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Lupus 2002Anti-dsDNA autoantibodies are the hallmark of systemic lupus erythematosus (SLE) and frequently correlate with disease activity. In this study we report the isolation...
Anti-dsDNA autoantibodies are the hallmark of systemic lupus erythematosus (SLE) and frequently correlate with disease activity. In this study we report the isolation and characterization of human anti-Id monoclonal antibody fragments as single-chain Fv fragments (scFv) against anti-dsDNA antibody. The anti-Id monoclonal antibodies, specific for anti-dsDNA antibodies, have been cloned from phage display antibody scFv libraries derived from a patient with SLE. The V gene repertoires were derived from the RNA obtained from the B cells of an SLE patient with anti-Ro/SSA and anti-La/SSB antibodies. Affinity-purified anti-dsDNA antibodies were used for selection of bacterial clones producing specific scFv antibody fragments against anti-dsDNA antibodies and little reactivity with normal IgG and other IgG antibodies by ELISA. The anti-Id antibody recognizes a public idiotope that is broadly cross-reactive with polyclonal and monoclonal anti-dsDNA antibodies. This binding was largely inhibited by dsDNA antigen. The anti-Id antibody inhibited anti-dsDNA binding to dsDNA antigen in immunoassays and in the Crithidia luciliae assay. The anti-Id scFv antibody fragments derived from human genes could modulate the pathogenicity of anti-dsDNA autoantibodies and may have therapeutic implications in SLE. They may also be used as probes in studies of the structure of the idiotype.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Antinuclear; Antibodies, Monoclonal; Crithidia; Fluorescent Antibody Technique, Indirect; Humans; Immunoglobulin Fragments; Immunotherapy; Lupus Erythematosus, Systemic; Peptide Library
PubMed: 12139374
DOI: 10.1191/0961203302lu207oa -
Acta Medica Okayama Jun 2002Anti-idiotype antibodies (Ab2) play an important role in the homeostasis of immune responses and are related to the development and the disease activity of certain...
Anti-idiotype antibodies (Ab2) play an important role in the homeostasis of immune responses and are related to the development and the disease activity of certain autoimmune diseases. The asialoglycoprotein receptor (ASGPR) is considered one of the target antigens in the pathogenesis of autoimmune chronic active hepatitis (AIH). We previously developed a mouse monoclonal antibody (clone 8D7) which recognizes rat and human ASGPR. In this study, to help investigate the anti-ASGPR antibody-anti-idiotype antibody network in patients with AIH, we developed a syngeneic mouse monoclonal Ab2 to the 8D7 anti-ASGPR antibody (Ab1). One clone, designated as 3C8, tested positive for specific reactivity to 8D7-Ab1 and did not bind to other irrelevant immunoglobulins. By competitive inhibition assays, the binding of 8D7-Ab1 to liver membrane extracts, i.e., the crude antigen preparation, was inhibited by 3C8-Ab2 in a dose-dependent manner, and the binding of 8D7-Ab1 to 3C8-Ab2 was inhibited by the liver membrane extracts. In the immunohistochemical analysis, 3C8-Ab2 blocked the specific staining of sinusoidal margins of rat hepatocytes by 8D7-Ab1. These results suggest that 3C8 anti-idiotype antibody recognizes the specific idiotypic determinants within the antigen-binding site of 8D7-Ab1.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Asialoglycoprotein Receptor; Hepatitis, Autoimmune; Hepatocytes; Humans; Hybridomas; Mice; Mice, Inbred BALB C; Rats
PubMed: 12108584
DOI: 10.18926/AMO/31715 -
Viral Immunology 1990One potential strategy for the control of human immunodeficiency virus (HIV) infection is immune network manipulation using anti-idiotypic antibodies: this study was...
Anti-idiotypic antisera raised against monoclonal antibody specific for a p24 gag region epitope detects a common interspecies idiotype associated with anti-HIV responses.
One potential strategy for the control of human immunodeficiency virus (HIV) infection is immune network manipulation using anti-idiotypic antibodies: this study was undertaken to demonstrate experimentally the potential of such an approach which, in a more highly evolved form, could be used for the treatment of the acquired immune deficiency virus (AIDS) and related disorders. Anti-idiotypic antibodies were generated in rabbits against a murine monoclonal antibody identifying an epitope on the p24 gag core protein of HIV. After extensive absorption on affinity columns to remove isotype- and allotype-specific antibodies, the purified anti-idiotypic antibody preparation was shown to have specific complementarity with the immunizing mouse monoclonal antibody. This anti-idiotypic antibody was also shown to recognize a common idiotype associated with HIV-specific antibodies from both humans and chimpanzees infected with the AIDS virus. In addition a group of rats immunized with the anti-Id responded with significant antibody titers to recombinant derived p24 gag. These data indicate that at least a subpopulation of these polyclonal anti-Id antibodies structurally mimics an HIV gag region epitope and suggest that immunoregulation by anti-idiotypic antibodies may have therapeutic utility for the AIDS epidemic.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Blotting, Western; Cross Reactions; Epitopes; Gene Products, gag; HIV Antibodies; HIV Core Protein p24; HIV Infections; Immunoglobulin Idiotypes; Male; Mice; Mice, Inbred BALB C; Pan troglodytes; Rabbits; Rats; Species Specificity; Viral Core Proteins
PubMed: 1694432
DOI: 10.1089/vim.1990.3.99 -
Indian Journal of Experimental Biology Dec 2000Ehrlich tumor expresses the ganglioside GT1b. The plasma of mice with Ehrlich ascites tumor burden also contains GT1b. The structural identity of plasma GT1b was...
Ehrlich tumor expresses the ganglioside GT1b. The plasma of mice with Ehrlich ascites tumor burden also contains GT1b. The structural identity of plasma GT1b was ascertained by a series of enzymatic degradation and mass spectral analysis. Mice were vaccinated with purified plasma GT1b admixed with Freund's adjuvant (FA). Sixty nine percent suppression of Ehrlich ascites tumor growth was observed in vaccinated mice. The suppression was dose-dependent. It is hypothesized that the tumor growth-suppression is a result of immune response to GT1b Humoral immune response to GT1b was demonstrated by passive hemagglutination assay of the sera of vaccinated mice. To test the hypothesis, the mice were administered with rabbit polyclonal anti-GT1b IgM antibody in varying doses and challenged with Ehrlich tumor. A significant reduction in tumor growth (65%) was observed in mice administered with anti-GT1b IgM antibody. Again, the suppression was dose-dependent. To verify further, another batch of mice was immunized with anti-idiotypic antibodies to rabbit anti-GT1b IgM raised in rat. The polyclonal anti-idiotype antibody is expected to carry the structural image of GT1b. In a dose-dependent manner, a maximum of 82% suppression of tumor growth was observed in mice immunized with the anti-idiotype antibody. This observation further strengthened the hypothesis that ganglioside mediated suppression of tumor growth may be a result of immunogenicity of the target ganglioside. This was also supported by positive reaction of the sera of anti-idiotype vaccinated mice with both anti-idiotype antibody and ganglioside GT1b in passive hemagglutination assay. The results favour the therapeutic potential of immunogenic tumor-associated gangliosides.
Topics: Animals; Antibodies, Anti-Idiotypic; Antigens, Neoplasm; Carbohydrate Sequence; Carcinoma, Ehrlich Tumor; Dose-Response Relationship, Immunologic; Gangliosides; Immunization; Immunoglobulin M; Male; Mice; Molecular Sequence Data; Rabbits
PubMed: 11411041
DOI: No ID Found -
Immunology Today Oct 1990Anti-metatype (Met) antibodies are anti-immunoglobulins that specifically recognize an antibody-liganded active site but lack specificity for either the ligand or the... (Review)
Review
Anti-metatype (Met) antibodies are anti-immunoglobulins that specifically recognize an antibody-liganded active site but lack specificity for either the ligand or the idiotype. As proposed here by Edward Voss, anti-metatype-metatype immunoglobulin interactions may serve as a model for the interaction of the T-cell receptor (TCR) with antigen-MHC (class I/class II) complexes: the anti-metatype immunoglobulin reagent simulates the TCR and the liganded antibody mimics the antigen-MHC complex. Such a model addresses the dilemma of two macromolecules interacting with the same antigenic determinant and may represent a rational approach to improve understanding of the initiation and regulation of the immune response.
