T-cell activation is an essential mechanism within the immune system and for the future of adoptive cell therapy. Activation triggers an immune response through exponential proliferation, growth, and differentiation.
Exposure to their specific antigen activates T cells, which bind to the antigen and generate a specialized and unique immune response message throughout the body.
The process of inducing T cell activation in a laboratory setting is a crucial aspect of therapeutic cell manufacturing. Successful and scalable in-lab T cell activation methods are essential to advancing cell therapy research and innovation. Modern therapeutic cell manufacturing products often struggle to achieve scalability due, in part, to the absence of reliable methods for T cell activation.
The success of adoptive cell therapies depends on the infusion of large amounts of highly specified T cells into patients. However, meeting this demand for cells presents a prominent challenge that limits the widespread use of this therapy. To overcome this obstacle, researchers extract small numbers of specific and ideal cells from localized tissues and then expand them in a laboratory setting.
By triggering apoptosis and anergy, the expansion process, though necessary, can also be damaging to the viability of the expanded cell population for downstream use. Considering this damage, T cell activation methods aim to gently stimulate T cells to activate and replicate while minimizing activation-induced death. Control over cell activation is instrumental to immunotherapy research and our ability to understand adaptive immunity, exhaustion, stem cell creation, and the overall immune response mechanism.
The T-cell receptor (TCR) stimulation triggers T cell activation by binding its unique antigen complement at the major histocompatibility complex (MHC). Co-stimulatory molecules that induce activation are also necessary at the MHC to protect cells from becoming anergic.
An antigen-presenting cell (APC) forms the MHC on its surface, triggering a wave of transduction cascades and cytokine release that fuels an inflammatory response targeted at the specific antigen that sparked the activation. T cells rapidly expand and release exponentially increasing amounts of cytokines and differentiation factors, equipped with the cellular information necessary to quell the infection. An explosion of rapid cell division and differentiation urges the immune response forward to eliminate the potential infection.
Co-stimulatory molecules CD3 and CD28 play a significant role in T cell activation and are crucial factors to consider and balance in ex vivo T cell activation methods. During T-cell activation, CD3 triggers a proliferation process that leads to T-cell differentiation. CD3 transmits a message into the nucleus for transcription, which is necessary for the process of proliferation to occur. In addition, CD28 binds to TCR to stimulate the activation cascade and the production of cytokines.
In the lab, finding a balance between CD3 and CD28 and the activation threshold of T cell starting material is an ongoing research challenge. Many solutions involve stabilizing one of the antibodies to a solid structure, such as a bead or plate, to reduce interaction between molecules. Different T cell types have varying requirements for stimulation and activation.
A typical solution to ex vivo activation needs is magnetic beads coated with antibodies to stimulate the activation of T cells and trigger proliferation. These beads are mixed within the T cell solution and suspended to induce activation.
Magnetic beads for activation are only micrometers in diameter and similar in size to antigen-presenting cells. The beads are covered in covalent bonds attached to anti-CD3 and anti-CD28 antibodies. Once introduced to the solution, the antibodies provide co-stimulation of TCR, prompting activation.
Magnetic beads yield many terminally differentiated CD8 cells, in addition to CD8 cells which become memory T cells shortly after infusion. This advantage is due to the importance of CD8 cells in the clearance of tumor cells. Because of this, magnetic beads are known to increase T cell immunity in patients with critically exhausted T cells.
In some cases, the robust effector cell population does not affect patients post-infusion. Because of this decreased effectiveness, magnetic beads are often not the best option for quality performance in adoptive cell therapies.
Similar to beads, plate-bound activation methods expose T cells to co-stimulatory factors and incubate them for activation in vitro. By coating the wells of a plate with CD3 and CD28, free antibodies provide activation signals to TCR.
Balancing proper co-stimulation exposure is integral to successful activation, and both bead and plate-based methods provide structure for the antibodies to stay secure. This method modulates the interaction between molecules and limits the risk of reaching the activation threshold.
Binding CD3 to the wells of a plate controls the interaction time between T cells, CD28, and CD3. This protects T cells from overstimulation, which induces apoptosis and cell population failures.
Plate-bound activation is better suited for small-scale experiments and studies that aim to explore the mechanisms of activation, rather than for large-scale therapeutic cellular manufacturing. The manual and non-scalable nature of plate-bound methods makes them unsuitable for meeting the high demand for T cells required for therapeutic purposes.
Compared to bead solutions, soluble antibody activation methods can yield d less differentiated CD8 and CD4 cells. Young, undifferentiated T cells make for the optimal starting material for ex vivo expansion and activation because of their high survival rate and adaptability. Robust throughout the manufacturing cycle, naive T cells provide an effective adoptive cell therapy and infusion success.
Surface-secured activation methods like plate-bound or bead-bound antibodies provide greater expansion for CD4 cells. Soluble antibody activation methods would not be the most efficient for CD4-based studies or applications.
Akadeum’s engineered naturally-buoyant microbubbles for targeting molecules in a solution are widely applicable to cell therapy and highly innovative. Fully customizable and scalable, our microbubbles can be modified to target antibodies, proteins, DNA, and other immune molecules. Providing a fixed anchor point for the antibodies by attaching them to microbubbles allows for efficient T cell activation with minimal exhaustion. Daughter cells fall to the bottom of the well, away from the activating conditions at the top of the culture. This reduction in interaction provides significant advantages by protecting the cells from overstimulation and activation-induced cell death.
Akadeum’s innovative microbubble technology has taken on a new application—T cell activation. Thanks to the gentle nature of our microbubbles, fragile T cell populations can be activated and expanded in a controlled manner and are ready for any downstream processes.
By utilizing the surface-binding properties of antibody-coated bubbles, Akadeum Life Sciences has found a proper balance between co-stimulatory molecule exposure and activation threshold time, resulting in a change to the industry. The result is a highly effective activation and expansion method that protects T cells from stimulation damage. Contact Akadeum today to discover how microbubbles can revolutionize ex vivo activation methods.