Conditioned suppression of a thymus-independent antibody response

An illness-induced taste aversion was conditioned in mice by pairing cyclophosphamide, an immunosuppressive drug, with the consumption of saccharin, a novel drinking solution. Two weeks after conditioning, animals were injected with the hapten trinitrophenyl (TNP) coupled to the thymus-independent carrier, lipopolysaccharide. Serum antibodies to TNP were titered 6 days later by passive hemagglutination. Relative to control groups, conditioned animals provided with saccharin at the time of antigenic stimulation and, again, 3 days later showed a significant attenuation of their anti-TNP antibody response. In a second experiment, the conditioned stimulus (CS) consisted of the novel saccharin drinking solution plus the noxious internal effects of an injection of LiCl. Conditioned animals reexposed to the CS again showed the lowest antibody titers, but differed significantly from only one of the control groups. Taken together, the results of these experiments confirm previous reports of conditioned immunosuppression and suggest that the effects of conditioning on a primary humoral antibody response can be observed in response to a T-cell independent antigen in the mouse.     

Carrier-induced suppression of the antibody response to a 'self' hapten.

Immunization of male rats and monkeys with gonadotropin-releasing hormone (GnRH) conjugated to a carrier results in a dramatic atrophy of the prostate. GnRH, linked to either diphtheria toxoid or tetanus toxoid as carrier, is now being evaluated for its use in the immunotherapy of hormone-dependent prostate enlargement in men. This report deals with the phenomenon of carrier-induced, epitope-specific regulation in the GnRH-carrier system. In experiments designed to assess the influence of the carrier on antibody responses to the 'self' hapten GnRH, we show that preimmunization with carriers diphtheria toxoid and tetanus toxoid results in a strain-dependent inhibition of anti-GnRH responses in mice. Results of adoptive transfer experiments indicate that T cells from carrier-presensitized mice are responsible for suppression of anti-haptenic antibodies and that T cells from conjugate-immunized mice, on the other hand, can actually help overcome hyporesponsiveness.     

Preservation of carrageenan-induced immune suppression with alleviation of toxicity in aprotinin-treated mice.

Although the immunosuppressive effect of carrageenan was unimpaired in aprotinin-treated mice, the extent of intravascular coagulation induced by this macrophage toxic agent was substantially reduced. Use of antiproteases such as aprotinin may prove beneficial in management of this adverse side effect of certain immunotherapeutic agents.     

Fc-dependent monoclonal IgG1-mediated suppression of antibody response

It has been known for a long time that passively administered antibodies (Abs) or immune complexes regulate the immune response to their specific antigen (Ag). IgG may sometimes suppress the humoral immune response against soluble antigens. The exact mechanism behind this phenomenon has not been understood yet and the requirement for the Fc part is still a matter of controversy. The present study was undertaken to clarify whether there is a true IgG-mediated Fc-dependent suppression of the immune response. Antigen and monoclonal antibody (mAb) used in this study were recombinant human interferon gamma (r-hIFN-gamma) and mouse monoclonal antibodies specific for human IFN-gamma [anti-hIFN-gamma mAb (CAy-IFNgamma38)] respectively. An intact IgG-free preparation of Fab plus various Fc fragments was prepared from papain-digested CAy-IFNgamma38. Ag/IgG and Ag/Fab complexes were prepared at various molar ratios. Keeping the Ag doses constant, mice were immunized either with Ag, Ag/IgG or Ag/Fab complexes. Primary immunization and the boosting were performed with the samples in complete and incomplete Freund's adjuvants respectively. Specific antibody levels were measured by an ELISA. Immunization performed with Ag/Fab complexes even at a molar ratio of 1:1.36 did not result in marked suppression of the response when compared to that of Ag only-immunization. In contrast, Ag/IgG complexes resulted in nearly 90% suppression of the antibody response. Our observations suggest that Fc part of IgG molecule plays a crucial role in suppression of the in vivo antibody response against the Ag when complexed with intact IgG.     

Age-related enhancement and suppression of a T-cell-dependent antibody response following stressor exposure.

