Use Of Anti-idiotypic Antibodies Research Articles

Generation of anti-idiotypic antibodies to detect anti-spacer antibody idiotopes in acute thrombotic thrombocytopenic purpura patients.

In autoantibody-mediated autoimmune diseases, autoantibody profiling allows patients to be stratified and links autoantibodies with disease severity and outcome. However, in immune-mediated thrombotic thrombocytopenic purpura (iTTP) patients, stratification according to antibody profiles and their clinical relevance has not been fully explored. We aimed to develop a new type of autoantibody profiling assay for iTTP based on the use of anti-idiotypic antibodies. Anti-idiotypic antibodies against 3 anti-spacer autoantibodies were generated in mice and were used to capture the respective anti-spacer idiotopes from 151 acute iTTP plasma samples. We next deciphered these anti-spacer idiotope profiles in iTTP patients and investigated whether these limited idiotope profiles could be linked with disease severity. We developed 3 anti-idiotypic antibodies that recognized particular idiotopes in the anti-spacer autoantibodies II-1, TTP73 or I-9, that are involved in ADAMTS13 binding; 35%, 24% and 42% of patients were positive for antibodies with the II-1, TTP73 and I-9 idiotopes, respectively. Stratifying patients according to the corresponding 8 anti-spacer idiotope profiles provided a new insight into the anti-spacer II-1, TTP73 and I-9 idiotope profiles in these patients. Finally, these limited idiotope profiles showed no association with disease severity. We successfully developed 3 anti-idiotypic antibodies that allowed us to determine the profiles of the anti-spacer II-1, TTP73 and I-9 idiotopes in iTTP patients. Increasing the number of patients and/or future development of additional anti-idiotypic antibodies against other anti-ADAMTS13 autoantibodies might allow idiotope profiles of clinical, prognostic value to be identified.

Cancer Antigen Prioritization: A Road Map to Work in Defining Vaccines Against Specific Targets. A Point of View

OPINION article Front. Oncol., 28 June 2012 | https://doi.org/10.3389/fonc.2012.00066

Handbook of Therapeutic Antibodies: Technologies, Emerging Developments and Approved Therapeutics Edited by Professor Stefan Dubel, Wiley-Blackwell. 2009. 1204pp ISBN: 978-3-527-32902-1 http://www.wiley.com/wileyCDA/wileyTitle/productCd-3527329021.html

Handbook of Therapeutic Antibodies: Technologies, Emerging Developments and Approved Therapeutics Edited by Professor Stefan Dubel, Wiley-Blackwell. 2009. 1204pp ISBN: 978-3-527-32902-1 http://www.wiley.com/wileyCDA/wileyTitle/productCd-3527329021.html

Anti-HER2 vaccines: new prospects for breast cancer therapy

Each year, breast cancer accounts for more than 400,000 new cancer cases and more than 130,000 cancer deaths in Europe. Prognosis of nonmetastatic breast cancer patients is directly related to the extent of the disease, mainly nodal spreading and tumor size, and to the molecular profile, particularly HER2 over-expression. In patients with HER2-over-expressing tumors, different studies have shown cellular and/or humoral immune responses against HER2 associated with a lower tumor development at early stages of the disease. These findings have led to the hypothesis that the generation of an anti-HER2 immune response should protect patients from HER2-over-expressing tumor growth. Taken together with the clinical efficiency of trastuzumab-based anti-HER2 passive immunotherapy, these observations allowed to envisage various vaccine strategies against HER2. The induction of a stable and strong immunity by cancer vaccines is expected to lead to establishment of immune memory, thereby preventing tumor recurrence. However, an immunological tolerance against HER2 antigen exists representing a barrier to effective vaccination against this oncoprotein. As a consequence, the current challenge for vaccines is to find the best conditions to break this immunological tolerance. In this review, we will discuss the different anti-HER2 vaccine strategies currently developed; considering the strategies having reached the clinical phases as well as those still in preclinical development. The used antigen can be either composed of tumoral allogenic cells or autologous cells, or specific to HER2. It can be delivered by dendritic cells or in a DNA, peptidic or proteic form. Another area of research concerns the use of anti-idiotypic antibodies mimicking HER2.

