IL-4 is mainly a cytokine produced by the activation of TH cells.CT.4S cells, an IL-4-dependent cell line, show hyporesponsiveness to IL-2, and their proliferative response is positively correlated with the activity of IL-4. Detection of the biological activity level of IL-4 provides indirect insight into the function of TH2 cells and some humoral immunity mediated with antibodies.

Operation method

IL-4 biological activity assay

Materials and Instruments

IL-4 induced
Saline Lymphocyte Stratification Solution FCS-IMDM Culture Solution PHA
Cell Culture Plate Incubator

Move

I. Measurement of IL-4 biological activityPreparation of CT.4S cell suspension①↓Inoculate cells in a 96-well culture plate②.↓
Add the sample to be tested and the standard ③
↓plus 3H-TdR④↓
Cells were collected and the amount of 3H-TdR adulteration was determined ⑤↓Plot the standard curve and calculate the units of activity of IL-4 for the sample to be tested ⑥

Caveat

①Digested with 0.25% trypsin and collected the vigorously growing adherent CT.4S cells; after centrifugation and washing twice with Hank's solution, 10% FCS-RPMI-1640 culture medium was modulated to a cell concentration of 1×105/ml.② Inoculate cells in 96-well cell culture plate, 0.1 ml per well.③ Add the appropriate dilution of the samples and standards to be tested, 0.1 ml per well. 3 replicate wells were set up for each sample, and a culture medium control was set up at the same time. 37 degrees, 5% CO2Incubate for 48 hours.④ and ⑤ were the same as IL-2.The standard curve was plotted on semi-log graph paper with cpm as the vertical coordinate and the corresponding concentrations of different dilutions of IL-4 standards as the horizontal coordinate, and then the IL-4 content in the samples to be tested was checked from the standard curve.Key Points In the process of IL-4 induction, a certain amount of IL-2 is often produced at the same time, which will affect the detection results of IL-4. Therefore, it is better to use IL-2mAb adsorbent to remove IL-2.

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