Combination therapy for use in cancer therapy
a cancer therapy and combination therapy technology, applied in the direction of antibody medical ingredients, drug compositions, dsdna viruses, etc., can solve the problems of genital warts and the tendency to get anal cancer
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example 1
LLO-Antigen Fusions Induce Anti-Tumor Immunity
Materials and Experimental Methods (Examples 1-2)
Cell Lines
[0190]The C57BL / 6 syngeneic TC-1 tumor was immortalized with HPV-16 E6 and E7 and transformed with the c-Ha-ras oncogene. TC-1, provided by T. C. Wu (Johns Hopkins University School of Medicine, Baltimore, Md.) is a highly tumorigenic lung epithelial cell expressing low levels of with HPV-16 E6 and E7 and transformed with the c-Ha-ras oncogene. TC-1 was grown in RPMI 1640, 10% FCS, 2 mM L-glutamine, 100 U / ml penicillin, 100 μg / ml streptomycin, 100 μM nonessential amino acids, 1 mM sodium pyruvate, 50 micromolar (mcM) 2-ME, 400 microgram (mcg) / ml G418, and 10% National Collection Type Culture-109 medium at 37° with 10% CO2. C3 is a mouse embryo cell from C57BL / 6 mice immortalized with the complete genome of HPV 16 and transformed with pEJ-ras. EL-4 / E7 is the thymoma EL-4 retrovirally transduced with E7.
[0191]L. monocytogenes Strains and Propagation
[0192]Listeria strains used were ...
example 2
LM-LLO-E7 Treatment Elicits TC-1 Specific Splenocyte Proliferation
[0211]To measure induction of T cells by Lm-E7 with Lm-LLO-E7, TC-1-specific proliferative responses, a measure of antigen-specific immunocompetence, were measured in immunized mice. Splenocytes from Lm-LLO-E7-immunized mice proliferated when exposed to irradiated TC-1 cells as a source of E7, at splenocyte: TC-1 ratios of 20:1, 40:1, 80:1, and 160:1 (FIG. 4). Conversely, splenocytes from Lm-E7 and rLm control-immunized mice exhibited only background levels of proliferation.
example 3
Fusion of E7 to LLO, actA, or a Pest Amino Acid Sequence Enhances E7-Specific Immunity and Generates Tumor-Infiltrating E7-Specific CD8+ Cells
Materials and Experimental Methods
[0212]500 mcl (microliter) of MATRIGEL®, comprising 100 mcl of 2×105 TC-1 tumor cells in phosphate buffered saline (PBS) plus 400 mcl of MATRIGEL® (BD Biosciences, Franklin Lakes, N.J.) were implanted subcutaneously on the left flank of 12 C57BL / 6 mice (n=3). Mice were immunized intraperitoneally on day 7, 14 and 21, and spleens and tumors were harvested on day 28. Tumor MATRIGELs were removed from the mice and incubated at 4° C. overnight in tubes containing 2 milliliters (ml) of RP 10 medium on ice. Tumors were minced with forceps, cut into 2 mm blocks, and incubated at 37° C. for 1 hour with 3 ml of enzyme mixture (0.2 mg / ml collagenase-P, 1 mg / ml DNAse-1 in PBS). The tissue suspension was filtered through nylon mesh and washed with 5% fetal bovine serum+0.05% of NaN3 in PBS for tetramer and IFN-gamma stain...
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