Imunoterapia no tratamento do câncer / Immunotherapy in cancer treatment

O avanço no conhecimento do funcionamento do sistema imunológico e de seu comportamento frente às neoplasias nos tem levado a novas modalidades de tratamento do câncer. Terapias como inibidores de checkpoint imunológico, transferência de células adotivas, anticorpos monoclonais e vacinas preventivas têm revolucionado o enfoque terapêutico para diversos tipos de neoplasias. Este documento traz uma breve descrição dos mecanismos de ação destas novas classes terapêuticas, com o objetivo de fornecer aos médicos alergistas e imunologistas noções de sua aplicação, ajudando-os e estimulando-os a aprofundar o conhecimento nas várias frentes de atuação no tratamento do câncer.

Advances in the knowledge of how the immune system works and its behavior in relation to neoplasms have led to new modalities of cancer treatment. Therapies such as immunological checkpoint inhibitors, adoptive cell transfer, monoclonal antibodies and preventive vaccines have revolutionized the therapeutic approach for various types of neoplasms. This paper provides a brief description of the mechanisms of action of these new therapeutic classes, with the aim of providing physicians and immunologists with notions of their application, helping and stimulating them to deepen the knowledge on the various fronts of action in cancer treatment.

Progress in immunotherapy for hepatocellular carcinoma / 生物工程学报

Hepatocellular carcinoma (HCC) is one of the malignant tumors with the highest morbidity and mortality in the world. The morbidity and mortality of HCC are increasing every year. Liver cancer is a serious threat to public health in China and the death rate of patients with liver cancer in China is the highest in the world. Beyond surgery, chemotherapy and radiotherapy, immunotherapy is an emerging treatment for cancer, which could control and kill tumor cells by relieving the inhibitory status of immune cells in the tumor microenvironment and activating the immune function of the body. Immune checkpoint inhibitors, adoptive immunotherapy and tumor vaccine are the major treatments of immunotherapy. Compared with traditional therapy methods, immunotherapy could enhance immune function, delay tumor progression, prolong the survival time of patients, and becomes a hotspot in the basic and clinical cancer research. This article reviews the research progress of immunotherapy for liver cancer.

Increasing the safety and efficacy of chimeric antigen receptor T cell therapy

Chimeric antigen receptor (CAR) T cell therapy is a promising cancer treatment that has recently been undergoing rapid development. However, there are still some major challenges, including precise tumor targeting to avoid off-target or "on-target/off-tumor" toxicity, adequate T cell infiltration and migration to solid tumors and T cell proliferation and persistence across the physical and biochemical barriers of solid tumors. In this review, we focus on the primary challenges and strategies to design safe and effective CAR T cells, including using novel cutting-edge technologies for CAR and vector designs to increase both the safety and efficacy, further T cell modification to overcome the tumor-associated immune suppression, and using gene editing technologies to generate universal CAR T cells. All these efforts promote the development and evolution of CAR T cell therapy and move toward our ultimate goal-curing cancer with high safety, high efficacy, and low cost.

Advancing chimeric antigen receptor T cell therapy with CRISPR/Cas9

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (CRISPR/Cas9) system, an RNA-guided DNA targeting technology, is triggering a revolution in the field of biology. CRISPR/Cas9 has demonstrated great potential for genetic manipulation. In this review, we discuss the current development of CRISPR/Cas9 technologies for therapeutic applications, especially chimeric antigen receptor (CAR) T cell-based adoptive immunotherapy. Different methods used to facilitate efficient CRISPR delivery and gene editing in T cells are compared. The potential of genetic manipulation using CRISPR/Cas9 system to generate universal CAR T cells and potent T cells that are resistant to exhaustion and inhibition is explored. We also address the safety concerns associated with the use of CRISPR/Cas9 gene editing and provide potential solutions and future directions of CRISPR application in the field of CAR T cell immunotherapy. As an integration-free gene insertion method, CRISPR/Cas9 holds great promise as an efficient gene knock-in platform. Given the tremendous progress that has been made in the past few years, we believe that the CRISPR/Cas9 technology holds immense promise for advancing immunotherapy.

