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Journal of Autoimmunity Feb 1993Idiotype-anti-idiotype interactions were investigated in the sera of patients with primary biliary cirrhosis (PBC), which is characterized by the presence of circulating...
Idiotype-anti-idiotype interactions were investigated in the sera of patients with primary biliary cirrhosis (PBC), which is characterized by the presence of circulating anti-mitochondrial antibodies (AMA). A mouse monoclonal antibody, CPZ674, has been raised to the E2 component of the pyruvate dehydrogenase complex (PDC), which is the target antigen of AMA. CPZ674 recognizes one of the epitopes recognizable by AMA, as demonstrated by competitive Western blotting. Anti-idiotypic antibodies in PBC sera were detected either by their specific binding to CPZ674 in an ELISA or by the formation of idiotype-anti-idiotype complexes with CPZ674, detected using chromatography on a Sephacryl-300 column and by a polyethylene glycol precipitation method. The specificity of the anti-idiotypic antibodies to AMA was shown by their ability to inhibit the binding of AMA to PDC, but not the binding of other autoantibodies to their relevant autoantigens. We have therefore produced evidence for the existence of idiotype-anti-idiotype interactions in PBC, but whether these anti-idiotypic antibodies are involved in the control of AMA is unknown.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibody Specificity; Autoantibodies; Humans; Liver Cirrhosis, Biliary; Mice; Mice, Inbred C57BL; Mitochondria; Pyruvate Dehydrogenase Complex
PubMed: 8457288
DOI: 10.1006/jaut.1993.1008 -
Journal of Immunology (Baltimore, Md. :... Sep 1984We previously described a monoclonal antibody, 9.1C3, which blocked natural killer (NK) cell-mediated cytolysis by acting on effector cells during a late step in the...
We previously described a monoclonal antibody, 9.1C3, which blocked natural killer (NK) cell-mediated cytolysis by acting on effector cells during a late step in the lethal hit stage. The present work describes the production in rabbits of anti-idiotypic (anti-id) antibodies to the 9.1C3 antibody. In addition to reacting specifically with the 9.1C3 antibody, the anti-id antibodies bound strongly to the K562 target cell. The anti-id antibodies blocked killing of K562 targets by NK, antibody-dependent cellular cytotoxicity, and NK-like cells but did not inhibit killing by cytotoxic T lymphocytes (CTL). Pretreatment of cells and washing before assay indicated that blocking occurred at the target cell level. Of particular interest, single cell assays with Percoll-enriched large granular lymphocytes demonstrated that the antibodies caused no reduction in binding. These data are consistent with a model for NK cell-mediated lysis that involves a secondary target cell receptor independent of the primary NK-target cell interaction. The anti-id antibodies immunoprecipitated cell surface proteins of relative m.w. 79K and 62K unreduced, and 94K and 79K reduced from K562 target cells. The development of anti-id antibodies may be a useful procedure to explore the structure and function of cellular receptors involved in NK cell-mediated cytolysis.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antigen-Antibody Reactions; Antigens, Surface; Antilymphocyte Serum; Binding Sites, Antibody; Binding, Competitive; Chemical Precipitation; Cytotoxicity, Immunologic; Humans; Immunity, Cellular; Immunoglobulin Idiotypes; Killer Cells, Natural; Rabbits
PubMed: 6611373
DOI: No ID Found -
MAbs 2012Multispecificity is not a well-understood property of some antibodies. Different functions have been attributed to multispecific natural antibodies, commonly associated...
Multispecificity is not a well-understood property of some antibodies. Different functions have been attributed to multispecific natural antibodies, commonly associated with the neutralization and clearance of antigens. Much less is known about the role of antibodies like these, based on their idiotypic connectivity. B7Y33 is a chimeric IgG1 version of a polyreactive α anti-idiotype antibody that is able to interact with different immunoglobulin and non-immunoglobulin antigens. Here we report the capacity of this antibody to enhance the immunogenicity of several autologous IgMs in adjuvant-free conditions. Our results suggest that the formation of immune complexes seems to be necessary, but not sufficient, to this activity. The potential involvement of the interaction of B7Y33 with the FcγRIIb is discussed.
Topics: Antibodies, Anti-Idiotypic; Autoantigens; Epitopes; Humans; Immunoglobulin G; Immunomodulation; Immunotherapy; Protein Binding; Receptors, IgG; Recombinant Fusion Proteins
PubMed: 22531446
DOI: 10.4161/mabs.19872 -
Cellular Immunology Jul 1989A rat monoclonal antibody (MoAb), termed KBA, against mouse lymphokine-activated killer (LAK) cells recognizes a LAK cell surface molecule termed LAA responsible for the...
