Partial enzymatic degradation of stroma allows enrichment and expansion of primary breast tumor cells.

The goal of this study was to isolate and expand tumor cells in culture that closely resemble invasive cells in primary breast carcinoma tissue. Based on the hypothesis that invasive tumor cells are released more readily upon digestion with connective tissue-degrading enzymes because they are not confined within a basement membrane, we have designed a novel procedure for their isolation. Using this method, we have successfully expanded in culture aneusomic tumor cells from several primary breast tumors. Twenty nine of 44 (66%) specimens processed yielded proliferative and passageable cultures of up to 2 x 10(7) cells. The original tumor tissue and cultures derived therefrom were compared for aneusomy and the abnormal expression of the erb-B2, p53, and bcl-2 gene products. Remarkable similarities were observed. However, some intratumor heterogeneity in chromosome content was found between touch preparations and cultured cells. Overexpression of erb-B2 was observed in the vast majority of cases (16 of 20), suggesting that this phenotype may be important for dysregulated proliferation in vitro. The simple and rapid method described in this report could enable routine expansion of primary breast tumors and provide adequate numbers of viable cells for studying and manipulating their functional characteristics.

Modulation of receptors and cytotoxic response of tumor necrosis factor-alpha by various lectins.

The effects of various lectins on the interaction of the human cervical carcinoma cell line ME-180 with recombinant human tumor necrosis factor-alpha (rTNF-alpha) was investigated. rTNF-alpha is known to have cytotoxic effects on this tumor cell line and has been reported to interact with these cells through a single class of specific high affinity receptors (Kd = 0.45 nM; approximately 1790 binding sites/cell). Exposure of cells to concanavalin A (ConA) causes an approximately 2-fold increase in rTNF-alpha receptors without any significant change in their affinity constant (Kd = 0.36 nM; approximately 3662 binding sites/cell). This increase in receptor number is dependent on temperature, the time of exposure and dose of ConA, and does not require the synthesis of new proteins. In spite of an increased binding of rTNF-alpha to cells, the cell killing induced by rTNF-alpha is totally blocked by ConA. Cells are also protected by this lectin from the synergistic cytotoxic effects of rTNF-alpha and recombinant human interferon-gamma. Furthermore, it was also found that ConA decreases the rate of internalization and dramatically inhibits the release and degradation of rTNF-alpha by the cells. These results, overall, demonstrate that ConA increases total number of binding sites for rTNF-alpha but blocks the transduction of the signal for the cytotoxic response.

MF59 adjuvant enhances the antibody response to recombinant hepatitis B surface antigen vaccine in primates.

The effect of the proprietary adjuvant MF59 on the immunogenicity of a recombinant hepatitis B virus (HBV) vaccine, PreS2+SAg, was investigated in baboons. The magnitude and duration of the antibody response to hepatitis B surface antigen (anti-HBs) induced by the HBV/MF59 vaccine was compared with the same antigen combined with alum and with two licensed alum vaccines. After one immunization, the HBV/MF59 vaccine generated anti-HBs titers >10 mIU/mL in a greater proportion of animals, and mean titers were 26- to 84-fold higher than titers from alum vaccines. After a third immunization, the HBV/MF59 vaccine generated titers 38- to 127-fold higher than alum vaccines. Seven months after the third immunization, HBV/MF59 elicited titers 75- to 472-fold higher than alum vaccines. The dramatic immune response elicited by HBV/MF59 in baboons suggests that MF59 may be a desirable adjuvant for use in improved HBV vaccines for humans.