Immunotherapy with natural killer cells: a possible approach for the treatment of Acute Myeloid Leukemia also in Brazil / Imunoterapia com células natural killer: um caminho possível para o tratamento da Leucemia Mielóide Aguda também no Brasil

SUMMARY The allogeneic hematopoietic stem cell transplantation (HSCT) can cure intermediate and high-risk acute myeloid leukemia. Even with the development of strategies to reduce HSCT toxicity, this is still a complex treatment with high morbidity and mortality. Knowledge of the graft versus leukemia effect of HSCT has prepared the way for the development of Adoptive Immunotherapy or in vitro expansion of activated lymphocytes without alloreactivity, with subsequent intravenous infusion. The infusion of genetically modified T lymphocytes and haploidentical natural killer cells has been tested as an alternative to HSCT with very interesting results worldwide and in Brazil, as we not only have the technology of in vitro expansion of clinical grade lymphocytes available, but also do it according to the Good Manufacturing Practices that have been determined internationally.

RESUMO O transplante de células-tronco hematopoéticas (TCTH) alogênico é curativo para leucemia mielóide aguda de risco intermediário e alto. Mesmo com o desenvolvimento de estratégias para minorar a toxicidade do TCTH, este ainda é um tratamento complexo com elevada morbi-mortalidade. O conhecimento sobre o efeito enxerto contra leukemia do TCTH pavimentou o caminho para o desenvolvimento da Imunoterapia Adotiva ou expansão in vitro de linfócitos ativados, sem alo-reatividade, com posterior infusão endovenosa. A infusão de Linfócitos T geneticamente modificados e de células Natural Killer haploidenticas tem sido testada como alternativa ao TCTH com resultados bastante interessantes no mundo e no Brazil já que não apenas dominamos a tecnologia de expansão in vitro de linfócitos em grau clínico, como o fazemos segundo as Boas Práticas de Manufatura determinadas internacionalmente.

Research progress on immunotherapy of nasopharyngeal carcinoma / 中国肿瘤临床

Nasopharyngeal cancer (NPC) is an Epstein–Barr virus-associated malignant tumor. Radiation alone or concurrent chemo-radiotherapy is the principal treatment method and can generally achieve excellent efficacy for NPC patients. However, the prognosis of locoregionally advanced disease or distant metastasis remains poor. Currently, immunotherapy has become a new treatment of sol-id tumors, and it is mainly activating the antitumor immune system of the body. The review aims to explore the progress of immuno-therapy, including adoptive immunotherapy, tumor vaccines, and immune checkpoint inhibitors in NPC treatment.

Effect of GDF11 on expansion of CD8 +memory stem T cells / 中国病理生理杂志

AIM:To investigate the effect of growth differentiation factor 11 ( GDF11 ) on the expansion of CD8 +memory stem T cells ( Tscm) and to further improve the effect of adoptive immunotherapy.METHODS:Healthy human peripheral blood mononuclear cells ( PBMCs) were isolated by density gradient centrifugation at first.Among the i-solated PBMCs, CD8 +T cells were further purified with MACS microbeads.The CD8 +T cells were then randomly divided into experimental groups and control group.The same volume of different concentrations of GDF11 were added into the ex-perimental groups, and the same volume of PBS solution was added into the control group.Finally, the expansion of Tscm in experimental groups and control group was measured by flow cytometry at several time points.RESULTS:GDF11 sig-nificantly increased the number of Tscm in CD8 +T cells in vitro expansion and also dramatically increased the ratio of Tscm in CD8 +T cells.Furthermore, 400 μg/L GDF11 treatment for 3 weeks was the optimal condition to induce CD8 +Tscm. CONCLUSION:GDF11 effectively increases the number and ratio of Tscm in the CD8 +T cells in cell culture growth, thereby creating a new strategy to further improve the efficiency of adoptive immunotherapy.

