Limits...
The murine nonclassical class I major histocompatibility complex-like CD1.1 molecule protects target cells from lymphokine-activated killer cell cytolysis.

Chang CS, Brossay L, Kronenberg M, Kane KP - J. Exp. Med. (1999)

Bottom Line: Passage of effector cells in recombinant interleukin (rIL)-2 enhanced protection by mCD1.1, suggesting an expansion of relevant A-LAK population(s) or modulation of A-LAK receptor expression.CD1.1 is by far the most divergent class I molecule capable of regulating NK cell activity.Finally, mCD1.1 expression rendered RMA/S cells resistant to lysis by A-LAK of multiple mouse strains.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology and Immunology, Faculty of Medicine, University of Alberta, Edmonton, Alberta T6G 2S2, Canada.

ABSTRACT
Classical class I major histocompatibility complex (MHC) molecules, as well as the nonclassical class I histocompatibility leukocyte antigen (HLA)-E molecule, can negatively regulate natural killer (NK) cell cytotoxicity through engagement of NK inhibitory receptors. We show that expression of murine (m)CD1.1, a nonpolymorphic nonclassical MHC class I-like molecule encoded outside the MHC, protects NK-sensitive RMA/S target cells from adherent lymphokine-activated killer cell (A-LAK) cytotoxicity. Passage of effector cells in recombinant interleukin (rIL)-2 enhanced protection by mCD1.1, suggesting an expansion of relevant A-LAK population(s) or modulation of A-LAK receptor expression. Murine CD1. 1 conferred protection from lysis by rIL-2-activated spleen cells of recombination activating gene (Rag)-1(-/-) mice, which lack B and T cells, demonstrating that mCD1.1 can protect RMA/S cells from lysis by NK cells. An antibody specific for mCD1.1 partially restored A-LAK lysis of RMA/S.CD1.1 transfectants, indicating that cell surface mCD1.1 can confer protection from lysis; therefore, mCD1.1 possibly acts through interaction with an NK inhibitory receptor. CD1.1 is by far the most divergent class I molecule capable of regulating NK cell activity. Finally, mCD1.1 expression rendered RMA/S cells resistant to lysis by A-LAK of multiple mouse strains. The conserved structure of mCD1.1 and pattern of mCD1.1 resistance from A-LAK lysis suggest that mCD1.1 may be a ligand for a conserved NK inhibitory receptor.

Show MeSH

Related in: MedlinePlus

mCD1.1 partially  inhibits the cytotoxic activity of  poly I:C–activated B6 NK cells.  Mice were injected with 200 μg  of poly I:C, and 18 h later the  spleen cells were harvested and  the plastic nonadherent spleen  cells were used as effector cells  for the lysis of 51Cr-labeled  RMA/S and RMA/S.CD1.1.  Each E/T ratio was used for triplicate determinations, and the results are expressed as means ±  SD. Spontaneous release values  were <10.3%. Similar results  were obtained in three separate  experiments.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2192909&req=5

Figure 1: mCD1.1 partially inhibits the cytotoxic activity of poly I:C–activated B6 NK cells. Mice were injected with 200 μg of poly I:C, and 18 h later the spleen cells were harvested and the plastic nonadherent spleen cells were used as effector cells for the lysis of 51Cr-labeled RMA/S and RMA/S.CD1.1. Each E/T ratio was used for triplicate determinations, and the results are expressed as means ± SD. Spontaneous release values were <10.3%. Similar results were obtained in three separate experiments.