Topics: Animals; Antibodies, Anti-Idiotypic; Histocompatibility Antigens; Models, Biological; Receptors, Antigen, T-Cell
PubMed: 2222759
DOI: 10.1016/0167-5699(90)90140-5 -
Anticancer Research 1991The murine monoclonal antibody HRS-3 (Ab1; isotype IgG1-Kappa), that defines the CD30 antigen (m.w. 120,000) expressed by Hodgkin-Reed Sternberg cells was used to...
Idiotype vaccine against Hodgkin's lymphoma: generation and characterization of an anti-idiotypic monoclonal antibody against the Hodgkin-associated (anti-CD 30) monoclonal antibody HRS-3.
The murine monoclonal antibody HRS-3 (Ab1; isotype IgG1-Kappa), that defines the CD30 antigen (m.w. 120,000) expressed by Hodgkin-Reed Sternberg cells was used to generate monoclonal anti-idiotype antibodies (Ab2) in syngeneic BALB/c mice. Ab2 were selected on the basis of their binding to HRS-3 immunoglobulin and F(ab')2 fragments and lack of reactivity with the whole immunoglobulin or F(ab')2 fragments of unrelated monoclonal antibodies of the same isotype and allotype. Such a putative anti-idiotypic Ab2, was designated antibody 12D3 and further characterized. 12D3 bound to the paratope of HRS-3, as determined by a 85% inhibition of binding of biotinylated HRS-3 to the cell surface of the CD30 positive Hodgkin cell line L450, and to semipurified CD30 positive cell lysates thereof at a concentration as low as 50 ng/well. These results demonstrate that 12D3 binds at or near the binding site of HRS-3 to the CD30 antigen. Purified 12D3 was coupled to keyhole limpet hemocyanine and used to immunize BALB/c mice and rabbits in order to obtain an Ab3 which binds to the CD30 antigen. These immune sera inhibited the binding of biotinylated 12D3 with HRS-3. Moreover, they showed binding activity with the CD30 positive L540 Hodgkin cell line as well as with the L540 cell lysates, indicating that an anti-anti-idiotopic antibody (Ab3) shares idiotopes with Ab1 (HRS-3). These data suggest that antibody 12D3 may be useful in the generation of an anti-idiotype vaccine against Hodgkin's lymphomas.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antigens, CD; Antigens, Neoplasm; Hodgkin Disease; Ki-1 Antigen; Mice; Mice, Inbred BALB C; Rabbits; Vaccines
PubMed: 1653553
DOI: No ID Found -
Radiotherapy and Oncology : Journal of... Jul 1992The 38C13 murine B cell lymphoma model was used to study the effect of the preinjection of unlabelled anti-idiotype monoclonal antibody (mAb) on the subsequent...
The 38C13 murine B cell lymphoma model was used to study the effect of the preinjection of unlabelled anti-idiotype monoclonal antibody (mAb) on the subsequent biodistribution of 131I-anti-idiotype mAb. Mice with established tumors received 0-500 micrograms of unlabelled anti-idiotype mAb 24 h prior to the administration of 131I-anti-idiotype (specific), or both 125I-anti-idiotype and 131I-isotype-matched irrelevant control (nonspecific) mAb. Mice were counted daily in a gamma counter and sacrificed at 2-144 h following injection. Mice were dissected and the weight and activity of the animals and organs were measured. Mice were bled periodically and circulating idiotype levels were measured using an ELISA assay. Five hundred micrograms of unlabelled anti-idiotype mAb increased the retention time of the specific but not the nonspecific mAb in all organs and tumor. Following pretreatment with unlabelled mAb, the cumulative tumor/whole body and tumor/normal organ ratios became similar to those of the nonspecific mAb, with concentration ratios (specific/nonspecific mAb) of approximately 1, which persisted until 96 h post injection when circulating idiotype reappears in antigen excess. In the absence of unlabelled mAb there was less retention in tumor and normal tissue. This is presumed to be due in part to decreased levels of circulating 131I-mAb secondary to rapid plasma clearance of antigen-antibody complexes and tumor cell mediated dehalogenation, which results when the specific mAb specifically binds the targeted antigen. Thus, the addition of unlabelled mAb increased the retention by decreasing the specific behavior of the anti-idiotypic antibody.
