Therapeutic Use Of Monoclonal Antibodies Research Articles

Krebs-Immuntherapie – wo stehen wir?

Cancer immunotherapy includes passive and active strategies. Passive immunotherapy such as the use of therapeutic monoclonal antibodies, and in a broader sense also of other immunological effector molecules, such as interferon-alpha is clinically established. The efficacy of passive immunotherapy attests to the fact that the immune system can successfully fight cancer. The logical next step is therefore to develop strategies for active immunotherapy, i.e. "vaccines against cancer". This review focuses on the current status of active immunotherapy with respect to clinical application. Although active immunotherapy is still in the experimental stage, the data are highly encouraging and it is expected that vaccination will soon become part of cancer management.

Principles of the therapeutic use of monoclonal antibodies in oncology

Monoclonal antibodies have reached the stage of therapeutic agents, mostly in oncology, as illustrated by their wide use in lymphoma, breast cancer or colorectal cancer. The unravelling of their mechanisms of action and their interactions with their cellular receptors allows us to engineer new classes of therapeutic antibodies with increased efficacy. The identification of some of the tumour escape mechanisms may also help to define new approaches for patient selection and immunomonitoring. The present review addresses these various aspects. To cite this article: W.H. Fridman, C. R. Biologies 329 (2006).

Neue Antikörper in der Krebsbehandlung

Since the development of hybridoma technology in 1975 monoclonal antibodies with pre-defined specificity can be produced. Only twenty years later did it become possible to make therapeutic use of monoclonal antibodies in oncology. To this end it was necessary to attach the antigen-binding site of a mouse antibody onto the scaffold of a human antibody molecule. Such chimeric or "humanized" antibodies may be used in passive immunotherapy without eliciting an immune response. Rituximab and trastuzumab are such humanized antibodies. They are used today routinely in the treatment of malignant lymphoma and breast cancer, respectively. These antibodies are usually used in combination with conventional cytostatic anticancer drugs.

César Milstein: October 8, 1927–March 24, 2002

César Milstein: October 8, 1927–March 24, 2002

César Milstein: October 8, 1927– March 24, 2002

César Milstein: October 8, 1927– March 24, 2002

Modern immunosuppressives

Eds HJ Schuurman , G Feutren & JF Bach . Published Birkhäuser Verlag AG 2001 . ISBN 3-7643-5981-1 This volume is a monograph in the series ‘Milestones in Drug Therapy’ and reviews present knowledge of modern immunosuppressive drugs. After an introductory chapter by Sir Roy Calne and David White, which reviews the history of immunosuppressive drugs, the book is divided into four sections: Immunosuppressive Drugs, Biologicals, New Avenues in Immunosuppression, and Perspectives. The chapter by Calne & White, who have collaborated over many years in the development and clinical application of new immunosuppressive drugs, nicely sets the scene for the remaining chapters. The section on immunosuppressive drugs covers cyclosporin, tacrolimus, sirolimus, and mycophenolate mofetil. The section on Biologicals covers the therapeutic use of monoclonal antibodies, and the targeting of IL-2 receptors with antibodies or chimeric toxins. New Avenues in Immunosuppression looks at emerging therapeutic strategies for the prevention and treatment of chronic allograft rejection, tolerance induction, and gene therapy approaches to immunosuppression. The final section on Perspectives looks at synergy among combinations of immunosuppressive therapies and the use of cyclosporin in the treatment of autoimmune diseases. The authors state in the preface that this monograph has been compiled for the use of clinicians working with immunosuppressive drugs in daily patient care and for researchers in preclinical and clinical studies. As a transplant surgeon, I found the chapters somewhat uneven, some being written with the clinician in mind and others very much for the research worker. On the whole, the chapter in the section on immunosuppressive drugs is well written indicating their pharmacological properties and their clinical benefits are clearly stated. However, the chapter dealing with tacrolimus ends with a short section comparing tacrolimus with cyclosporin, questioning which agent to use but does not satisfactorily answer this question and does not indeed fully discuss the incidence of side-effects of each drug in comparison with the other. The chapter on emerging therapeutic strategies for the prevention and treatment of chronic allograft rejection is well worth reading being crisply written and extremely well referenced. The chapter by Chou & Kahan on the approach to synergy in combination therapy contains complicated mathematics and is clearly written for the research worker although the last few pages of the chapter provide a summary of drug interactions which will be of value to clinicians. The final chapter is an overview on the use of cyclosporin in the treatment of autoimmune disease. This looks at the incidence of adverse effects when using cyclosporin in autoimmune disease, which apparently differ from those observed in clinical transplantation. I found this chapter somewhat out of place in this volume and I think it would also have benefited from some linguistic revision by the editors. Overall, this is a well-referenced and up-to-date volume and there is something in this book for everybody involved in the development and use of immunosuppressive agents. However, there are several other books available to clinicians on the subject of immunosuppressive agents and I suspect that this book will find its niche with the research worker rather than with the clinician.