Mentions: We first carried out in vitro killing assays of RMA/S and RMA/S.CD1.1, using poly I:C–activated B6 NK cells. As expected, the class I MHC–deficient cell line, RMA/S, is susceptible to NK cell lysis. However, a modest but reproducible reduction in lysis of the RMA/S.CD1.1 transfectant was detected at higher E/T ratios where cytotoxicity is substantial, but not apparently at plateau, and can be readily compared between the experimental groups (Fig. 1). This suggested that mCD1.1 is able to inhibit NK cell cytotoxic activity, but perhaps only a small fraction of the total poly I:C–activated NK cell population is negatively regulated by mCD1.1. Therefore, we performed cytolytic assays using B6 A-LAKs to test the possibility that mCD1.1-regulated killer cells could be expanded when cultured in the presence of rIL-2. The RMA/S cell line and two independent RMA/S.CD1.1 transfectant clones were compared for their sensitivity to lysis by nylon wool nonadherent B6 spleen cells that become plate adherent in succeeding days of culture in rIL-2. RMA/S and the two RMA/S.CD1.1 transfectants were susceptible to lysis by the day 3 and 4 A-LAKs. Nevertheless, a moderate reduction in lysis by these A-LAKs of both RMA/S.CD1.1 transfectants relative to RMA/S was detected (Fig. 2). Interestingly, day 5 and 6 A-LAK cytotoxic activities against the RMA/S.CD1 transfectants was substantially reduced relative to the highly sensitive RMA/S target cell (Fig. 2). Resistance to lysis of the RMA/S.CD1.1 transfectants was found to be enhanced despite an increase of lysis observed for the untransfected RMA/S target cell at lower E/T ratios by the day 5 and 6 A-LAKs (Fig. 2). The two CD1 transfectants, RMA/ S.CD1.1A and RMA/S.CD1.1B, express comparable densities of CD1.1 (data not shown) and show a similar level of resistance to A-LAK lysis, indicating that the results are reproducible, occurring with more than one CD1.1 transfectant. Subsequent experiments described in this report were carried out with the RMA/S.CD1.1A transfectant, nominally referred to as RMA/S.CD1.1. Taken together, the preceding results obtained with IL-2–activated A-LAKs strongly argue that mCD1.1, like classical class I MHC, may negatively regulate A-LAK cytotoxic activity. In agreement with the poly I:C data, it seems that a small proportion of the initial A-LAK population may be negatively regulated by mCD1.1, which possibly can be expanded by passage and culture in the presence of rIL-2.


The murine nonclassical class I major histocompatibility complex-like CD1.1 molecule protects target cells from lymphokine-activated killer cell cytolysis.

Chang CS, Brossay L, Kronenberg M, Kane KP - J. Exp. Med. (1999)

mCD1.1 partially  inhibits the cytotoxic activity of  poly I:C–activated B6 NK cells.  Mice were injected with 200 μg  of poly I:C, and 18 h later the  spleen cells were harvested and  the plastic nonadherent spleen  cells were used as effector cells  for the lysis of 51Cr-labeled  RMA/S and RMA/S.CD1.1.  Each E/T ratio was used for triplicate determinations, and the results are expressed as means ±  SD. Spontaneous release values  were <10.3%. Similar results  were obtained in three separate  experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2192909&req=5