A rat monoclonal antibody (MoAb), termed KBA, against mouse lymphokine-activated killer (LAK) cells recognizes a LAK cell surface molecule termed LAA responsible for the binding between LAK and target cells. In order to identify a target molecule of LAK cells, we prepared anti-KBA idiotype antibodies (anti-KBA-Id) from rabbit anti-KBA sera. Immunoglobulins were separated by ammonium sulfate precipitation followed by sequential affinity column chromatographies using Affi-gel coupled with rat MoAbs other than KBA and KBA-coupled gel. An immunoglobulin(s) in a KBA-gel-bound fraction showed the selective reactivity to KBA, comprising anti-KBA-Id character. This anti-KBA-Id inhibited the binding of KBA to LAK. Moreover, it bound with a portion of mouse leukemia cells sensitive to LAK cells, but not with normal mouse cells, and inhibited the binding of LAK cells to a target leukemia. These findings indicate that the anti-KBA-Id contain anti-Id which possess a three-dimensional structure that mimics a mirror image of the antigen (LAA)-combining site in KBA or the structure of LAA. The antigen reactive with anti-KBA-Id was characterized as a glycoprotein.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antigens, Surface; Immunoglobulin Idiotypes; Killer Cells, Natural; Lymphokines; Male; Mice; Mice, Inbred C57BL; Rabbits
PubMed: 2786755
DOI: 10.1016/0008-8749(89)90020-8 -
Journal of Immunological Methods Dec 2013Human/mouse chimeric monoclonal antibody (mAb) ch14.18 is directed against disialoganglioside GD2 and has demonstrated activity and efficacy in high-risk neuroblastoma...
Human/mouse chimeric monoclonal antibody (mAb) ch14.18 is directed against disialoganglioside GD2 and has demonstrated activity and efficacy in high-risk neuroblastoma (NB). For the purpose of industrial production, ch14.18 was manufactured in Chinese hamster ovarian cells (ch14.18/CHO) in order to facilitate clinical trials in Europe. To determine immunopharmacological effects of ch14.18 in preclinical models and clinical trials, a validated method of quantitative detection of ch14.18/CHO in serum is an important tool. We recently described the generation and characterization of ganglidiomab, a monoclonal anti-idiotype Ab (AIT) of ch14.18 (Lode et al., 2013), which was used to establish quantitative and validated enzyme-linked immunosorbent assay (ELISA) methods using ganglidiomab as a capture mAb. With these ELISA methods, we first demonstrated binding of ch14.18/CHO to ganglidiomab to a similar extent as to the nominal antigen GD2 and in contrast to GD1b and GM2 precursor and metabolite gangliosides, used as negative controls. In order to determine both low (0.5-3.1 μg/ml) and high levels of ch14.18/CHO (1.0-25 μg/ml) in the serum of NB patients treated with ch14.18/CHO, we established two ELISA methods with high and low sensitivity using 1/1001, and 1/5126 sample dilutions, respectively. For validation, we used a set of tailored quality controls (QC) containing distinct concentrations of ch14.18/CHO (1.0, 2.0, 7.0, and 20.0 μg/ml). We determined the limit of detection (LOD) for both ELISA methods to be 0.50 μg/ml for the high sensitivity and 1.02 μg/ml for low sensitivity ELISA. The within-assay precision was 12% for high and 4% for low sensitivity ELISA, and the coefficients of variation (CV) were under 20% for all assays (3% for QC-1.0, 5% for QC-2.0, 7% for QC-7, and 3% for QC-20). With this method, we showed that neither eight freeze-thaw cycles nor storage at room temperature for up to 168 h affected ch14.18/CHO stability in serum. Finally, we analyzed ch14.18 Ab serum levels in selected NB patients receiving ch14.18/CHO as a continuous or bolus infusion with a peak concentration at the last day of Ab application (17.14 ± 7.20mg/ml with continuous and 19.78 ± 2.26 mg/ml with bolus infusion). In summary, we describe validated ELISA methods using ganglidiomab as a capture mAb suitable for the pharmacological evaluation of ch14.18/CHO in NB patients.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Cricetinae; Enzyme-Linked Immunosorbent Assay; Gangliosides; Humans; Neuroblastoma; Sensitivity and Specificity
PubMed: 24055592
DOI: 10.1016/j.jim.2013.09.008 -
Annals of the Rheumatic Diseases Jan 2013Millions of patients worldwide are treated with therapeutic monoclonal antibodies. These biological therapeutics can be immunogenic, resulting in anti-drug antibody...
OBJECTIVES
Millions of patients worldwide are treated with therapeutic monoclonal antibodies. These biological therapeutics can be immunogenic, resulting in anti-drug antibody formation which leads to loss of response. Fully human biological agents, such as the anti-tumour necrosis factor α (anti-TNFα) antibody adalimumab, are considered to be weakly immunogenic, but anti-adalimumab antibodies (AAA) were recently detected in more than half of treated patients with rheumatoid arthritis (RA) within 28 weeks of treatment. A study was undertaken to determine the mechanism by which AAA lead to loss of response.
