IgE Antibody Response In Mice Research Articles

Protection of Factor VIII-Specific IgG Antibody Response and IgE Formation By Pretreatment with Liposomal α-Galactosylceramide in the Enzyme Replacement Therapy Mouse Model

▪Enzyme replacement therapy (ERT) is indispensable for patients of hemophilia or lysosome storage disease (LSD) lifetime. However, the efficacy of ERT is diminished or lost in some patients who acquired the immune responses by the repeated enzyme infusions. If reduced the immune responses such as neutralizing IgG antibody formation and/or anaphylaxis, one enzyme for ERT would be continuously administered without loss of efficacy and/or anaphylaxis risk. We here show that the pre-treatment with α-galactosylceramide (αGC), a representative ligand for invariant natural killer T (iNKT) cells prior to the ERT could remarkably suppress not only the enzyme-specific IgG antibody response but also total IgE formation in a mice model.In our previous studies, liposomal, but not aqueous, αGC formulations were delivered to the splenic marginal zone (MZ) B cells which could enhance IL-10 production after the interaction with iNKT cells, and then the expansion of tolerogenic dendritic cells (DCs) and Foxp3-positive regulatory T cells (Tregs) (Ishii et al. Front Biosci 2008). The pretreatment with a liposomal αGC (lipo-αGC) formulation resulted in the remarkable suppression of both IgG and IgE antibody responses in mice immunized with alum-absorbed ovalbumin. In addition, administration of RGI-2001, a clinical study grade of lipo-αGC formulation, at the timing of bone marrow transplantation could induce the expansion of tolerogenic DCs and/or allo-specific Tregs in mouse models of either acute graft versus host disease (GvHD) or the mixed chimera formation (Duramad et al. Biol Blood Marrow Transplant 2011, Hirai et al. Am J Transplant 2014 and Hirai et al. Am J Transplant2016). Taken together, it was suggested that administration of lipo-αGC formulations prior to the ERT could work for the protection of the enzyme-specific antibody responses.To test the suppressive capability of a lipo-αGC for ERT application, a mouse system to monitor a factor VIII-specific IgG antibody titer and IgE level in bloods was established. In BALB/c mice given the repeated intravenous injections of recombinant factor VIII, the factor VIII-specific IgG titer and total IgE concentration were substantially elevated (ERT-mice). The mice pretreated with the intravenous single injection of the lipo-αGC showed that not only the factor VIII-specific IgG but also total IgE was significantly suppressed compared to the untreated control mice. The antibody-suppressive efficacy by multiple injection of the lipo-αGC was comparable with that by single injection in the ERT-mice. However, cytokine profile of splenic iNKT cells was dramatically changed in BALB/c mice pretreated with either single or multiple lipo-αGC injections. The iNKT cells derived from mice pretreated with the single the lipo-αGC injection expressed high level of IFN-γ, IL-4 and IL-13 mRNAs but not IL-10 mRNA. Conversely, the iNKT cells pretreated with multiple the lipo-αGC injections expressed higher level of IL-10 mRNA more than IFN-γ mRNA, but remarkably diminished both IL-4 and IL-13 mRNAs. As known that αGC-activated iNKT cells could rapidly produce cytokines within a few hours, we then examined the last injection timing of multiple injections into the ERT-mice. The last injection at the timing of the ERT start did not show stronger efficacy than that at three day before the ERT start. Results collectively suggested that the suppression of IgG/IgE formations in the ERT-mice might not be caused by high IL-10 expression of iNKT cells by pretreatment with multiple the lipo-αGC injections, but other phenomena generated in three days after the interaction of activated iNKT cells with αGC-presenting MZ B cells.We conclude that the lipo-αGC pretreatment is capable of protecting harmful formation of both the enzyme-specific neutralizing IgG and anaphylaxis-inducing IgE antibodies during ERT. DisclosuresNo relevant conflicts of interest to declare.

