Dataset Information

Cytotoxicity effect of trastuzumab on canine peripheral blood mononuclear cells.

ABSTRACT:

Background

Trastuzumab is an antibody drug used to treat human epidermal growth factor receptor 2 (HER2) overexpressing human metastatic breast cancer. Antibody-dependent cellular cytotoxicity (ADCC) is considered to be the major mechanism of cytotoxicity of the drug. However, its ability to induce an ADCC response in canine peripheral blood mononuclear cells (PBMCs) is not well established.

Aims

We aimed to evaluate the ability of trastuzumab in enhancing the cytotoxicity of PBMCs against canine tumor cells.

Methods

We used canine tumor cell lines isolated from metastatic mammary gland tumors (CHMm and CIPm) and thyroid adenocarcinoma (CTAC). The binding of trastuzumab to the cells was confirmed using flow cytometry analysis. Peripheral blood mononuclear cells obtained from healthy beagles and lymphokine-activated killer (LAK) cells, generated by interleukin-2 (IL-2) stimulation of PBMCs, were used as effector cells. Standard lactate dehydrogenase (LDH) release assay was used to measure the cytotoxicity of the LAK cells against tumor cell lines in the presence of trastuzumab.

Results

Trastuzumab enhanced the cytotoxicity of PBMCs against CHMm. Moreover, LAK cells killed CHMm synergistically in the presence of trastuzumab. However, the presence of trastuzumab did not produce such a synergistic effect when LAK cells acted against CIPm and CTAC.

Conclusion

We confirmed the ability of trastuzumab to induce an ADCC response in canine PBMCs and determined its synergistic effect with LAK cells. Although the in vitro system in the present study did not show the induction of trastuzumab-mediated ADCC response against all canine tumor cell lines, the results of this study indicate the potential antitumor activity of trastuzumab in canines.

SUBMITTER: Oyamada T

PROVIDER: S-EPMC7871734 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Cytotoxicity effect of trastuzumab on canine peripheral blood mononuclear cells.

Oyamada T T Okano S S

Iranian journal of veterinary research 20200101 4

<h4>Background</h4>Trastuzumab is an antibody drug used to treat human epidermal growth factor receptor 2 (HER2) overexpressing human metastatic breast cancer. Antibody-dependent cellular cytotoxicity (ADCC) is considered to be the major mechanism of cytotoxicity of the drug. However, its ability to induce an ADCC response in canine peripheral blood mononuclear cells (PBMCs) is not well established.<h4>Aims</h4>We aimed to evaluate the ability of trastuzumab in enhancing the cytotoxicity of PBMC ...[more]

PMID: 33584838

Similar Datasets

Project description:Abstract Introduction: Despite state-of-the-art surgical methods combined with radio- and chemotherapy the clinical prognosis of brain cancer patients remains poor. The brain is an immune privileged site and brain cancers are known to form a highly immunosuppressive microenvironment, suggesting that patients could benefit from immunomodulatory drugs. Multiple therapies targeting the immune system are currently under development, including application of a potent activator of immune responses, interleukin (IL)-12. This cytokine stimulates the cytotoxic function of lymphocytes and can trigger rejection of gliomas in preclinical studies. Even though IL-12 has a relatively short half-life, systemic IL-12 can quickly lead to severe adverse reactions. Local application or targeting is therefore mandatory. Local IL-12 therapy is currently tested in early stage clinical trials of intratumoral genetherapy using viral vectors (ClinicalTrials.gov identifiers NCT02026271 and NCT02062827). An alternative approach is the use of convection-enhanced delivery (CED). This way IL-12 could be dosed precisely to the tumour focus with minimal systemic leakage. Furthermore, to increase the local half-life of IL-12, we have fused it with the Fc part of human IgG4 (IL-12Fc). Using CED, we are aiming to validate efficacy and safety of local IL-12Fc monotherapy in a preclinical setting with canine brain tumours. Materials and Methods: Canine peripheral blood mononuclear cells (PBMCs) were stimulated with increasing concentrations of human IL-12Fc or with recombinant human or canine IL-12 in presence or absence of concanavalin A. Interferon-γ (IFN-γ) production was measured by ELISA. Phosphorylation of STAT4 was measured by western blot. Porcine PBMCs were stimulated in the presence of recombinant porcine IL-2 and IL-18. IFN-γ production was measured by ELISA. Results: Human IL-12Fc is able to stimulate canine and porcine PBMCs to secrete IFN-γ in a dose-dependent manner. Furthermore, stimulation with hIL-12Fc leads to phosphorylation of STAT4. Conclusions: Human IL-12 immunotherapeutics are able to stimulate canine PBMCs and thus therapy of spontaneous brain tumours in dogs can be used for preclinical testing. To estimate leakage and brain perfusion in the dog and ultimately in humans, we will next test CED of hIL-12Fc in large white landrace pigs. We are aiming to initiate a trial to assess the efficacy of local delivery of human IL-12Fc as therapy for canine brain tumours.

Dataset Information

Cytotoxicity effect of trastuzumab on canine peripheral blood mononuclear cells.

Background

Aims

Methods

Results

Conclusion

Publications

Cytotoxicity effect of trastuzumab on canine peripheral blood mononuclear cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets