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

Ascione, Alessandro; Arenaccio, Claudia; Mallano, Alessandra; Flego, Michela; Gellini, Mara; Andreotti, Mauro; Fenwick, Craig; Pantaleo, Giuseppe; Vella, Stefano; Federico, Maurizio

doi:10.1186/s12896-019-0559-x

Cited by 16 publications

(10 citation statements)

References 44 publications

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“…. As expected, neither E6 nor E7 associated with EVs, as shown by the western blot comparative analysis including a His-tagged single-chain antibody (scFv) fused with Nef mut (26) (Fig. 3A).…”

Section: Hpv16-e6 and -E7-specific Cd8 + T Cell Immune Response In MIsupporting

confidence: 70%

Long-term antitumor CD8⁺ T cell immunity induced by endogenously engineered extracellular vesicles

Ferrantelli

Manfredi

Chiozzini

et al. 2021

Preprint

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We developed a novel approach to induce antigen-specific CD8+ T cytotoxic lymphocyte (CTL) immunity based on in vivo engineering of extracellular vesicles (EVs). This is an innovative vaccination approach employing a DNA vector expressing a mutated HIV-1 Nef protein (Nefmut) that has lost the anti-cellular effects typical of the wild-type isoform, meanwhile showing an unusual efficiency of incorporation into EVs. This function persists even when foreign antigens are fused to its C-terminus. In this way, Nefmut traffics large amounts of antigens fused to it into EVs spontaneously released by cells expressing the Nefmut-based DNA vector. We previously provided evidence that the inoculation in mice of a DNA vector expressing the Nefmut/HPV16-E7 fusion protein induced an E7-specific CTL immune response as detected 2 weeks after the second immunization. In an effort to optimize the anti-HPV16 CD8+ T cell immune response, we found that the co-injection of DNA vectors expressing Nefmut fused with E6 and E7 generated a stronger anti-HPV16 immune response compared to that we observed in mice injected with the single vectors. When TC-1 cells, i.e., a tumor cell line co-expressing E6 and E7, were implanted before immunization, all mice survived until day 44, whereas no mice injected with either void or Nefmut-expressing vectors survived until day 32 after tumor implantation. A substantial part of mice (7 out of 12) cleared the tumor. When cured mice were re-challenged with a second sub cute implantation of TC-1 cells, and followed for additional 135 days, whereas none of them developed tumors. Both E6- and E7-specific CD8+ T immunity was still detectable at the end of the observation time. Hence, the immunity elicited by engineered EVs, besides curing already developed tumors, is strong enough to guarantee the resistance to additional tumor attack. This results is of relevance for therapy against both metastatic and relapsing tumors.

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Section: Hpv16-e6 and -E7-specific Cd8 + T Cell Immune Response In MIsupporting

confidence: 70%

Long-term antitumor CD8⁺ T cell immunity induced by endogenously engineered extracellular vesicles

Ferrantelli

Manfredi

Chiozzini

et al. 2021

Preprint

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“…( Figure 4 A and Figure S9 ). In this assay, a DNA vector expressing a His-tagged single-chain antibody (scFv) fused with Nef mut [ 27 ] was used as control.…”

Section: Resultsmentioning

confidence: 99%

Long-Term Antitumor CD8+ T Cell Immunity Induced by Endogenously Engineered Extracellular Vesicles