Phycocyanin Enhances Secretary IgA Antibody Response and Suppresses Allergic IgE Antibody Response in Mice Immunized with Antigen-Entrapped Biodegradable Microparticles

In the present study, we have investigated the effects of phycocyanin, a biliprotein of Spirulina platensis, on mucosal and systemic immune responses and allergic inflammation in C3H/HeN and BALB/cA mice. To induce the antigen-specific antibodies in the peripheral lymphoid tissues such as Peyer's patches and mesenteric lymph nodes, biodegradable ovalbumin-entrapped poly (DL-lactide-co-glycolide) particles were used as an antigen. Two weeks after the onset of phycocyanin ingestion, mice were immunized with an aqueous ovalbumin (OVA) solution. Starting at one week after the primary immunization, the mice were subjected to oral immunization with the biodegradable OVA microparticles twice a week. IgA, IgE and IgG1 antibodies were determined by ELISA. The OVA microparticles of 4-microm diameter successfully induced antigen-specific antibodies. In the mice that received phycocyanin treatment for 6 wk, a marked increase in the antigen-specific, as well as the total, IgA antibody level was observed in the Peyer's patches, mesenteric lymph nodes and intestinal mucosa as well as in the spleen cells. Both antigen-specific IgG1 and IgE antibody levels in the serum were suppressed by ingestion of phycocyanin for 8 wk. However, inflammation of the small intestine, monitored as vascular permeability by the Evans blue-leaking method was reduced by phycocyanin at 6 wk, which preceded the suppression of antigen-specific IgG1 and IgE antibody production by 2 wk. These results suggest that phycocyanin enhances biological defense activity against infectious diseases through sustaining functions of the mucosal immune system and reduces allergic inflammation by the suppression of antigen-specific IgE antibody.

SUPPRESSION OF IgE ANTIBODY RESPONSE IN MICE BY A POLYSACCHARIDE, AZ9, PRODUCED BY KLEBSIELLA OXYTOCA STRAIN TNM3

ABSTRACTA soil bacterium, Klebsiella oxytoca TNM3 was found to produce a polysaccharide named AZ9 that shows suppressive effects on IgE antibody response in mice. When mice were administered with 50∼ 100 mg/kg AZ9 subcutaneousely for 4 consecutive days after immunization with trinitrophenyl (TNP)-keyhole limpet hemocyanin, anti-TNP IgE production was significantly suppressed, while the level of anti-TNP IgM was affected marginally. In AZ9-administered mice, IL-4 secretion from splenic cells was reduced to ∼30% of the untreated control. Thus, AZ9 suppression of IgE production may be due to attenuating effects on the Th2-type response. Although oral administration of AZ9 alone had no effects on IgE production, ovalbumin (OVA)-induced oral tolerance of anti-TNP IgE response to TNP-OVA was markedly augmented when a suboptimal dose of OVA was administered orally in combination with AZ9. Collectively, our data suggest that AZ9 has beneficial suppressive effects on IgE-dependent allergic responses.

Induction of IgE Antibody Response in Mice by Continuous Administration of Excretory/secretory Products of the Nematode Nippostrongylus brasiliensis.

IgE-inducing potential of excretory/secretory products (ES) of the nematode Nippostrongylus brasiliensis was examined in mice immunized with ES without use of adjuvant. Repeated i.p. injections or continuous i.p. or s.c. infusion of ES induced significant increases in total serum IgE. Continuous i.p. or s.c. infusion of ES also induced ES-specific IgE antibody, while repeated i.p. injections did not. IL-4 production was increased by ex vivo stimulation with ES of mesenteric lymph node cells from repeatedly immunized mice.On the other hand, continuous intraduodenal infusion of ES induced neither total nor specific IgE responses. These results suggest that continuously secreted ES from the nematode are relevant for the up-regulation of IL-4 and IgE responses, although additional factor (s) may be necessary for the uptake of ES across the intestinal mucosal barrier.

Selective augmenting effects of nitric oxide on antigen-specific IgE response in mice

Here, we report the enhancing effects of nitric oxide (NO) on an IgE antibody response in mice. Anti-trinitrophenyl (TNP) IgE production induced in vitro in TNP keyhole limpet hemocyanin (KLH)-primed spleen cells was inhibited by approximately 70% when an NO synthase (NOS) inhibitor, l-NG-monomethyl-l-arginine, was added at 10−7–10−6 M to the lymphocyte culture. On the other hand, addition of NO-generating agents to the culture resulted in a marked enhancement of the IgE production. In contrast, anti-TNP IgM and IgG1 responses were affected only marginally when the IgE production was either suppressed or augmented by these agents. NO did not directly augment IgE class switching in normal B cells stimulated with lipopolysaccharide and interleukin (IL)-4. NO-mediated augmentation of the IgE response is considered to be of a physiological significance because administration of aminoguanidine (AG), an inhibitor of inducible NOS, to immunized mice resulted in a preferential suppression of anti-TNP IgE production in vivo. This may be explained by the observation that AG-administration increased interferon-γ expression without changing that of IL-4 in the immunized mice. Taken together, these observations suggest a pathophysiological role of NO in the development of IgE-mediated allergic diseases.

Evaluation of Allergenicity of Egg Yolk Immunoglobulin Y and Other Egg Proteins by Passive Cutaneous Anaphylaxis

The allergenicity of egg yolk immunoglobulin (IgY) was studied using passive cutaneous anaphylaxis. It was observed that the intact yolk, water soluble fraction of the yolk, whole IgY molecule or its antigen binding (Fab′) and crystallizable (pFc′) fragments produced by pepsin digestion, elicited low IgE antibody response in mice when compared to egg white, a known allergen. The pFc′ fragment of IgY was more antigenic than the Fab′ fragment as determined by ELISA. Low cross-reactivity was also observed between the egg yolk proteins and egg white antibodies. The low IgE antibody response to the egg yolk proteins was associated with a higher suppressor T-lymphocytes or lower helper T-lymphocytes, or both, in comparison to egg white derived lymphocytes.

Suppressive Effect Of Withania Somnifera Root Extract on the Induction of Anti-Ovalbumin IgE Antibody Response in Mice

Withania somnifera Dunal (Solanaceae) (WS), a well known Indian herbal drug, was examined for its effect on downregulation of antigen-specific IgE antibody response in mice. The extract prepared from the dried roots of WS (WSE) was administered intraperitoneally along with ovalbumin (OVA), a classical allergen, in the presence of aluminum hydroxide as an adjuvant. It exerted a significant suppression of OVA-specific IgE antibody production in BALB/c mice (H-2 d) as determined by passive cutaneous anaphylaxis (PCA). The extract also inhibited the production of OVA-specific IgE antibody, when administered 24 h prior to, and 6 h after, immunization. Further, WSE not only down-regulated OVA-specific IgE antibody response in other haplotypes of mice such as C57Bl/6 (H-2 b) and SWR/J (H-2 q), but it also suppressed the antigen-specific IgE antibody response against other allergens tested in BALB/c mice. Thus, the basic concept for its appli cation in the alleviation of different IgE mediated immunopathological conditions is supported.

Effects of cyclosporine and dexamethasone on IgE antibody response in mice, and on passive cutaneous anaphylaxis in the rat.

The suppressive effects of cyclosporine A (CsA) and dexamethasone (Dxt) on antigen-specific IgE responses to ovalbumin (OA) were studied in BALB/c mice. The effects upon other isotypes were also analyzed. The antiovalbumin IgE response did not change when low doses of CsA [8 mg/kg intraperitoneally (i.p.)] were administered; IgA also remained unchanged, while IgG and IgG1 decreased significantly. At higher CsA doses (16 mg/kg i.p. or orally), a decrease was noted for all the ispotypes assayed. Dxt administered orally at 0.3 mg/kg selectively inhibited IgE and IgA but did not influence IgG or IgG1 levels. Delayed hypersensitivity reactions to OA were not modified by CsA, but were depressed by Dxt. Although CsA had not effect on passive cutaneous anaphylaxis in the rat, Dxt significantly reduced this reaction when it was administered 6 h before challenge. These results suggest that Dxt has more specific antiallergic activity than CsA.