METHODS
The specificity of the repertoire of AAA was investigated in a cohort of 50 AAA-positive RA patients. Inhibition experiments using TNFα and patient-derived anti-adalimumab monoclonal antibodies were performed.
RESULTS
The antibody response against adalimumab is highly restricted: Fab fragments of a single monoclonal antibody specific for the idiotype of adalimumab inhibited 98.65% (25th-75th percentiles: 98.25-99.90) of the total anti-adalimumab reactivity in serum from 50 AAA-positive patients. The anti-adalimumab response was confined to the TNFα binding region of adalimumab, thereby neutralising its therapeutic efficacy. In line with this restricted specificity, small immune complexes were found in the circulation of AAA-forming patients.
CONCLUSIONS
The humoral immune response against adalimumab is highly restricted and limited to the idiotype of the therapeutic antibody. All antibodies result in functional neutralisation of the drug, thereby providing a mechanism by which AAA formation leads to clinical non-response.
Topics: Adalimumab; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal, Humanized; Antibodies, Neutralizing; Antibody Specificity; Antigen-Antibody Complex; Antirheumatic Agents; Arthritis, Rheumatoid; Enzyme-Linked Immunosorbent Assay; Humans
PubMed: 22759910
DOI: 10.1136/annrheumdis-2012-201445 -
Journal of Immunotherapy (Hagerstown,... Jan 1998The anti-idiotype (Id) monoclonal antibody (mAb) 1A7 immunoglobulin G1 (IgG1, kappa), raised in syngeneic mice against the murine anti-ganglioside GD2 mAb 14G2a mimics a...
The anti-idiotype (Id) monoclonal antibody (mAb) 1A7 immunoglobulin G1 (IgG1, kappa), raised in syngeneic mice against the murine anti-ganglioside GD2 mAb 14G2a mimics a carbohydrate epitope on GD2 and serves as a surrogate protein antigen for this disialoganglioside. Immunization of allogeneic C57BL/6 mice and rabbits with 1A7 induced anti-GD2 antibodies of IgG isotype that recognize purified GD2 by enzyme-linked immunosorbent assay (ELISA) and GD2-positive human melanoma cells (M21/P6) by fluorescence-activated cell sorter (FACS) analysis. The specificity of the antisera for GD2 was further confirmed by dot-blot analysis. These antisera also specifically lyse GD2-positive M21/P6 target cells in an antibody-dependent cellular cytotoxicity assay. Taken together, these results suggest that the anti-Id 1A7 can induce GD2-specific IgG antibodies that can recognize cell surface-associated as well as soluble disialoganglioside GD2.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibody-Dependent Cell Cytotoxicity; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Gangliosides; Humans; Immunization; Immunoglobulin G; Melanoma; Mice; Mice, Inbred BALB C; Rabbits
PubMed: 9456440
DOI: 10.1097/00002371-199801000-00010 -
Journal of Immunological Methods Nov 1990A new immunohistochemical assay was developed for the detection of human monoclonal antibody (HuMAb) bound to human biopsied tumor tissues. A murine anti-idiotype... (Comparative Study)
Comparative Study
A new immunohistochemical assay was developed for the detection of human monoclonal antibody (HuMAb) bound to human biopsied tumor tissues. A murine anti-idiotype monoclonal antibody, alpha type, 18C6 (IgGl), was raised against an IgM HuMAb, L612, defining a tumor-associated ganglioside antigen (GM3) and used as a probe in a three step cell-binding assay (HuMAb + anti-id + biotinylated anti-mouse Ig). Anti-id 18C6 has an exclusive binding specificity for HuMAb L612, but does not interfere with the binding of L612 to antigen positive melanoma cell lines or to a purified antigen, GM3. The applicability of 18C6 in the three step cell-binding assay was tested first using a melanoma cell line, UCLASO-M12. L612 bound to M12 cells was specifically detected by 18C6 without any background reactivity in ELISA. When this assay was compared with the standard two-step cell-binding assay (HuMAb + peroxidase-conjugated anti-human IgM) using various cultured tumor cell lines, parallel reactivity was observed. The three-step cell-binding assay was then applied to various fresh-frozen human tumor sections. Positive reactivity was demonstrated on various histologic types of human tumor tissues: primary melanoma (10/10), metastatic melanoma (4/4), nevus (10/10), lung cancer (3/6), breast cancer (2/6), and colon cancer (1/1). Adjacent normal tissues were unstained. Control experiments included the cell-binding assay with L612 alone, 18C6 alone. L612 + unrelated mouse IgG, and unrelated IgM HuMAb (L72) + 18C6; but biotinylated anti-mouse IgG did not react with these control preparations. The results indicate that anti-id 18C6 is a highly specific probe to assess the expression of the ganglioside antigenic epitope recognized by the L612 HuMAb on biopsied human tumor tissues.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Biopsy; Enzyme-Linked Immunosorbent Assay; G(M3) Ganglioside; Humans; Immunoenzyme Techniques; Immunohistochemistry; Mice; Mice, Inbred BALB C; Neoplasms; Sensitivity and Specificity; Tumor Cells, Cultured
PubMed: 2230146
DOI: 10.1016/0022-1759(90)90120-k -
Chinese Medical Journal Sep 2001To generate and characterize anti-idiotypic monoclonal antibody (Ab2) that bears the internal image of nasopharyngeal carcinoma (NPC) associated antigen.
