Fig 1: Drug efficacy of anti-PD1 antibody combined with CEACAM6/4-1BBL genetic vaccine. (A) Statistical result of the therapeutical effect of CEACAM6/4-1BBL vaccine, and PD1 antibody (Nivolumab) combined with vaccine, on the growth of MC38 heterotopic tumor in mice (averages of tumor volume ratios, and SD). Treatments started at 2 weeks after heterotopic tumor transplantation. Control, given vehicle. Vaccine, given the bivalent vaccine of pIRES-CEACAM6-4-1BBL/SL3261 alone. Combined, treated with bivalent vaccine and PD1 antibody (Nivolumab, 20 mg/kg) together. (B) Representative images of isolated MC38 tumors of the control (2490 mm3), vaccine (2147), and combined immunotherapy (1181) groups. N = 4 per time point. ∗∗∗P < 0.001, Combined vs. Control; #P < 0.05, Combined vs. Vaccine.
Fig 2: Identification and expression of CEACAM6/4-1BBL double genes in COS7 cells. (A) The result of reverse-transcription PCR for identification of CEACAM6/4-1BBL double genes expression in COS7 cells 48 h after transfection. Lane 1, COS7 cells transfected with pIRES-4-1BBL; 2, COS7 cells transfected with pIRES-CEACAM6; 3–4, COS7 cells transfected with pIRES-CEACAM6/4-1BBL; 5, COS7 cells transfected with pIRES vector plasmids; 6, COS7 cells transfected with nothing (negative); 7–8, positive control of pIRES-CEACAM6/4-1BBL plasmids. 4-1BBL: 930 bp; forward primer: 5-GCTCTAGAGCCACCATGGACCAGCACGCACTTG-3; reverse primer: 5-GGCGGCCGCGTCATCCCTGAGGGGGGTC-3. CEACAM6: 801 bp; forward primer: 5-CAGAGCCAAACAACAGAT-3; reverse primer: 5-CATTATTACTTATGCTGACCT-3. Primary uncropped image of (A) is shown in supplementary Figure S1. (B) Representative Western blots showing the successful expression of CEACAM6/4-1BBL proteins in COS7 cell line 48 h after transfection. Control: COS7 cells transfected with vector plasmids. Double-gene plasmid: COS7 cells transfected with pIRES-CEACAM6/4-1BBL plasmids. (C) Western blotting images showing the expression of 4-1BBL and CEACAM6 proteins in the intestine and tumor tissues of mice treated or not treated with the bivalent genetic vaccine of pIRES-CEACAM6-4-1BBL/SL3261. GAPDH was used as an internal reference. Primary uncropped images of (B) and (C) are shown in supplementary Figure S2.
Fig 3: Tumor tissue PD1/PD-L1 expression after treatment with CEACAM6/4-1BBL vaccine and anti-PD1 immunotherapy. (A) Representative immunohistochemical images showing the expression of PD-1 and PD-L1 in MC38 tumor tissue after treatment. Control, given vehicle; Vaccine alone, given the bivalent vaccine; Combined immunotherapy, given bivalent vaccine and anti-PD1 drug (Nivolumab). Tissue samples were collected at 2 weeks after treatments. Purple, hematoxylin staining for nucleus; brown, anti-PD1 or anti-PD-L1 staining. Scale bars, 100 μm. (B) Statistical results of the mean integral optical density (MIOD, IOD/Area) of the IHC images. N = 3 per group. ∗P < 0.05, ∗∗∗P < 0.001 vs. Control; #P < 0.05, ###P < 0.001 vs. Vaccine alone.
Supplier Page from Abcam for Anti-CEACAM6 antibody [EPR4403] - BSA and Azide free