International Immunology, Vol 10, 1093-1101, Copyright © 1998 by Oxford University Press
Q Vos, JR Ortaldo, M Conan-Cibotti, MD Vos, HA Young, SK Anderson, K Witherspoon, I Prager, CM Snapper and JJ Mond
NK cells not only function as cytotoxic effector cells, but also have
immunoregulatory roles including the enhancement of Ig secretion. To have a
stable and uniform population of NK cells to study their role in Ig
secretion, we generated murine NK clones. Thus, culture of splenocytes from
mice that were homozygous for a mutation in the p53 tumor suppressor gene
(p53-KO) with IL-2 and poly(IC) resulted in a long-term NK line, from which
four stable clones were derived. This approach also yielded a long-term NK
line from splenocytes of normal C57BL/6 mice. Identification of the clones
as members of the NK lineage was based on large granular morphology,
expression of NK-TR and absence of TCR gene rearrangement. Flow cytometry
revealed that all clones expressed IL-2R alpha and beta, chains and B220,
but no CD3, NK1.1, DX5 or Ly-49. RT-PCR analysis showed heterogeneity in
NK1.1 gene expression, and demonstrated expression of perforin and several
granzymes in all clones. Three out of four clones lysed YAC-1, but not P815
target cells, corresponding to a pattern of NK specificity. All NK clones
enhanced Ig secretion in an in vitro model for T cell- independent type 2
antigens, albeit to varying degrees. We found no correlation between the
degree of helper activity of the NK clones and the level of their cytotoxic
activity on YAC-1 targets. Thus, we established murine NK clones, and show
that they mediate both cytotoxicity and enhancement of Ig secretion.
ARTICLES
Phenotypic and functional characterization of a panel of cytotoxic murine NK cell clones that are heterogeneous in their enhancement of Ig secretion in vitro
Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA.
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