Tetanus Toxoid Antibodies Research Articles

Serum antibody responses of juvenile and infant rhesus monkeys injected with Haemophilus influenzae type b and pneumococcus type 6A capsular polysaccharide-protein conjugates

Juvenile and infant rhesus monkeys were injected subcutaneously with saline solutions of Haemophilus influenzae type b (Hib) and pneumococcus type 6A (Pn6A) capsular polysaccharides conjugated to either tetanus toxoid (TT), horseshoe crab hemocyanin, or cholera toxin (CT), and the antibody responses of the monkeys to both bacterial components were measured. All three Hib conjugates were immunogenic and elicited booster responses; their comparative immunogenicity was Hib-CT greater than Hib-TT greater than Hib-horseshoe crab hemocyanin. Hib alone did not elicit antibodies in the juveniles. Juveniles responded earlier and with higher levels of antibodies than did infants. TT, as well as diphtheria-tetanus toxoids-pertussis vaccine adsorbed injected concurrently at a separate site, increased both Hib and TT antibody responses in juveniles (P less than 0.05). Concurrent injection of 5 Lf of fluid TT with a nonimmunogenic 5-micrograms dose in infants elicited levels of Hib antibodies comparable to those elicited by 50 micrograms of Hib-TT. Hib antibodies elicited by the conjugates remained at protective levels in both juveniles and infants 2 months after the last injection, were bactericidal, and conferred passive immunity against bacteremia in infant rats. Passive immunization of juveniles with tetanus immune globulin before each injection of Hib-TT did not suppress Hib antibodies. Hib-TT and Hib-CT elicited increases of Hib antibodies of the immunoglobulin M and G isotypes in the infants. The Pn6A-TT conjugate was considerably less immunogenic than the Hib-TT conjugate; only a few of the juveniles or infants responded with protective levels of Pn6A antibodies. Pn6A antibodies from responders conferred protection in mice against intraperitoneal challenge with Pn6A organisms. TT antibodies were elicited in both juvenile and infant animals after one injection of 50 micrograms of Hib-TT and in the infants injected with 5 micrograms of Hib-TT plus 5 Lf of TT; 5 micrograms of Hib-TT and Pn6A-TT in combination alone did not elicit TT antibodies. Hib-CT elicited CT antibodies in both juveniles and infants.

Binding and activation of the first complement component by soluble immune complexes: effect of complex size and composition.

The interaction between soluble immune complexes and the first component of complement (C1) was studied. Complexes were prepared from purified bovine thyroglobulin (BTg) or tetanus toxoid (TT) and immunospecific IgG antibodies. Purified human precursor C1 was incubated with dilutions of the preparations, and the inhibition of C1 haemolytic activity was determined as a measure of C1-binding. The activation of C1 was assessed by measuring the amount of C4 consumed by generated C1. The molar antibody/antigen (Ab/Ag) ratio of BTg--anti-BTg mixtures strongly influenced their C1-binding and C1-activating capacities: mixtures with high Ab/Ag ratios were by far the most efficient. On the other hand, the Ab/Ag ratio had only a limited influence on the activity of TT--anti-TT complexes. The effect of complex size was investigated by ultracentrifugation of antibody-antigen mixtures on calibrated sucrose density gradients followed by C1-binding and -activation experiments with the fractions obtained. For both types of immune complex, the C1-binding and -activating capacities increased markedly with increasing complex size. Thus, both the size and the Ab/Ag ratio of soluble immune complexes influence their capacity to activate the classical complement pathway. The effect of the Ab/Ag ratio, however, may also be dependent on the antigen molecule(s) present in the complexes.

The percentages of six immunoglobulin isotypes in human antibodies to tetanus toxoid: standardization of isotype-specific second antibodies in solid-phase assay.

The solid-phase radioimmunoassay for human antibodies was improved so that it gave the total concentration as well as the concentrations of IgM, IgA, IgG1, IgG2, IgG3 and IgG4 antibodies, all seven in the same "L units"/ml. This was accomplished by standardization of monoclonal antibodies with monoisotypic antibodies rather than myelomas; myelomas were found unsatisfactory for this purpose. For the first time it was possible to determine the isotype composition of an antibody population in percent terms. The weight equivalent of the L-unit in one hyperimmune tetanus antitoxin preparation was approximately 7.4 ng. The new solid-phase radioimmunoassay was applied to tetanus toxoid antibodies of the booster response. Total concentrations varied from 1700-26 000 L units/ml (13-200 micrograms/ml). Concentrations of IgG1 antibodies were 1600-25 000 units/ml (average 91% of the total) and concentrations of IgG4 antibodies 12-6900 units/ml (average 6.9% of the total). Antibodies of the other four isotypes were not detected in all sera and together they never exceeded 3% of the total.

