Recruitment of alloreactive natural killer cells to the rat peritoneum by a transfected cell line secreting rat recombinant interleukin-2.
In order to obtain large numbers of natural killer (NK) cells from single rats for functional studies, we have devised a method for the generation of IL-2-activated NK cells in vivo. Rats were implanted intraperitoneally with cell-impermeable diffusion chambers (DC) containing cultures of transfected Chinese hamster ovary (CHO) cells secreting rat recombinant interleukin-2 (rIL-2). This resulted in a dramatic increase in the peritoneal exudate cell (PEC) number with a peak (300-1000 x 10(6)) 1 week after implantation. The majority were mononuclear cells of which a large proportion were CD3-NKR-P1+ NK cells, but with substantial numbers of macrophages (M phi) and CD3+8+NKR-P1+ T cells also. The NK activity against standard tumor target cells was high among PEC from six different inbred rat strains tested. However, the NK cell-mediated reactivity against concanavalin A (ConA)-activated T cell blasts from a panel of major histocompatibility complex (MHC) congenic strains differed widely. PEC from some strains (PVG, LOU/C, and AO) efficiently lysed all the MHC-disparate lymphoblasts. In other strains (BN and LEW) more restricted allorecognition repertoires were observed, whereas PEC from one strain (DA) were unresponsive. The secretion of rat rIL-2 intraperitoneally did not lead to a significant increase in the IL-2 level in the blood or in the total number or activity of NK cells in blood and spleen. The present method represents a most potent technique for generating large numbers of functional rat NK cells and shows the high efficiency with which IL-2 can induce NK cell recruitment in vivo.
Løvik
G
G
Department of Anatomy, University of Oslo, Norway.
Vaage
J T
JT
Naper
C
C
Benestad
H B
HB
Rolstad
B
B
D000818:Animals; D001202:Ascitic Fluid / Q000166:cytology / Q000276:immunology; D016466:CHO Cells / Q000276:immunology*; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D006224:Cricetinae; D003602:Cytotoxicity, Immunologic; D015194:Diffusion Chambers, Culture / Q000379:methods*; D005434:Flow Cytometry; D007376:Interleukin-2 / Q000378:metabolism*; D007694:Killer Cells, Natural / Q000276:immunology*; D010529:Peritoneal Cavity / Q000166:cytology*; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
D007376:Interleukin-2; D011994:Recombinant Proteins