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Fig. 7 | BMC Biotechnology

Fig. 7

From: Development of a novel human phage display-derived anti-LAG3 scFv antibody targeting CD8+ T lymphocyte exhaustion

Fig. 7

The treatment with the divalent scFvF7-Fc Ab increases the activation of peptide-stimulated antigen-specific CD8+ T lymphocytes. a Both Nef- and Mart1-specific CD8+ T lymphocytes were put in co-culture with HLA-matched B-LCLs previously treated with appropriate peptides, and in the presence of either IgGs, divalent scFvF7-Fc Ab, or the murine anti-LAG3 17B4 mAb. As control, both B-LCLs (either untreated or treated with appropriate peptides) and CD8+ T cells were cultivated either alone or in co-culture. After o.n. incubation, supernatants were harvested, clarified, and tested for IFN-γ contents. The mean values ±SEM are shown as calculated from triplicate wells from a representative of three independent experiments. b Dose-response effect of the divalent scFvF7-Fc Ab and inhibition by recombinant LAG3. Co-cultures comprising B-LCLs and Nef-specific CD8+ T lymphocytes were set up as reported for panel A, albeit in the presence of different amounts of the reconstituted F7 Ab. Conditions comprising the highest concentration of the divalent scFvF7 and increasing doses of recombinant LAG3 were also run. After o.n. cultivation, supernatants were tested for IFN-γ contents. The mean values ±SEM are shown as calculated from triplicate wells from a representative of two independent experiments. c Effects of the divalent scFvF7-Fc as sensed by IFN-γ ELISPOT assay. IFN-γ-specific CD8+ T cell response as detected by ELISPOT assay after o.n. co-cultures of Nef-specific CD8+ T cells with B-LCLs pre-incubated or not with the specific peptide. The co-cultures were run in the presence or absence of either the divalent scFvF7-Fc Ab or the murine anti-LAG3 17B4 mAb. The former co-cultures were carried also out also in the presence of either recombinant LAG3, or the same amounts of an unrelated recombinant protein (i.e., HIV-1 gp120). Results are expressed as spot-forming units (SFU)/105 CD8+ T cells, and are representative of two experiments performed with triplicate wells

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