Production d'anticorps antitétaniques humains par des hybridomes

Peripheral blood lymphocytes from healthy people recently immunized against tetanus toxoid (TT) were fused with human malignant B-cell lines or mouse myeloma cells (X63 Ag8.653) in an attempt to establish stable B cell hybridomas secreting anti TT antibodies. Human-human fusion experiments were not successful. In contrast, the five heterospecific fusion experiments yielded between 60 and 100% of wells that contained growing hybrids. Five of these hybrids repeatedly secreted anti-TT antibodies. One of the hybrids was cloned and secreted 10-20 micrograms/ml of human IgM, lambda anti-TT antibody. Heterospecific hybridization thus appears as an interesting method to obtain human monoclonal antibodies, allowing the study of their properties.

EBV transformed B cell lines present to antigen specific T cell clones determinants different from those recognized by antibody in an HLA-DR linked restricted fashion

Human B cells nonspecifically activated by Epstein Barr virus (EBV) can present tetanus toxoid (TT) antigen to TT specific helper T cell lines and T cell clones. Presentation of TT antigen by EBV-B cells did not require the presence of TT specific B cells and did not involve B cell surface immunoglobulins. Immunosorbent purified antibody to TT added in great excess to EBV-B cells pulsed with TT for 18 hr did not inhibit the capacity of the EBV-B cells to present TT antigen. Furthermore, EBV-B cells induced proliferation in T cells in the presence of urea denatured TT which contained less than 1% TT reactive with serum anti TT antibody. Studies of TT presentation by a panel of EBB-B cells obtained from HLA typed donors indicated that T cell recognition of TT presented by EBV-B cells was MHC restricted by products of the HLA-D region some of which differed from serologically defined HLA-DR. These results indicate that the immunogenic moiety presented by EBV activated human B cells consists of antigenic determinants, which differ from those recognized by serum antibody, and of HLA-DR linked MHC determinants. This moiety is similar to that presented by monocytes. The implications of these findings for the amplification of the immune response by activated B cells are discussed.

The effect of albumin resuscitation for shock on the immune response to tetanus toxoid

A prior prospective random study showed that supplemental albumin led to a significant ( P ⩽ 0.05) fall in immunoglobulin (Ig) content of IgG, IgM, and IgA; no measurement was made on the immune response to a known antigen. Therefore, the immune response to tetanus toxoid, measured by antibody precipitation, was studied in 43 nonalbumin patients and 22 albumin patients from whom frozen sera were available. The 65 patients received an average of 16 units blood during resuscitation and 0.5 ml tetanus toxoid. Using single radial immunodiffusion technique, duplicate double-blind measurements of precipitants formed by the patients tetanus toxoid antibody and purified tetanus toxoid (TTA) incorporated in 1% agarose gel were made. TTA levels in the 22 albumin patients were compared with all nonalbumin patients and with two subgroups of 22 patients—one randomly chosen; the other matched for number of transfusions. TTA was also correlated with time after injury. TTA was significantly less in albumin patients (215 ± 100 units) than in the nonalbumin patients (387 ± 233 units) and both subgroups of nonalbumin patients. TTA correlated directly with time after injury in the nonalbumin patients (slope = −0.4). Albumin resuscitation for shock results not only in a fall in Ig content but also a reduction in the immune response to tetanus toxoid.

The selective role of membrane IgG in the antigen-induced inhibition of human in vitro antibody synthesis.

IgG-anti-tetanus toxoid-producing B cell precursors from recently immunized individuals can be stimulated by pokeweed mitogen and T cells to produce IgG-Tet antibodies in vitro. Treatment of these cells with tetanus toxoid selectively inhibits the synthesis of IgG-Tet. Because the IgG-Tet precursors display heterogeneity in their surface isotypes (mu, delta, gamma), the aim of this study was to determine the surface isotype responsible for transmitting this antigen-induced inhibitory signal to the B cell. B cells were fractionated by rosetting techniques on the basis of surface IgM or IgD. The 4 resulting B cell subsets (SIgM+, sIgM-, sIgD+, sIgD-) were found to be equally susceptible to antigen-induced inhibition. Experiments were then performed with anti-isotype antibodies to investigate further the roles of sIgM, sIgD, and sIgG in antigen inhibition. Treatment of peripheral blood lymphocytes with 10 micrograms/ml of anti-gamma antibody inhibited IgG-Tet antibody production, whereas the addition of up to 100 micrograms of anti-mu or anti-delta antibodies did not. Tetanus toxoid and anti-gamma antibody inhibition of IgG-Tet synthesis in vitro followed similar temporal kinetics, with the IgG-Tet precursors being sensitive to inhibition by either tetanus toxoid or anti-gamma antibodies during the first 3 days of culture and not becoming totally resistant to these agents until day 5 of culture. Capping off of sIgG receptors, with the subsequent addition of tetanus toxoid, abolished the antigen-induced inhibition of IgG-Tet seen previously, whereas capping off of sIgD or sIgM did not affect antigen inhibition of IgG-Tet antibodies. These results implicate sIgG molecules as those responsible for rendering IgG-Tet precursors susceptible to antigen-induced inhibition.