Ferrantelli

Manfredi

Chiozzini

et al. 2021

Cancers

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We developed an innovative method to induce antigen-specific CD8+ T cytotoxic lymphocyte (CTL) immunity based on in vivo engineering of extracellular vesicles (EVs). This approach employs a DNA vector expressing a mutated HIV-1 Nef protein (Nefmut) deprived of the anti-cellular effects typical of the wild-type isoform, meanwhile showing an unusual efficiency of incorporation into EVs. This function persists even when foreign antigens are fused to its C-terminus. In this way, Nefmut traffics large amounts of antigens fused to it into EVs spontaneously released by the recipient cells. We previously provided evidence that mice injected with a DNA vector expressing the Nefmut/HPV16-E7 fusion protein developed an E7-specific CTL immune response as detected 2 weeks after the second immunization. Here, we extended and optimized the anti-HPV16 CD8+ T cell immune response induced by the endogenously engineered EVs, and evaluated the therapeutic antitumor efficacy over time. We found that the co-injection of DNA vectors expressing Nefmut fused with E6 and E7 generated a stronger anti-HPV16 immune response compared to that observed in mice injected with the single vectors. When HPV16-E6 and -E7 co-expressing tumor cells were implanted before immunization, all mice survived at day 44, whereas no mice injected with either void or Nefmut-expressing vectors survived until day 32 after tumor implantation. A substantial part of immunized mice (7 out of 12) cleared the tumor. When the cured mice were re-challenged with a second tumor cell implantation, none of them developed tumors. Both E6- and E7-specific CD8+ T immunities were still detectable at the end of the observation time. We concluded that the immunity elicited by engineered EVs, besides counteracting and curing already developed tumors, was strong enough to guarantee the resistance to additional tumor attacks. These results can be of relevance for the therapy of both metastatic and relapsing tumors.

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“…The critical roles of PD1 and LAG3 checkpoints in CD8+ T cell responses were explored by functional evaluation of CD8+ T cells. CD8+ T lymphocytes are specific for tumor antigens, and their driven immune reactivation holds promise for significant therapeutic benefit against exhaustion of lymphocyte subsets in cancer patients [36].…”

Section: Computational and Mathematical Methods In Medicinementioning

confidence: 99%

LAG3 and PD1 Regulate CD8+ T Cell in Diffuse Large B-cell Lymphoma Patients

Liu

Zhan

et al. 2021

Computational and Mathematical Methods in Medicine

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Background. Diffuse large B-cell lymphoma (DLBCL) is a clinically and genetically heterogeneous lymphoid malignancy. The unsatisfactory outcome for refractory patients has prompted efforts to explore new therapeutic approaches for DLBCL. However, the mechanisms involved in treatment associated with immune checkpoints remain unclear. This study is aimed at investigating the potential roles of programmed cell death protein 1 (PD1) and lymphocyte activation gene 3 (LAG3) in CD8+ T cells for treatment in DLBCL. Methods. Utilizing flow cytometry, we examined the content of T cells, the levels of cytokines, and the expression of PD1 and LAG3 in patients with DLBCL as well as in healthy controls. Levels of cytokines in CD8+ T cells from DLBCL patients before and after treatment were compared by blocking of PD1 and LAG3 in magnetic bead-sorted CD8+ T cells. Results. We found that the proportion of CD4+ T cells and CD8+ T cells was increased in DLBCL patients after treatment. The levels of cytokines trended toward those of healthy controls in treatment. PD1 (+), LAG3 (+), or PD1 (+) LAG3 (+) were all expressed in lower amounts in CD4+ T cells and CD8+ T cells after treatment than in untreated DLBCL patients. In addition, blockade of PD1 and LAG3 in sorted CD8+ T cells markedly inhibited cytokine production in response to treatment. Conclusion. PD1 and LAG3 in CD8+ T cells may be important targets of therapy and play therapeutic role in patients with DLBCL.

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

Cited by 16 publications

References 44 publications

Long-term antitumor CD8⁺ T cell immunity induced by endogenously engineered extracellular vesicles

Long-term antitumor CD8⁺ T cell immunity induced by endogenously engineered extracellular vesicles

Long-Term Antitumor CD8+ T Cell Immunity Induced by Endogenously Engineered Extracellular Vesicles

LAG3 and PD1 Regulate CD8+ T Cell in Diffuse Large B-cell Lymphoma Patients

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

Cited by 16 publications

References 44 publications

Long-term antitumor CD8+ T cell immunity induced by endogenously engineered extracellular vesicles

Long-term antitumor CD8+ T cell immunity induced by endogenously engineered extracellular vesicles

Long-Term Antitumor CD8+ T Cell Immunity Induced by Endogenously Engineered Extracellular Vesicles

LAG3 and PD1 Regulate CD8+ T Cell in Diffuse Large B-cell Lymphoma Patients

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Long-term antitumor CD8⁺ T cell immunity induced by endogenously engineered extracellular vesicles

Long-term antitumor CD8⁺ T cell immunity induced by endogenously engineered extracellular vesicles