Suppression of IgE antibody response in mice by a naphthalene derivative, TEI-6472

In the present report, we investigated suppressive effects of a naphthalene derivative, (7E)- N-(2-carboxyphenyl)-8-(2-naphthyl)-5,6-trans-5, 6-methano-7-octenamide L-lysine salt (TEI-6472), on in vitro and in vivo antigen-specific IgE response. Anti-trinitrophenyl (TNP) IgE response induced in vitro in TNP-keyhole limpet hemocyanin (KLH)-primed murine spleen cells was suppressed by about 60% in the presence of 10 −6 M TEI-6472. On the other hand, anti-TNP IgG1 and IgM response were not significantly suppressed under the same conditions. Proliferative responses of BALB/c spleen cells stimulated by lipopolysaccharide, concanavalin A or allogeneic spleen cells were not inhibited by TEI-6472 at 10 −6-10 −5 M. Interleukin 4 production from helper T-cell clone, D10.G4.1 was suppressed only slightly (less than 20%) at 10 −6 M TEI-6472. This compound was also effective in suppressing secondary anti-TNP IgE response in mice that were immunized twice with TNP-KLH and alum. When 10–20 mg/kg/day TEI-6472 was administered s.c. for 5 consecutive days starting from one day before the first and the second immunization, secondary anti-TNP IgE response was inhibited most strongly (40–45%). Anti-TNP IgG1 response was also inhibited but to a smaller extent (20–24%), while anti-TNP IgM response was suppressed only slightly (0–15%). These results suggest that, under appropriate conditions, TEI-6472 can suppress IgE responses more preferentially both in vitro and in vivo.

Factors in ruminant colostrum that influence cell growth and murine IgE antibody responses.

Bovine colostrum was investigated as a source of biologically active molecules capable of stimulating the growth of mammalian cells in culture and modifying the immune response in a murine model. An extract prepared from bovine colostral whey by cation exchange and reversed-phase chromatography stimulated the growth of L6 rat myoblasts, Balb/c-3T3 mouse fibroblasts and BHK-21 baby hamster kidney cells with equal or greater potency than fetal bovine serum. Fractionation of the bovine colostral extract by gel-permeation chromatography in M-acetic acid identified a number of cell-growth factors for each cell type. Bovine colostral extract was compared with an ovine colostral whey preparation for its ability to modulate IgE antibody responses in mice. Doses of 8 and 4 mg/d of ovine colostral whey or bovine colostral extract specifically suppressed IgE antibody responses, whereas at lower doses suppression did not occur. We conclude that bovine colostrum contains cell-growth factors as well as immunomodulatory factors that are able to regulate the IgE response in a heterologous species.

Effect of anti-DNP IgG1- and IgG2a-secreting hybridomas in vivo on the development of an anti-DNP IgE antibody response in mice.

We reported previously that CBA mice pretreated with dinitrophenyl-Bordetella pertussis (DNP-BP) conjugates exhibited sharply decreased anti-DNP IgE, and increased IgG2a antibodies following immunization with DNP-ovalbumin (DNP-OA) in alum. The objective of the present experiments was to determine whether the decrease in anti-DNP IgE was attributed to a regulatory effect exerted by IgG2a antibodies. Anti-DNP monoclonal antibodies (Mab) of the IgG1 or IgG2a isotype were passively transferred to mice, 24 h before a primary immunization with DNP-OA in alum. Anti-DNP IgE production was drastically suppressed in recipients of IgG1 but not of IgG2a Mab. Similar results were obtained when the Mab were endogenously produced by intraperitoneal implantation of anti-DNP-secreting hybridomas into (BALB/cxCBA)F1 (BCF1) mice. However, neither IgG1 nor IgG2a isotypes suppressed IgE antibody production if the hybridoma implantation took place 10 days after hapten priming. These results are, to our knowledge, the first to show a clear dissociation between the effect of either passively transferred or endogenously secreted IgG1 and IgG2a antibodies in their ability to inhibit a primary anti-hapten IgE antibody response.

Isolation of Ascaris suum components which suppress IgE antibody responses.