New progress of adoptive immunotherapy in the treatment of hematologic malignancies based on chimeric antigen receptor T-cells: reports from the 56th American Society of Hematology annual meeting / 白血病·淋巴瘤

T cells redirected to specific antigen targets with engineered chimeric antigen receptors (CARs) are emerging as powerful therapies in hematologic malignancies.Various CAR designs,manufacturing processes,and study populations,among other variables,have been studied and reported in clinical trials.The accumulating data supporting CAR-T cells therapy for disease such as chronic lymphocytic leukemia (CLL)and acute lymphocytic leukemia (ALL) highlight what may well become the next sea change in the care of patients with all types of hematologic neoplasia.The scientific symposium on CAR-T cells therapy inspires the mounting enthusiasm regarding this new treatment strategy.Here,the results of the reported clinical trials and the outlook for CAR-T cells therapies were reviewed and discussed.Many questions remain in the field of CAR-T cells directed to hematologic malignancies,but the encouraging response rates pave a wide road for future investigation.

Research progress of application of immunotherapy by chimeric antigen receptor-engineered T cells / 肿瘤研究与临床

The technique of chimeric antigen receptor-engineered T cells (CAR-T) is combined single chain antibody and T-cell activation modification and has specific and durable killing effect on tumors.It had achieved stunning results on clinical trials of B lymphocytic leukemias,B lymphomas and solid tumors.Meanwhile it also had potential risk on the off target effects,cytokine release syndrome,graft versus host disease and so on.This paper briefly reviews the principal of CAR-T cellular mechanism,advantages and clinical applications.

Changes of regulatory T cells in peripheral blood after gemcitabine chemotherapy for pancreatic cancer patients / 药学实践杂志

Objective To investigate the influence of gemcitabine chemotherapy on levels of regulatory Tcells (Tregs) in peripheral blood for patients with pancreatic cancer and provide evidence and reference for improving the efficacy of adoptive im-munotherapy .Methods 32 patients were enrolled in this study from January 2012 to October 2014 ,among whom 16 received gemcitabine chemotherapy combined with adoptive immunotherapy (gemcitabine group) ,the other 16 patients received adoptive immunotherapy only(control group) .The level of Tregs in peripheral blood ,side effect and overall survival were observed be-fore and after the therapy .Results The number of Tregs in peripheral blood was significantly decreased after gemcitabine chemotherapy ,and it was also lower than that of the control group .The overall survival time of the gemcitabine group was 1.3 mo longer than the control group(10 .0 mo vs 8 .7 mo) .Conclusion Therapeutic regimen of gemcitabine can remarkly de-plete Tregs in peripheral blood of patients with pancreatic cancer ,effectively regulate tumor immune tolerance ,and improve the efficacy of adoptive immunotherapy .

Effect of DC-CIK cell on the proliferation, apoptosis and differentiation of leukemia cells

OBJECTIVE@#To observe the effect of co-culture cytokine-induced killer cells (CIK) and homologous dendritic cells (DC) on the proliferative activity and phenotype change of the DC-CIK cell and the cell killing activity of leukemia HL-60.@*METHODS@#50 mL cord blood sample was obtained from infants delivered by full term healthy woman and the cord blood mononuclear cells were isolated by density gradient centrifugation. Non-adherent cells were collectedfor the induction culture of CIK, adherent cells were differentiated into mature DC; cultured mature DC was mixed with and CIK in the proportion of 1:5 for 12 d. Killing activity of DC-CIK co-cultured cell on leukemia HL-60 was detected by MTT assay.@*RESULTS@#Compared with CIKs, the co-cultured DC-CIKs presented a markedly higher proliferation and killing activity.@*CONCLUSIONS@#Co-culture of DC-CIK cells led to a significant increase of the proliferation and cytotoxicity of CIK.

Silencing itch by small speciifc interfering RNA enhance immune activity of mouse T lymphocyte to kill MFC stomach neoplasms cells in vitro / 中国癌症杂志

