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Immunomodulatory effects of Diospyros peregrina fruit preparation (DFP) in non-small cell lung cancer (NSCLC) by utilizing dendritic cell-mediated antigen presentation and T helper (TH) cell differentiation

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Abstract

Diospyros peregrina is a dioecious plant which is native to India. It belongs to the family of Ebenaceae and is extensively used to treat various ailments, such as leucorrhoea and other uterine-related problems. Though few studies have been on D. peregrina for their anti-tumour response, little is known. Therefore, this intrigued us to understand its immunomodulator capabilities on various types of cancer extensively. Our primary focus is on NSCLC (Non-Small Cell Lung Cancer), which is ranked as the second largest form of cancer in the world, and the treatments demand non-invasive agents to target NSCLC effectively. In an objective to generate an efficient Lung Cancer Associated Antigen (LCA) specific anti-tumour immune response, LCA was presented using dendritic cells (DCs) in the presence of D. peregrina fruit preparation (DFP). Moreover, we also investigated DFP’s role in the differentiation of T-helper (TH) cells. Therefore, this study aimed at better LCA presentation mediated by DFP by activating the LCA pulsed DCs and T helper cell differentiation for better immune response. DCs were pulsed with LCA for tumour antigen presentation in vitro, with and without DFP. Differentially pulsed DCs were irradiated to co-culture with autologous and allogeneic lymphocytes. Extracellular supernatants were collected for the estimation of cytokine levels by ELISA. LDH release assay was performed to test Cytotoxic T lymphocytes (CTLs) mediated lung tumour cell cytotoxicity. Thus, DFP may be a potential vaccine to generate anti-LCA immune responses to restrict NSCLC.

Graphical Abstract

A graphical representation of the cascade of events pertaining to the augmentation of the process of cancer antigen presentation, TH cell differentiation and subsequent tumour cell cytotoxicity in presence or absence of DFP

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Data availability

Data are available on request from the authors.

Abbreviations

NSCLC:

Non-small cell lung cancer

LCA:

Lung cancer associated antigens

DFP:

Diospyros peregrina Fruit preparation

DC:

Dendritic cell

TH :

T helper

ELISA:

Enzyme-linked immunosorbent assay

LDH:

Lactate dehydrogenase

CTL:

Cytotoxic T lymphocytes

Treg:

Regulatory T cells

TLR:

Toll-like receptors

IL:

Interleukin

PBMC:

Peripheral blood mononuclear cells

rhGM-CSF:

Recombinant human granulocyte–macrophage colony-stimulating factor

PBS:

Phosphate buffer saline

TMB:

3,3′,5,5′-Tetramethylbenzidine

HRP:

Horseradish peroxidase

RT-qPCR:

Real-time quantitative polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

BSA:

Bovine serum albumin

PBST:

Phosphate buffer saline tween-20

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Acknowledgements

We thank DST-FIST Facility of Department of Biotechnology at SRM Institute of Science and Technology, Kattankulathur, India for allowing us to perform flow cytometry on BD FACS Melody™ cell sorter.

Funding

This research was supported by Science and Engineering Research Board (SERB), Department of Science and Technology (DST), Govt. of India (sanction order No. ECR/2016/000965).

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Authors and Affiliations

  1. Department of Biotechnology, SRM Institute of Science and Technology, Kattankulathur, Chennai, Tamil Nadu, 603203, India

    Nawaneetan Sriraman, Ankita Sarkar, Sohom Naskar, Nitika Mahajan, Oishi Mukherjee, R. Pradeep & Koustav Sarkar

  2. Department of Clinical Pharmacology, SRM Medical College Hospital and Research Centre, Kattankulathur, Tamil Nadu, 603203, India

    Melvin George

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Contributions

NS and AS: investigation, data curation, writing–original draft. SN and NM: investigation, data curation, writing–original draft. OM and PR: standardization of techniques. MG: resource person. KS: conceptualization, writing–review & editing, project administration, supervision, funding acquisition.

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Correspondence to Koustav Sarkar.

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The authors have no relevant financial or non-financial interests to disclose.

Ethical approval

For molecular studies involved in the current research project, blood samples of normal subjects and NSCLC patients were collected and handled under the protocol approved by the institutional ethics committee at SRM Medical College Hospital, Kattankulathur, Chennai, Tamil Nadu, India (Ethics Clearance Number 2909/IEC/2021).

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Signed informed consent was collected from each NSCLC patient and normal individuals who took part in the current study.

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Sriraman, N., Sarkar, A., Naskar, S. et al. Immunomodulatory effects of Diospyros peregrina fruit preparation (DFP) in non-small cell lung cancer (NSCLC) by utilizing dendritic cell-mediated antigen presentation and T helper (TH) cell differentiation. Med Oncol 41, 107 (2024). https://doi.org/10.1007/s12032-024-02331-7

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