer's disease: pyroglutamate amyloid beta.

Pyroglutamate amyloid beta3-42 (pGlu-Abeta3-42), a highly amyloidogenic and neurotoxic form of Abeta, is N-terminally truncated to form a pyroglutamate and has recently been proposed as a key target for immunotherapy. Optimized ACI-24, a vaccine in development for the treatment and prevention of Alzheimer's disease, focuses the antibody response on the first 15 N-terminal amino acids of Abeta (Abeta1-15). Importantly, clinical data with an initial version of ACI-24 incorporating Abeta1-15, established the vaccine's safety and tolerability with evidence of immunogenicity. To explore optimized ACI-24's capacity to generate antibodies to pGlu-Abeta3-42, pre-clinical studies were carried out. Vaccinating mice and non-human primates demonstrated that optimized ACI-24 was well-tolerated and induced an antibody response against Abeta1-42 as expected, as well as high titres of IgG reactive with pyroGlu-Abeta. Epitope mapping of the polyclonal response confirmed these findings revealing broad coverage of epitopes particularly for Abeta peptides mimicking where cleavage occurs to form pGlu-Abeta3-42. These data are in striking contrast to results obtained with other clinically tested Abeta targeting vaccines which generated restricted and limited antibody diversity. Taken together, our findings demonstrate that optimized ACI-24 vaccination represents a breakthrough to provide a safe immune response with a broader Abeta sequence recognition compared to previously tested vaccines, creating binders to pathogenic forms of Abeta important in pathogenesis including pGlu-Abeta3-42.

Classical gene amplifications in human breast cancer are not associated with distant solid metastases.

To determine the relationship between breast cancer progression and gene amplification, we screened 62 distant metastases and 122 primary breast tumours for the amplification of the proto-oncogenes MYC and ERBB2 and the 11q13 chromosomal region. Surprisingly, solid metastases showed an absence of gene amplification. These results suggest that the amplification of the proto-oncogenes MYC and ERBB2 and the 11q13 chromosomal region seem to be involved mainly in the genesis of the primary breast tumour rather than its progression.

Loss of heterozygosity on 7q31 occurs early during breast tumorigenesis.

Although human breast tumorigenesis is associated with the accumulation of mutations both in oncogenes and in tumor suppressor genes, the identity of the genetic alterations that are critical in the early stages of the breast tumorigenic process remains obscure. A high frequency (27-41%) of loss of heterozygosity (LOH) occurrence has been shown at the MET locus on chromosome band 7q31 and this specific alteration is associated with poorer survival. Here, we report that restriction fragment length polymorphism (RFLP) analysis on 221 informative (heterozygous) primary breast tumors and 57 informative relapses (13 local recurrences and 44 distant metastases) revealed a similar frequency of 7q31 LOH as tumors progress from primary cancer to relapse, in marked contrast to other changes such as 11p15.5 LOH. This finding suggests that inactivation of a putative tumor suppressor gene located in 7q31 is a very early event in breast tumorigenesis. Our results also show that metastatic potential is an induced phenomenon that occurs at a relatively early stage, rather than a marker of tumor progression.

Detection of a soluble antigen defined by monoclonal antibody 83D4 in serous effusions associated with breast carcinoma.

Monoclonal antibody (MoAb) 83D4 was generated by immunization with cell suspensions obtained from sections of formol-fixed paraffin embedded human breast cancer. It recognized an antigen expressed in breast carcinomas but not in normal breast tissue. Pleural and ascitic fluids from 66 patients were studied by an 83D4 heterologous sandwich radioimmunoassay (SRIA) using solid-phase immobilized wheat germ agglutinin to detect the 83D4 soluble antigen. Using a cutoff level of 5 units/ml of 83D4 antigen, higher values were found in 22 of 27 breast cancer-associated effusions (mean = 10.72 +/- 6.80 units/ml). The 20 nonmalignant effusion fluids tested showed lower values (mean = 1.16 +/- 1.49 units/ml, P less than 0.001). The antigen was undetectable or present in low levels in effusions from patients with hematologic malignancies. When SRIA results were compared with conventional cytologic diagnosis in breast-cancer effusions, elevated levels of 83D4 soluble antigen were found in all patients (8 of 8) in whom malignant cells had been detected, in 4 of 8 patients with the diagnosis of "suspected malignancy," and in 10 of 11 patients with negative cytologic findings. Using an immunoglucosidase method on cell smears of various origins, MoAb 83D4 stained metastatic cells of breast and ovary carcinomas but did not reactive with mesothelial cells and other normal or malignant cell types. These results suggest that quantitation of the 83D4 soluble antigen may be used to improve the diagnosis of cancer in serous effusions.