Anti-Idiotypic Antibodies and “Tumor-Only” Antigens: An Update

The use of anti-idiotypic antibodies for cancer treatment continues to be a major field of investigation. One ma- jor challenge for tumor immunotherapy is the identification of antigens associated only or preferably with malignant cells. We summarize here some of the most recent preclinical and clinical advances using two targets that can be considered tumor-specific antigens: N-glycolyl (NeuGc)-gangliosides and the idiotype of B cell lymphomas. Recent developments with tumor-associated protein antigens and the role of anti-idiotypic antibodies in autoimmune diseases are also discussed.

Immunomanipulation of appetite and body temperature through the functional mimicry of leptin.

Although current obesity therapies produce some benefits, there is a need for new strategies to treat obesity. A novel proposal is the use of anti-idiotypic antibodies as surrogate ligands or hormones. These anti-idiotypic antibodies carry an internal motif that imitates or mimics an epitope in the antigen (i.e., hormone or ligand). Thus, anti-idiotypic antibodies to several ligands may mimic them in transducing signals when binding to their receptors. We developed an anti-idiotypic polyclonal antibody against the region of a leptin monoclonal antibody that competitively binds leptin, mimicking the active site structure of leptin. To test whether our anti-idiotype could also reproduce leptin functions, we examined food intake, body weight, and colonic temperature in male Wistar rats (n = 9) in response to intracerebroventricular administration of the leptin anti-idiotype. Our leptin anti-idiotype induced a significant reduction in food intake coupled with an increase in body temperature comparable to that of leptin. That is, the intracerebroventricular administration of 8.0 microg of leptin anti-idiotype or 5.0 microg leptin significantly increased colonic temperature (Delta 1.9 +/- 0.11 degrees C and Delta 1.7 +/- 0.12 degrees C, respectively). In addition, both decreased 24-hour food intake (-26.4 +/- 2.4% and -21.9 +/- 2.2%) compared with the control. The gain in body weight was also decreased by acute administration of the anti-idiotype (-1.4 +/- 0.28%) and leptin (-1.1 +/- 0.17%) vs. the phosphate-buffered saline control (1.3 +/- 0.15%). These studies revealed that the leptin anti-idiotype inhibited food intake and enhanced heat production, mimicking leptin's central actions.

Immunologic quantitation of the carcinoma specific human carcinoma antigen in clinical samples.

Based on the cross-reactivity of the human carcinoma antigen, HCA, with epiglycanin, a mouse mammary carcinoma cell surface glycoprotein, HCA has been detected in the tissue and blood of patients with every type of epithelium-derived cancer tested. Competitive binding assays utilized the following antiepiglycanin antibodies: a polyclonal rabbit antiserum (immunoglobulin [Ig] G and IgM) in a radioimmunoassay; mouse monoclonal antibodies (Ab-1, IgM) on immunoplates; anti-idiotypic (Ab-2) and anti-anti-idiotypic (Ab-3) monoclonal antibodies (both IgG) from spleen cells of C57BL mice immunized, respectively, with Ab-1 and Ab-2, and utilized on immunoplates. IgG and IgM antibodies were evaluated for their ability to detect HCA and to distinguish between the blood of patients with, or without, carcinomas. Assays with the rabbit antiserum distinguished plasmas of metastatic breast carcinoma patients from those of patients with benign breast disease with a sensitivity of approximately 93% (specificity 90%). Antiepiglycanin IgM monoclonal antibodies (i.e., AE3) showed high specificity and sensitivity (> 90%) with sera from advanced carcinoma patients when compared with normal sera. The IgG anti-anti-idiotypic (Ab-3) monoclonal antibodies (i.e., AF2), which bind the same epitope as Ab-1, appear to possess less nonspecific binding capacity, however, than the Ab-1 (IgM) antibodies. Anti-Ab-1 (i.e., C8F2) anti-idiotypic monoclonal antibodies, which bear an idiotope equivalent to the epitope present in epiglycanin and the HCA, demonstrated greater consistency as a standard calibrator and for coating wells than epiglycanin. The concentration of the HCA in the body fluids of patients with carcinomas may be accurately determined by competitive binding assays. It is suggested that the use of anti-idiotypic antibodies (IgG), rather than epiglycanin/HCA, and Ab-3 anti-anti-idiotypic antibodies (IgG), rather than Ab-1 (IgM), will improve the consistency, as well as the sensitivity and specificity, of the assay.