Figure 1: mCD1.1 partially inhibits the cytotoxic activity of poly I:C–activated B6 NK cells. Mice were injected with 200 μg of poly I:C, and 18 h later the spleen cells were harvested and the plastic nonadherent spleen cells were used as effector cells for the lysis of 51Cr-labeled RMA/S and RMA/S.CD1.1. Each E/T ratio was used for triplicate determinations, and the results are expressed as means ± SD. Spontaneous release values were <10.3%. Similar results were obtained in three separate experiments.
Mentions: We first carried out in vitro killing assays of RMA/S and RMA/S.CD1.1, using poly I:C–activated B6 NK cells. As expected, the class I MHC–deficient cell line, RMA/S, is susceptible to NK cell lysis. However, a modest but reproducible reduction in lysis of the RMA/S.CD1.1 transfectant was detected at higher E/T ratios where cytotoxicity is substantial, but not apparently at plateau, and can be readily compared between the experimental groups (Fig. 1). This suggested that mCD1.1 is able to inhibit NK cell cytotoxic activity, but perhaps only a small fraction of the total poly I:C–activated NK cell population is negatively regulated by mCD1.1. Therefore, we performed cytolytic assays using B6 A-LAKs to test the possibility that mCD1.1-regulated killer cells could be expanded when cultured in the presence of rIL-2. The RMA/S cell line and two independent RMA/S.CD1.1 transfectant clones were compared for their sensitivity to lysis by nylon wool nonadherent B6 spleen cells that become plate adherent in succeeding days of culture in rIL-2. RMA/S and the two RMA/S.CD1.1 transfectants were susceptible to lysis by the day 3 and 4 A-LAKs. Nevertheless, a moderate reduction in lysis by these A-LAKs of both RMA/S.CD1.1 transfectants relative to RMA/S was detected (Fig. 2). Interestingly, day 5 and 6 A-LAK cytotoxic activities against the RMA/S.CD1 transfectants was substantially reduced relative to the highly sensitive RMA/S target cell (Fig. 2). Resistance to lysis of the RMA/S.CD1.1 transfectants was found to be enhanced despite an increase of lysis observed for the untransfected RMA/S target cell at lower E/T ratios by the day 5 and 6 A-LAKs (Fig. 2). The two CD1 transfectants, RMA/ S.CD1.1A and RMA/S.CD1.1B, express comparable densities of CD1.1 (data not shown) and show a similar level of resistance to A-LAK lysis, indicating that the results are reproducible, occurring with more than one CD1.1 transfectant. Subsequent experiments described in this report were carried out with the RMA/S.CD1.1A transfectant, nominally referred to as RMA/S.CD1.1. Taken together, the preceding results obtained with IL-2–activated A-LAKs strongly argue that mCD1.1, like classical class I MHC, may negatively regulate A-LAK cytotoxic activity. In agreement with the poly I:C data, it seems that a small proportion of the initial A-LAK population may be negatively regulated by mCD1.1, which possibly can be expanded by passage and culture in the presence of rIL-2.

Bottom Line: Passage of effector cells in recombinant interleukin (rIL)-2 enhanced protection by mCD1.1, suggesting an expansion of relevant A-LAK population(s) or modulation of A-LAK receptor expression.CD1.1 is by far the most divergent class I molecule capable of regulating NK cell activity.Finally, mCD1.1 expression rendered RMA/S cells resistant to lysis by A-LAK of multiple mouse strains.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology and Immunology, Faculty of Medicine, University of Alberta, Edmonton, Alberta T6G 2S2, Canada.

ABSTRACT
Classical class I major histocompatibility complex (MHC) molecules, as well as the nonclassical class I histocompatibility leukocyte antigen (HLA)-E molecule, can negatively regulate natural killer (NK) cell cytotoxicity through engagement of NK inhibitory receptors. We show that expression of murine (m)CD1.1, a nonpolymorphic nonclassical MHC class I-like molecule encoded outside the MHC, protects NK-sensitive RMA/S target cells from adherent lymphokine-activated killer cell (A-LAK) cytotoxicity. Passage of effector cells in recombinant interleukin (rIL)-2 enhanced protection by mCD1.1, suggesting an expansion of relevant A-LAK population(s) or modulation of A-LAK receptor expression. Murine CD1. 1 conferred protection from lysis by rIL-2-activated spleen cells of recombination activating gene (Rag)-1(-/-) mice, which lack B and T cells, demonstrating that mCD1.1 can protect RMA/S cells from lysis by NK cells. An antibody specific for mCD1.1 partially restored A-LAK lysis of RMA/S.CD1.1 transfectants, indicating that cell surface mCD1.1 can confer protection from lysis; therefore, mCD1.1 possibly acts through interaction with an NK inhibitory receptor. CD1.1 is by far the most divergent class I molecule capable of regulating NK cell activity. Finally, mCD1.1 expression rendered RMA/S cells resistant to lysis by A-LAK of multiple mouse strains. The conserved structure of mCD1.1 and pattern of mCD1.1 resistance from A-LAK lysis suggest that mCD1.1 may be a ligand for a conserved NK inhibitory receptor.

Show MeSH
Related in: MedlinePlus