scholarly journals P09.03 Hyaluronic acid as a new immunologic adjuvant in cancer: design of effective preventive and therapeutic vaccination strategies for HER2/neu-positive breast tumors

2021 ◽  
Vol 9 (Suppl 1) ◽  
pp. A28.2-A29
Author(s):  
D Carpanese ◽  
I Montagner ◽  
A Dalla Pietà ◽  
V Rossi ◽  
A Penna ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Cancer Vaccines ◽  
Tumor Regression ◽  
Vaccination Strategy ◽  
Cellular Responses ◽  
List Type ◽  
Therapeutic Vaccination ◽  
Tlr Agonist ◽  
Humoral Responses

BackgroundThe use of proteins as immunogens is attractive for the development of vaccines, but requires efficient adjuvants to overcome their weak immunogenicity. Recently, we investigated the potential of the TLR2/4 agonist hyaluronan (HA) as an immunological adjuvant for protein-based vaccines.1 2 Conjugation of HA to antigens strongly increased their immunogenicity and promoted their rapid translocation to draining lymph nodes, resulting in robust and long-lasting humoral responses.1 On these bases, we investigated the potentiality of HA-based technology in the design of cancer vaccines. To this aim, HA was conjugated to the extracellular domain of rat HER2/neu (rHER2/neu) and validated in the preventive and therapeutic vaccination settings.Materials and MethodsFemale BALB/c or BALB-neuT mice were immunized with rHER2/neu-HA. In vivo depletion of CD4+, CD8+ T and B cells was performed, and sera and spleens were collected to characterized antigen-specific humoral and cellular responses. Vaccinated BALB/c mice were challenged and re-challenged with rHER2/neu-overexpressing TUBO cells to assess the protective or therapeutic activity of rHER2/neu-HA vaccination strategy, as well as immunological memory.ResultsHA performed efficiently as robust and long-lasting humoral (IgG1, IgG2a, and IgG2b) and cellular responses were detected using very low antigen doses and number of boosters. Outstandingly, at 1-year post-vaccination, anti-rHER2/neu specific antibodies showed even improved effector functions (maturation of affinity for the receptor and increased complement-derived cytotoxicity functions). HA vaccination turned out effective in both the prophylactic (100% mice survived) and therapeutic (tumor regression in 2/12 mice) settings, and broke tolerance against rHER2/neu, delaying spontaneous tumor growth in BALB-neuT mice. Both humoral and cellular responses contributed to the success of HA-based vaccination, but CD8+ T cells played only a marginal role.ConclusionsCancer vaccines have not yet achieved significant clinical efficacy due to their poor immunogenicity, and the validation of more effective adjuvants occurred sometimes at the expense of safety. HA combines the unique immunomodulatory features of a TLR agonist with the tolerability of a fully natural polymer, proving to be a promising adjuvant for the creation of effective and safe cancer vaccines with the potential for rapid clinical translation.ReferencesDalla Pietà A, Carpanese D, et al. Hyaluronan is a natural and effective immunological adjuvant for protein-based vaccines. Cell Mol Immunol 2021;18(5):1197–1210.Rosato A, Montagner IM, Carpanese D, Dalla Pietà A. Hyaluronic acid as a natural adjuvant for protein and peptide-based vaccines. 30.04.2020. WO/2020/084558, PCT/IB2019/059122.Disclosure InformationD. Carpanese: None. I. Montagner: None. A. Dalla Pietà: None. V. Rossi: None. A. Penna: None. G. Zuccolotto: None. G. Pasut: None. A. Grigoletto: None. A. Rosato: None.

scholarly journals Recent Advances in the Use of Galleria mellonella Model to Study Immune Responses against Human Pathogens

2018 ◽  
Vol 4 (4) ◽  
pp. 128 ◽  
Author(s):  
Thais Pereira ◽  
Patrícia de Barros ◽  
Luciana Fugisaki ◽  
Rodnei Rossoni ◽  
Felipe Ribeiro ◽  
...  
Keyword(s):  
Immune Response ◽  
Immune Responses ◽  
Galleria Mellonella ◽  
Cellular Responses ◽  
Innate Immune ◽  
In Vivo Studies ◽  
Human Pathogens ◽  
Radical Production ◽  
Humoral Responses