Specific detection of myeloma plasma cells using anti-idiotypic single chain antibody fragments selected from a phage display library.

Bone marrow plasma cells constitute the bulk of malignant cells in multiple myeloma patients. B-lymphocytes having immunoglobulin heavy chain gene rearrangements identical to those of the malignant clone (clonally related B-lymphocytes) may function as malignant plasma cell precursors. We and others proposed the use of anti-idiotypic antibodies to isolate and study clonally related B-lymphocytes. This strategy failed until now because anti-idiotypic antibodies raised by conventional hybridoma techniques proved to react frequently with epitopes shared by different idiotypes. Recently, we succeeded in selecting specific single chain Fv antibodies from phage libraries. To select single chain Fv bearing phages specifically directed against the immunoglobulin idiotype expressed by myeloma tumor cells we panned a semisynthetic phage library against purified myeloma paraprotein Fab fragments. The selection was performed in the presence of soluble polyclonal immunoglobulin as a competitor. Three independent selections for three myeloma patients yielded 10-26 clones. Between two and seven of the selected clones were reactive with patient Fab and not with polyclonal immunoglobulin in enzyme-linked immunosorbent assays. Five out of six anti-idiotypic single chain Fvs were able to specifically stain fixed monoclonal plasma cells in myeloma bone marrow. Idiotype specificity of these single chain Fvs was confirmed by flow cytometry since they did not react with monoclonal plasma cells of other patients, a panel of nine myeloma cell lines, isolated polyclonal bone marrow plasma cells and cultured B-lymphocytes. Using these anti-idiotypic reagents we were able to detect 25 myeloma plasma cells in a background of 50000 immunoglobulin isotype-matched cells of the myeloma cell line UM-1 or 50000 donor bone marrow cells (sensitivity 0.05%). This paper shows that highly specific anti-idiotypic single chain Fv antibody fragments selected from a phage display library can be used to detect rare idiotypic cells in patient samples.

The use of anti-idiotypic antibodies for the correction of autoimmune reactions to brain protein S-100

The possibility of eliminating hyperproduction of antibodies to brain protein S-100 in immunized mice through specific immunocorrection by anti-idiotypic antibodies to this protein or their Fab fragments is explored. Anti-idiotypic antibodies and their Fab fragments inhibit humoral immune response to S-100, which correlates with normalization of several behavior responses.

The Treatment of Factor VIII Inhibitors ‐ A General Overview

The management of patients with classic hemophilia who develop inhibitory antibodies against transfused factor VIII has been the frequent subject of international and national conferences. While treatment of such patients cannot be considered as standardized, several therapeutic approaches are currently available. These include the induction of immune tolerance, removal or eradication of inhibitory antibodies by immune adsorption and/or immune suppression, neutralization of the inhibitor by porcine factor VIII or high doses of human factor VIII, and bypassing of the inhibitor using recombinant factor VIIa or other bypassing agents. In addition to currently available therapeutic strategies, other approaches are under investigation, including the use of selected factor VIII peptides that block the inhibitor; the use of immunologically mute factor VIII products, for example a human-porcine factor VIII chimera not recognized by factor VIII inhibitors, and the use of anti-idiotypic antibodies. The ability to recognize those patients destined to develop inhibitors is also under active investigation.

The Treatment of Factor VIII Inhibitors - A General Overview

The management of patients with classic hemophilia who develop inhibitory antibodies against transfused factor VIII has been the frequent subject of international and national conferences. While treatment of such patients cannot be considered as standardized, several therapeutic approaches are currently available. These include the induction of immune tolerance, removal or eradication of inhibitory antibodies by immune adsorption and/or immune suppression, neutralization of the inhibitor by porcine factor VIII or high doses of human factor VIII, and bypassing of the inhibitor using recombinant factor VIIa or other bypassing agents. In addition to currently available therapeutic strategies, other approaches are under investigation, including the use of selected factor VIII peptides that block the inhibitor; the use of immunologically mute factor VIII products, for example a human-porcine factor VIII chimera not recognized by factor VIII inhibitors, and the use of anti-idiotypic antibodies. The ability to recognize those patients destined to develop inhibitors is also under active investigation.