The effects of uncontrollable footshock on the peak splenic plaque-forming cell (PFC) response and serum antibody titers to sheep red blood cells (10(6) cells ip) were assessed in 3-month-old and 9-month-old male CD-1 mice. Exposure to uncontrollable footshock provoked an immunosuppression in mice of both age groups. The critical period for the induction of the suppression (i.e., 72 hr after inoculation) did not differ between the 3-month-old and 9-month-old mice; however, the suppression could be provoked more readily in the older animals. In the 9-month-old mice, the variations of immune activity were dependent on the severity of the stressor and the time of stressor application. Specifically, in contrast to the suppression induced by footshock, a relatively mild stressor such as exposure to a novel environment effectively increased the PFC response. A marked enhancement of the PFC response and antibody titers was evident in older animals that were shocked immediately or 24 hr after inoculation. The possibility exists that stressor application in older mice may influence regulatory processes that are associated with an immune response and that the nature of these regulatory mechanisms may vary with the time after antigenic challenge.     

Complement activation is not required for IgG-mediated suppression of the antibody response

Feedback suppression of the antibody response by IgG is known to be dependent on intact Fc regions. However, it is not clear which of the Fc-mediated effector functions is required. In the present report we have studied whether ability or inability of the IgG antibodies to activate the complement system was of consequence for their immunosuppressive effect. First, a monoclonal IgG1-anti-2,4,6-trinitrophenyl (TNP) antibody, unable to activate complement via the classical or alternate pathway, was shown to be able to inhibit more than 90% of the in vivo sheep erythrocyte-specific antibody response in mice when TNP coupled to sheep erythrocytes was used as antigen. Second, we investigated the immunosuppressive ability of a non-complement-activating mutant IgG2a-anti-TNP monoclonal antibody. The mutant differs from the wild type by a single amino acid substitution in the CH2 domain leading to inability to fix complement factor C1q. However, the mutant has the same affinity for antigen and the same Fc receptor-binding capacity as the wild type antibody. It is demonstrated that the mutant was as efficient as the wild type antibody in inhibiting an in vitro antibody response to TNP-coupled sheep erythrocytes. These findings confirm the non-determinant specificity and Fc dependence of IgG-mediated suppression, and show that the Fc-mediated effector mechanism is independent of complement activation. The results instead suggest binding to Fc receptors as a necessary step in feedback immunosuppression and favor inactivation of B cells by cross-linking of Fc and antigen receptors on their surface rather than elimination of antigen by complement-dependent phagocytosis as the effector mechanism.     

Genetic susceptibility to carrageenan-induced innate inflammatory response in inbred strains of rats.

Rat models are useful for the genetic dissection of the biology of innate immunity. Inbred rat strains were evaluated for carrageenan-induced innate inflammatory responses. Results indicated that the genetic control of innate immune responses is polygenic and influenced by gender, and may not necessarily be consistent with the genetics of experimental arthritis. The newly identified susceptible strains, in order of decreasing susceptibility, include Dahl salt-sensitive (S), Dahl salt-resistant (R), Milan normotensive strain (MNS) and Wistar Kyoto (WKY) rats. Similarly, the newly identified relatively resistant strains, in decreasing order of resistance, include DA rats, spontaneously hypertensive rats (SHRs) and Brown Norway (BN) rats. Linkage analyses using combinations of these susceptible and resistant strains are proposed.     

Prevention of carrageenan-induced pleurisy in mice by anti-CD30 ligand monoclonal antibody

CD30 ligand (CD30L) and its receptor CD30 are members of the tumor necrosis factor (TNF) and TNF receptor superfamilies that play a major role in inflammation and immune regulation. To gain insight into the in vivo role of CD30L/CD30 in inflammatory diseases, we have used carrageenan (CAR)-induced pleurisy in mice, a preclinical model of airway inflammation where type 1 proinflammatory cytokines such as interleukin (IL)-1 and TNF-alpha play a key pathogenic role. The data show that prophylactic treatment with anti-CD30L mAb markedly reduces both laboratory and histological signs of CAR-induced pleurisy. These data suggest involvement of CD30-mediated signals in acute immunoinflammatory pathways induced by CAR.     

Effects of tolerogenic conjugates in a canine model for reaginic hypersensitivity. I. suppression of hapten-specific IgE antibody response

Intraperitoneal administration to dogs of conjugates consisting of the 2,4-dinitrophenyl (DNP) groups coupled to nonimmunogenic macromolecules such as the copolymer of D-glutamic acid and D-lysine (DNP16-DGL) prior to sensitization with DNP2-ovalbumin led to the development of hapten-specific tolerance with respect to the IgE antibody response. Administration of these same conjugates to sensitized dogs resulted in complete abrogation of the ongoing anti-DNP IgE antibody production. A similar hapten-specific suppression of the ongoing anti-DNP response was also observed using the conjugates of DNP9-canine gamma globulins, and the tolerogenic effect was dose-dependent. The state of hapten-specific immunosuppression induced by these two types of tolerogenic conjugates was maintained despite repeated booster injections of the sensitizing antigens at biweekly inervals.     