Antigen-specific detection of soluble immune complexes by a solid phase specific antibody system

An antigen-specific immune complex assay based on the following principle has been developed: xenogeneic, antigen-specific antibodies are attached to a solid phase. During first incubation with patient's serum, immune complexes in antigen excess are bound to the xenogeneic antibody by their free antigenic determinants. In a second incubation a labeled antibody, specific for human immunoglobulins, is combined with the antibody part of the immune complex. Quantitation of the label allows the determination of the immune complexes. The principle of the method has been varied using artificial soluble immune complexes of tetanus toxoid and human anti-tetanus toxoid antibody, and immune complexes prepared by mixing patient sera containing either HBsAg or anti-HBsAg antibodies. The reliability of the results is shown by their coefficient of variation (2.5%). With the method described soluble immune complexes predominantly in slight antigen excess, which are thought to be responsible for development of immune complex disease, have been detected.

A rapid automated latex screen for tetanus toxoid antibodies.

This report describes the sensitisation of latex particles and the use of this reagent on the Autotape system to select blood donations containing greater than or equal to 4 IU/ml of tetanus toxoid antibody. Plasma containing tetanus toxoid antibodies is processed to provide immunoglobulin for passive protection against tetanus. The techniques used for screening include immunoelectrophoresis [3], tetanus toxoid sensitised latex particles [5], haemagglutination [1] and automated haemagglutination [6]. The technique used by the Sheffield Regional Transfusion Centre until recently, was immunoelectrophoresis, selecting donors with 4 or more IU/ml. The Autotape system, previously described for brucellosis [4] and syphilis testing [2], provides a system for automated slide testing and a 5-min tetanus toxoid coated latex test has been developed to select suitable donations for the production of specific immunoglobulin.

A Rapid Automated Latex Screen for Tetanus Toxoid Antibodies

A Rapid Automated Latex Screen for Tetanus Toxoid Antibodies

Combined immune deficiency, autoantibody formation, and mucocutaneous candidiasis

A 6-year-old girl with chronic mucocutaneous candidiasis (MCC) and an immunologic deficiency syndrome was studied in detail over a period of 2 years. The patient had a severe defect in cell-mediated immunity characterized by failure to mount a 48-hr delayed response to a battery of intradermal skin tests, persistent lymphopenia, and failure to be sensitized with 1-chloro, 2,4-dinitro-benzene (DNCB). Her lymphocytes neither produced migratory inhibitory factor nor consistently responded with an increased 3H-thymidine uptake when placed in tissue culture medium with Candida antigen. There was no evidence of a circulating factor in the patient's plasma capable of inhibiting the incorporation of 3H-thymidine by homologous lymphocytes cultured in the presence of Candida albicans and other antigens. The patient had defective antibody production to some but not all antigens. She failed to produce protective tetanus toxoid antibody titers and remained Schick test positive despite repeated diphtheria, pertussis, and tetanus (DPT) injections. She also had defective antibody responses to repeated paratyphoid-typhoid injections and was found to have a low serum IgA level. This patient possessed autoantibodies against smooth muscle and parietal cells and an inconstant positive antilymphocytotoxic assay. Immunologically unrelated precipitin antibody systems against human and calf thymus were also demonstrated in the patient's serum. Eradication of the Candida infection with intravenous amphotericin B, although resulting in marked clinical improvement, did not alter the immunologic abnormalities.

Enhanced Antitoxin Responses in Irradiated Mice Elicted by Complexes of Tetanus Toxoid and Specific Antibody

Summary Enhanced primary antitoxin responses were obtained in mice immunized by intravenous injection with complexes of tetanus toxoid and mouse antitoxin presumably formed either in vivo or prepared in vitro in antigen-antibody ratios of antibody excess, equivalence and antigen excess. The demonstration of the enhancement phenomenon elicited by complexes of toxoid and isologous mouse antitoxin provide conclusive evidence that the antibody portion of the complex does not need to be of heterologous origin in order to elicit enhanced primary antibody responses in mice. Intravenous immunization with the above complexes elicited enhanced primary responses in irradiated animals, whereas minimal responses were obtained with antigen only. Little difference was observed in primary responses in nonirradiated mice when antigen only or antigen complexed with specific antibody was given by subcutaneous injection. However, enhanced primary antitoxin responses were obtained in irradiated mice (400 rads) immunized with the various complexes over the responses observed in irradiated animals immunized with antigen only. The greatest degree of enhancement occurred when the complexes were prepared in the region of equivalence and antigen excess. Secondary antitoxin responses were repressed when the same complexes of antigen and antibody were injected to elicit secondary responses. A corresponding repression of secondary responses was observed in irradiated mice when radiation doses of 300 rads were delivered 24 hr before the second injection of antigen complexed with specific mouse antitoxin.