Advancing Oncology Research with Immune Response Studies: The Role of xMAP-Based Bio-Plex Multiplex Immunoassays

In the rapidly evolving field of cancer research, understanding the complex interplay between tumors and the immune system is essential for developing effective therapies. Bio-Plex Multiplex Immunoassays, for research use only and powered by Luminex xMAP technology, provide a cutting-edge tool for simultaneously measuring multiple immune biomarkers. This technology enables scientists to profile the immune landscape of cancer more comprehensively, aiding in the prediction of immunotherapy outcomes, surveillance of the tumor microenvironment, and identification of biomarkers of adverse immune reactions. These advancements in research tools are paving the way for development of personalized cancer treatments in the future.

Introduction

The human immune system is a complex network of cells, tissues, and organs that defend the body against pathogens. Beyond infection control, the immune system plays a pivotal role in the body’s defense against cancer, influencing both tumor progression and therapeutic outcomes. Cancer immunology, the study of immune system interactions with tumors, has been instrumental in revealing crucial insights that give rise to treatment options such as cell therapy, immune checkpoint inhibitors, monoclonal antibodies, and cancer vaccines (Waldman et al. 2020). A deeper understanding of these immune responses through oncology research can potentially help scientists predict therapeutic outcomes and improve future patient care.

Cancer immunotherapy, a type of cancer treatment that stimulates the patient’s immune system to fight cancer cells, has revolutionized oncology, yet its success is hindered by the complexity of the immune system and the tumor microenvironment (TME). One of the main challenges in this area of research is the need for precise, high-throughput assays that can measure multiple immune biomarkers simultaneously. The research use only (RUO) Bio-Plex Multiplex Immunoassays, powered by Luminex xMAP technology, are well-suited to measure multiple cytokines, chemokines, and other immune-related proteins, allowing scientists to obtain a comprehensive profile of the immune landscape of cancer.

The Immune Response in Oncology

Bio-Plex Multiplex Immunoassays enable high-throughput analysis of multiple biomarkers simultaneously.

Cancer cells evade immune detection through multiple mechanisms, including downregulating antigen presentation and secreting immunosuppressive molecules in the tumor microenvironment (Hanahan and Weinberg 2011, Spranger and Gajewski 2018). Therapies using immune checkpoint inhibitors, such as anti-PD-1 and anti-CTLA-4, have been successful in reactivating the immune system against tumors (Pardoll 2012). However, the effectiveness of these therapies varies, with some patients experiencing significant tumor regression and others seeing little benefit (Costantini et al. 2021, Ribas and Wolchok 2018).

This variability in treatment response highlights the importance of studying relevant immune biomarkers simultaneously. Measuring multiple immune biomarkers is critical for understanding those biomarkers’ interactions with cancer and for understanding patient responses to therapies. Bio-Plex Multiplex Immunoasssays are particularly useful research tools for this because they offer high sensitivity, specificity, and throughput coupled with a high level of multiplex capability.

Bio-Plex Multiplex Immunoassays in Immune Response Studies

Traditional ELISA assays measure one target at a time, require larger sample volumes, and are time consuming. In contrast, Bio-Plex Multiplex Immunoassays allow simultaneous measurement of multiple biomarkers in a single well with a small sample volume. This approach allows researchers to save precious samples while gaining a comprehensive picture of immune biomarker responses to cancer and to therapies. For instance, the Bio-Plex Pro Human Cytokine 27-Plex Assay quantifies 27 cytokines, including upregulating inflammatory cytokine IL-6 and downregulating multifunctional cytokines IP-10, eotaxin, IL-4, and IL-7, all of which are critical mediators cited in literature for inflammation and immune regulation in cancer (Torkildsen et al. 2024). Cytokine, chemokine, and growth factor biomarkers serve as indicators for scientists studying disease development and progression, potential metastasis, and response to treatment (Kartikasari et al. 2021).

Role of Bio-Plex Multiplex Immunoassays in Advancing Oncology

Bio-Plex Multiplex Immunoassays have the potential to help advance oncology studies in three important ways:

Predicting Response to Immunotherapy

Identifying which types of patients are most likely to benefit from immunotherapy remains a key challenge for scientists. While immune checkpoint inhibitors and CAR T-cell therapy have shown remarkable success in some patients, other patients fail to respond (Kim and Cho 2022). Cytokine profiling using Bio-Plex Multiplex Immunoassays, intended for research use only, can assist researchers in categorizing patients by immune status and can support the prediction of therapeutic outcomes in translational and preclinical studies. For instance, elevated levels of proinflammatory cytokines such as IL-6 and TNF-α have been linked by scientists to poor prognosis, potentially signaling resistance to immunotherapy. Conversely, increased levels of IFN-γ and IL-12 are linked by researchers in the literature to a stronger anti-tumor immune response, suggesting a better response to immune checkpoint inhibitors (Spranger and Gajewski 2018). In these instances, multiplexing allows researchers to gather multiple biomarker data from a single well to correlate immune status with therapy response and better predict treatment outcomes, guiding future clinical strategies (Michalaki et al. 2004).

