CN-121227629-B - Regulated γδ T cells and methods of their therapeutic intervention and use

Abstract

The invention provides a gamma delta T cell subjected to regulation treatment and an intervention regulation method and application thereof, relating to the field of biotechnology, the methods of interventional modulation include the application of inhibition to the JAK1-STAT5 signaling pathway of γδ T cells to reduce IL-18 induced secretion of IL-5 and/or IL-13. The intervening regulation method can inhibit the JAK1-STAT5 channel of gamma delta T cells, and can remarkably reduce IL-18 induced secretion of IL-5 and IL-13 with tumor migration invasion promoting effect. This eliminates negative regulation of γδ T cells, thereby enhancing their overall antitumor capacity and improving safety.

Inventors

• XIANG ZHENG
• CHEN YAN
• YIN ZHINAN
• XU YAN
• CHEN MEIJI

Assignees

• 暨南大学

Dates

Publication Date

20260508

Application Date

20251201

Claims (3)

1. 1. A method of interventional modulation of γδ T cells for non-therapeutic purposes comprising: co-culturing peripheral blood mononuclear cells containing gamma delta T cells with zoledronic acid, IL-2 and IL-18 in vitro to obtain gamma delta T cells amplified in vitro; Gamma delta T cells are used in contact with JAK inhibitors Abrocitinib to reduce IL-18 induced secretion of IL-5 and/or IL-13.
2. 2. The method of interventional modulation of γδ T-cells of non-therapeutic interest according to claim 1, wherein the contacting with γδ T-cells with JAK inhibitor Abrocitinib comprises: Plating γδ T cells; Adding the JAK inhibitor Abrocitinib to the gamma delta T cells plated, and And co-culturing the gamma delta T cells and the JAK inhibitor Abrocitinib for a preset time, wherein the preset time is the time capable of inhibiting the JAK1-STAT5 signal pathway.
3. 3. Use of the method for the interventional modulation of γδ T-cells of non-therapeutic interest according to claim 1 or 2 for the preparation of an anti-tumor cell therapy product, wherein the tumor is lung cancer.

Description

Regulated γδ T cells and methods of their therapeutic intervention and use Technical Field The invention relates to the technical field of biology, in particular to a gamma delta T cell subjected to regulation treatment and an intervention regulation method and application thereof. Background In recent years, tumor immunotherapy techniques have been developed in breakthrough, and Adoptive cell therapy (Adoptive CELL THERAPY, ACT) has become a very promising cancer treatment strategy by expanding immune cells in vitro and then reinfused into patients. In ACT therapy, γδ T cells are of great interest for their broad spectrum antitumor potential independent of the Major Histocompatibility Complex (MHC), and show good safety in clinic. To enhance its killing activity, the prior art often uses cytokines for stimulus amplification in vitro. Among them, interleukin-18 (IL-18) is widely used to promote secretion of interferon-gamma (IFN- γ) by γδ T cells to enhance type I immune response, exerting antitumor effect. Despite the promise of γδ T cell therapies, the overall anti-tumor effect in patients with advanced tumors remains limited. How to effectively improve the therapeutic efficacy is a key technical problem to be solved in the field. Currently, the understanding of γδ T cell function regulation is not complete, and particularly when using strong stimulatory factors such as IL-18, its complex biological effects are not fully elucidated. The prior art mostly focuses on the positive effect of IL-18 on IFN-gamma promotion, but ignores its potential bi-directionality. The mechanism by which IL-18 regulates type II cytokines (e.g., IL-5 and IL-13) in γδ T cells is not yet known. In view of the potential to promote tumor progression reported by IL-5 and IL-13, it is highly desirable to elucidate whether the prior art also inadvertently activates negative regulatory pathways that promote tumor immune escape when utilizing IL-18. This lack of knowledge of the gamma delta T cell function regulatory mechanisms results in blindness in existing stimulation protocols, which prevents the improvement of therapeutic efficacy. In view of this, the present invention has been made. Disclosure of Invention The invention aims to provide a gamma delta T cell subjected to regulation treatment, an intervening regulation method and application thereof, wherein the method inhibits a JAK1-STAT5 channel of the gamma delta T cell, can reduce IL-18-induced secretion of IL-5/IL-13 with tumor promotion activity, further eliminates negative regulation, enhances the overall anti-tumor efficacy of the cell and improves the safety. In order to achieve the above object of the present invention, the following technical solutions are specifically adopted: In a first aspect, the invention provides a method for the interventional modulation of γδ T cells comprising: inhibition of the JAK1-STAT5 signaling pathway of γδ T cells is applied to reduce IL-18 induced secretion of IL-5 and/or IL-13. In alternative embodiments, the inhibition of the JAK1-STAT5 signaling pathway of γδ T cells employs at least one of the following methods: A. contacting a JAK inhibitor with γδ T cells; B. a gene editing technique; C. RNA interference technology. In an alternative embodiment, the contacting with a JAK inhibitor using γδ T cells comprises: Plating γδ T cells; the JAK inhibitor is added to the gamma delta T cells plated, and the gamma delta T cells are co-cultured with the inhibitor for a preset time period, the preset time period being a time period sufficient to inhibit the JAK1-STAT5 signaling pathway. In alternative embodiments, the JAK inhibitor comprises a JAK1 inhibitor. In an alternative embodiment, the method further comprises, prior to plating the γδ T cells: And (3) co-culturing peripheral blood mononuclear cells containing gamma delta T cells with zoledronic acid, IL-2 and IL-18 in vitro to obtain the gamma delta T cells cultured in vitro. In a second aspect, the invention provides a regulatory treated γδ T cell obtained by an interventional regulatory method for γδ T cells according to any one of the preceding embodiments. In a third aspect, the invention provides a pharmaceutical composition comprising a modulated γδ T cell as described in the previous embodiments, and a pharmaceutically acceptable carrier. In a fourth aspect, the invention provides a kit comprising a JAK inhibitor of the JAK1-STAT5 signaling pathway, at least one γδ T cell stimulators, and instructions; Wherein the stimulating factor is at least one selected from IL-18, IL-2 and zoledronic acid; the instructions indicate performing an interventional modulation method of γδ T cells according to any one of the preceding embodiments, or applying the JAK inhibitor and γδ T cell stimulating factor to γδ T cells. In a fifth aspect, the invention provides the use of a regulatory treated γδ T cell according to the previous embodiments for the preparation of an anti-tumor cell therapy produ