Balb c mice

Murine colon carcinoma cell lines MC38-OVA and CT26 and T cell lymphoma cell line EG7-OVA were purchased from ATCC, and cells were cultured in RPMI 1640 supplemented with 10% FBS, 100 U/mL penicillin and 100 mg/mL streptomycin. C57BL/6J and Balb/c mice were obtained from SPF Biotechnology Co Ltd. C57BL/6J;CD45.1 mice were bought from Peking University Health Science Center Animal Science Department. OT-I mice were obtained from Jackson Laboratory. MC38-OVA cells (1.5 × 10⁵) and EG7-OVA cells (1 × 10⁶) were harvested and washed twice with PBS then injected s.c. into the right flank in 100 μL of PBS. When tumor volume was about 100 mm³, tumor-bearing mice were randomly assigned to receive PBS or decitabine (Sigma-Aldrich, 0.2 mg/kg/day) i.p. for 3 days. For the DP group, 2 days after decitabine treatment, anti–PD-1 antibody (Clone RMP1-14, Jiangsu Hengrui Pharmaceuticals Co Ltd, i.p. 200 μg per mouse) was administered every 3 days (2 or 4 doses). Tumor volume was estimated every other day and the tumor volume was calculated according to the formula (length × width²/2). For CD8⁺ or CD4⁺ T cell depletion, 200 μg anti-CD8 (Clone 2.43, BioXCell) or anti-CD4 (Clone GK1.5, BioXCell) was given i.p. twice weekly beginning 1 day prior to DP therapy.

Animal experiments were performed according to the guidelines of the Institutional Experimental Animal Welfare and Ethics Committee. BALB/c mice aged 42–56 days were purchased from SPF Biotechnology (Beijing, CN) and used for all experimental groups. 100 TCID₅₀ of virus in 10 μL volume was injected intracerebrally into mice under anesthetic conditions (Mishra and Byrareddy, 2020 (link)). Mice in the 4-HT⁺ group and PBS group were administered (intraperitoneal) daily of 0.4 μg 4-HT/g weight of the animal. All mice from each group were sacrificed on day 5 post-inoculation, and their brains were harvested, homogenized, and titered by TCID₅₀ assay or qPCR.

Five-week-old, specific pathogen-free female BALB/c mice were purchased from SPF Biotechnology (Beijing, China) and reared in the Laboratory Animal Centre of the Academy of Military Medical Sciences. Mice were housed with a constant ambient temperature (23 ± 3 °C) and humidity (55 ± 5%). Food, bedding, and water were changed every 4 days. All animal experiments were approval by the Ethics Committee of the Academy of Military Medical Sciences. Subcutaneous immunization was conducted on Days 0, 14, and 28 in mice. The immunization dose was 100 µL (2 µg of pertussis antigen and 100 µg of aluminum hydroxide adjuvant (General Chemical Corp, Brighton, MI, USA) per mouse, and the control group was immunized with an equal volume of PBS (Table S1). Ten days after the last immunization, blood was collected to test for antibody potency. A bacterial challenge was performed 14 days after the third immunization to evaluate the vaccine-induced protection.

BALB/c mice (female, 4–6 weeks) were purchased from SPF Biotechnology (Beijing, China) and kept in the animal facility of the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences. All animal studies followed the Institute of Laboratory Animal Science’s Institutional Animal Care and Use Committee’s guidelines and were approved by the Institutional Animal Care and Use Committee.
Mice (n = 5) were immunized subcutaneously either three times at 0, 4, and 8 weeks with 1 µg L1 VLPs alone or 10 µg L1-L2 cVLPs formulated with 50 µg Aluminium hydroxide gel (InvivoGen) and 5 µg monophosphoryl lipid A (MPLA) (InvivoGen), respectively. Sera were collected 2 weeks after the last boost and heat inactivated at 56°C for 30 min.

Male Wistar rats (80-100 g), 4 to 6-week-old female BALB/c mice, and 6 to 8-week-old female C57BL/6 mice were purchased from SPF Biotechnology Co., Ltd (Beijing, China). All animals were bred at the TMU Animal Care Facility. The blood-stage P. berghei ANKA strain and P. falciparum 3D7/Dd2/803 strains were stored as stabilates in liquid nitrogen. Parasitemia was counted by light microscope examination of Giemsa-stained thin blood smears.