One of the Bio-Plex Assay Kits, the Bio-Plex Pro Human Immunotherapy 20-Plex Panel, includes 20 cytokines, growth factors, and macrophage-associated proteins that are highly relevant in the study of various immunotherapy responses. This multiplex panel can help researchers understand how cytokines, chemokines, and growth factors are expressed at different stages of cancer progression — before, during, and after treatment. The multiplexed dataset can provide critical insights into disease mechanisms, treatment responses, and potential therapeutic targets in a shorter time as opposed to assaying one target at a time using traditional ELISA. These biomarkers and their significance in immunotherapy shown in research studies published in the literature are listed in the table below.  

Cytokines, growth factors, and macrophage-associated proteins included in the Bio-Plex Immunotherapy 20-Plex Panel and their significance in immunotherapy.

Monitoring the Tumor Microenvironment

The tumor microenvironment (TME) refers to the environment surrounding a tumor, including immune cells, blood vessels, signaling molecules, and the extracellular matrix. Researchers have demonstrated that the TME can significantly impact the immune response to cancer. Tumors often create an immunosuppressive environment that promotes their growth and inhibits immune cell activity by releasing certain cytokines and chemokines (Tormoen et al. 2018).

Cytokines such as IL-10 and TGF-β have been cited in publications as inhibitors of the immune response. By analyzing cytokine levels before treatment, scientists can determine if a tumor is likely to avoid detection by the immune system. The ability of researchers to study multiple specific cytokines can help them identify which ones are markers of poor treatment outcomes or more aggressive forms of cancer.

Bio-Plex Multiplex Assays allow the concurrent measurement of multiple immune biomarkers in the TME — in addition to profiling cytokines, Bio-Plex Assays enable assessment of chemokines. The Bio-Plex Pro Human Chemokine 40-Plex Panel can aid scientists in measuring chemokines, such as CCL2, CCL5, and CXCL10, that recruit immune cells to the tumor site. Monitoring and analyzing these chemokines in tumor tissue enables scientists to assess the extent of immune infiltration to determine whether the tumor is actively recruiting or repelling immune cells. 

Investigating Biomarkers of Immune-Related Adverse Events

While immunotherapy has shown great promise in treating cancer, it can also cause immune-related adverse events (irAEs) such as autoimmune disorders and cytokine storm syndrome. Understanding the immune biomarkers associated with irAEs can support scientists in improving the safety of immunotherapy (Wojtukiewicz et al. 2021). For example, cytokine dysregulation — particularly elevated IL-2Rα, IL-6, IL-10, IFN-γ, CXCL2, and CCL17 — has been linked by researchers to inflammatory conditions in patients receiving immune checkpoint inhibitors and CAR-T cell therapy (Gonugunta et al. 2021, Fajgenbaum and June 2020). Many cytokines, chemokines, and growth factors found in the Bio-Plex Pro Human Cytokine Screening Panel, 48-Plex have been shown by researchers to be critically important in the evaluation of inflammation biomarker regulation in cancer patient research (Rocha et el. 2023).

The following biomarkers are included in the Bio-Plex Pro Human Cytokine Screening Panel, 48-Plex:

Conclusion

The study of immune responses in oncology is crucial for advancing cancer research that enables deeper understanding of cancer biology and supports the improvement of therapeutic outcomes. Bio-Plex Multiplex Immunoassays, for research use only, offer a powerful solution for measuring immune biomarkers simultaneously, providing researchers with a detailed view of the immune landscape in cancer. Whether predicting response to immunotherapy, monitoring the tumor microenvironment, or investigating biomarkers of adverse events, Bio-Plex Assays can play a critical role for scientists in oncology research.

By leveraging the multiplexing capabilities of Bio-Plex Multiplex Immunoassays, translational and preclinical scientists can guide the development and advancement of more personalized and effective treatment strategies for patients in the future.

Research Use Only.  Not for use in diagnostic procedures.

References

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Kartikasari AER et al (2021). Tumor-induced inflammatory cytokines and the emerging diagnostic devices for cancer detection and prognosis. Front Oncol 11, 692142.

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Michalaki V et al. (2004). Serum levels of IL-6 and TNF-alpha correlate with clinicopathological features and patient survival in patients with prostate cancer. Br J Cancer 90, 2312–2316. 

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Rocha Martins P et al. (2023). Linking tumor immune infiltrate and systemic immune mediators to treatment response and prognosis in advanced cervical cancer. Sci Rep 13, 22634.

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Torkildsen CF et al. (2024). New immune phenotypes for treatment response in high-grade serous ovarian carcinoma patients. Front Immunol 15, 1394497.

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