Regulation of anti-hapten antibody response by chemically modified carrier antigen preferentially provoking delayed-type hypersensitivity: I. Possible T—T cell interaction in the suppression of antibody response

The i.p. immunization with chemically modified antigen (dodecanoyl-bovine serum albumin, d-BSA) emulsified in Freund's incomplete adjuvant (FIA) of CBA mice provoked delayed-type hypersensitivity (DTH), but not any detectable formation of antibody to the original antigen (BSA). Furthermore, it was found that immunization with d-BSA could generate T cells capable of inhibiting the antibody response to hapten on BSA, and the immunosuppressive effects of these T cells were presumably not due to direct action on hapten-primed and antibody producing B cells. These results were obtained from the following experiments: (1) anti-hapten antibody response to dinitrophenylated-BSA (DNP-BSA) was inhibited when the mice had been primed previously with d-BSA in FIA. This inhibition was regulated by the specificity of the carrier, since the mice treated with d-BSA did not inhibit the anti-DNP antibody response after the immunization with DNP-heterologous carrier, i.e. DNP-keyhole limpet haemocyanin (DNP-KLH). (2) The passive transfer of spleen cells, which had been obtained from donors primed with d-BSA in FIA, inhibited the primary anti-DNP antibody response of syngeneic mice after immunization with DNP BSA. (3) Injection of d-BSA-primed spleen cells suppressed an adoptive anti-DNP antibody response in mice which had been irradiated and had previously had their immunocompetence reconstituted by the cell transfers with both DNP-primed and BSA-primed spleen cells. This in vivo immunosuppressive effect of d-BSA-primed spleen cells did not act on hapten-primed B cells, since d-BSA-primed spleen cells could not suppress the adoptive secondary antibody response reconstituted by DNP-primed cells and bacterial alpha-amylase (BαA)-primed cells. This finding suggests that a T—T cell interaction exists for the suppression of the anti-DNP antibody response to DNP-BSA by d-BSA-primed cells.     

Characteristics of immunological memory in mice. II. Resistance of nonrecirculating memory cells to antigen-mediated suppression of the secondary antibody response.

Mice were primed and subsequently challenged at various times with subcutaneous injections of sheep erythrocytes, and some characteristics of the secondary responses in the draining brachial and axillary lymph nodes were investigated. It was found that the secondary response within primed nodes was resistant to immunological preemption, a competition-like phenomenon which severely depresses primary responses. Since it was also shown that circulating memory cells could be inhibited by preempting injections of antigen, it was concluded that the resistance of primed nodes to preemption was due to the presence within them of a nonrecirculating subpopulation of memory cells. The size of this population was dependent both on the amount of priming antigen and the time after priming. The observation that the response given by these cells remained unaffected by doses of antigen which could depress a primary response does not favor the view that suppression of immune responses by preemption or antigenic competition is due to a factor which acts directly and indiscriminately on all immunologically competent cells.     

Regulation of cellular antibody synthesis: Selective suppression by antibody of the immune response induced by a high antigen dose

Mice immunized with a high dose of sheep red blood cells and 24–48 hours later given specific antiserum produced on the average only 1 per cent of the direct and indirect number of plaque-forming cells (PFC) found in the non-antibody treated controls, whereas animals given a low antigen dose followed by antiserum produced 30 per cent of the number of PFC found in the controls. It is suggested that the low antigen dose preferentially stimulated antigen-sensitive cells having a receptor for the antigen of high affinity, whereas the high antigen dose in addition stimulated cells having lower affinity receptors and that, therefore, the passively transferred antibody would compete more efficiently for the antigen with the antigen-sensitive cells triggered by a high antigen dose than with the higher affinity cells stimulated by a low antigen dose.

When antigen and antibody were introduced simultaneously the selective suppression of the immune response to a high antigen dose did not occur. Possible mechanisms for this finding are discussed.