The use of invertebrates for in vivo studies in microbiology is well established in the scientific community. Larvae of Galleria mellonella are a widely used model for studying pathogenesis, the efficacy of new antimicrobial compounds, and immune responses. The immune system of G. mellonella larvae is structurally and functionally similar to the innate immune response of mammals, which makes this model suitable for such studies. In this review, cellular responses (hemocytes activity: phagocytosis, nodulation, and encapsulation) and humoral responses (reactions or soluble molecules released in the hemolymph as antimicrobial peptides, melanization, clotting, free radical production, and primary immunization) are discussed, highlighting the use of G. mellonella as a model of immune response to different human pathogenic microorganisms.

scholarly journals Artificially cloaked viral nanovaccine for cancer immunotherapy

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Manlio Fusciello ◽  
Flavia Fontana ◽  
Siri Tähtinen ◽  
Cristian Capasso ◽  
Sara Feola ◽  
...  
Keyword(s):  
Cancer Immunotherapy ◽  
Cancer Vaccines ◽  
Tumor Antigens ◽  
Vaccination Strategy ◽  
Therapeutic Setting ◽  
Anti Cancer ◽  
Interesting Approach ◽  
Personalized Cancer

AbstractVirus-based cancer vaccines are nowadays considered an interesting approach in the field of cancer immunotherapy, despite the observation that the majority of the immune responses they elicit are against the virus and not against the tumor. In contrast, targeting tumor associated antigens is effective, however the identification of these antigens remains challenging. Here, we describe ExtraCRAd, a multi-vaccination strategy focused on an oncolytic virus artificially wrapped with tumor cancer membranes carrying tumor antigens. We demonstrate that ExtraCRAd displays increased infectivity and oncolytic effect in vitro and in vivo. We show that this nanoparticle platform controls the growth of aggressive melanoma and lung tumors in vivo both in preventive and therapeutic setting, creating a highly specific anti-cancer immune response. In conclusion, ExtraCRAd might serve as the next generation of personalized cancer vaccines with enhanced features over standard vaccination regimens, representing an alternative way to target cancer.

scholarly journals In vivo single-cell analysis using calcofluor - white staining detects high expression phenotype in L. lactis cultures engineered for hyaluronic acid production

2020 ◽  
Author(s):  
Anantha-Barathi Muthukrishnan ◽  
Antti Häkkinen ◽  
Velvizhi Devi Rajendran ◽  
Anupama Kozhiyalam ◽  
Guhan Jayaraman
Keyword(s):  
Hyaluronic Acid ◽  
Single Cell ◽  
Fine Tuning ◽  
Host Cells ◽  
High Expression ◽  
Significant Heterogeneity ◽  
List Type ◽  
Calcofluor White ◽  
Expression Phenotype

ABSTRACTHyaluronic acid (HA) is a biopolymer with wide applications in the field of medicine and cosmetics. Bacterial production of HA has a huge market globally. Certain species of Streptococcus are native producers of HA but they are pathogenic. Therefore, safer organisms such as L. lactis are engineered for HA production. However, there are challenges such as low yield, low molecular weight and polydispersity of HA obtained from these cultures. Optimisation of bioprocess parameters and downstream purification parameters are being addressed to overcome these challenges. We explore these problems from the perspective of microbial heterogeneity, since variations in phenotype affect the yield and properties of the product in a bioreactor. For this perspective, a method to quantitatively assess the occurrence of heterogenous phenotypes depending on the amount of HA produced at the single-cell level is required. Here, we evaluated for the first time the use of calcofluor white staining method combined with in vivo fluorescence confocal microscopy to quantify the heterogeneity in phenotypes of L. lactis cells engineered for HA production.From the microscopy image analysis, we found that the population harbours significant heterogeneity with respect to HA production and our novel approach successfully differentiates these phenotypes. Using the fluorescence intensity levels, first we were able to confidently differentiate cells not expressing HA (Host cells without HA genes for expression) from cells with genes for HA production (GJP2) and induced for expression, as there is a consistently two-fold higher level of expression in the GJP2 cells independently of the cell size. Further, this method revealed the occurrence of two different phenotypes in GJP2 cultures, one of a high-expression phenotype (40% of the population) and the other one of a low-expression (remaining 60% of the population), and it is the high expression phenotype that contributes to the increase in the HA expression of the GJP2 population compared with the host cells. Thus, it is essential to identify the extrinsic and intrinsic factors that can favour most of the cells in the population to switch and stabilise into the high-expression phenotype state in a bioreactor, for higher yield and possibly reduced heterogeneity of the product, such as polydispersity in chain lengths. For such optimisation studies, this in vivo method serves as a promising tool for rapid detection of phenotypes in the bioreactor samples under varying conditions, allowing fine tuning of the factors to stabilise high-expression phenotypes thereby maximizing the yield.Graphical Abstractdone.Key PointsCalcofluor staining successfully differentiated the phenotypes based on HA levels.This study revealed the occurrence of significant heterogeneity in HA expression.This method will aid for rapid optimization of factors for improved HA production.

