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Feline hybridoma growth factor/interleukin-6 activity
1Department of Veterinary Internal Medicine, Faculty of Agriculture, University of Tokyo, Japan.
Insights
A new assay using a mouse-rat hybridoma clone (B3B1) can measure feline hybridoma growth factor (HGF)/interleukin-6 (IL-6) activity. This method is specific for feline IL-6 and reveals species-specific properties.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Interleukin-6 (IL-6) is a pleiotropic cytokine involved in immune responses and cell proliferation.
- Measuring feline IL-6 activity is crucial for understanding feline immunology and disease.
- Existing assays may not be specific or sensitive enough for feline samples.
Purpose of the Study:
- To develop a novel assay for quantifying feline hybridoma growth factor (HGF)/interleukin-6 (IL-6) activity.
- To characterize the specificity and physicochemical properties of feline HGF/IL-6.
Main Methods:
- Established a mouse-rat hybridoma clone (B3B1) responsive to feline HGF/IL-6.
- Assessed the proliferative response of B3B1 to various cytokines.
- Utilized gel filtration and anion exchange chromatography for physicochemical characterization.
- Tested the neutralizing capacity of anti-human IL-6 antiserum.
Main Results:
- The B3B1 clone exhibited specific proliferation in response to feline HGF/IL-6, unaffected by other tested cytokines.
- Anti-human IL-6 antiserum did not neutralize feline HGF/IL-6, indicating antigenic differences.
- Feline HGF/IL-6 displayed a molecular weight of 30,000-40,000 and eluted at 0.2-0.3 M NaCl.
- Physicochemical properties of feline HGF/IL-6 differed slightly from murine and human IL-6.
Conclusions:
- A specific and reliable assay for feline HGF/IL-6 activity was successfully developed.
- The study identified distinct antigenic and physicochemical characteristics of feline IL-6 compared to other species.
- This assay provides a valuable tool for feline cytokine research.
Abstract:
An assay system was developed to measure feline hybridoma growth factor (HGF)/interleukin-6 (IL-6) activity in biological samples containing many kinds of cytokines by using the proliferation of the newly established mouse-rat hybridoma clone, B3B1. The proliferative response of this B3B1 clone was IL-6-specific, and could not be promoted by other cytokines including IL-1, IL-2, IL-3, and granulocyte-colony-stimulating factor (G-CSF). The anti-human B-cell stimulatory factor 2 (BSF-2)/IL-6 antiserum did not neutralize feline HGF/IL-6 activity in conditioned media prepared from feline con A-stimulated splenocytes and unstimulated alveolar macrophages, indicating antigenic differences between species. Feline HGF/IL-6 was eluted into the fractions corresponding to a molecular weight of 30,000-40,000 in gel filtration, and into the fractions at a salt concentration of 0.2-0.3 M NaCl in anion exchange chromatography. The physicochemical properties of feline HGF/IL-6 were slightly different from those of murine and human IL-6.
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