Infusion reactions associated with the therapeutic use of monoclonal antibodies in the treatment of malignancy.

With the FDA approval of Rituximab in 1998 for the treatment of lymphoma, and Trastuzumab in 1999 for the treatment of breast cancer, monoclonal antibodies were officially added to the therapeutic armamentarium against malignancy. Most of the side effects associated with these agents are due to antigen-antibody interactions on specific cells and tissues. One of the most predictable side effects of these products is a constellation of various systemic effects including flu-like symptoms such as headache, fever, sweats, skin rash, shortness of breath, hypotension, nausea, and asthenia that occurs with the first infusion of such products. Rarely severe hypotension, bronchospasm, and hypoxia and even death have occurred. The pathophysiology of these reactions appears to be secondary to the release of cytokines as the antibodies bind do circulating antigen-expressing cells that are then removed in the reticuloendothelial system of the lungs, spleen and liver. In patients with large numbers of antigen-dense cells that have a high mitotic index, such as prolymphocytic leukemia, mantle cell lymphoma, or lymphosarcoma cell leukemia, there is a risk of true tumor lysis syndrome. One should be particularly cautious when treating patients with high numbers of circulating antigen-expressing cells in the setting of underlying cardiovascular or respiratory disease.

Pharmacology and safety assessment of humanized monoclonal antibodies for therapeutic use.

The humanization of monoclonal antibodies has generated a class of therapeutic products with improved safety, longer half-lives, and greatly diminished immunogenicity. These engineered proteins are highly species specific and in many cases only cross-react in humans. Where there is cross-reactivity in nonhuman primates or other species, it is not always clear that the pharmacologic effects reflect the potential actions in human volunteers or patients. As with other biologic products, the profile of humanized monoclonal antibodies dictates the preclinical strategy. The preclinical programs for the 2 humanized monoclonal antibodies described here, anti-HLA-DR (Hu1D10) and anti-CD3 (HuM291), demonstrate several unique aspects that affected their preclinical development strategy. Hu1D10 binds to a posttranslational form of HLA-DR and recognizes this antigen in some but not all human and nonhuman primates. The second antibody, HuM291, cross-reacts with CD3 only in the chimpanzee, which is not an optimal test species. In addition, a marketed anti-CD3 product exists (OKT3), and in the preclinical development of our antibody during testing of efficacy and safety, we needed to focus on adverse effects that might be similar to those of OKT3. In these studies, the safety, pharmacokinetics, immunogenicity, and pharmacology (B- and T-cell depletion and recovery) of the 2 antibodies were evaluated. The focus in this review is on the safety and pharmacology testing and the current status of each drug.