scholarly journals Cellular responses of hyaluronic acid-coated chitosan nanoparticles

2018 ◽  
Vol 7 (5) ◽  
pp. 942-950 ◽  
Author(s):  
Abdulaziz Almalik ◽  
Ibrahim Alradwan ◽  
Majed A. Majrashi ◽  
Bashayer A. Alsaffar ◽  
Abdulmalek T. Algarni ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Hyaluronic Acid ◽  
Biomedical Applications ◽  
Chitosan Nanoparticles ◽  
Cellular Responses

In recent years, nanotechnology has been proven to offer promising biomedical applications for in vivo diagnostics and drug delivery, stressing the importance of thoroughly investigating the biocompatibility of potentially translatable nanoparticles (NPs).

LACK OF EFFECT OF CORTICOSTEROIDS ON THE IN VIVO DISAPPEARANCE OF SULFHYDRYL-INHIBITED AND ANTIBODY-COATED ERYTHROCYTES

1964 ◽  
Vol 47 (3_Suppl) ◽  
pp. S28-S36
Author(s):  
Kailash N. Agarwal
Keyword(s):  
Red Cells ◽  
List Type ◽  
Red Cell

ABSTRACT Red cells were incubated in vitro with sulfhydryl inhibitors and Rhantibody with and without prior incubation with prednisolone-hemisuccinate. These erythrocytes were labelled with Cr51 and P32 and their disappearance in vivo after autotransfusion was measured. Prior incubation with prednisolone-hemisuccinate had no effect on the rate of red cell disappearance. The disappearance of the cells was shown to take place without appreciable intravascular destruction.

STUDIES IN CONGENITAL GENERALIZED LIPODYSTROPHY

1973 ◽  
Vol 72 (3) ◽  
pp. 495-505 ◽  
Author(s):  
Oddmund Søvik ◽  
Svein Oseid
Keyword(s):  
Biological Activity ◽  
Insulin Response ◽  
Epididymal Adipose Tissue ◽  
Large Individual ◽  
Immunoreactive Insulin ◽  
List Type ◽  
Glucose Load ◽  
Congenital Generalized Lipodystrophy ◽  
The One

ABSTRACT The biological activity of plasma insulin from 4 cases of congenital generalized lipodystrophy has been studied, using rat diaphragm and epididymal adipose tissue in vivo. The results are compared with previous data on plasma immunoreactive insulin obtained in these patients. 2 of the 4 cases exhibited unusually high biological insulin activities during the fasting state as well as after an intravenous (iv) glucose load. In the fat pad assay activities as high as 10 000 μU insulin per ml were observed. During childhood the biological insulin activities were generally high, although there were large individual variations. However, in the one case studied after the age of puberty, the insulin response to a glucose load was negligible. Taken together, the biological and immunological activities observed strongly suggest the presence of pancreatic insulin in these patients. It appears that the circulating insulin has a fully biological activity. The decreasing insulin activities after cessation of growth are in agreement with the appearance of frank diabetes at this time.

