Neoadjuvant immunotherapy represents a pioneering approach in the preoperative treatment of cancer, providing new strategies for tumor reduction and improved patient outcomes by modulating the immune response. This study investigated neoadjuvant immunotherapy using intratumoral administration of mannan-BAM, Toll-like receptor ligands, and anti-CD40 antibody (MBTA therapy) followed by surgery in murine models of MTT pheochromocytoma, B16-F10 melanoma, and 4T1 and E0771.lmb mammary carcinomas. In the MTT pheochromocytoma model, it was found that neoadjuvant MBTA therapy followed by surgery could prevent the development of distant metastases in 100% of treated animals, compared to a 60% mortality rate in the control group due to metastatic disease after surgery. These outcomes were achieved even in tumors three times larger than those in the control group. In the aggressive 4T1 model, neoadjuvant MBTA therapy resulted in slower tumor progression and a significant prolongation of survival. In the B16-F10 and E0771.lmb models, neoadjuvant MBTA therapy also protected animals from metastases development and tumor recurrence upon rechallenge with tumor cells after surgery. Transcriptomic analysis revealed enhanced effector immune cell infiltration, cytotoxicity, and antigen presentation in retransplanted tumors from MBTA-treated mice, indicating robust immune memory. Notably, the exclusion of the anti-CD40 antibody from the neoadjuvant MBTA therapy (MBT therapy) yielded comparable outcomes in protection against metastases development. These findings advocate for further investigation of intratumoral neoadjuvant MBTA therapy for immunologically "cold" tumors, including those at high risk of metastases or recurrence.
- MeSH
- antigeny CD40 imunologie antagonisté a inhibitory MeSH
- imunoterapie * metody MeSH
- lokální recidiva nádoru * prevence a kontrola MeSH
- mannany farmakologie MeSH
- melanom experimentální * imunologie terapie patologie MeSH
- metastázy nádorů MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- neoadjuvantní terapie * metody MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Immunotherapy emerges as a fundamental approach in cancer treatment. Up to date, the efficacy of numerous different immunotherapies has been evaluated. The use of microorganisms or their parts for immune cell activation, referred to as Pathogen-Associated Molecular Patterns (PAMPs), represents highly promising concept. The therapeutic effect of PAMPs can be further amplified by suitable combination of different types of PAMPs such as Toll like receptor (TLR) agonists and phagocytosis activating ligands. Previously, we used the combination of phagocytosis activating ligand (mannan) and mixture of TLR agonists (resiquimod (R-848), poly(I:C), inactivated Listeria monocytogenes) for successful treatment of melanoma in murine B16-F10 model. In the present study, we optimized the composition and timing of previously used mixture. Therapeutic mixture based on well-defined chemical compounds consisted of mannan anchoring to tumor cell surface by biocompatible anchor for membranes (BAM) and TLR agonists resiquimod, poly(I:C), and lipoteichoic acid (LTA). The optimization resulted in (1) eradication of advanced stage progressive melanoma in 83% of mice, (2) acquisition of resistance to tumor re-transplantation, and (3) potential anti-metastatic effect. After further investigation of mechanisms, underlying anti-tumor responses, we concluded that both innate and adaptive immunity are activated and involved in these processes. We tested the efficacy of our treatment in Panc02 murine model of aggressive pancreatic tumor as well. Simultaneous application of agonistic anti-CD40 antibody was necessary to achieve effective therapeutic response (80% recovery) in this model. Our results suggest that herein presented immunotherapeutic approach is a promising cancer treatment strategy with the ability to eradicate not only primary tumors but also metastases.
