Association of polyclonal antibodies and Anti-pd1 or Anti-pdl1 antibodies for the treatment of cancer
A polyclonal antibody, monoclonal antibody technology, applied in the field of immunology
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Embodiment 1
[0185] Example 1: ELISA control of polyclonal antibodies obtained by immunizing rabbits with cancer cell lines
[0186] Rabbits were immunized with cells from the murine tumor cell line Hepa 1.6 (hepatocellular carcinoma) to obtain the corresponding anti-tumor polyclonal antibodies.
[0187] The tumor cells were subcutaneously injected twice with an interval of two weeks.
[0188] 7 days after the first subcutaneous injection (Rabbit–I1+7) and 7 days after the second subcutaneous injection (Rabbit–I2+7), sera were actually collected in pre-immunized rabbits (Rabbit–J0) for testing The ability of the antibodies to recognize tumor cells used to immunize rabbits.
[0189] More specifically, the antibody response of the immunized rabbits was measured using a cellular ELISA assay based on the Hepa 1.6 cells used to immunize the rabbits. The collected serum was serially diluted 1:20; 1:60; 1:180; 1:540; 1:1620; 1:4860; 1:14580; Incubate for 2 hours at room temperature. After t...
Embodiment 2
[0193] Example 2: In vivo use of pooled polyclonal antibodies in syngeneic and orthotopic mouse models
[0194] The Hepa1.6 hepatocellular carcinoma mouse model has been used and described in Gauttier et al., Int J Cancer. 2014 Dec 15;135(12):2857-67.
[0195] This model is syngeneic and in situ in immunocompetent C57 / BL6J mice.
[0196] 2.5.10 resuspended in PBS 6 Individual Hepa1.6 cells were injected into the portal vein of 8-week-old male mice, which, if left untreated, resulted in hepatic hypertension and death of the animals within 12-15 days.
[0197] Four days later (D4) different treatments were administered:
[0198] - Group 1 (Group 1; n=7; Control), receiving: control isotype (3G8: irrelevant monoclonal antibody (mAb) - isotype IgG1) (8 mg / kg, 2 times a week, 28 days) + rabbit polyclonal antibody before immunization (20mg / kg, 2 times a week, 28 days).
[0199] - Second group (group 2; n=8; comparison), receiving: control isotype (3G8: irrelevant monoclonal an...
Embodiment 3
[0215] Example 3: Polyclonal antibodies of the invention allow conversion of cold tumors to hot tumors
[0216] Rabbits were immunized with mouse B16F10 melanoma cells to obtain corresponding anti-tumor polyclonal antibodies.
[0217] It is well known that the B16F10 melanoma cell line is a good example of a "cold" tumor because, for example, it is resistant to anti-PDL1 antibodies, where such antibodies have very limited effect on tumor regression (Chen et al., Cancer Immunol res. 2014; 3(2):149-60; Ueha et al., 2015; 3(6):631-40).
[0218] These IgG fractions of polyclonal antibodies raised against B16F10 cells from rabbits were infused into subcutaneous implants every two weeks (12.5 mg / kg twice a week) from day 4 (D4) to day 30 (D30) B16F10 melanoma cells (D0) in Balb / C mice (Group 2 - n=8).
[0219] The same animal control group (group 1-n=7) subcutaneously implanted with B16F10 melanoma cells received only rabbit polyclonal antibody (20 mg / kg, twice a week, 28 days) ...
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