Topics: Animals; Antibodies, Anti-Idiotypic; Enzyme-Linked Immunosorbent Assay; Female; Iodine Radioisotopes; Lymphoma, B-Cell; Mice; Mice, Inbred C3H; Radioimmunotherapy; Tissue Distribution
PubMed: 1410571
DOI: 10.1016/0167-8140(92)90376-6 -
Journal of Immunology (Baltimore, Md. :... Sep 1985Rabbit reagents previously thought to display specificity for a cross-reactive idiotype on anti-VHa allotype antibody from all tested rabbits have recently been shown to...
A reevaluation of rabbit anti-allotype antibody for the presence of cross-reactive idiotypes. I. A species-specific idiotype on rabbit anti-a1 antibody is recognized by guinea pig anti-IdX antibody.
Rabbit reagents previously thought to display specificity for a cross-reactive idiotype on anti-VHa allotype antibody from all tested rabbits have recently been shown to be contaminated with an induced (latent) molecule similar or identical to the original antigen (rabbit a1 or a2 allotype). In an attempt to circumvent this problem, we have immunized guinea pigs with rabbit anti-a1 allotype antibody to produce heterologous anti-idiotype antibody. The resulting guinea pig antibody (GP anti-R IdX) recognizes anti-a1 antibody from each of 17 immunized rabbits, and in four tested samples reacts with 22 to 100% of the molecules. Neither goat nor guinea pig anti-a1 reacts with the guinea pig anti-R IdX antibody, even though the goat, guinea pig, and rabbit anti-a1 Ab all recognize a similar set of a1 determinants. The reaction between IdX-bearing rabbit anti-a1 and guinea pig anti-R IdX is inhibited by the original antigen (a1 IgG), demonstrating that the IdX is at or near the antigen combining site of anti-a1 antibody. Immunoelectron microscopy of immune complexes supports this conclusion and demonstrates that the reactive site on the GP anti-R IdX is at or near its antigen combining site.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibody Specificity; Antigen-Antibody Complex; Cross Reactions; Epitopes; Guinea Pigs; Immunoglobulin Allotypes; Immunoglobulin Idiotypes; Microscopy, Electron; Rabbits; Species Specificity
PubMed: 2410508
DOI: No ID Found -
Cancer Immunology, Immunotherapy : CII May 2006A murine monoclonal anti-idiotype (Id) antibody, 3H1 has been developed and characterized previously. Anti-Id 3H1 mimics a specific epitope of carcinoembryonic antigen...
A murine monoclonal anti-idiotype (Id) antibody, 3H1 has been developed and characterized previously. Anti-Id 3H1 mimics a specific epitope of carcinoembryonic antigen (CEA) and can be used as a surrogate antigen for CEA. 3H1 induced anti-CEA immunity in different species of animals as well as humans and showed promise as a potential vaccine candidate in phase I/II clinical trials for colon cancer patients. One area of interest to us has been the development of new immune adjuvants that may augment the potency of 3H1 as a tumor vaccine. Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are potent immunostimulatory agents capable of enhancing the Ag-specific Th1 response when used as immune adjuvants. In this study, we have evaluated the efficacy of 3H1 as a tumor vaccine when admixed with a select CpG ODN 1826 in transgenic mice that express human CEA. The vaccine potential of 3H1 was also assessed in the presence of another widely used adjuvant, QS-21. 3H1 coupled to keyhole limpet hemocyanin (KLH) and mixed with Freund's adjuvant (FA) was used as a gold standard in this system. 3H1 vaccination with different adjuvants induced both humoral and cellular anti-3H1, as well as anti-CEA immunity in CEA transgenic mice. The immune sera could lyse CEA-transfected murine colon carcinoma cells, C15 effectively in an antibody-dependent cellular cytotoxicity assay. The anti-CEA antibody responses were somewhat comparable in each adjuvant-treated group of mice, whereas cellular immune responses were significantly greater when CpG was used as an adjuvant. Splenocytes obtained from 3H1-CpG-immunized mice showed an increased proliferative CD4(+) Th1-type T-cell response when stimulated in vitro with 3H1 or CEA and secreted elevated levels of Th1 cytokines (IL-2, IFN-gamma). This vaccine also induced MHC class I antigen-restricted CD8(+) T-cell responses. In a solid tumor model, C15 tumor growth was significantly inhibited by 3H1 vaccinations. In 3H1-CpG-vaccinated mice, the duration of survival was, however, longer compared to the 3H1-QS21-vaccinated mice. These findings suggest that 3H1-CpG vaccinations can break peripheral tolerance to CEA and induce protective antitumor immunity in this murine model transgenic for human CEA.
