Augmentative effects of phytohemagglutinin-P on proliferation and cytotoxicity of interleukin-2-activated canine peripheral blood lymphocytes.

Abstract

Canine peripheral blood lymphocytes (PBL) were simulated with recombinant human interleukin-2 (rhIL-2) alone, or with phytohemagglutinin-P (PHA) and subsequent rhIL-2 in order to obtain large numbers of lymphokine-activated killer (LAK) cells. Incubation of PBL with rhIL-2 alone allowed proliferation of large granular lymphocyte (LGL)-like lymphocytes, and the cytotoxic activity of the lymphocytes against canine transmissible venereal sarcoma cells was detected 5-7 days after the culture onset. However, the lymphocytes died within 2 weeks of culture, resulting in limited generation of functional LAK cells. Thus, PBL pretreated with PHA are subjected to rhIL-2-dependent culture. Small- or middle-sized lymphocytes predominantly proliferated in response to rhIL-2, and proliferation of the lymphocytes was sustained for longer than 30 days by repeated stimulations with PHA and subsequent rhIL-2. Cytotoxicity reached significant levels from 2 weeks after the culture onset and thereafter remained almost constant for at least 2 weeks, leading to large-scale production of the LAK cells. Pretreatment of PBL and PHA seems to enhance the LAK cell functions through modification of the precursors of the effector LAK cells.