Background and purpose: Itch protein is an established regulator of T cell immune response thresholds, belong to a class of E3 ubiquitin-transferring enzymes, widely involve in the ubiquitination of several key signaling molecules, such as ZAP70, P85, VAV, PLC-γ, PKC-θ, etc, plays a critical role in tumor induced immu-nosuppression. Itch ligase activity regulate T-cell anergy and development of regulatory T cells in the periphery by modulating key components of T-cell receptor and transforming growth factor-βsignaling. Therefore, manipulation of Itch activities may provide the opportunities to develop future therapies for immune disorders such as autoimmunity and cancer. speciifc small interfering RNA(siRNA) was utilized to silence the expression of Itch gene of T-lymphocytes and investigate the cytotoxicity activity of transfected T lymphocytes against MFC stomach neoplasms cells in vitro. Methods:T lymphocytes were isolated from the spleen of 615 mice and transfected by speciifc siRNA to silence the expression of Itch gene, The expression of Itch protein were examined by Western bolt in each group;72 hours after transfection, The secretion level of IL-2, INF-γwere measured by enzyme-linked immunosorbent assay (ELISA). At the end, the cytotoxicity activity changes against MFC stomach neoplasms cells was compared between transfected T lym-phocytes, negative control and blank control in vitro. Results:Compared with control group, the expression rate of Itch protein of transfected T-lymphocytes was decreased to 16%after transfection 48 hours;72 hours after transfection, the secretion level of IL-2 in transfection group, negative control and blank control respectively were (1 891.96±141.91)pg/mL, (1 241.69±91.67)pg/mL and (1 175.03±89.14)pg/mL (P<0.001), the secretion level of INF-γin transfection group, negative control and blank control respectively were (958.33±75.46)pg/mL, (683.33±66.67)pg/mL and (691.72±68.72) pg/mL (P<0.05). Transfected T lymphocyte also showed more efifcient killing ability against MFC stomach neoplasms cells than negative control and blank control in vitro, the highest killing rate has reached (54.18±2.96)%. Conclusion:Silencing Itch gene can signiifcantly promoted the secretion level of IL-2, INF-γof mice T lymphocyte, enhanced the cytotoxicity activity of T lymphocyte against MFC stomach neoplasms cells in vitro.

Experimental study on in vitro preparation of adoptive immunotherapy cells induced by CD40 in combination with cytokines / 中国免疫学杂志

Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.

Cellular immune therapy for viral infections in transplant patients.

Stem cell and organ transplantation are considered as the major advances of modern medicine. Unfortunately the success of transplantation is limited by its toxicity and infectious complications as a result of profound immunosuppression. Viral infections are an extremely common and predictable problem in these patients. Antiviral drugs given either prophylactically or as early therapy for patients with detectable viral loads appear to be an effective strategy for reducing viral infections. However, long-term treatment with these drugs is associated with significant toxicity, expense and the appearance of drug resistant virus isolates ultimately resulting in treatment failure. Over the last few years, there is increasing evidence that cellular immune therapies can reverse the outgrowth of haematological malignancies and can also provide therapeutic benefit against lethal viral infections. While the expansion and adoptive transfer of virus-specific T-cells from the healthy donor can be an effective strategy to control viral replication, this is not possible when donors are seronegative or are subsequently inaccessible. Recent studies have demonstrated successful expansion of virus-specific T-cells from seropositive stem cell transplant recipients of a seronegative graft with active virus disease and the long term reconstitution of protective anti-viral immunity following their adoptive transfer back into the patients. Furthermore, this immunotherapeutic strategy has also been extended for multiple pathogens including cytomegalovirus, Epstein-Barr virus, adenovirus and BK polyoma-virus. This approach can be employed to rapidly expand multiple pathogens-specific T cells that can be used for adoptive immunotherapy. Finally, new assays to monitor T cell immunity have been developed which will allow to identify the high risk transplant patients who may develop virus-associated complications post-transplantation and can be given adoptive T cell therapy prophylactically.

Inhibition of Human Pancreatic Tumor Growth by Cytokine-Induced Killer Cells in Nude Mouse Xenograft Model

Pancreatic cancer is the fourth commonest cause of cancer-related deaths in the world. However, no adequate therapy for pancreatic cancer has yet been found. In this study, the antitumor activity of cytokine-induced killer (CIK) cells against the human pancreatic cancer was evaluated in vitro and in vivo. Human peripheral blood mononuclear cells were cultured with IL-2-containing medium in anti-CD3 for 14 days. The resulting populations of CIK cells comprised 94% CD3+, 4% CD3-CD56+, 41% CD3+CD56+, 11% CD4+, and 73% CD8+. This heterogeneous cell population was called cytokine-induced killer (CIK) cells. At an effector-target cell ratio of 100:1, CIK cells destroyed 51% of AsPC-1 human pancreatic cancer cells, as measured by the 51Cr-release assay. In addition, CIK cells at doses of 3 and 10 million cells per mouse inhibited 42% and 70% of AsPC-1 tumor growth in nude mouse xenograft assays, respectively. This study suggests that CIK cells may be used as an adoptive immunotherapy for pancreatic cancer patients.