An extract from adult Ascaris suum was fractionated on a Sephadex G-200 column and individual elution fractions assessed for their ability to induce or suppress an IgE antibody response in mice immunized simultaneously with a heterologous antigen (ovalbumin). The results showed that, whilst fractions eluted in the first peak suppressed the anti-ovalbumin IgE antibody production, those eluted in the third peak induced a significant anti-Ascaris IgE response. The mixture of the two kinds of fractions resulted in suppression of anti-Ascaris IgE antibodies. The apparent molecular weight (MW) of the first and third peaks was 530,000 and 29,000 daltons, respectively. After delipidation, the extract still retained most of its biological activities. SDS-PAGE of this material showed selective loss of high-MW components when compared to the whole extract. No bands were observed in SDS-PAGE of peak I in contrast to peak III, which displayed several bands. Immunoblots of all these samples showed at least 3 bands above MW 150,000, which reacted with anti-peak I antiserum, in the unfractionated extracts and in peak I, but not in peak III. When the extract was fractionated by affinity chromatography using anti-peak I antibodies as ligands, the immunogenic components were present in the effluent, whereas the suppressive components were recovered in the eluate.

Enhancing effect of suspended particulate matter on the IgE antibody production in mice.

Suspended particulate matter (SPM), suspended in the polluted environmental atmosphere, are perpetually inhaled into the human body and are considered to have profound effects on human health. This study investigated the enhancing effect of SPM on the IgE antibody production in mice. The IgE antibody responses in mice immunized with intranasal administration of ovalbumin (OA) plus SPM at 3-week intervals were higher than responses in the animals immunized with OA alone. When the dose of OA administered as an antigen was 0.25 microgram, the time course and magnitude of enhancement by SPM was comparable to those by killed Bordetella pertussis, a common adjuvant. SPM had an enhancing effect on IgE antibody production even in a small dose such as 0.25 microgram administered at 3-week intervals. The possibility cannot be excluded that the natural exposure of humans to SPM in the environmental atmosphere may explain the high prevalence rate of allergic rhinitis caused by pollens in polluted districts in Japan.

Diesel-exhaust particulates inoculated by the intranasal route have an adjuvant activity for IgE production in mice

Our previous study indicated that the IgE antibody responses in mice immunized with intraperitoneal injection of the antigens mixed with diesel-exhaust particulates (DEP) were higher than those in the animals immunized with the antigens alone. We examined the adjuvant activity of DEP inoculated by the intranasal route, i.e., the natural entrance of DEP. In 3-week interval immunization, the IgE antibody responses in mice immunized with intranasal inoculation of ovalbumin (OA) mixed with DEP were higher than responses in the animals immunized with OA alone. DEP had an adjuvant activity for anti-OA IgE antibody production, even in a small dose such as 1 micrograms administered with a 3-week interval. Also in 1-week interval immunization, the enhancing effect of DEP on anti-OA IgE antibody production was demonstrated when mice were immunized with intranasal inoculation of OA and DEP. The possibility cannot be excluded that DEP, which are kept buoyant in the environmental atmosphere of urban districts, may exert an adjuvant activity for IgE antibody production after being inhaled into the human body and have some relation to the mechanism of the outbreak of allergic rhinitis caused by pollens in Japan.

Adjuvant activity of diesel-exhaust particulates for the production of IgE antibody in mice

The prevalence rate of allergic rhinitis caused by pollen has strikingly increased in Japan in the last three decades. The number of diesel cars in use has also rapidly increased in the country. This fact urged us to study the effects of particulates emitted from diesel cars on the production of IgE antibody. The primary IgE antibody responses in mice immunized with intraperitoneal injection of ovalbumin (OA) mixed with diesel-exhaust particulates (DEP) were higher than those in the animals immunized with OA alone. This effect of DEP on the production of IgE antibody in mice was also demonstrated when mice were immunized with repeated injections of dinitrophenylated-OA. In addition, persistent IgE-antibody response to major allergen of Japanese cedar pollen (JCPA), a most common pollen causing allergic rhinitis in Japan, was observed in mice immunized with JCPA mixed with DEP but not in the animals immunized with JCPA alone. The results do indicate that the adjuvant activity of DEP can not be excluded as a possible cause of the associated change in the number of diesel cars and allergic rhinitis caused by pollen in Japan.

