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. 2019 Jul 10;9:9998. doi: 10.1038/s41598-019-46329-y

Figure 1.

Figure 1

Association between HCMV infection and SLE through UL44 and characterisation of an anti-UL44 antibody. Plasma anti-HCMV and anti-UL44 IgG levels were determined in a cohort of HCMV IgG seropositive 32 SLE patients and 69 controls. (a) Plasma anti-HCMV IgG levels were significantly higher in SLE patients than controls (mean of 3.251 vs 2.208; ****P < 0.0001). Results shown are from a representative experiment. The experiment was repeated for a total of 3 times. Data represent mean ± SD. Statistical significance was determined using two-tailed t test. (b) Among HCMV(+) individuals with anti-UL44 IgG antibodies, SLE patients had significantly higher plasma concentrations of anti-UL44 IgG antibodies than controls (median of 2.152 vs 0.575 µg/ml; **P = 0.0016). Each data point is an average from 3 independent experiments. Data represent median ± interquartile range. Statistical significance was determined using two-tailed Mann-Whitney U test. (c) ROC curve was plotted for anti-UL44 IgG for anti-UL44 IgG(+) SLE and controls (AUC = 0.696; P = 0.00160). (d) Schematic illustration of the panning process. (e) Computational modelling of the HCMV UL44 protein. The epitope targeted by 3A11 is indicated in red. (f) Co-immunoprecipitation (Co-IP) was performed using 3A11 (left panel) and anti-dsDNA antibody (right panel) on uninfected (−) or RV1305-infected (+) ARPE-19 cell lysates. Immunoprecipitation products were electrophoresed and immunoblotted separately using monoclonal antibodies against UL44, nucleolin, ku70 and pp65. Co-IP using 3A11 revealed that nucleolin, ku70 and pp65 complexes with UL44. dsDNA was observed to be part of these complexes when co-IP was performed with anti-dsDNA antibody. Full-length blots are presented in Supplementary Fig. 5.