CLONING, FUNCTIONAL ACTIVITIES AND IN-VIVO TISSUE DISTRIBUTION OF RATNKR-P1(-ALPHA-BETA(+) CELLS() TCR)

We have successfully cloned nine NKR-P1(+) TCR alpha beta(+) cells fro m PVG rat spleens, utilizing murine macrophage inflammatory protein-1 alpha (MIP-1 alpha) and IL-2. These clones are either double negative (DN, CD4(-)CD8(-)), which included clones 3.31, 3.71, 4.19, 4.59 and 4 .65, or single positive (SP, CD4(+)CD8(-)), which included clones 1.64 , 3.8, 3.76 and 3.78. No CD8(+) clone was recovered. All nine clones a re restricted in terms of their expression of the Vp antigens, since t hey express V(beta)8.2 but not V(beta)8.5, V(beta)10 or V(beta)16. The se clones are agranular and they fail to generate NK or LAK activity u pon incubation with IL-2, IL-12 or their combination. On the basis of their production of intracellular cytokines they can be divided into t hree categories: (i) SP clones (1.64, 3.8, 3.76 and 3.78) do not produ ce IL-2 or IL-4, but produce IFN-gamma and IL-12, and they vary in the ir production of IL-1, RANTES or tumor necrosis factor (TNF)-alpha; (i i) DN clones 4.59 and 4.65 produce IL-la and IFN-gamma only, and fail to produce other cytokines; and (iii) DN clones 3.31, 3.71 and 4.19 pr oduce IL-1 alpha, IL-1 beta, IL-2, IL-12, IFN-gamma RANTES and TNF-alp ha. From all the clones examined only DN clones 3.31 and to a lesser d egree 4.19 produce IL-4. In vivo tissue localization of clones 3.8, 3. 31 and 4.59 shows that these cells distribute into the liver and bone marrow 24 h post i.v. administration. Their accumulation in the liver and bone marrow along with their ability to secrete various cytokines, suggest that these cells may influence the generation, differentiatio n or apoptosis of immune or hematopoietic cells.