Monoclonal antibody therapy in rheumatoid arthritis

Monoclonal antibodies bind to their targets with high specificity and therefore have excellent potential as therapeutic agents. Biotechnological advances have allowed the production of large quantities of engineered monoclonal antibodies for therapeutic use. Recent research in rheumatoid arthritis has identified important mediators of synovitis. Monoclonal antibodies targeting these have been tested in clinical trials over the last decade. Anti-cytokine therapies, in particular anti-tumour necrosis factor alpha monoclonal antibodies, suppressed inflammation and produced rapid symptomatic improvement. Anti-lymphocyte monoclonal antibodies produced long-lasting disease suppression in animal models of rheumatoid arthritis. The use of depleting anti-lymphocyte monoclonal antibodies in rheumatoid arthritis had been disappointing as they did not penetrate the synovial joint in sufficient quantity to suppress disease without producing severe and protracted peripheral blood lymphopenia. Consequently, their use in rheumatoid arthritis had been abandoned. In contrast, clinical trials of non-depleting anti-CD4 monoclonal antibodies in rheumatoid arthritis showed that they could suppress synovitis. However, it remains unclear whether they could lead to prolonged disease improvement.

Protein A mimetic peptide ligand for affinity purification of antibodies.

A peptide mimicking protein A for its ability to recognize the Fc immunoglobulin portion has been identified through screening of a synthetic multimeric peptide library. Screening of the multimeric library, composed of randomized synthetic tripeptide tetramers, has been carried out using a very simple assay, measuring the library ability to interfere with the interaction between protein A and biotinylated immunoglobulins, monitored on solid phase using an enzyme-linked immunosorbent assay format. The tetrameric tripeptide identified after three screening cycles was produced in larger amounts and then immobilized in high yield on preactivated solid support for the preparation of affinity columns, which proved useful for a very convenient one-step purification of antibodies directly from crude sera. Antibody purity after affinity purification was close to 95 per cent, as determined by densitometric scanning of sodium dodecyl sulphate-polyacrylamide gel electrophoresis gels of purified fractions, and up to 2 mg of antibody could be purified from 1 ml of peptide-derivatized affinity support. The ligand was stable to treatment with a vast array of sanitation agents, such as ethanol and 0.1 M sodium hydroxide, and to repeated use, thus making the ligand applicability extremely attractive for the purification of monoclonal antibodies for therapeutic use. Column binding selectivity was similar to that of protein A-affinity columns, since immunoglobulin G from several sources (rabbit, goat, sheep, mouse) was conveniently purified, with no detection of leaked ligand fragments in the purified preparations.

Comparison of the production of a human monoclonal antibody against HIV-1 by heterohybridoma cells and recombinant CHO cells: A flow cytometric study

The production of human monoclonal antibodies for therapeutic use is of increasing importance for treatment of viral infections such as AIDS. As human x mouse heterohybridomas rarely reach the growth rates and cell specific production rates of mouse hybridomas the transfection of standard cell lines, such as CHO or BHK, is a promising alternative. This has the additional advantage that the IgG subtype can be changed to suit the desired application. However, the use of a cell line that has not originally developed to produce antibodies, as lymphocytes and myeloma cells have, might have unrecognised drawbacks. This will be especially significant in the case of antibodies as each molecule consists of 4 chains linked by disulphide bonds which require specific intracellular factors to be properly folded and processed (Heavy chain binding protein, Protein Disulfide Isomerase a.o.). In this study we have therefore compared two cell lines: a human x mouse heterohybridoma producing IAM-2F5, a human IgG(3) antibody specific for HIV-1 with neutralising properties and a Chinese Hamster Ovary cell transfected with dihydrofolate reductase and with the heavy and light chain genes of IAM-2F5 modified to IgG(1). From each cell line three subclones were selected with low, medium and high specific production rates. Batch cultures were performed and the following cellular parameters analysed by flow cytometry; 1) total RNA content (translational activity); 2) total protein content; 3) cell cycle phase distribution; 4) concentration of light and heavy chains; 5) concentration of helper proteins such as BiP and PDI. The production rate of heterohybridoma cells was best reflected in the intracellular concentration of kappa chain, while the gamma chain concentration was comparable for all three subclones. In the CHO cells the gamma chain expression and thus gene copy number appeared to be the limiting factor. The GRP78/BiP concentration in CHO remained unchanged in spite of a 5-fold higher concentration of gamma chain in the high producing subclone. The PDI concentration in CHO cells was much lower compared to the heterohybridoma cells, irrespective of production rates.

