Production Of Anti-idiotypic Antibodies Research Articles

Induction of allergen-specific IgE and IgG responses by anti-idiotypic antibodies.

In order to explore idiotypic, anti-idiotypic, and anti-anti-idiotypic responses to allergens, BALB/c mice were immunized with affinity-purified human idiotypic antibodies directed against a highly purified shrimp allergen. This resulted in the production of anti-idiotypic antibodies which were quantitated by using rabbit idiotypic antibodies raised against the same purified allergen. The mouse anti-idiotypic antibodies recognized shrimp-specific human idiotypic antibodies of the IgE isotype from 18 of 20 individuals, and IgG antibodies from 14 of 20 shrimp-sensitive patients. Immunization of BALB/c mice with affinity-purified, allergen-specific anti-idiotypic antibodies induced anti-allergen IgE and IgG responses in the absence of the allergen. This paper thus presents evidence that anti-idiotypic antibodies raised against allergen-specific idiotypic antibodies may substitute for the original allergen in the induction of allergen-specific idiotypic antibodies. The demonstration of shared idiotopes on IgG and IgE antibodies in the sera of shrimp-sensitive patients supports the use of allergen-specific anti-idiotypic antibodies as surrogate allergens.

Protective effects of anti-antiidiotypic IgE antibodies obtained from an IgE monoclonal antibody specific for a 26-kilodalton Schistosoma mansoni antigen.

A rat IgE mAb specific for larval Ag (26 kDa, 56 kDa) has been shown to protect rats against Schistosoma mansoni infection. Immunizations of Lou/M rats performed with this IgE (Ab1) induced the production of antiidiotypic antibodies (Ab2). Moreover, after this Ab2 production, anti-antiidiotypic antibodies (Ab3) were revealed. The screening of Ab3 isotypes showed the presence of IgG Ab3 and more interestingly of IgE Ab3, i.e., the same isotype as Ab1. These IgE and IgG antibodies recognized predominantly the 26-kDa Ag and were cytotoxic for schistosomula in the presence of platelets for IgE Ab3 and eosinophils for IgG Ab3. Both IgE and IgG Ab3 conferred by passive transfer protective immunity to infected rats (up to 50%). Thus the immunization with an IgE mAb led in part to the production of Ab3 of the same isotype as Ab1. In conclusion, these results suggest that the isotype selection of the antibodies of the third generation (Ab3) might be influenced by the Ab1. The respective role of the idiotope and isotype of Ab1 in isotype regulation is discussed.

Inhibition of pregnancy before and after implantation in rats with monoclonal antibody against progesterone

A monoclonal antibody against progesterone completely blocked pregnancy when rats were injected intraperitoneally at 0.53 μmol/kg/day on days 1 and 2, or on day 11 of a first pregnancy. The antibody was equally effective when injected again during a second pregnancy. There was no evidence for anti-idiotypic antibody production even when the injection of a heterologous (mouse) antibody was repeated in a second pregnancy.

In vitro T-cell functions specific to an anti-DNA idiotype and serological markers in patients with systemic lupus erythematosus (SLE)

The human monoclonal autoantibody 16/6 is a common anti-DNA idiotype found to have clinical relevance in patients with systemic lupus erythematosus (SLE). Therefore the ability of peripheral blood T cells of SLE patients and healthy controls to proliferate and to produce helper T-cell factors following stimulation with this idiotype was tested. It was found that T cells of 75% of healthy donors proliferated to the 16/6 idiotype, whereas only 22% of SLE patients responded to this idiotype by proliferation. On the other hand, the capability to produce T-cell helper factors specific to the 16/6 idiotype was found in a higher percentage of SLE patients (48%) as compared to healthy controls (31%). The low frequency of proliferative responses in SLE patients might be due either to the chronic exposure to the 16/6 idiotype or to the production of antiidiotype antibodies against the 16/6 idiotype, which interfere with the response to the latter stimulator.

In vitro production of antiidiotypic antibodies by EBV-transformed B cell lines from renal transplant recipients

In vitro production of antiidiotypic antibodies by EBV-transformed B cell lines from renal transplant recipients

Immunization with Leishmania-specific T cell not B cell lines or hybridomas can modulate the response of susceptible mice infected with viable parasites.

