Helper innate lymphoid cells as cell therapy for cancer

Although the first cancer immunotherapy was given in the clinic more than a century ago, this line of treatment has remained more of a distant goal than a practical therapy due to limited understanding of the tumour microenvironment and the mechanisms at play within it, which led to failures of numerous clinical trials. However, in the last two decades, the immune checkpoint inhibitors (ICIs) and chimeric antigen receptor-T cell therapies have revolutionized the treatment of cancer and provided proof-of-concept that immunotherapies are a viable option. So far, immunotherapies have majoritarily focused on utilizing T cells; however, T cells are not autonomous but rather function as part of, and therefore are influenced by, a vast cast of other immune cells, including innate lymphoid cells (ILCs). Here, we summarize the role of ILCs, especially helper ILCs, in tumour development, progression and metastasis, as well as their potential to be used as immunotherapy for cancer. By reviewing the studies that used helper ILCs as adoptive cell therapy (ACT), we highlight the rationale behind considering these cells as novel ACT for cancer as well as identify open questions and areas for future research

The IL-25/ILC2 axis promotes lung cancer with a concomitant accumulation of immune-suppressive cells in tumors in humans and mice

Background: Group 2 innate lymphoid cells (ILC2) can be activated by interleukin (IL)-33 or IL-25. IL-25-activated ILC2 cells help protect the host against helminth infection while exacerbating allergic-like inflammation and tissue damage in the lung. In the context of cancer, IL-33-activated ILC2 cells were found to bear anti-tumoral functions in lung cancer while IL-25-activated ILC2 cells promoted tumorigenesis in colorectal cancer. The role of IL-25-activated ILC2 cells in lung cancer remains to be addressed. Methods: We examined the overall survival of human non-small cell lung cancer (NSCLC) patients according to IL25 expression as well as the distribution of ILC2 cells and regulatory T cells (Tregs) in various NSCLC patient tissues and peripheral blood (PB) of healthy donors (HDs). We analyzed the effect of adoptive transfer of IL-25-activated ILC2 cells on tumor growth, metastasis and survival in a heterotopic murine model of lung cancer. Results: We report that human NSCLC patients with high IL-25 expression have reduced overall survival. Moreover, NSCLC patients bear increased frequencies of ILC2s compared to HDs. Frequencies of Tregs were also increased in NSCLC patients, concomitantly with ILC2s. In mice bearing heterotopic lung cancer, adoptive transfer of IL-25-activated ILC2s led to increased tumor growth, increased metastasis and reduced survival. The frequencies of monocytic myeloid-derived suppressor cells (M-MDSCs) were found to be increased in the tumors of mice that received ILC2s as compared to controls. Conclusion: Overall, our results indicate that the IL-25/ILC2 axis promotes lung cancer potentially by recruiting immune-suppressive cells to the tumors both in humans and in mice, and that it may therefore represent a suitable novel target for NSCLC immunotherapeutic development

DEVELOPMENT and CHARACTERIZATION OF MONOCLONAL ANTIBODY AGAINST A DISEASE MARKER PROTEIN

Sunulan bu tez çalışmasının amacı model olarak seçilen bir hastalık belirteci proteine karşı yüksek afiniteli monoklonal antikor geliştirilmesi, antikorun geniş ölçekte üretimi, saflaştırılması ve karakterizasyonudur. Bu amaçla kistik fibrozisde hastalık belirteci olan PAP (Pankreatit ile İlişkili Protein)’a karşı monoklonal antikor geliştirmek amacıyla 6/8 haftalık toplam 10 fare intraperitonal enjeksiyon ile immunize edilmiştir. ELISA testi ile bağışıklık yanıtları belirlenen farelerin dalak hücreleri ile myeloma hücrelerinin hibridizasyonu yapılmış, elde edilmiş antikorların karakterizasyon çalışmaları yapılmıştır. Çalışmada kullanılan 10 fareden 3’ünde diğerlerine göre daha yüksek immun yanıt oluştuğu belirlenmiş, bunlardan biri ile yapılan füzyon çalışması sonucu antikor aktivitesine sahip 32 hibrit klon elde edilmiştir. Tekrarlanan ELISA kontrol çalışmalarında antikor aktivitesine sahip hücrelerin %25’i PAP antijenine karşı spesifik antikor yanıtı vermiş ve bunlarında %50’si bu yanıtı sürdürmüştür. Sonuç olarak bu hücrelerden 4 hybridoma hücresinin daha sonra tanı kiti geliştirmek amacıyla kullanılabilecek yüksek duyarlılıkta monoklonal antikor ürettiği belirlenmiştir. Elde edilen antikorlar özgüllük, afinite, altizotip olarak değerlendirilmiş 4 antikorun her birinin özgül olduğu iki antikorun afinitesinin daha yüksek olduğu görülmüş ve elde edilen 4 antikor da saflaştırılmıştır. Bu tez çalışması gelecekte hayvan hastalıklarının tanısı veya daha başka sebeplerle farklı proteinlerin tespitine yönelik antikorların üretim çalışmaları için önemli bir kaynak oluşturacaktır.In this study, the production of monoclonal antibody against a disease marker selected as a targeted model, the characterization, large-scale production and purification of the produced antibodies are intended to work. For this purpose, Pancreatitis - associated protein which is a Cystic fibrosis marker used as an antigen were intraperitoneally injected to 6-8 weeks old 10 Balb/C mice at specific doses and time intervals. The antibody titers of mice were determined by indirect ELISA, and the best immune response in mice was selected for the fusion. The fusion was carried out using the mouse spleen cell and the F0 myeloma cells in the presence of polyethylene glycol (PEG). A higher immunoreactivity was detected in 3 of 10 mice used in the study. Fusion study with one of these resulted 32 hybrid clones with antibody activity. In repeated ELISA control studies, 25% of the cells with antibody activity gave a specific antibody response to the PAP antigen and 50% of them maintained this response. As a result, it has been determined that four hybridoma cells from these cells produce monoclonal antibody with high sensitivity which can be used later to develop a diagnostic kit. The obtained monoclonal antibodies were found to have a high specificity, the immunoglobulin type and the light chain type of the obtained monoclonal antibodies were determined as IgG1 and ? (kappa), respectively. Monoclonal antibodies were characterized in terms of their affinities and 2 of the fourth antibodies were found to have a high affinity. This thesis study will be an important source for future studies of the production and characterization of antibodies for the detection of animal diseases or detection of proteins for different purposes

