Differential susceptibility of murine T and B lymphocytes to freeze-thaw and hypotonic shock

Experiments were carried out to determine if thymic-derived lymphocytes (T-cells) could be differentially damaged by hypotonic and/or freeze-thaw stress. The uptake of ³H-thymidine after stimulation of murine spleen cells with phytohemagglutinin-P (PHA-P) or bacterial endotoxin (LPS) was used as an indicator of recovery. Optimal freezing and thawing techniques showed that 100% of LPS-responsive cells could be recovered, compared to 65% of PHA-responsive cells. These differences could be increased by treatment of spleen cells with 0.17 m NH₄Cl prior to freezing and thawing. This represented a recovery of 50% of LPS-responsive cells and less than 10% of PHA-responsive cells. A similar effect could be obtained by treating NH₄Cl-treated spleen cells with distilled water prior to culture. It is hypothesized that T-cells are more susceptible to osmotic damage than B-cells due to their differences in membrane characteristics.