| Objective: To evaluate the inhibitory effects of anti-Marburg monoclonal antibody LZ01 alone and in combination with Tim-1 recombinant protein on viral infection in vitro.Methods: Marburg pseudovirus was prepared by transfecting MARV-GP plasmid for use in subsequent experiments.The affinity of MRAV GP Δ mucin protein to LZ01 was measured using ELISA,BLI.The neutralizing activity of LZ01 was detected by antibody neutralization assay.NK cell activation mediated by LZ01 and LZ01-M1 was detected by flow cytometry.Tim-1-overexpressing HEK-293 T cells were prepared as target cells by transient transfection of Tim-1 expression plasmid.The bioluminescence method was used to evaluate the promoting effect of Tim-1 and its polymorphisms on MARV infection,and to detect the blocking effect of Tim-1-ECD recombinant protein on the promoting effect of Tim-1.Finally,the synergistic effect of Tim-1-ECD recombinant protein combined with LZ01 was detected.Results:(1)The Marburg pseudovirus was successfully prepared and had high infectivity to HEK-293 T cells.(2)The binding activity of MRAV GP Δ mucin protein to LZ01 was very well.(3)LZ01 has prominent neutralizing activity.(4)Both LZ01 and LZ01-M1 had ability to trigger ADCC,and the activity of LZ01-M1 was significantly stronger than LZ01,which was antibody concentration-dependent.(5)Tim-1abundance-dependently enhanced the infectivity of Marburg pseudovirus.(6)Tim-1-359 aa and-364 aa had a promoting effect,and the activity of Tim-1-359 aa was stronger than Tim-1-364 aa.(7)Tim-1-ECD recombinant protein significantly inhibited Marburg pseudovirus infection in Tim-1 introduced HEK293 T cells.(8)Tim-1-ECD recombinant protein in combination with LZ01 significantly enhanced the inhibition of viral infection in HEK-293 T cells.Conclusion: The anti-Marburg monoclonal antibody LZ01 exhibits prominent neutralizing activity alone and shows synergistic effects with Tim-1-ECD recombinant protein. |
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