     

In vivo suppression and enhancement of the murine homocytotropic antibody response by staphylococcal enterotoxin A

Effects of staphylococcal enterotoxin A (SEA) on the mouse homocytotropic antibody (HCA) system were studied. Groups of BDF mice received 10 micrograms SEA either orally or intraperitoneally at 0, 24, 48 h before or after immunization with 100 micrograms ovalbumin in 1 mg A1(OH)3 gel. Primary and secondary HCA responses were determined by 48-hour passive cutaneous anaphylactic reactions in genetically hairless mice. It was found that effects of SEA on HCA responses were dependent on the time and route of SEA administration. In general, early administration (48 h before immunization) of SEA showed suppression, while later administration (either 24 h before or after immunization) of SEA demonstrated enhancement. A further delay of SEA administration (48 h after immunization) exerted suppressive effects except when it was given intraperitoneally in the anamnestic HCA experiments. The mouse HCA system proved to be a suitable in vivo correlate of in vitro plaque-forming cell responses modulated by SEA.     

Regulation of allergic reaction by aerobic Corynebacterium equi extract, CEF. I. Antigen-nonspecific suppression of reaginic antibody response in mice

The effect of a water-soluble fraction (CEF) that was prepared from an extract of Corynebacterium equi on primary reaginic antibody formation was studied in Balb/c mice. Mice were immunized with a hapten carrier (DNP-OVA) and received intraperitoneal injections of CEF 7 and 2 days prior to, or 2 and 7 days after the immunization. PCA titers of both antihapten (DNP) and anticarrier (OVA) antibodies of IgE class were reduced significantly by the CEF treatment. Evidence was presented in adoptive transfer experiments that the number of IgE-producing cells in the CEF-treated mice was lower than that of controls. Suppression of IgG1 anti-DNP antibody formation was also achieved by the CEF treatment. Formation of IgG1 anti-OVA antibodies, however, was not suppressed significantly by the treatment. The suppressive activities of CEF were shown to be dose-dependent, but timing of CEF administration did not appear critical.     

Alteration of clearance function by group A streptococcal pyrogenic exotoxin and its relation to suppression of the antibody response.

The effect of purified group A streptococcal pyrogenic exotoxin (SPE) type A on the processing of and antibody response to sheep erythrocytes (SRBC) was studied in BALB/cWat mice. The rate of clearance of 51Cr-labeled SRBC from the bloodstream was decreased 3 or 24 h following a single intravenous injection of 1 or 10 microgram of SPE. Delayed uptake of label was observed in both the livers and spleens of SPE-treated mice, suggesting an inhibitory effect of the toxin on phagocytic cells of the reticuloendothelial system. Three daily intravenous injections of 0.1 or 1 microgram of purified SPE type A suppressed the early immunoglobulin response to SRBC. The role of altered macrophage function in producing the immunosuppression was tested in macrophage transfer experiments. SPE treatment suppressed the antibody response to SRBC transferred by normal macrophages, indicating that the immunosuppressive effect of the toxin was not due solely to altered antigen processing by macrophages.     

Plasmodium berghei: suppression of antibody response to sporozoite stage by acute blood stage infection

Mice infected with the erythrocytic stages of Plasmodium berghei show an impaired host response to immunization with irradiated sporozoites of the same malarial parasite. The stage-specific anti-sporozoite response was measured by indirect immunofluorescence upon adsorption of the sera with parasitized red blood cells. P. berghei-infected mice, immunized with irradiated sporozoites on the fourth day of a blood-induced malaria infection, developed a normal anti-sporozoite antibody response. However, this antibody response was more short-lived compared with the antibody response in normal mice immunized with a similar dose of irradiated sporozoites. The immune response was severely depressed when the animals were immunized on day 7 or later after malaria infection. None of the sporozoite-immunized animals, including those which responded to the first immunization, developed a secondary antibody response on reinoculation with irradiated sporozoites. A fully established anti-sporozoite immune response, obtained after multiple immunizations with irradiated sporozoites and which resulted in stage-specific protection of the animals, was not affected by a superimposed blood stage malaria infection. The titres of the anti-sporozoite antibodies in these animals were unaltered, in spite of their high parasitaemias. Reduction of the malaria parasitaemia by chloroquine treatment abolished the immunosuppressive effects of the disease.

These observations are discussed in relation to anti-sporozite immunity and immunosuppression in man in malaria endemic areas.