Monoclonal antibodies (mAb), T cell lines, and T or B cell hybridomas were prepared from BALB/c, CBA, or E1 mice infected with Leishmania mexicana. Various mAb were produced which inhibited the growth and motility of parasites in vitro. T cell lines (hybridomas) were screened for their ability to release interleukin 2 on specific antigen exposure. Passive transfer of mAb or T cell lines to infected adult mice caused little perturbation of parasite growth. Recipient naive mice were immunized with purified Ig or irradiated cells from these sources and were subsequently infected with viable parasites. Only preimmunization with T cell lines (hybridomas) led to exacerbation of parasite growth, although enzyme-linked immunosorbent assays could detect the production of anti-idiotype antibodies in mAb (B cell hybridoma)-immunized mice. Either nylon wool-purified T cells or serum Ig from T cell-immunized mice could be used to immunize further naive recipients for protection against parasite growth. These data have implications for the development of anti-idiotype vaccines for Leishmania antigens.

Induction of Immunoglobulin Secretion in Follicular Non-Hodgkin’s Lymphomas: Role of Immunoregulatory T Cells

B cell neoplasms are clonal expansions of B lymphocytes thought to be frozen at various points along the normal B cell differentiation pathway. We studied cell suspensions from lymph nodes involved by follicular (nodular) non-Hodgkin's lymphoma to determine the capacity of the malignant B cells to secrete immunoglobulin (Ig). Neoplastic B cells from all 14 follicular lymphomas secreted monoclonal immunoglobulin in culture when appropriate signals were provided. In most cases, maximal Ig secretion occurred when autologous T cells were removed by E rosette depletion, replaced with allogeneic normal T cells, and the cultures were exposed to 12-O-tetradecanoylphorbol-13-acetate. Autologous T cells exerted a suppressor effect on Ig secretion in 8/8 cases studied, diminishing the response of the malignant B cells to allogeneic T cells. This suppressor effect did not correlate with the percentage of cells staining with anti-Leu-2a or with "helper-suppressor" (Leu-3a-Leu-2a) ratios of the lymph node T cells. Our findings demonstrate that the arrested differentiation of most follicular lymphomas is reversible and implicate a T cell-mediated host immunoregulatory mechanism affecting Ig secretion in vivo. An additional contribution of our results is the demonstration of a cell culture system for synthesis of sufficient monoclonal Ig for use as an immunogen in production of anti-idiotype antibodies.

Induction of immunoglobulin secretion in follicular non-Hodgkin's lymphomas: role of immunoregulatory T cells

B cell neoplasms are clonal expansions of B lymphocytes thought to be frozen at various points along the normal B cell differentiation pathway. We studied cell suspensions from lymph nodes involved by follicular (nodular) non-Hodgkin's lymphoma to determine the capacity of the malignant B cells to secrete immunoglobulin (Ig). Neoplastic B cells from all 14 follicular lymphomas secreted monoclonal immunoglobulin in culture when appropriate signals were provided. In most cases, maximal Ig secretion occurred when autologous T cells were removed by E rosette depletion, replaced with allogeneic normal T cells, and the cultures were exposed to 12-O-tetradecanoylphorbol-13- acetate. Autologous T cells exerted a suppressor effect on Ig secretion in 8/8 cases studied, diminishing the response of the malignant B cells to allogeneic T cells. This suppressor effect did not correlate with the percentage of cells staining with anti-Leu-2a or with “helper- suppressor” (Leu-3a-Leu-2a) ratios of the lymph node T cells. Our findings demonstrate that the arrested differentiation of most follicular lymphomas is reversible and implicate a T cell-mediated host immunoregulatory mechanism affecting Ig secretion in vivo. An additional contribution of our results is the demonstration of a cell culture system for synthesis of sufficient monoclonal Ig for use as an immunogen in production of anti-idiotype antibodies.

Anti-idiotypic antibodies to HLA-DR4 and DR2.

