Functional nuclei of LPS-nonresponder C3H/HeJ mice after transfer into LPS-responder C3H/HeN cells by cell fusion

Publisher Website
Google Scholar
Abstract
Splenic lymphocytes of C3H/HeJ mice are defective in responding to the mitogenic activity of bacterial lipopolysaccharide (LPS), which was attributed to a mutation at a single gene locus on chromosome. Apparently, the genetic defect in C3H/HeJ mice is related to triggering mechanisms rather than to LPS binding. A method for separating mouse splenic lymphocytes into karyoplasts (minicells) and cytoplasts (enucleated cells) with high purity is known. This method makes possible the exchange of nuclei from 1 type of lymphocyte to another. Karyoplasts were purified from LPS-nonresponder C3H/HeJ mice and transferred to mitomycin C-treated or X ray-irradiated splenic lymphocytes taken from LPS-responder C3H/HeN mice by fusion with polyethyleneglycol. These reconstituted hybrid cells could respond to LPS, suggesting that nuclei from C3H/HeJ mice could be activated by LPS if proper signals were provided from cell-surface receptors. Moreover, responsiveness to concanavalin A of mitomycin C-treated or X-ray-irradiated lymphocytes was also recovered by the karyoplasts microinjection from untreated lymphocytes.