Subpopulations of tau interact with microtubules and actin filaments in various cell types.

It has been demonstrated that microtubule-associated proteins (MAPs) interact with tubulin in vitro and in vivo. However, there is no clear evidence on the possible roles of the interactions of MAPs in vivo with other cytoskeletal components in maintaining the integrity of the cell architecture. To address this question we extracted the neuronal cytoskeleton from brain cells and studied the selective dissociation of specific molecular isospecies of tau protein under various experimental conditions. Tau, and in some cases MPA-2, were analysed by the use of anti-idiotypic antibodies that recognize epitopes on their tubulin binding sites. Fractions of microtubule-bound tau isoforms were extracted with 0.35 M NaCl or after the addition of nocodazole to allow microtubule depolymerization. Protein eluted with this inhibitor contained most of the assembled tubulin dimer pool and part of the remaining tau and MAP-2. When the remaining cytoskeletal pellet was treated with cytochalasin D to allow depolymerization of actin filaments, only tau isoforms were extracted. Immunoprecipitation studies along with immunolocalization experiments in cell lines containing tau-like components supported the findings on the roles of tau isospecies as linkers between tubulin in the microtubular structure with actin filaments. Interestingly, in certain types of cells, antibody-reactive tau isospecies were detected by immunofluorescence with a discrete distribution pattern along actin filaments, which was affected by cytochalasin disruption of the actin filament network. These results suggest the possible in vivo roles of subsets of tau protein in modulating the interactions between microtubules and actin filaments.

Immunomodulation of the growth hormone — IGF-I axis

Abstract Immunomodulation of the growth hormone/insulin-like growth factor-I (GH/IGF-I) axis has huge potential for increasing animal production. Treatment with GH is known to increase somatic growth, muscle protein synthesis, efficiency of food utilisation and serum IGF-I levels, and to decrease carcass fat content in laboratory rodents and/or farm animals. In addition GH is also known to substantially enhance milk production in both cattle and sheep. All of these actions of GH are substantially enhanced if complexed with certain monoclonal antibodies (MAbs). The phenomenon of Mab-mediated enhancement of GH action offers three potential advantages over simple hormone treatment. Firstly, it would be difficult for GH alone to achieve the level of response promoted by GH/MAb complexes without repeated injections (three or four times per week) of massive amounts of hormone. Secondly, it is possible to produce a site-directed antiserum capable of enhancing endogenous GH activity by immunisation of livestock with an appropriate peptide, this would require only two or three injections of minute amounts of a biologically inactive peptide. Thirdly, using an active immunisation approach means that both the active constituents, namely, hormone and antibody are produced endogenously. Peptides capable of producing GH enhancing antisera have been successfully selected on the basis of computer prediction programmes and by mapping the epitope defined by a GH enhancing MAb. Current work involves investigating analogues of this epitope peptide in order to increase antigenicity without loss of specificity. Whether it will be possible to apply this approach to enhancement of IGF-I activity is as yet unknown. The mechanism(s) underlying this phenomenon is not known, although the observation that a MAb can enhance the effects of GH on chondrocyte proliferation in vitro suggests a cellular rather than a systemic mechanism. The one report of antibody-mediated enhancement of IGF-I activity utilised a polyclonal antiserum, thus it is possible that this may operate via a different mechanism. Alternative strategies such as immunoneutralisation of somatostatin and the use of antiidiotypic antibodies as GH mimics have also been proposed as a means of increasing animal production. However, to date MAb-mediated enhancement of hormone action seems to offer the best alternative to hormone treatment, whether this promise is fulfilled remains to be seen.