Suppression of Murine IgE Responses with Amino Acid Polymer/Allergen Conjugates

Conjugates of poly-N-methylglycine (polysarcosine) and grass pollen extracts, previously found to be capable of suppressing immature IgE antibody responses in mice, were shown to be highly effective at inhibiting the capacity of immune splenocytes to produce a secondary response in sub-lethally irradiated recipient animals. Anamnestic IgE responses in mice primed without adjuvant were also suppressed, but the effects were modest and of short duration. The predictive value of murine models for selecting clinically appropriate specific IgE suppressive agents and treatment schedules that might be successfully employed for clinical use are discussed.

Melittin-specific monoclonal and polyclonal IgE and IgG1 antibodies from mice.

Melittin, a bee venom peptide consisting of 26 amino acid residues, elicited high IgG and IgE antibody responses in mice of BALB/c and CAF1 strains, but not in mice of A/J, AKR, and C57BL/6 strains. Greater than 80% of the melittin-specific antibodies in sera of responder mice were found to bind the hydrophilic carboxyl-terminal heptapeptide of melittin. Three melittin-specific monoclonal antibodies were obtained from responder mice by the hybridoma technique. Two are of the IgG1 isotype and one is of the IgE isotype. One monoclonal antibody of the IgG1 isotype binds the carboxyl-terminal heptapeptide of melittin, while the other two monoclonal antibodies do not. However, competitive binding studies suggest that all three monoclonal Ig bind at the same, or adjacent, site of melittin. These findings, together with the known amphiphilic property of melittin, suggest that the immunogenicity of this peptide is a consequence of its binding to cell surface phospholipids.

Role of antigen structure in the regulation of IgE isotype expression.

Tobacco glycoprotein (TGP) is a glycoprotein containing rutin-like polyphenol groups that is purified from cured tobacco leaves and can be detected in condensates of tobacco smoke. One-third of normal humans have been shown to manifest immediate, IgE-mediated, wheal and flare reactions to an intradermal injection of TGP. Rutin-like moieties are also found in a wide variety of vegetable foods. The possible importance of sensitivity to TGP in the pathogenesis of the vascular and pulmonary complications of tobacco smoking has stimulated us to study the immune response of mice to TGP and the role of rutin groups in influencing isotype expression. A series of three intradermal injections of TGP elicits a long-lasting IgE antibody response in mice. However, no hemagglutinating antibodies are produced. Similarly, immunization with a rutin derivative of bovine serum albumin stimulates IgE antibodies to bovine serum albumin but little hemagglutinating antibodies. In contrast, mice injected in the same manner with bovine serum albumin produce both IgE and hemagglutinating antibodies. Thus, the rutin moiety is implicated as exerting a regulatory effect on isotype expression by suppressing the production of serum antibodies of isotypes other than IgE. The immunization procedure employed (which involves an initial injection of 100 micrograms of antigen in phosphate-buffered saline, followed, at monthly intervals, by two intradermal injections of 100 micrograms of antigen precipitated on alum) apparently fails to stimulate the normal "down-regulation" of the IgE response so that a persisting high-titered response is obtained.

105 Enhancement of IgE antibody response in mice by a pollen mitogen

105 Enhancement of IgE antibody response in mice by a pollen mitogen

Boosterable IgE antibody response in mice without the use of adjuvant.

A mouse model is presented giving boosterable IgE antibody responses preceding the IgG antibody responses without using adjuvants. Groups of CBA mice were injected subcutaneously with minute doses of penicilloyl bovine gammaglobulin, 3.8-94 ng daily for one or two 10-day periods. Some groups of animals were simultaneously given Bordetella pertussis bacteria intraperitoneally. The IgE antibody responses were recorded with passive cutaneous anaphylaxis (PCA) in rats as well with RAST. IgG antibody responses were determined with a double antibody assay. An overall good agreement between the antibody activity in the PCA and RAST was seen, both tests showing boosterable IgE antibody responses. Some discrepancies between the two tests are discussed.