Oncolytic reovirus enhances rituximab-mediated antibody-dependent cellular cytotoxicity against chronic lymphocytic leukaemia

Parrish, Christopher; Scott, Gina B.; Migneco, G; Scott, Karen J.; Steele, Lynette P.; Ilett, Elizabeth J.; West, Emma; Hall, Kathryn; Selby, Peter J.; Buchanan, David L.; Varghese, Abraham; Cragg, Mark S.; Coffey, Matt; Hillmen, Peter; Melcher, Alan; Errington-Mais, Fiona

doi:10.1038/leu.2015.88

Cited by 37 publications

(50 citation statements)

References 62 publications

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“…As a control, the experiment was therefore repeated with Rituximab, another therapeutic mAb also targeting the CD20 antigen. Rituximab gave similar results, which correspond well with the ongoing search for possibilities to enhance natural killer cell (NK-cell) activity and ADCC response of Anti-CD20 mAbs [31][32][33] and confirms that the assay in principle works.For both, Erbitux and Herceptin, a considerably lower EC 50 value was measured for the basic charge variant fraction compared to the main and acidic counterparts, which corresponds to a better performance in the assay. For Herceptin the EC 50 value of fraction B was almost 70% lower compared to fraction M. This means that for this fraction, only one third of the protein concentration is necessary in order to achieve similar lysis activity.…”

supporting

confidence: 63%

Microheterogeneity of therapeutic monoclonal antibodies is governed by changes in the surface charge of the protein

Hintersteiner

Lingg

Janzek

et al. 2016

Biotechnology Journal

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It has previously been shown for individual antibodies, that the microheterogenity pattern can have a significant impact on various key characteristics of the product. The aim of this study was to get a more generalized understanding of the importance of microheterogeneity. For that purpose, the charge variant pattern of various different commercially available therapeutic mAb products was compared using Cation-Exchange Chromatography with linear pH gradient, antigen affinity, Fcreceptor affinity, antibody dependent cellular cytotoxicity (ADCC) and conformational stability. For three of the investigated antibodies, the basic charge variants showed a stronger binding affinity towards FcγRIIIa as well as an increased ADCC response. Differences in the conformational stability of antibody charge variants and the corresponding reference samples could not be detected by differential scanning calorimetry. The different biological properties of the mAb variants are therefore governed by changes in the surface charge of the protein and not by an altered structure. This can help to identify aspects of microheterogeneity that are critical for product quality and can lead to further improvements in the development and production of therapeutic antibody products. Keywords: ADCC · Bio-better · Cation exchange chromatography · Effector functions · FcγRIIIa · Therapeutic mAbsCorrespondence: Prof. Alois Jungbauer, Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Muthgasse 18, 1190 Vienna, Austria E-mail: alois.jungbauer@boku.ac.at Abbreviations: ADCC, antibody dependent cellular cytotoxicity; ApHGE, analytical ionexchange chromatography with pH gradient elution; CEX, Cation exchange chromatography; EGFR, epidermal growth factor receptor; mAb, monoclonal antibody; DSC, differential scanning calorimetry; SPR, surface plasmon resonance ants it is still difficult to decide, how precisely a product produced by different methods must match [14][15][16][17][18]. Cation exchange chromatography (CEX) with pH gradient elution has repeatedly been reported as a suitable method for high-resolution separation of antibody variants exhibiting different surface charge characteristics [19][20][21][22]. In this study we used this technique for the analysis and the large-scale separation of several highselling therapeutic antibodies available on the market today, to obtain one acidic (A), one main (M) and one basic (B) charge variant fraction, a methodology previously developed in our group [23,24]. This did not only provide new information on the charge variant pattern of individual monoclonal antibody products, it also gave us the chance to characterize the charge variants found in these antibody products with a wide variety of additional analytical methods and thereby study the overall effect of the presence of charge variants on the stability and biological activity of therapeutic mAbs in general.As starting material, we obtained clinical doses of the seven monoclonal antibodies listed in Table 1, all of w...