Monoclonal antibody development for quantitative analysis of pancreatitis-associated protein

European Biotechnology Congress -- MAY 25-27, 2017 -- Dubrovnik, CROATIA[No Abstract Available

DataSheet_2_The IL-25/ILC2 axis promotes lung cancer with a concomitant accumulation of immune-suppressive cells in tumors in humans and mice.pdf

BackgroundGroup 2 innate lymphoid cells (ILC2) can be activated by interleukin (IL)-33 or IL-25. IL-25-activated ILC2 cells help protect the host against helminth infection while exacerbating allergic-like inflammation and tissue damage in the lung. In the context of cancer, IL-33-activated ILC2 cells were found to bear anti-tumoral functions in lung cancer while IL-25-activated ILC2 cells promoted tumorigenesis in colorectal cancer. The role of IL-25-activated ILC2 cells in lung cancer remains to be addressed.MethodsWe examined the overall survival of human non-small cell lung cancer (NSCLC) patients according to IL25 expression as well as the distribution of ILC2 cells and regulatory T cells (Tregs) in various NSCLC patient tissues and peripheral blood (PB) of healthy donors (HDs). We analyzed the effect of adoptive transfer of IL-25-activated ILC2 cells on tumor growth, metastasis and survival in a heterotopic murine model of lung cancer.ResultsWe report that human NSCLC patients with high IL-25 expression have reduced overall survival. Moreover, NSCLC patients bear increased frequencies of ILC2s compared to HDs. Frequencies of Tregs were also increased in NSCLC patients, concomitantly with ILC2s. In mice bearing heterotopic lung cancer, adoptive transfer of IL-25-activated ILC2s led to increased tumor growth, increased metastasis and reduced survival. The frequencies of monocytic myeloid-derived suppressor cells (M-MDSCs) were found to be increased in the tumors of mice that received ILC2s as compared to controls.ConclusionOverall, our results indicate that the IL-25/ILC2 axis promotes lung cancer potentially by recruiting immune-suppressive cells to the tumors both in humans and in mice, and that it may therefore represent a suitable novel target for NSCLC immunotherapeutic development.</p

DataSheet_1_The IL-25/ILC2 axis promotes lung cancer with a concomitant accumulation of immune-suppressive cells in tumors in humans and mice.pdf

BackgroundGroup 2 innate lymphoid cells (ILC2) can be activated by interleukin (IL)-33 or IL-25. IL-25-activated ILC2 cells help protect the host against helminth infection while exacerbating allergic-like inflammation and tissue damage in the lung. In the context of cancer, IL-33-activated ILC2 cells were found to bear anti-tumoral functions in lung cancer while IL-25-activated ILC2 cells promoted tumorigenesis in colorectal cancer. The role of IL-25-activated ILC2 cells in lung cancer remains to be addressed.MethodsWe examined the overall survival of human non-small cell lung cancer (NSCLC) patients according to IL25 expression as well as the distribution of ILC2 cells and regulatory T cells (Tregs) in various NSCLC patient tissues and peripheral blood (PB) of healthy donors (HDs). We analyzed the effect of adoptive transfer of IL-25-activated ILC2 cells on tumor growth, metastasis and survival in a heterotopic murine model of lung cancer.ResultsWe report that human NSCLC patients with high IL-25 expression have reduced overall survival. Moreover, NSCLC patients bear increased frequencies of ILC2s compared to HDs. Frequencies of Tregs were also increased in NSCLC patients, concomitantly with ILC2s. In mice bearing heterotopic lung cancer, adoptive transfer of IL-25-activated ILC2s led to increased tumor growth, increased metastasis and reduced survival. The frequencies of monocytic myeloid-derived suppressor cells (M-MDSCs) were found to be increased in the tumors of mice that received ILC2s as compared to controls.ConclusionOverall, our results indicate that the IL-25/ILC2 axis promotes lung cancer potentially by recruiting immune-suppressive cells to the tumors both in humans and in mice, and that it may therefore represent a suitable novel target for NSCLC immunotherapeutic development.</p