Interleukin-2-Huaqiang electronic network

L06ESDL5V0H4-2

C-2

Other names: T cell growth factor (TCF)

Definition:

Interleukin-2 (IL-2) is a cytokine with broad biological activity, primarily produced by activated CD4+ and CD8+ T cells. It plays a key role in promoting the proliferation of all T cell subsets and enhances B cell activation. IL-2 is essential in regulating immune responses, contributing to antibody production, hematopoiesis, and tumor surveillance.

[Chinese name]: Interleukin-2

[Short Pinyin]: BXBJS-2

[English name]: Interleukin-2

[Category]: Immuno-enhancement agent || Other anti-tumor drugs and adjuvant medicines

[Alias]: Leucovorin, Hinjier, T cell growth factor, interleukin-2

[Foreign name]: Interleukin-2, IL-2

[Indications]: IL-2 is a glycoprotein with a molecular weight of 14,500 that stimulates the proliferation of antigen- or mitogen-activated T cells. Recombinant IL-2 is used in clinical research, mainly for kidney cancer, melanoma, and non-Hodgkin’s lymphoma. LAK cells, which are heterogeneous and include macrophages, B, and T lymphocytes, show specific anti-cancer activity without MHC restriction.

1. Materials and Methods:

1.1 Preparation of Crude IL-2:

Aseptic blood collection using heparin as an anticoagulant, followed by dilution with RPMI 1640 at 1:1 ratio. The sample is layered on Ficoll-Paque, centrifuged at 1500 rpm for 30 minutes, and the leukocyte layer is collected. After washing twice with RPMI 1640, cells are suspended in 10% FBS-RPMI 1640 at 1–3×10⁶ cells/ml. Cells are incubated for 4 days at 37°C, then centrifuged, and supernatant is filtered. PHA (1–2 μg/ml) and B lymphoblastoid cells (10⁶ cells/ml) are added, and the mixture is cultured for 18–24 hours. The supernatant is filtered through a 0.45 μm filter to obtain crude IL-2.

2. Purification of IL-2:

2.1 Ammonium sulfate precipitation and dialysis: Crude IL-2 is treated with 50% ammonium sulfate at 4°C for 2 hours, then centrifuged. The supernatant is further saturated with 80% ammonium sulfate overnight, and precipitate is dissolved in PBS containing 1% PEG6000. The solution is concentrated via vacuum dialysis.

2.2 Sephadex G-100 gel filtration: The sample is passed through a 2 × 180 cm column at 80 ml/h, and fractions are collected. OD280 values are measured, and standard proteins are used to generate a calibration curve. Biological activity is tested, and active fractions are pooled.

2.3 BlueSepharose chromatography: The pooled sample is loaded onto a BlueSepharose column at 15 ml/h. Elution is performed with a NaCl gradient (0.075 mol/L to 0.6 mol/L). Fractions are collected, and OD280 and NaCl concentration are measured. Active fractions are combined and stored at -20°C.

3. Verification:

3.1 Activity determination: IL-2 is diluted in 10% FBS-RPMI 1640 at 50%, 25%, 12.5%, and 6.25%. Each dilution is tested in a multiwell plate with CT6 cells. After incubation, H3-thymidine is added, and radioactivity is measured. A dose-response curve is plotted to determine activity units.

3.2 Protein content: Measured using UV spectrophotometry at 280 nm and 260 nm. Protein concentration is calculated as: (144 × OD280 – 0.75 × OD260) × dilution factor.

3.3 Toxicity and bacterial testing follow guidelines similar to those for thymosin.

Application:

[Pharmacological action]: IL-2 enhances T-cell proliferation, activates cytotoxic killer cells, and boosts the function of T cells, monocytes, and macrophages.

[Clinical application]: Used in chronic active hepatitis B infection and certain cancers.

[Usage and dosage]: Intravenous infusion at 25,000–50,000 U per dose, once daily.

[Adverse reactions]: Fever, chills, nausea, vomiting, diarrhea, respiratory issues, neurological changes, cardiovascular symptoms, and blood abnormalities may occur.

[Specifications]: Available as powder injection in 1,000U, 10,000U, 100,000U, and 1,000,000U doses.

[Note]: Use with caution in patients with epilepsy, severe hypotension, heart or kidney failure, high fever, or those over 60 years old. Regular monitoring of liver and kidney function is required during treatment.