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. 2014 May 15;10(5):e1004131. doi: 10.1371/journal.ppat.1004131

Figure 7. Soluble ectodomains of HCMV vFcγRs interfere with FcγR activation.

Figure 7

(A) SKOV-3 cells were opsonized with 100 ng/ml trastuzumab for 30 minutes and washed three times with D-MEM 10% FCS (vol/vol) before soluble proteins were added in graded concentrations concomitantly with BW:FcγRIIIA-ζ transfectants. mIL-2 was determined in supernatants (which were harvested after 16 h of co-cultivation of responder cells with target cells) by ELISA. (B) As in (A) but SKOV-3 cells were opsonized with 500 µg/ml trastuzumab for 30 minutes and washed three times with D-MEM 10% FCS (vol/vol) before soluble proteins were added in graded concentrations concomitantly with BW:FcγRI-ζ cells. (C) MRC-5 cells were infected with HCMV HB5ΔIRLΔgp68/Δgp34 (2 PFU/cell) for 72 h before soluble proteins were added in graded concentrations concomitantly with BW:FcγRIIIA-ζ responder cells. MRC-5 fibroblasts were opsonized with 1∶50 diluted Cytotect for 30 min. After removing of unbound antibodies by washing, soluble proteins and BW:FcγR-ζ transfectants were added and co-cultivated overnight. (D) as in (C), but HCMV HB5ΔIRLΔgp68/Δgp34 infected target cells were opsonized with 1∶10 diluted Cytotect and BW:FcγRIIA-ζ cells were used as responders. n = 3 replicates, means with standard deviations (error bars) are shown for 2 independent experiments. Significance of results (Student's t-test) are presented in Table S1 as *: p<0.05 **: p<0.01 ***: p<0.001.