The immunobiology of cord blood transplantation

Despite significant recent advances in the applicability and outcome following unrelated cord blood transplantation (UCBT), infections remain a major cause of mortality associated with poor immune recovery in the first 6 months after UCBT. Enhanced immune reconstitution not only could improve survival by reduced transplant related mortality, but may also favorably impact on relapse incidence by improved graft-versus-leukemia effects. This review will summarize our current understanding of the biology of immune recovery post-UCBT with an emphasis on adaptive T cell dependent immunity. New efforts to boost immunity will be also highlighted including our own laboratory, where ex vivo T cell expansion is pursued towards adoptive immunotherapy.

Phase II study of adoptive immunotherapy for advanced melanoma / 肿瘤

Objective: To observe the efficacy and safety of chemotherapy combined with cytokine-induced killer (CIK) cells in the treatment of metastatic melanoma (MM). Methods: Fifty three chemotherapy-naive MM patients were administered fotemustine at 100 mg/m2 on d 1-5; dacarbazine were given at 400 mg/d on d 2-6; CIK were infused on d 7, d 14, d 16. Twenty-eight days were regarded as one cycle. The clinical efficay was evaluated every 2 cycles. Results: Thirty-four out of 53 patients were eligible to be evaluated. The overall response rate (ORR) was 23.5% including 1 case with complete response (CR) (2.9%) and 7 cases with partial response (PR) (20.6%). Fourteen cases had stable disease (44%). The clinical benefit response was 67.5%. Median progression free survival (PFS) was 8 months. Median overall survival (OS) was 11 months. The patients with normal lactate dehydrogenase (LDH) had longer OS; those with stable disease had longer PFS and OS. Adverse reaction included grade III/IV thrombopenia (41%), decrease in the number of WBC (23.5%), and hyperreactivity (2 cases). No treatment-related death occurred. Conclusion: Chemotherapy followed by the infusion of CIK was tolerable. The response rate was higher than common chemotherapy. It could prolong the survival time of tumor patients with elevated LDH and non progression disease. Phase III study is needed to confirm the results.

Proliferation of antigen specific T cells in vitro and analysis of phenotype and function / 中华微生物学和免疫学杂志

Objective To explore a new peptide-based approach independent of HLA to generate antigen-specific CD+ CD8+T cells. Methods Peripheral blood mononuclear cells(PBMC) were stimula- ted for 6 h with IE-1 peptide pool. Then the activated IFN-γsecreting ceils were tested by immunomagnetic selection. And the selected cells were cultured with radio-inactivated PBMC in medium with 100 IU/ml IL-2 for 4 weeks. Results The generated T cell lines consisted of IE-1 specific CD4+ T (6.88%) and CD8+ T cells 92.99%, which demonstrated antigen-specific killing and cytokine secretion. Conclusion T ceils can be proliferated with this new procedure, and maintain its phenotype and antigen-specific function.

Anti-tumor mechanisms of lymphopenic mice transferred with tumor-specific lymphocytes and immunized with mIL-21 tumor vaccine / 中华微生物学和免疫学杂志

Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.

Immuno-cell Therapy / 日本補完代替医療学会誌

The growing knowledge of cancer immunology during the past 20 years has led to the current implementation of immunotherapy. Immuno-cell therapy, in which ex vivo processed T lymphocytes and dendritic cells are used as agents, has developed and spread and is now accepted as a common treatment with the identification of a number of cancer peptide antigens. The response rate to immuno-cell therapy is reported to be around 10-20%. Some clinical studies have reported that immuno-cell therapy as a postoperative adjuvant therapy improved survival rates. This paper outlines the historic background and the current medical scene of immuno-cell therapy.<br>