Production and Preliminary In Vivo Evaluations of a Novel in silico-designed L2-based Potential HPV Vaccine

2020 ◽  
Vol 21 (4) ◽  
pp. 316-324
Author(s):  
Manica Negahdaripour ◽  
Navid Nezafat ◽  
Reza Heidari ◽  
Nasrollah Erfani ◽  
Nasim Hajighahramani ◽  
...  
Keyword(s):  
In Silico ◽  
Group Versus ◽  
Tlr Agonists ◽  
E Coli ◽  
Metal Affinity ◽  
Th1 And Th2 Cytokines ◽  
Ifn Γ ◽  
Humoral Responses ◽  
Future Work

Background: L2-based Human Papillomavirus (HPV) prophylactic vaccines, containing epitopes from HPV minor capsid proteins, are under investigation as second-generation HPV vaccines. No such vaccine has passed clinical trials yet, mainly due to the low immunogenicity of peptide vaccines; so efforts are being continued. A candidate vaccine composed of two HPV16 L2 epitopes, flagellin and a Toll-Like Receptor (TLR) 4 agonist (RS09) as adjuvants, and two universal T-helper epitopes was designed in silico in our previous researches. Methods: The designed vaccine construct was expressed in E. coli BL21 (DE3) and purified through metal affinity chromatography. Following mice vaccination, blood samples underwent ELISA and flow cytometry analyses for the detection of IgG and seven Th1 and Th2 cytokines. Results: Following immunization, Th1 (IFN-γ, IL-2) and Th2 (IL-4, IL-5, IL-10) type cytokines, as well as IgG, were induced significantly compared with the PBS group. Significant increases in IFN-γ, IL-2, and IL-5 levels were observed in the vaccinated group versus Freund’s adjuvant group. Conclusion: The obtained cytokine induction profile implied both cellular and humoral responses, with a more Th-1 favored trend. However, an analysis of specific antibodies against L2 is required to confirm humoral responses. No significant elevation in inflammatory cytokines, (IL-6 and TNF-α), suggested a lack of unwanted inflammatory side effects despite using a combination of two TLR agonists. The designed construct might be capable of inducing adaptive and innate immunity; nevertheless, comprehensive immune tests were not conducted at this stage and will be a matter of future work.

NF-κB Down–regulation and PARP Cleavage by novel 3-α-butyryloxy-β-boswellic Acid Results in Cancer Cell Specific Apoptosis and in vivo Tumor Regression

2013 ◽  
Vol 13 (5) ◽  
pp. 777-790 ◽  
Author(s):  
Yasrib Qurishi ◽  
Abid Hamid ◽  
Parduman R. Sharma ◽  
Zahoor A. Wani ◽  
Dilip M. Mondhe ◽  
...  
Keyword(s):  
Cancer Cell ◽  
Tumor Regression ◽  
Parp Cleavage ◽  
Boswellic Acid ◽  
Down Regulation

scholarly journals An ARF GTPase module promoting invasion and metastasis through regulating phosphoinositide metabolism

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Marisa Nacke ◽  
Emma Sandilands ◽  
Konstantina Nikolatou ◽  
Álvaro Román-Fernández ◽  
Susan Mason ◽  
...  
Keyword(s):  
Metastatic Cancer ◽  
Cellular Responses ◽  
Signalling Pathways ◽  
External Signal ◽  
Phosphoinositide Metabolism ◽  
Invasion And Metastasis ◽  
3 Dimensional

AbstractThe signalling pathways underpinning cell growth and invasion use overlapping components, yet how mutually exclusive cellular responses occur is unclear. Here, we report development of 3-Dimensional culture analyses to separately quantify growth and invasion. We identify that alternate variants of IQSEC1, an ARF GTPase Exchange Factor, act as switches to promote invasion over growth by controlling phosphoinositide metabolism. All IQSEC1 variants activate ARF5- and ARF6-dependent PIP5-kinase to promote PI(3,4,5)P3-AKT signalling and growth. In contrast, select pro-invasive IQSEC1 variants promote PI(3,4,5)P3 production to form invasion-driving protrusions. Inhibition of IQSEC1 attenuates invasion in vitro and metastasis in vivo. Induction of pro-invasive IQSEC1 variants and elevated IQSEC1 expression occurs in a number of tumour types and is associated with higher-grade metastatic cancer, activation of PI(3,4,5)P3 signalling, and predicts long-term poor outcome across multiple cancers. IQSEC1-regulated phosphoinositide metabolism therefore is a switch to induce invasion over growth in response to the same external signal. Targeting IQSEC1 as the central regulator of this switch may represent a therapeutic vulnerability to stop metastasis.

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