Development and function of CD94-deficient natural killer cells - PubMed (original) (raw)

Development and function of CD94-deficient natural killer cells

Mark T Orr et al. PLoS One. 2010.

Abstract

The CD94 transmembrane-anchored glycoprotein forms disulfide-bonded heterodimers with the NKG2A subunit to form an inhibitory receptor or with the NKG2C or NKG2E subunits to assemble a receptor complex with activating DAP12 signaling proteins. CD94 receptors expressed on human and mouse NK cells and T cells have been proposed to be important in NK cell tolerance to self, play an important role in NK cell development, and contribute to NK cell-mediated immunity to certain infections including human cytomegalovirus. We generated a gene-targeted CD94-deficient mouse to understand the role of CD94 receptors in NK cell biology. CD94-deficient NK cells develop normally and efficiently kill NK cell-susceptible targets. Lack of these CD94 receptors does not alter control of mouse cytomegalovirus, lymphocytic choriomeningitis virus, vaccinia virus, or Listeria monocytogenes. Thus, the expression of CD94 and its associated NKG2A, NKG2C, and NKG2E subunits is dispensable for NK cell development, education, and many NK cell functions.

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Conflict of interest statement

Competing Interests: The authors have read the journal's policy and have the following conflicts: J.H.P. is an employee of Merck; P.S. and T.E. are employees of Novo Nordisk. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials. The other authors have no financial conflicts of interest.

Figures

Figure 1

Figure 1. Splenic CD94-deficient and CD94Tg/– NK cells are phenotypically normal.

(A) Splenocytes from B6, CD94-deficient, CD94Tg/–, and 129/SvJ mice were analyzed for CD94-NKG2 expression. CD94-NKG2+ cells were analyzed for NKp46 and TCRβ expression as shown in the second column. (B) NK cells (NKp46+ TCRβ–) were analyzed for expression of CD94, NKG2A/C/E, NK1.1, Ly49C/I, and Ly49H. (C) CD19– bone marrow cells were analyzed for NK cell precursors (NKG2D+ CD122+) and the developmental markers DX5, αV, CD27, and CD11b. Data are representative of three to five experiments each.

Figure 2

Figure 2. CD94-deficient NK cells are educated and efficiently kill YAC-1 targets and MHC I-deficient splenocytes.

(A) Splenocytes from B6, CD94-deficient, CD94Tg/–, and 129/SvJ mice were assayed for degranulation as measured by surface staining for CD107a and intracellular IFN-γ production upon stimulated with plate-bound anti-NKp46 mAb. (B) Splenocytes from CD94-deficient mice and wildtype 129/SvJ mice primed with poly I:C in vivo were assayed for NK cell-mediated cytotoxicity against the YAC-1 target cell. (C) CD94-deficient mice and B6 mice either depleted of NK cells with anti-NK1.1 mAb or treated with PBS received a mixture of CFSE-labeled wildtype and B2m−/− B6 splenocytes. Twenty-four hours later the frequency or CFSE-labeled donor cells in the spleen that were _B2m−/−_was determined by expression of H-2Kb. Data are representative of three to five experiments each.

Figure 3

Figure 3. CD94 is not necessary for control of MCMV.

(A) Two days after infection with MCMV splenic NK cells from B6 and CD94-deficient mice were analyzed for expression of CD69 and intracellular IFN-γ and granzyme B. (B) CD94-deficient and CD94Tg/– mice, (C) B6 mice treated with the non-depleting, blocking anti-NKG2A/C/E chimeric rat-mouse monoclonal antibody 20D5HCmIgG1-Q, anti-NK1.1 depleting antibody or PBS (D), or BALB/c mice receiving the 20D5HCmIgG1-Q antibody or PBS were infected with 5×104 pfu MCMV and then analyzed for viral titers three days later. Graphs represent the average ± s.e.m. of four or five animals per group. Not statistically significant (n.s.).

Figure 4

Figure 4. CD94 is not necessary for clearance of LCMV, vaccinia virus or Listeria infection.

(A) B6 and CD94-deficient mice were infected with 2×105 pfu of LCMV (Armstrong strain). Spleens and livers were analyzed for LCMV titers on the indicated days. (B) B6 and CD94-deficient mice were challenged with 1×105 cfu Listeria monocytogenes. Spleens and livers were analyzed for bacterial burdens three days later. (C) B6 and CD94-deficient mice were challenged with 5×106 pfu vaccinia virus. Spleens and ovaries were analyzed for viral titers seven days later. Graphs represent the average ± s.e.m. of three to five animals per group per time point.

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