- MeSH
- adenokarcinom imunologie patologie terapie MeSH
- fagocytóza * MeSH
- imidazoly terapeutické užití MeSH
- imunoterapie MeSH
- kyseliny teichoové terapeutické užití MeSH
- lipopolysacharidy terapeutické užití MeSH
- mannany terapeutické užití MeSH
- melanom experimentální imunologie patologie terapie MeSH
- myši inbrední C57BL MeSH
- nádorové buněčné linie MeSH
- nádory slinivky břišní imunologie patologie terapie MeSH
- neutrofily imunologie MeSH
- poly I-C terapeutické užití MeSH
- toll-like receptory agonisté MeSH
- tumor burden účinky léků MeSH
- tumor infiltrující lymfocyty imunologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Interleukin (IL)-2 has been approved for treatment of metastatic renal cancer and malignant melanoma. However, its unfavorable pharmacologic properties, severe side effects and the negative role of IL-2 in maintaining T regulatory cells are severe drawbacks. It has been shown that immunocomplexes of IL-2 and certain anti-IL-2 mAbs possess selective and high stimulatory activity in vivo. Here, we show that IL-2/S4B6 mAb immunocomplexes expand not only CD122(high) subsets and newly activated CD8(+) T cells but also natural killer T cells and γδ T cells. Further, we demonstrate that natural killer (NK) cells expanded by IL-2/S4B6 mAb immunocomplexes in vivo have high cytolytic activity, which can be further increased by coadministration of IL-12. We also demonstrate that IL-2/S4B6 mAb immunocomplexes possess noticeable antitumor activity in two syngeneic mouse tumor models, namely BCL1 leukemia and B16F10 melanoma, but only if administered early in tumor progression. To effectively treat established tumors, we administered the tumor-bearing mice first with N-(2-hydroxypropyl)methacrylamide copolymer-bound doxorubicin conjugate, and subsequently with IL-2/S4B6 mAb immunocomplexes alone or with IL-12 to induce an efficient antitumor immune response. Importantly, we show that the conjugate has significantly lower immunosuppressive activity than free doxorubicin when using dosage with comparable antitumor activity, thus eliminating the majority of tumor cells while leaving the immune system mostly unimpaired for stimulation with IL-2/S4B6 mAb immunocomplexes. Indeed, we demonstrate that the conjugate and IL-2/S4B6 mAb immunocomplexes together have synergistic antitumor activity.
- MeSH
- akrylamidy aplikace a dávkování MeSH
- buňky NK imunologie MeSH
- CD8-pozitivní T-lymfocyty imunologie MeSH
- doxorubicin aplikace a dávkování MeSH
- experimentální leukemie terapie MeSH
- imunokonjugáty terapeutické užití MeSH
- imunosupresiva aplikace a dávkování MeSH
- interleukin-12 aplikace a dávkování MeSH
- interleukin-2 imunologie terapeutické užití MeSH
- melanom experimentální terapie MeSH
- monoklonální protilátky aplikace a dávkování MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Photodynamic therapy (PDT) has been approved as proper and effective kind of treatment for certain types of cancer and non-malignant diseases. We tested photodynamic effects on G361 human melanoma cells sensitized by zinc-5,10,15,20-tetrakis(4-sulphonatophenyl) porphyrine (ZnTPPS(4)), chloraluminium phtalocyanine disulfonate (ClAlPcS(2)) and 5-aminolevulinic acid (ALA). In particular, we examined the PDT efficiency depending on applied light dose (0.8; 1.7; 3.3; 6.6; 13.2; 26.4Jcm(-2)). The DNA gel electrophoresis, methylthiazol tetrazolium bromide (MTT) viability test, fluorescent microscopy using calcein AM and propidium iodide (PI) staining, and rhodamine 123 mitochondrial membrane potential assay were performed to detect and evaluate the cell death process. We also measured the time course of reactive oxygen species (ROS) production and its dependence on sensitizer concentration within continuously irradiated sensitized cells. In conclusion, these results demonstrate most significant phototoxic effect of ClAlPcS(2)-PDT in spite of significantly higher ROS production induced by ZnTPPS(4)-PDT on G361 cells. On the other hand, ALA-PDT has a minimal photoeffect and induces negligible ROS formation in G361 cells at the conditions described below.
- MeSH
- barvicí látky MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- fluorescenční barviva MeSH
- fluorescenční mikroskopie MeSH
- fotochemoterapie MeSH
- fotosenzibilizující látky farmakologie MeSH
- fragmentace DNA účinky léků účinky záření MeSH
- indoly farmakologie MeSH
- kyselina aminolevulová farmakologie MeSH
- lidé MeSH
- melanom experimentální terapie MeSH
- membránové potenciály účinky léků MeSH
- metaloporfyriny farmakologie MeSH
- mitochondriální membrány účinky léků MeSH
- nádorové buněčné linie MeSH
- organokovové sloučeniny farmakologie MeSH
- propidium MeSH
- reaktivní formy kyslíku metabolismus MeSH
- rhodamin 123 MeSH
- světlo MeSH
- tetrazoliové soli MeSH
- thiazoly MeSH
- vztah dávky záření a odpovědi MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this article, we evaluated a planar microwave applicator for in vivo superficial hyperthermia treatments on small tumors in the mouse mimicking treatments for human neoplasms. The design of the applicator, was challenged by the small dimensions of the tumors and unwanted diffusion of heating in the tumor-bearing animals. The required solution was to limit the penetration of microwaves in the depth of the tissue maintaining the full efficacy of hyperthermia. The study was firstly performed by computer simulations of SAR distribution inside a flat homogeneous phantom, considering various thicknesses of the integrated water bolus. Simulations, validated by the measurements, were also used to evaluate the impedance matching. Further tests were performed on homogeneous agar phantom to simulate the temperature distribution in the biological tissue and to preliminary assess the possible modality and schedule of microwave hyperthermia delivery. The in vivo experiments showed the evidence of direct microwave-induced heating and damage of the melanoma tissue in a range of penetration coherent both with computer simulations and phantom studies. The described approach appears perspective for designing limited-microwave-delivery applicators tailored for treatments of human superficial tumors and pre-tumoral lesions.