The aim of our study was to determine whether antibodies recognizing epitopes of HLA-DR antigens (idiotypic antibodies or Ab1) induce the production of anti-idiotypic antibodies (Ab2). We tested the capacity of the F(ab')2 fragment obtained from two sera, one with no anti-HLA antibodies (serum ES) and one depleted by absorption of anti-HLA lymphocytotoxins (serum FH), to block the anti-DR antibodies reacting with the HLA-DR antigens of the immunizing donor. The F(ab')2 fragment obtained from serum ES inhibited the anti-DR2 activity of an earlier post-delivery bleeding obtained from the same woman. The anti-idiotypic antibodies contained by this serum also inhibited the anti-DR2 activity of a reference anti-DR2 antiserum 8W907 and of an anti-MT1 antiserum 8W1231. Similarly, the F(ab')2 fragment obtained from serum FH, after absorption of her anti-DR4 antibody, inhibited the anti-DR4 activity of autologous and homologous antisera. These data suggest that sera of parous women contain anti-idiotypic antibodies directed against regulatory idiotypes of anti-DR antibodies.

Induction of multi-specific antibodies to bovine serum albumin after production of anti-idiotype antibodies to an albumin-specific monoclonal antibody.

Induction of multi-specific antibodies to bovine serum albumin after production of anti-idiotype antibodies to an albumin-specific monoclonal antibody.

UC 729-6, a human lymphoblastoid B-cell line useful for generating antibody-secreting human-human hybridomas.

UC 729-6, a 6-thioguanine-resistant human lymphoblastoid B-cell line, was fused with normal and malignant human lymphocytes. Parent UC 729-6 cells were diploid with a 21p+ marker chromosome, expressed surface and cytoplasmic IgM kappa, and doubled every 17 hr. The resulting human-human hybridomas were pseudotetraploid containing the 21p+ marker, doubled every 20-30 hr, and secreted 3-9 micrograms of human Ig per ml per 10(6) hybrid cells for greater than 9 months in continuous culture. Human Ig-secreting hybridomas were generated in 88% (14/16) of the fusions carried out and were cloned by limiting dilution (one cell per three wells) without the use of feeder layers. The mean fusion frequencies (number of wells, plated at 10(5) cells per well, showing hybrid growth per 10(6) lymphocytes fused) of UC 729-6 with normal lymphocytes ranged from 0.45 to 2.9 and with malignant B lymphocytes, from 3 to 10. Analysis of the human-human hybridomas derived from lymphocytes isolated from regional draining lymph nodes of cancer patients revealed several that secreted human monoclonal antibody that reacted by an enzyme immunoassay with some carcinoma cell lines but not with normal fibroblast cell lines. These data suggest that (i) UC 729-6 can be fused with human lymphocytes to generate stable human-human hybridomas, some of which secrete antibody reactive to human cell surface antigens, and (ii) UC 729-6 can be used to rescue Ig from nonsecretory malignant B cells and thereby allow for the production of anti-idiotype antibodies.

Subclass restriction of the antiidiotypic antibody response in the rat

Antiidiotypic antibodies were induced in LOU/M rats by immunization with two myeloma proteins of LOU origin: IR-162 (IgE) and IR-418 (IgG2a). Antibodies to IR-162 were easily obtained after a limited number of immunizations with protein in soluble form; polymerization with glutaraldehyde did not enhance immunogenicity. Antibodies to IR-418 appeared only after a large number of immunizations with protein in polymerized form or with protein copolymerized with rabbit IgG. All of the antibodies, either to IR-162 or to IR-418, were found to be idiotype-specific. In every case for which significant levels of antiidiotypic antibodies were produced, most or all of the antibodies belonged to rat IgG1 subclass. Since, in mice, antiidiotypic antibodies are restricted to the IgG1 subclass, our results indicate a functional analogy between rat and mouse IgG1. Our studies also suggest that the rat IgG1 subclass may be predominantly expressed in T-cell-dependent antibody responses, such as production of antiidiotypic antibodies.

Cancer and depression: cancer presenting with depressive illness: an autoimmune disease?

It is proposed that some cases of depressive illness in cancer patients may be caused by immunological interference with the activity of serotinin, one of the neurotransmitters thought to be implicated in depression. This interference could be mediated in two ways. Antibody induced against a protein released from cancer cells could, on the basis of cross-reactivity with CNS tissue, bind to receptors for serotonin and block them. Such primary antibodies could stimulate the production of anti-idiotypic antibodies, which would act as an alternative receptor for serotonin and reduce its synaptic availability.