Immunoregulatory circuits and potential treatment of connective tissue diseases.

Connective tissue diseases are generated by different immunoregulatory alterations. Their better knowledge may lead to new treatment modalities. In systemic lupus erythematosus (SLE), increased IL-10 production by non-T cells might exert an inhibitory effect on Thl CD4+ T cells which would explain the decreased T cell functions observed in these patients. In rheumatoid arthritis (RA) patients, there may be a balance within the synovium, where the local production of IFN-gamma may limit the anti-inflammatory properties of IL- 10, thus leading to chronic damage. This article shows that rational approaches to therapy need to be individualized. In SLE, the potential therapeutic use of monoclonal antibodies to IL-10 seems to be gathering strength, whereas in RA exactly the opposite is contemplated: IL-10 is tried for its potential therapeutic use.

Fibronectine plasmatique humaine. Comparaison de méthodes de préparation d'un concentré à usage thérapeutique à partir de différentes sources

Three methods, successive precipitations, affinity chromatography on immobilized gelatin and immunoaffinity chromatography with monoclonal anti-fibronectin antibodies were optimized and compared in order to be used for large scale preparation of human plasma fibronectin (Fn). The functional properties of the various Fn preparations were investigated by means of two assays: quantitation of the gelatin-binding activity by ELISA and quantitation of the Fn-mediated attachment of fibroblasts on plastic. Functional alterations of the purified Fn were observed when it was isolated by successive precipitations. Both chromatographic methods provide a rapid and convenient way for isolation of pure and functional Fn. Mass production of monoclonal antibodies is too expensive and legislative requirements for the therapeutic use of monoclonal antibodies are limiting factors for the choice of immunopurification as large scale isolation procedure. Plasma Fn can be isolated from different sources: fresh frozen plasma, cryoprecipitate supernatant or by-products from factor VIII preparation. When gelatin-Sepharose chromatography is performed under optimized conditions, fibronectins isolated from these sources show similar properties. Large scale purifications of Fn from a by-product of factor VIII preparation were performed either by gelatin affinity chromatography or by successive precipitations. These two purification methods can be easily scaled-up since the data obtained closely correlate with analytical results. The chromatographic method supplies a higher purified (98 vs 75%) and functional (95 vs 50%) material when compared with successive precipitations. Yield is also higher (50 vs 26%). The starting material undergoes viral inactivation and the affinity purified Fn, sterile, atoxic, apyrogen, which can be freeze-dried without additives fulfils all requirements for an injectable product.

Recombinant gamma interferon provokes resistance of human breast cancer cells to spontaneous and IL-2 activated non-MHC restricted cytotoxicity

Natural and lymphokine activated killer cells (NK and LAK) are believed to play an important role in the control of tumour progression and metastasis. Their specific receptors on tumours cells are still unknown. Several studies suggest that these cells recognise and eliminate abnormal cells with deleted or reduced expression of MHC class I molecules. Previous reports suggest that interferons (IFN), by increasing MHC class I expression on target cells, induce resistance to killing by NK cells. We investigated the role of MHC molecule expression by two human breast cancer cell lines T47D and ZR75-1 in their susceptibility to NK and LAK cells. These two cell lines spontaneously express low levels of HLA class I antigens but no HLA class II molecules. After IFN-gamma treatment they both overexpressed MHC class I and de novo expressed class II molecules as detected by flow cytometry, quantified by a radioimmunoassay and analysed by two-dimensional gel electrophoresis. Opposed to untreated cells these IFN-gamma treated cells were resistant to NK and LAK lysis. Furthermore, preincubation of IFN-gamma treated breast cancer cells with F(ab')2 fragments of monoclonal antibodies to HLA class I and HLA class II molecules was unable to restore lysis. In contrast, several complete monoclonal antibodies including anti-HLA class I and HLA class II induced the lysis of target cells whether or not they had been treated by IFN-gamma. The therapeutic use of monoclonal antibodies directed against antigens expressed on tumour cells (ADCC) in conjunction with interferon therapy should be discussed in lymphokine-based strategies for treatment of cancer patients.