- MeSH
- biologické modely MeSH
- design vybavení MeSH
- fantomy radiodiagnostické MeSH
- indukovaná hypertermie přístrojové vybavení metody MeSH
- melanom experimentální patologie terapie MeSH
- mikrovlny terapeutické užití MeSH
- myši MeSH
- nádory kůže patologie terapie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Local recurrences at the site of tumor resection or after chemotherapy, as well as distant micrometastases represent major problems in oncology. Therapeutic strategies based on insertion of immunostimulatory genes into the genome of tumor cells followed by vaccination with the resulting genetically modified and irradiated cellular vaccines represent a new potential prospect for the treatment of cancer patients. These strategies are based on the presumption that many, if not all tumors, possess cell surface antigens capable of being recognized by defence effectors of the immune system, as well as on the presumption that local treatment of primary tumors can, due to its immunizing potential, result also in the inhibition of distant metastases. Genetically modified cellular vaccines were found to be efficient against cancer both in experimental models and in tumor-bearing patients. It was also shown in various systems that the efficacy of conventional therapeutic modalities can be supported by adjuvant administration of genetically modified vaccines, as well as by depletion of immunosuppressive immunocyte subsets. The purpose of this review was to summarize and evaluate the results obtained with the administration of genetically modified cellular vaccines as well as with depletion of immunosuppressive immunocytes performed as treatment of minimal residual disease after surgery / chemotherapy in the experimental model of murine tumors mimicking human HPV16-associated neoplasms. The prospects and limitations of these adjuvant immunotherapeutic modalities are discussed.
- MeSH
- infekce papilomavirem farmakoterapie imunologie chirurgie MeSH
- lidé MeSH
- lidský papilomavirus 16 genetika imunologie MeSH
- melanom experimentální genetika imunologie terapie MeSH
- myši MeSH
- protinádorové vakcíny terapeutické užití MeSH
- regulační T-lymfocyty imunologie MeSH
- reziduální nádor genetika imunologie terapie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Our study was aimed to characterize the phenotype and functional endpoints of local microwave hyperthermia (LHT, 42 degrees C) on tumor infiltrating and spleen leukocytes. The effectiveness of LHT applied into the tumor of B16F10 melanoma-bearing C57/BL6 mice was compared with anesthetized and non-treated animals. Subpopulations of leukocytes were analyzed using the flow cytometry, and the cytotoxic activity of splenocytes against syngeneic B16F10 melanoma and NK-sensitive YAC-1 tumor cell lines was evaluated in (51)Cr-release assay. Similarly, the in vitro modification of the heat treatment was performed using healthy and melanoma-bearing splenocytes. We found a 40 % increase of activated monocytes (CD11b+CD69+) infiltration into the tumor microenvironment. In the spleen of experimental animals, the numbers of cytotoxic T lymphocytes (CTLs-CD3+CD8+) and NK cell (CD49b+NK1.1+) raised by 22 % and 14 %, respectively, while the NK1.1+ monocytes decreases by 37 %. This was accompanied by an enhancement of cytotoxic effector function against B16F10 and YAC-1 targets in both in vivo and in vitro conditions. These results demonstrate that LHT induces better killing of syngeneic melanoma targets. Furthermore, LHT evokes the homing of activated monocytes into the tumor microenvironment and increases the counts of NK cells and CTL in the spleen.
- MeSH
- buňky NK imunologie MeSH
- CD antigeny analýza MeSH
- cytotoxicita imunologická MeSH
- cytotoxické T-lymfocyty imunologie MeSH
- diatermie MeSH
- fenotyp MeSH
- financování organizované MeSH
- imunofenotypizace MeSH
- melanom experimentální imunologie terapie MeSH
- mikrovlny terapeutické užití MeSH
- monocyty imunologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- pohyb buněk MeSH
- průtoková cytometrie MeSH
- slezina imunologie MeSH
- tumor infiltrující lymfocyty imunologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- melanom experimentální imunologie terapie MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- kongresy MeSH