The production of anti-idiotypic antibodies and of idiotype-anti-idiotype immune complexes after polyclonal activation induced by bacterial LPS.

The production of anti-idiotypic antibodies and of idiotype-anti-idiotype immune complexes after polyclonal activation induced by bacterial LPS.

Induction or elimination of tumor-specific immunity against a chemically-induced rat tumor using auto-anti-idiotypic immunity.

DA rat sarcoma P1 and P2 were induced by dimethylbenz(a)anthracene. A tumor-specific immune response of DA rats against P1-tumor cells could be demonstrated at the humoral and cellular level. DA anti-P1 antibodies were purified on fixed P1-tumor cells and used as auto-immunogen in DA rats for the production of anti-idiotypic antibodies. Such anti-idiotypic antibodies could be demonstrated by using a solid-phase radioimmunoassay and by their ability to induce secondary type of DA anti-P1 response in vitro. In addition, such antibodies were able to induce cytotoxic T lymphocytes capable of eliminating P1-tumor cells but not control tumor cells. In some of the auto-immunized DA rats enhanced P1-tumor growth could be observed, indicating that the anti-idiotypic immune response had led to a selective ablation of idiotypic, potential anti-P1 reactive T cells.

Immunoglobulin secretion by mouse X human hybridomas: an approach for the production of anti-idiotype reagents useful in monitoring patients with B cell lymphoma.

Tumor cells were obtained from a patient with nodular lymphoma, a monoclonal B cell malignancy. These non-Ig-secreting lymphoma cells were hybridized to mouse myeloma cells. Hybridomas secreting human Ig were obtained in high frequency. One clone, which was a stable producer of human heavy and light chains, but which had lost production of mouse light chains, was chosen for study. It secreted a human IgM pentamer. The Ig produced by this clone was purified and used as an immunogen for the production of anti-idiotype antibodies. These antibodies recognized the specific cell surface Ig of the lymphoma cells and have been used to detect idiotype-bearing cells in the blood of the original patient, even at times when the disease was in clinical remission. Moreover, free idiotype could be detected in the patient's serum. Both the levels of Id+ cells and free idiotype correlated with disease activity, rising in relapse and falling in remission.

The Production of Anti-Idiotypic Antibodies to BALB/c Plasmacytoma Globulins in BALB/c Mice

Abstract Slater, Ward and Kunkel (1) were the first to draw attention to the individual antigenic characteristics of myeloma globulins. Later it was determined that these globulins have the immunochemical properties of idiotypic antibodies with specific antigenic groupings in the variable portion of the Fab fragment (2–4). Antibodies to these determinants have been developed by prolonged immunization of rabbits with myeloma globulins and absorption of these antibodies with large amounts of normal γ globulin (2, 3), thus removing antibodies to the non-idiotypic components of the molecules. Absorption would not be required if immunization could be accomplished in a recipient with the same allotype as the donor. The following report deals with the production of anti-idiotypic antibodies to BALB/c plasmacytoma globulins in normal BALB/c mice. The presence of such antibodies was demonstrated by hemagglutination, hemagglutination inhibition, and also in some instances by immunodiffusion. Materials and Methods.

Immunological analysis of rabbit anti-antibody systems.

A study has been made of the production of antiidiotypic antibodies (anti-antibodies) arising during the immunization of 39 rabbits with 19 individual samples of rabbit anti-Proteus vulgaris x 19 antibodies adsorbed onto bacilli. In addition to the regular demonstration of antiidiotypic antibodies attention is drawn to the frequent occurrence of antiallotypic antibodies against molecules of immunoglobulin classes other than IgG, especially macroglobulins, which may arise during such immunization. In four cases attempts to raise autoantiidiotypic antibodies were unsuccessful, as expected. The idiotypic specificities (antigenic determinants) have been found mainly on IgG but also sometimes on IgM molecules. The individual specificity of the antiidiotypic antibodies appears to be absolute, as long as the antiallotypic antibodies are recognized and excluded; e.g., each idiotypic specificity is characteristic of only one single rabbit and of a single kind of antibody within that rabbit. These principles hold even when antibodies of rabbit families are examined: the parental idiotypic determinants could not be found in the offspring. In two samples tested the idiotypic specificities were found on the Fab fragment of IgG molecule but not on its antibody-combining site.