Determination of the immunoreactivity of radiolabelled monoclonal antibody fragments by binding to immobilised antiidiotypic antibodies

The feasibility of using immobilised anti-combining site (anti-idiotypic) antibodies as targets for assessing the immunoreactivity of radiolabelled anti-tumour monoclonal antibodies has been assessed. With two anti-tumour monoclonal antibodies (anti-CEA and anti-gp72) it was possible to quantify binding of their 125I labelled Fab fragment preparations to their anti-idiotypic monoclonal antibodies immobilised on cyanogen bromide activated Sepharose. Binding was specific for immobilised anti-idiotypic antibodies reactive with the anti-tumour antibody fragments. Moreover binding was inhibited by unlabelled Fab or intact monoclonal antibody, but not by an irrelevant antibody or its Fab fragment. The use of antiidiotypic antibodies for quantifying immunoreactions of radiolabelled antibodies has advantages over the use of initial target antigen, which may be available only in inconvenient forms, such as cultured tumour cells.

Induction of delayed-type hypersensitivity responses by monoclonal anti-idiotypic antibodies to tumor cells expressing carcinoembryonic antigen and tumor-associated glycoprotein-72.

The use of anti-idiotypic antibodies as immunogens represents one potential approach to active specific immunotherapy of cancer. Two panels of syngeneic monoclonal anti-idiotypic antibodies were generated. One panel was directed against mAb CC49 and the other to mAb COL-1. mAb CC49 recognizes the pancarcinoma antigen (Ag), tumor-associated glycoprotein-72 (TAG-72), and mAb COL-1 recognizes carcinoembryonic antigen (CEA). Seven anti-idiotypic (AI) antibodies (Ab2) designated AI49-1-7 were generated that recognize the variable region of mAb CC49. These mAb were shown to inhibit the interaction of mAb CC49 (Ab1) with TAG-72 (Ag). Five anti-idiotypic antibodies designated CAI-1-5 were also generated to the anti-CEA mAb, COL-1 (Ab1). These Ab2 were shown to inhibit the interaction between COL-1 (Ab1) and CEA (Ag). Immunization of mice, rats, and rabbits with Ab2 directed against CC49 or COL-1 could not elicit specific Ab3 humoral immune responses, i.e., antibody selectively reactive with their respective target antigens. However, immunization of mice with the CC49 anti-idiotypic antibody (Ab2), designated AI49-3, could induce a delayed-type hypersensitivity response (DTH) specific for tumor cells that express TAG-72. Similarly, immunization of mice with an anti-idiotypic antibody directed against COL-1, designated CAI-1, could induce specific DTH cell-mediated immune responses to murine tumor cells that express human CEA on their surface. These results thus demonstrate that while some anti-idiotype mAb may not be potent immunogens in eliciting Ab3 humoral responses, they are capable of eliciting specific cellular immune responses against human carcinoma-associated antigens. This type of mAb may ultimately be useful in active immunotherapy protocols for human carcinoma.

Idiotype and anti-anti-idiotype antibodies to Neisseria gonorrhoeae lipooligosaccharides with bactericidal activity but no cross-reactivity with red blood cell antigens.

A panel of murine mAb against three different epitopes of Neisseria gonorrhoeae lipooligosaccharides (LOS) was developed. Only one of these, 3G5, displays bactericidal activity against all in vitro serum-resistant strains of N. gonorrhoeae. Evidence suggests that sialylation, which could occur in vivo, modifies some LOS epitopes in such a way that the strains become resistant to bactericidal activity and are no longer recognized by specific antibodies. The epitope recognized by our bactericidal mAb is not affected by sialylation as shown by immunoblot analysis. We also provided evidence that the 3G5 epitope is different from RBC precursor Ag, since our mAb did not induce RBC agglutination. Since LOS induce an immune response in a T cell-independent fashion and are highly toxic, they cannot be used for immunization. Use of anti-idiotypic antibodies (aId) could be a way to bypass these difficulties. Therefore, in the present study, aId were produced in rabbits and rendered idiotype-specific by appropriate adsorption. These aId specifically bind to the relevant Id but not to LOS, and inhibit only the binding of anti-LOS mAb (3G5) to LOS preparations from N. gonorrhoeae in a dose-response fashion. The specificity of our aId for the binding site of anti-LOS mAb is suggested by the binding inhibition of affinity-purified aId to Id by LOS. In addition, the capacity of aId to inhibit bactericidal activity of this anti-LOS mAb and the idiotypic cross-reactivity between rat and mouse anti-LOS antibodies support this point. Finally, the elicitation of anti-LOS activity with bactericidal activity upon immunization of naive mice with aId confirms the internal image properties of the aId. These data suggest that a bactericidal mAb suitable for immunoprotection was obtained, and the production of aId opens the door for development of a vaccine.

