Combination product for use in tumor vaccination
a combination product and tumor technology, applied in the field of tumor immunotherapy, can solve the problems of avoiding vaccination approaches, unable to employ tumor-specific antigens that are not also self-antigens, and no longer mount effective t cell responses from vaccination with tumor antigens
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example 1
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[0161]Uptake of Subunit KLH-FITC by DCOne mDC
[0162]DCOne mDC were loaded with fluorochrome-conjugated subunit KLH during maturation process for 4 hours and 24 hours as described in the material and methods section. The internalization of subunit KLH was analysed using flow cytometer. Trypan blue quenches the cell surface-bound antigen and was used to distinguish between surface-bound antigen and internalized antigen. The observed internalization of subunit KLH-FITC by DCOne mDC after 4 hrs was 54.6±8.8%, and 49.9±9.8% with trypan blue and after 24 hours 86.1±7.0%, and 81.7±8.0% with trypan blue indicating effective antigen uptake by DCOne mDC (FIG. 1A).
[0163]Labelling of OV90 Tumor Cells with Subunit KLH
[0164]OV90 ovarian carcinoma cells were cultured with subunit KLH-FITC for 4 hours and 24 hours as described in the material and methods section. We observed 13.5±2.6%, labelling of OV90 for 4 hours of which 7.5±1.0% of cells with intracellular antigen as demonstrated...
example 2
wth Inhibition in a Humanized U87-MG Glioblastoma Mouse Model and in a Humanized A375 Melanoma Mouse Model
[0175]The U87-MG mice received one i.p. vaccination of either PBS or KLH and one i.t. injection to mark the tumor either with KLH or KLH-loaded DCOne mDCs. Tumor growth was monitored three times per week using a digital caliper. Tumor volumes (in mm3) were calculated according to the following formula: Volume=(width×length{circumflex over ( )}2) / 2. At day 18, we observed a slowed tumor growth in vaccinated mice as compared to the mice injected with PBS, with i.t. injection of KLH-loaded DCOne having the strongest effect (FIG. 4).
[0176]The A375 melanoma mice received two i.p. vaccinations of either PBS or KLH and one i.t. injection to mark the tumor either with KLH or KLH-loaded DCOne mDCs. Tumor growth was monitored three times per week using a digital caliper. Tumor volumes (in mm3) were calculated according to the following formula: Volume=(width×length{circumflex over ( )}2) / ...
example 3
diated KLH-Specific Antibody Production
[0178]Possible antibody responses against KLH resulting from intratumoral injection and / or vaccination was quantified by ELISA (D0, D14 and sacrifice). We observed that both in U87-MG and A375 mice, there was no significant difference regarding anti-KLH IgM concentration between treated groups and the PBS control group over time. However, we surprisingly observed that mice from the KLH / KLH loaded DCOne group produced significantly more anti-KLH IgG than the PBS control group at sacrifice (FIGS. 6A-6B). This implies a T cell dependent IgM to IgG switch (Geha et al., NEJM, 330:1008-1009 (1994)).
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