In vivo administration of lymphocyte-specific monoclonal antibodies in nonhuman primates. V. Evidence that humoral immune response to monoclonal antibodies and immunotoxin conjugates abrogates their cytotoxic activity.

Monoclonal antibodies, either alone or conjugated to toxins, hold promise as important therapeutic agents. However, the immune response to these foreign protein agents may markedly limit their therapeutic utility in vivo. We have administered both an interleukin-2 receptor-specific monoclonal antibody (anti-IL-2R) and a CD2-specific monoclonal antibody linked to the ribosome-inactivating protein gelonin to macaque monkeys. The monkeys developed high-titer antibody responses to mouse Ig and, when immunotoxin was administered, to the toxin gelonin. Their antimouse Ig antibody responses were broadly reactive with mouse Ig of differing idiotypes and isotypes. Furthermore, sera from these monkeys blocked the in vitro cytotoxic effect of anti-IL-2R or immunotoxin. This blocking was mediated by both the antimouse Ig and the antigelonin antibodies. Serum from a monkey infused with one CD2-specific monoclonal antibody blocked the in vitro cytotoxicity of two other isotypically different CD2-specific monoclonal antibody conjugates. In addition, this serum blocked the in vitro cytotoxicity of a gelonin-monoclonal antibody conjugate of an unrelated specificity. These data indicate that the immune response to some monoclonal antibodies and toxins might preclude the later use of this class of substances in an individual. Therefore, strategies for the parental therapeutic use of monoclonal antibodies and immunotoxins must take into consideration the possible limiting effects of the humoral immune response to these agents.

Monoclonal antibodies. Future potential in cancer chemotherapy.

Evaluation of the results obtained this far in the therapeutic use of monoclonal antibodies, either unconjugated, or conjugated to radioactive isotopes, chemotherapeutic agent and toxins

Viral inactivation strategies for monoclonal antibodies: DRY heating

The use of hybridoma cell lines to produce monoclonal antibodies for therapeutic use carries with it the risk of transmitting viruses known to contaminate such cell lines. It is shown here that a murine monoclonal IgM can be heated at 80 °C for 72 hours without loss of activity and with significant killing of two types of test virus.

The Diagnostic and Therapeutic Use of Monoclonal Antibodies in Colorectal Cancer

Preliminary trials have illustrated that monoclonal antibodies can be administered safely, tumor binding can be achieved, and sensitive radioimaging may be possible. Additionally, clinical responses in patients with colorectal carcinoma can occur. The limitations of murine monoclonal therapy are now being addressed and in time will be overcome. Monoclonal antibodies offer great promise in the diagnosis and therapy of malignant disease. Further investigation using innovative approaches and in combination with other therapies may help to reach this goal.

The therapeutic use of monoclonal antibodies

The therapeutic use of monoclonal antibodies

Monoclonal antibody analysis of purified and cell-associated tissue factor

Tissue factor (TF), one of the cell-surface initiators of blood coagulation, has been implicated as the major molecule of this type and as a critical controlling molecule in hemostasis, thrombosis and inflammation. Analysis of the expression of human TF by cells has been hampered by the lack of suitable molecular probes. We have prepared a library of twenty-four murine hybridomas which stably secrete monoclonal antibodies to human TF. Based on their characteristics, these monoclonals can be categorized into a minimum of five distinct groups. Twenty-three of the hybridoma antibodies strongly inhibited TF activity, which was attributable to blocking of formation of the bimolecular complex of TF and factor VII. We have used these antibodies to demonstrate directly that TF is the sole high affinity factor VII receptor on an intact cell. We have also demonstrated the immunologic relationship between constitutive and induced expression of the protein responsible for TF-like activity by several cells and tissues. Most of the antibodies were found to inhibit TF activity expressed by other primate species, and the potential in vivo therapeutic use of monoclonal antibodies of differing intramolecular specificity is discussed.