Induction of an immune response through the idiotypic network with monoclonal anti-idiotype antibodies in the carcinoembryonic antigen system.

Anti-idiotype antibodies can mimic the conformational epitopes of the original antigen and act as antigen substitutes for vaccination and/or serological purposes. To investigate this possibility concerning the tumor marker carcinoembryonic antigen (CEA), BALB/c mice were immunized with the previously described anti-CEA monoclonal antibody (MAb) 5.D11 (AB1). After cell fusion, 15 stable cloned cell lines secreting anti-Ids (AB2) were obtained. Selected MAbs gave various degrees of inhibition (up to 100%) of the binding of 125I-labeled CEA to MAb 5.D11. Absence of reactivity of anti-Id MAbs with normal mouse IgG was first demonstrated by the fact that anti-Id MAbs were not absorbed by passage through a mouse IgG column, and second because they bound specifically to non-reduced MAb 5.D11 on Western blots. Anti-5.D11 MAbs did not inhibit binding to CEA of MAb 10.B9, another anti-CEA antibody obtained in the same fusion as 5.D11, or that of several anti-CEA MAbs reported in an international workshop, with the exception of two other anti-CEA MAbs, both directed against the GOLD IV epitope. When applied to an Id-anti-Id competitive radioimmunoassay, a sensitivity of 2 ng/ml of CEA was obtained, which is sufficient for monitoring circulating CEA in carcinoma patients. To verify that the anti-Id MAbs have the potential to be used as CEA vaccines, syngeneic BALB/c mice were immunized with these MAbs (AB2). Sera from immunized mice were demonstrated to contain AB3 antibodies recognizing the original antigen, CEA, both in enzyme immunoassay and by immunoperoxidase staining of human colon carcinoma. These results open the perspective of vaccination against colorectal carcinoma through the use of anti-idiotype antibodies as antigen substitutes.

The use of anti-idiotypic antibodies as a strategy for studying glutamate receptors.

The use of anti-idiotypic antibodies as a strategy for studying glutamate receptors.

Chapter 1 The Use of Antiidiotype Antibodies for the Characterization of Protein—Protein Interactions

Publisher Summary This chapter outlines the theoretical basis for the analysis of protein–protein interactions using network antibodies, considers the advantages and limitations of the approach, and discusses two experimental systems that illustrate many of these points. An immunological approach to problems of protein–protein interactions is presented. Two successful uses of this approach to explore protein–protein interactions, which are difficult to characterize by classic methods, are described in detail. The paired in vitro immunization approach has the advantages of speed and simplicity and results in the production of monoclonal antibodies, rather than highly variable polyclonal antiidiotype antisera. The antiidiotype approach is particularly suited to exploring the interactions of the defined sorting and localization signals that have been characterized by the studies of protein targeting. Although there are limitations imposed by the structure of the antigen-combining site of an immunoglobulin, the method is a valuable and widely applicable tool for understanding the cell biology of signals in proteins.

Anti-idiotypic antibodies to feline leukemia virus: an approach for retroviral immunization strategies.

The present study describes an approach to the development and use of anti-idiotypic antibodies as a possible immunization strategy to prevent retroviral infection. The rationale for using anti-idiotypes (anti-Ids) to try to elicit an antigenic-specific immune response is examined, and the production and characterization of polyclonal and monoclonal anti-Ids are described. Several techniques were used to determine antigenic mimicry and anti-Id subtypes. The potential use of anti-Ids in feline leukemia virus (FeLV) receptor studies and vaccine trials in vivo were investigated. Results from these studies suggest that the anti-Id strategy is feasible for the FeLV model. Polyclonal Ab2 reagents were developed that blocked virus-receptor binding and thus inhibited viral infection in vitro and induced humoral immune responses in 6- to 8-week old kittens characterized by production of Ab3 with the ability to bind the original FeLV envelope protein gp70 as assessed by Western blot analysis.