Anti cd8 pe

For memory T cell analyses, splenocytes from various vaccinated groups were harvested, lysed with 1x RBC lysis buffer, washed and 1 × 10⁶ cells per well was utilized for each reaction followed by flow cytometry (BD FACSCaliber, Hampton, NH, USA) analysis. Splenocytes were stained with anti-CD44-APC (Biolegend, San Diego, CA, USA), anti-CD4-FITC (Biolegend, San Diego, CA, USA), anti-CD8-PE (Biolegend, San Diego, CA, USA), anti-CD25-APC (Biolegend, San Diego, CA, USA), or anti-Foxp3-APC (Biolegend, San Diego, CA, USA).

To analyse chemokine receptor expression by FACS, cells were incubated first with blocking solution (mouse serum in PBS) during 20 minutes, followed by a second incubation with a biotinylated mouse anti-CCR9 antibody (1 μg/10⁶ cells) (eBioscience, CA, USA) in FACS buffer (PBS containing 2% fetal bovine serum and 0.1% NaN₃) for 30 min at 4°C. Finally cells were incubated with a solution of streptavidin-APC or streptavidin-PerCP during 30 min at 4°C in the dark.
For phenotypic detection of other cell populations, single cell suspensions were preincubated with a mixture of anti-CD16/CD32 antibodies (BioLegend, CA, USA) during 10 min at room temperature. Following first incubation and several washes, cells were incubated with either anti-CD4-PeCy7, anti-CD8-PE, anti-Gr1-PeCy5 (BioLegend, CA, USA), or anti-Siglec (BD, CA, USA) antibodies in FACS buffer for 30 min at 4°C (according to manufacturer's suggestions). Finally, samples were analyzed in Attune® Acoustic flow cytometer (Life Technologies, Thermo Fisher Scientific, MA, USA). Analysis of data was performed with FlowJo 8.7 software (Tree Star, Inc.).

Silicon phthalocyanine dichloride (SiPCCl₂), 3‐aminopropyltrimethoxysilane (APTES), Hoechst 33342, tetraethylorthosilicate (TEOS), and 2, 7‐dichlorodihydroflfluorescein diacetate (DCFH‐DA) were obtained from Sigma‐Aldrich. Hexadecyl trimethylammonium chloride (CTAC), triethanolamine (TEA), and FeCl₃ were obtained from Sinopharm Chemical Reagent Co. Ltd. Dylight550‐NHS was purchased from Thermo Fisher Scientific (Waltham, USA). ICG‐NHS was obtained from Xi'an Rixi Biological Technology Co. Ltd (Xi'an, China). Calcein‐AM/propidium iodide (PI), and 1, 3‐diphenylisobenzofuran (DPBF) were obtained from KeyGen BioTech (Shanghai, China). Annexin V‐FITC/PI and CCK8 were obtained from Dojindo Laboratories. Annexin V‐APC/PI was obtained from Jiangsu KeyGEN BioTECH Corp., Ltd (Jiangsu, China) Neutrophil (mouse) isolation Kit was purchased from Cayman Chemical (Michigan, USA). Anti‐CD11c‐APC, anti‐CD137‐APC, anti‐CD11b‐APC, anti‐CD86‐PE‐Cy7, antiCD3‐APC, anti‐CD80‐PE, anti‐CD8‐PE, anti‐Ly‐6G/Ly‐6C‐PE, CD54, CD95, anti‐CD8 antibody, and anti‐CD69‐FITC was purchased from BioLegend, Inc (San Diego, CA, USA). ELISA kits from Neobioscience Technology (Shenzhen, China) were obtained to detect the IFN‐γ, TNF‐α, IL‐12, and IL‐2. H22 mouse liver cancer cell‐specific neoantigen (sequence: HTDAHAQAFAALFDSMH) was obtained from GenScript USA Inc.

Ethylenediamine tetraacetic acid (EDTA), 4 % paraformaldehyde, HSA, Coomassie brilliant blue G250 and hematoxylin and eosin (H&E) staining kit were purchased from Beijing Solarbio Technology Co., Ltd (China). 4′,6-Diamidino-2-phenylindole Dihydrochloride (DAPI), trypan blue and glutathione (GSH) were obtained from Sigma-Aldrich Chemical Co. (USA). SMCC and 5,5′-dithiobis (2-nitrobenzoic acid) (DTNB) were purchased from Adamas-beta (Shanghai, China). BCA protein quantitation kit and cell freezing medium were purchased from Beyotime Biotechnology Co., Ltd. (China). DMEM medium, RPMI 1640 medium, phosphate buffer saline (PBS), fetal bovine serum (FBS), penicillin and streptomycin were purchased from Gibco (USA). Interleukin-4 (IL-4) and Granulocyte-macrophage colony-stimulating factor (GM-CSF) were purchased from Wolcavi Biotech (Beijing, China). Mouse interferon-gamma (IFN-γ) enzyme-linked immunospot (ELISpot) plus kit was purchased from Mabtech (Sweden). Anti-CD11c-APC, anti-CD80-PE and anti-CD86-PE-Cy7 were purchased from Thermo Fisher Scientific (USA). Anti-CD3-APC and anti-CD8-PE were purchased from BioLegend, Inc (USA). Mouse IFN-γ enzyme-linked immunosorbent assay (ELISA) kit, mouse interferon beta (IFN-β) ELISA kit and mouse tumor necrosis factor alpha (TNF-α) ELISA kit were purchased from Fankewei (Shanghai, China).

Magnesium (Mg) rods were obtained from JingJun Materials Technology Co. Ltd. (Suzhou, China). Chloroplatinic acid (H₂PtCl₆) and 2′,7′-dichlorofluorescein diacetate (DCFH-DA) were purchased from Sigma-Aldrich Co., Ltd. Sodium hydroxide (NaOH) was purchased from Sinopharm Chemical Reagent Co., Ltd. Magnesium chloride (MgCl₂), magnesium hydrate [Mg(OH)₂], and MB were purchased from Aladdin Co., Ltd. All chemicals and reagents were of analytical grade and used without further purification. Anti-CD3-FITC (Biolegend, clone 17A2, Catalog: 100204), anti-CD4-APC (Biolegend, clone GK1.5, Catalog: 100412), anti-CD8-PE (Biolegend, clone 53–6.7, Catalog: 100708), anti-CD45-FITC (Biolegend, clone I3/2.3, Catalog: 147710), anti-CD11b-PE (Biolegend, clone M1/70, Catalog: 101208), anti-Gr-1-APC (Biolegend, clone RB6-8C5, Catalog: 108412) were obtained from Biolegend and diluted at 1:300 for cell staining.