OBJECTIVE
To generate and characterize anti-idiotypic monoclonal antibody (Ab2) that bears the internal image of nasopharyngeal carcinoma (NPC) associated antigen.
METHODS
Using NPC monoclonal antibody (Ab1) as immunogen, hybridoma cells were obtained by fusion of SP2/0 myeloma cells with immunized murine spleen cells. Positive clones were screened by Sandwich ELISA and a binding inhibition test. To determine whether Ab2 possess the internal image of the original antigen or not, mice were immunized with Ab2. ELISA and the competitive inhibition assay tested anti-anti-idiotypic antibodies (Ab3) in anti-sera. Cell-mediated immunity to tumors induced by Ab2 was investigated by a delayed-type hypersensitivity response and the mouse T-cell proliferation assay.
RESULTS
Anti-idiotypic monoclonal antibodies against the monoclonal anti-NPC antibodies FC2 and HNL5 were generated that recognize NPC associated antigens. These Ab2, which were designated 2H4 and 5D3, could inhibit the binding of FC2 or HNL5 to NPC cell lines. Anti-sera from the immunized mice, which contained Ab3, could compete with FC2 or HNL5 for binding with NPC cell by a competitive inhibition assay. Mice immunized with 2H4 or 5D3 coupled with keyhole limpet hemocyanin (KLH), showed a positive and specific delayed-type hypersensitivity (DTH) reaction after stimulation by NPC cells. The mouse T cell proliferative assay indicated that there was a significantly higher proliferative response of the splenocytes in the experimental groups than that in control groups.
CONCLUSIONS
Anti-idiotypic antibodies 2H4 and 5D3 are Ab2 beta bearing the internal image of the epitope of NPC associated antigen. Either 2H4 or 5D3 expressing three-dimensional shapes that resemble the structure of natural antigens could induce humoral and cellular immune response.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibody Specificity; Antigens, Neoplasm; Binding, Competitive; Humans; Hypersensitivity, Delayed; Immune Sera; Immunity, Cellular; Mice; Mice, Inbred BALB C; Nasopharyngeal Neoplasms; Tumor Cells, Cultured
PubMed: 11780392
DOI: No ID Found -
MAbs 2010Recombinant antibodies may be engineered to obtain improved functional properties. Functional mapping of the residues in the binding surfaces is of importance for...
Recombinant antibodies may be engineered to obtain improved functional properties. Functional mapping of the residues in the binding surfaces is of importance for predicting alterations needed to yield the desired properties. In this investigation, 17 single mutation mutant single-chain variable fragments (scFvs) of the anti-idiotypic antibody anti-TS1 were generated in order to functionally map amino acid residues important for the interaction with its idiotype TS1. Residues in anti-TS1 determined to be very important for the interaction were identified, Y32L, K50L, K33H, and Y52H, and they were distributed adjacent to a centrally located hydrophobic area, and contributed extensively to the interaction energy (≥2.5 kcal/mol) in the interaction. Quantitative ELISA assays, BIAcore technologies and three-dimensional surface analysis by modeling were employed to visualize the consequences of the mutations. The expression levels varied between 2 - 1,800 nM as determined by ELISA. All the 17 scFvs displayed higher dissociation rates (60 - 1,300 times) and all but two of them also faster association rates (1.3 - 56 times). The decrease in affinity was determined to be 1.6 - 12,200 times. Two of the mutants displayed almost identical affinity with the wild type anti-TS1, but with a change in both association and dissociation rates. The present investigation demonstrates that it is possible to generate a large panorama of anti-idiotypic antibodies, and single out a few that might be of potential use for future clearing and pre-targeting purposes of idiotypic-anti-idiotypic interactions.
Topics: Amino Acid Sequence; Antibodies, Anti-Idiotypic; Antibody Affinity; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Humans; Hybridomas; Immunoglobulin Variable Region; Keratin-8; Kinetics; Molecular Sequence Data; Mutagenesis, Site-Directed; Protein Structure, Quaternary; Single-Chain Antibodies
PubMed: 21124071
DOI: 10.4161/mabs.2.6.13275