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supporting

confidence: 63%

Microheterogeneity of therapeutic monoclonal antibodies is governed by changes in the surface charge of the protein

Hintersteiner

Lingg

Janzek

et al. 2016

Biotechnology Journal

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“…There is increasing preclinical data that the efficacy of OVs may be increased by a combination with other anti-cancer drugs, as has been shown for reovirus enhanced rituximab mediated antibody dependent cellular cytotoxicity against chronic lymphocytic leukemia [14]. Considering these results, Pexa-Vec may potentially be used to treat lymphoid malignancies with other modalities to overcome the resistance or improve the outcome.…”

Section: Discussionmentioning

confidence: 99%

Gene expression profiling of hematologic malignant cell lines resistant to oncolytic virus treatment

Lee

Kim

et al. 2016

Oncotarget

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Pexa-Vec (pexastimogene devacirpvec; JX-594) has emerged as an attractive tool in oncolytic virotherapy. Pexa-Vec demonstrates oncolytic and immunotherapeutic mechanisms of action. But the determinants of resistance to Pexa-Vec are mostly unknown. We treated hemoatologic malignant cells with Pexa-Vec and examined the gene-expression pattern of sensitive and resistant cells. Human myeloid malignant cell lines (RPMI-8226, IM-9, K562, THP-1) and lymphoid cancer cell lines (MOLT4, CCRF-CEM, Ramos, U937) were treated with Pexa-Vec. Pexa-Vec was cytotoxic on myeloid cell lines in a dose-dependent manner, and fluorescent imaging and qPCR revealed that Pexa-Vec expression was low in RAMOS than IM-9 after 24 hrs and 48 hrs of infection. Gene expression profiles between two groups were analyzed by microarray. Genes with at least 2-fold increase or decrease in their expression were identified. A total of 660 genes were up-regulated and 776 genes were down-regulated in lymphoid cancer cell lines. The up- and down-regulated genes were categorized into 319 functional gene clusters. We identified the top 10 up-regulated genes in lymphoid cells. Among them three human genes (LEF1, STAMBPL1, and SLFN11) strongly correlated with viral replication. Up-regulation of PVRIG, LPP, CECR1, Arhgef6, IRX3, IGFBP2, CD1d were related to resistant to Pexa-Vec. In conclusion, lymphoid malignant cells are resistant to Pexa-Vec and displayed up-regulated genes associated with resistance to oncolytic viral therapy. These data provide potential targets to overcome resistance, and suggest that molecular assays may be useful in selecting patients for further clinical trials with Pexa-Vec.

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“…Therefore, reoNAb complexes were generated using either live or UV-inactivated reovirus; both activate monocytes ( Supplementary Fig. S3) but UVinactivated reovirus is unable to infect and kill tumor cells directly (13). Monocytes loaded with UV-reoNAb complexes abrogated melanoma cell death following co-culture ( Fig.…”

Section: Infectious Reovirus Mediates the Killing Of Tumor Cells By Rmentioning

confidence: 99%

Antibody-Neutralized Reovirus Is Effective in Oncolytic Virotherapy

Berkeley

Steele

Mulder

et al. 2018

Cancer Immunology Research

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Immunotherapy is showing promise for otherwise incurable cancers. Oncolytic viruses (OVs), developed as direct cytotoxic agents, mediate their antitumor effects via activation of the immune system. However, OVs also stimulate antiviral immune responses, including the induction of OV-neutralizing antibodies. Current dogma suggests that the presence of preexisting antiviral neutralizing antibodies in patients, or their development during viral therapy, is a barrier to systemic OV delivery, rendering repeat systemic treatments ineffective. However, we have found that human monocytes loaded with preformed reovirus-antibody complexes, in which the reovirus is fully neutralized, deliver functional replicative reovirus to tumor cells, resulting in tumor cell infection and lysis. This delivery mechanism is mediated, at least in part, by antibody receptors (in particular FcγRIII) that mediate uptake and internalization of the reovirus/antibody complexes by the monocytes. This finding has implications for oncolytic virotherapy and for the design of clinical OV treatment strategies. .

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Oncolytic reovirus enhances rituximab-mediated antibody-dependent cellular cytotoxicity against chronic lymphocytic leukaemia

Cited by 37 publications

References 62 publications

Microheterogeneity of therapeutic monoclonal antibodies is governed by changes in the surface charge of the protein

Microheterogeneity of therapeutic monoclonal antibodies is governed by changes in the surface charge of the protein

Gene expression profiling of hematologic malignant cell lines resistant to oncolytic virus treatment

Antibody-Neutralized Reovirus Is Effective in Oncolytic Virotherapy

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