Topics: Adjuvants, Immunologic; Animals; Antibodies, Anti-Idiotypic; Antigens, Neoplasm; Cancer Vaccines; Carcinoembryonic Antigen; Cell Cycle Proteins; Colonic Neoplasms; Cytoskeletal Proteins; Female; Hemocyanins; Humans; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neoplasm Proteins; Oligodeoxyribonucleotides; Saponins
PubMed: 16044253
DOI: 10.1007/s00262-005-0009-6 -
Clinical Cancer Research : An Official... Aug 1997Carcinoembryonic antigen (CEA) is expressed in a wide variety of adenocarcinomas, and it is well recognized that cancer patients are immunologically "tolerant" to CEA.... (Clinical Trial)
Clinical Trial Randomized Controlled Trial
Carcinoembryonic antigen (CEA) is expressed in a wide variety of adenocarcinomas, and it is well recognized that cancer patients are immunologically "tolerant" to CEA. The purpose of this study was to determine whether we could break immune tolerance to CEA by vaccinating patients with a monoclonal anti-idiotype antibody that is the internal image of CEA and to determine what impact this might have on patient survival. Twenty-four patients with advanced CEA-positive colorectal cancer who failed standard therapies except for two were entered into this Phase Ib trial. One patient was considered not assessable, because on the day of entering into the study, she was diagnosed with acute myelogenous leukemia. Patients were treated with 1, 2, or 4 mg of aluminum hydroxide-precipitated 3H1 anti-idiotype antibody every other week for four injections and then monthly until tumor progression was observed. Immunological monitoring included humoral and cellular idiotypic and CEA responses, and all patients were evaluated for toxicity, response, and survival. Hyperimmune sera from 17 of 23 patients demonstrated an anti-anti-idiotypic Ab3 response, and 13 of these responses were demonstrated to be true anti-CEA responses (Ab1'). The antibody response was polyclonal, and 11 mediated antibody-dependent cellular cytotoxicity. Ten patients had idiotypic T-cell responses, and five had specific T-cell responses to CEA. None of the patients had objective clinical responses, but overall median survival for the 23 evaluable patients was 11.3 months, with 44% 1-year survival (95% confidence interval, 23-64%). Toxicity was limited to local swelling and minimal pain. Anti-idiotype monoclonal antibody 3H1 that mimics CEA was able to break immune tolerance in the majority of treated patients. Overall survival of 11.3 months was comparable to other phase II data with advanced colorectal cancer patients treated with a variety of chemotherapy agents, including irinotecan, with considerably less toxicity. Although it is not clear that the vaccine itself had an impact on survival, this should be determined in a Phase III randomized trial.
Topics: Adult; Aged; Antibodies, Anti-Idiotypic; Antibodies, Heterophile; Antibodies, Monoclonal; Antibody-Dependent Cell Cytotoxicity; Autoantibodies; Cancer Vaccines; Carcinoembryonic Antigen; Colorectal Neoplasms; Disease Progression; Disease-Free Survival; Female; Humans; Lymphocyte Activation; Male; Middle Aged; Neoplasm Metastasis; Patient Selection; Survival Analysis; T-Lymphocytes; Time Factors
PubMed: 9815809
DOI: No ID Found