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Article on Recombinant Human Hexamer-Dominant IgM Monoclonal Antibody to Ganglioside GM3 for Treatment of Melanoma

View larger version (17K): [in this window] [in a new window] [Download PPT slide]   Fig. 2. Fluorescence-activated cell sorting analysis of the binding of L612, CA19, CJ45, and LA74 to GM3-positive or GM2-positive cancer cell lines. GM3-positive mouse B16F10 melanoma cells (A), GM3-positive human M1 melanoma cells (B), and GM3-negative but GM2-positive human A549 lung cancer cells (C) were incubated with 100 µg/mL CA19, CJ45, L612, LA74, control human IgM (Cont. IgM), or no antibody (No 1st Ab) and analyzed as described in Materials and Methods.

View larger version (24K): [in this window] [in a new window] [Download PPT slide]   Fig. 3. CDC from L612, CA19, and CJ45 against GM3-positive or GM3-negative culture cells. Antibody-induced CDC against B16F10 mouse melanoma cells, M1 human melanoma cells, and A549 human lung cancer cells was tested using human serum complement (A) or nude rat serum complement (B). B16F10, M1, and A549 cells that had been labeled with 51Cr-sodium chromate were incubated with 0.04, 0.1, 0.2, 1, 5, and 10 µg/mL L612, CA19, CJ45, LA74, or control human IgM. Human serum was added to B16F10 cells for a final concentration of 25% and to M1 cells for a final concentration of 25%. Nude rat serum was added to the B16F10, M1, and A549 cells for a final concentration of 25%. Cytotoxicity (%) was determined from the radioactivity of the supernatants. Points, mean of three independent experiments; bars, SD.

View larger version (12K): [in this window] [in a new window] [Download PPT slide]   Fig. 4. Survival of B16F10 melanoma-bearing nude rats after administration of antibodies. On the same day after i.p. injection of 5 x 106 B16F10 cells, rats received an i.v. injection of CA19, CJ45, L612, or LA74. A, survival (%) associated with CA19 and LA74, both at a total dose of 30 mg/kg (three injections of 10 mg/kg). B, survival (%) associated with CA19, CJ45, and L612, each at a total dose of 30 mg/kg (three injections of 10 mg/kg). Control animals received commercially available human IgM (vehicle). Each group consisted of six to nine animals. *, significant difference from the vehicle-treated group.

View larger version (10K): [in this window] [in a new window] [Download PPT slide]   Fig. 5. Correlation between serum 5-S-CD levels and survival of B16F10 melanoma bearing nude rats after administration of CA19 or LA74. After i.p. injection of 5 x 106 B16F10 melanoma cells, nude rats received 30 mg/kg (three i.v. injections of 10 mg/kg) of CA19 (blue), LA74 (red), or vehicle control (black). Blood concentrations of 5-S-CD on day 15 (A) and day 22 (B) were analyzed as described in Materials and Methods and plotted against survival. Each group consisted of nine animals.

View larger version (41K): [in this window] [in a new window] [Download PPT slide]   Fig. 6. Immunohistochemistry of human normal skin and melanoma tissues with CA19. Frozen sections of normal human skin (A) and two melanoma tissue specimens from patients with stage IV disease (B and C). Tissues were incubated with 2 µg/mL of biotinylated CA19. Bar, 20 µm.

1. Azuma Y, Ishikawa Y, Kawai S, et al. Recombinant human hexamer-dominant IgM monoclonal antibody to ganglioside GM3 for treatment of melanoma. Clin Cancer Res 2007;13:2745–50.[Abstract/Free Full Text]

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