In vitro Biological

• Core Biomarkers: CFSE Dilution Intensity, Ki-67 (PCNA)

• Positive control: ImmunoCult^TM Human CD3/CD28 Activator (STEMCELL)

• CFSE flow cytometry tracks dye dilution during cell division to distinguish proliferation generations. Combined with Ki-67 staining, it accurately reflects T cell proliferation activity.

• Vaccine R&D: Immunogenicity

• Tumor Immunotherapy: TIL/TCR-T

• Autoimmune: Pathogenesis

• Core Biomarkers: dual marker of CD69 and CD25

• Positive control: PMA + Ionomycin (cocktail)

• Quantify early stimulation marker CD69 and mid-term marker CD25 expression via flow cytometry to distinguish activated T cell proportion for evaluating immune function status.

• Drug R&D Screening

• Basic immunology research

• Immunotherapy: CAR-T evaluation

• Core Biomarkers: focus on key effectors IL-2, IFN-γ, TNF-α to reflect immune activation

• Positive control: PMA + Ionomycin (cocktail) as gold standard

• Dual detection of cytokines in supernatant (ELISA/MSD) and single cell level (i.e. intracellular staining).

• Vaccine R&D: Immunogenicity

• Tumor Immunotherapy: TIL/TCR-T

• Autoimmune: Pathogenesis

• Core Biomarkers: Granzyme B, LDH release

• Gold Std Positive Ctrl: Immobilized Anti-CD3 (clone OKT3)

• Evaluate killing potency by detecting Granzyme B release, and/or measuring LDH release in supernatant.

• CAR-T/TCR-T Therapy QC

• NK Cell Therapy Evaluation

• Tumor Immunology

• Core Biomarkers: PD-1, PD-L1

• Gold Std Positive Ctrl: Anti-cd3/CD28 co-stimulation

• Activated PD-1 on T cells behave as immune “brake”, quantitative detection of PD-1/PD-L1 levels reveals their correlation with tumor immune evasion.

• Predict immunotherapy efficacy & prognosis

• Monitor CAR-T cell exhaustion

• Basic immunology & drug R&D

• Core Biomarkers: CD80 (M1) / CD163 (M2)

• Positive control: LPS+IFN-γ(M1) | IL-4+IL-13(M2)

• Macrophage polarize to pro-inflammatory M1 or anti-inflammatory M2 phenotype, which can be distinguished by flow cytometry along with characteristic cytokine secretion.

• Tumor microenvironment

• Immunotherapy R&D

• Autoimmune/Inflammatory disease

• Core Biomarkers: CD83, CD86, CCR7, MHC-I, MHC-II

• Positive control: LPS (E.Coli O111:B4, Sigma), IFN-γ, TNF-α

• Dendritic cells are the “sentinels” of immune cells, only mature dendritic cell expressing CD83/CD86 are potent to activate naïve T cells.

• Vaccine R&D

• Immunotherapy

• Core Biomarkers: Citrullinated histone H3, myeloperoxidase

• Positive control: PMA

• Activated neutrophile releases Neutrophil Extracellular Trap to detain pathogens. NET is composed of DNA backbone and antimicrobial proteins (e.g. histone, myeloperoxidase, and neutrophile elastase. This sticky web immobilizes and kills invading pathogens.

• Infectious diseases (e.g. sepsis, bacterial/fungal infection)

• Autoimmune disease

• Core Biomarkers: Transcription factor Foxp3; Surface receptor CTLA-4, LAG-3, PD-1; Enzyme CD39, CD73; Suppressive cytokine: IL-10, IL-35, TGF-β

• Gold std. positive ctrl: Rh TGF-β + IL-2 cocktail

• T-regs are specialized subset of T cells that function as immune “brakes”, preventing excessive immune responses. T-regs suppress immune functions of T, B, and antigen-presenting cells (APC) by direct cell-to-cell contact and release inhibitory molecules IL-10 and TGF-β.

• Tumor immune evasion

• Autoimmune disease

• Core Biomarkers: cell migrate rate, CXCR3, CCR4, CD103, CD69

• Positive control: Rh. CXCL10, CXCL12, CXCL8, CCL2

• Simulate the in vivo environment via Transwell assay to quantify immune cells’ directional migration to chemokines, and analyze chemokine receptor (e.g. CXCR3) by flow.

• Tumor immune infiltration

• Autoimmune and inflammatory disease

• Immunomodulatory drug R&D