Antibody Formation In Mice Research Articles

Potentiation of the tolerogenicity of benzylpenicilloylated eicosa-L-lysine by conjugation with 4-methoxybenzyl 3.beta.-cholestanyl succinate

It was previously found that amino acid polymers such as oligolysines bearing haptenic groups in high densities efficiently suppress anti-hapten IgE antibody formation. Such conjugates are strong elicitors of anaphylaxis and therefore may not be used for desensitization of drug allergic patients. Here we report on the synthesis and immunological evaluation of benzylpenicilloylated (BPO) eicosa-L-lysines containing none, one, or two lipophilic p-(hydroxymethyl)benzyl cholestan-3 beta-yl succinate (OSuco) groups. The lipophilic derivatives suppress primary as well as ongoing anti-BPO IgE antibody formation in mice much more efficiently than their hydrophilic counterpart. The lipophilic but not the hydrophilic derivatives form stable micelles in water and suppress the antibody formation according to different cellular mechanisms. The relationship between structure, hydrophobicity, and mode of action is discussed.

Further evidence against random polyclonal antibody formation in mice with lupus-like graft-vs-host disease.

The pathologic symptoms in F1 mice with chronic graft-vs-host disease (GVHD) (GVH F1) strongly resemble those of systemic lupus erythematosus (SLE). Mice with SLE-like GVHD do not produce antibodies to a number of non-self and self antigens. This finding is inconsistent with the widely accepted view that the (auto)-antibody formation in SLE is polyclonal in the sense that B cells are triggered at random, i.e., irrespective of their specificity. In the present study, therefore, we performed a systematic study of the kinetics of total IgM- and IgG-secreting splenic B cells and tested their specificities. The total IgM-secreting B cell population was increased only in the first week after the initiation of SLE-like GVHD; it seemed to reflect a random, but self-limited, polyclonal B cell stimulation. In contrast, the total number of IgG-secreting cells in the GVH F1 mice was increased to a much higher extent than that of the IgM-secreting cells and remained increased. At no time during GVHD was there an increase in the number of plaque-forming cells (PFC) spontaneously secreting IgG antibodies to non-self antigens. The GVH reaction (GVHR) did, however, lead to the appearance of PFC that secreted IgG antibodies to DNA. Similarly, the GVH F1 mice showed high serum titers of antibodies to self antigens characteristic of SLE and to endogenous viruses, but during the entire observation period they failed to develop serum antibodies to non-self antigens and insulin. Hence, the enhanced production of Ig, especially that of IgG, that occurs in SLE-like GVHD is not a random process, because it requires the presence of antigen, or signal 1. The data support our hypothesis that only certain kinds of self antigen, such as DNA and cell membrane epitopes, can cross-link the Ig receptors on the corresponding B cells and thus provide an adequate signal 1. Given the increase in help, or signal 2, in chronic GVHD, only the B cell clones that simultaneously receive an adequate signal 1 seem to be driven into clonal proliferation and IgG secretion.

Effect of vitamin E on IgE antibody formation in mice.

Effect of vitamin E (alpha-tocopheryl acetate and alpha-tocopheryl nicotinate) on IgE antibody formation in mice was investgiated . Female BALB/c mice were immunized with dinitrophenylated ascaris protein (DNP-As) and aluminium hydroxide gel (alum). Supplementation of vitamin E in diets or oral administration of vitamin E mixed with sesame oil resulted in a suppression of IgE antibody formation. On the contrary to IgE antibody formation, IgM or IgG (hemagglutinin; HA) formation was significantly enhanced. These results indicate that vitamin E is capable of suppressing IgE antibody formation and enhancing non-IgE antibody formation.

Effect of E-64, a thiol protease inhibitor, on antibody formation in mice

E-64, L- trans -epoxysuccinyl-leucylamido (4-guanidino) butane, a specific inhibitor of thiol proteases originally isolated from a culture broth of fungi, and its synthetic analogues, were examined for immune responses to the splenocytes of BDF 1 mice. In the cultures of 2-day-primed splenocytes of the mice, E-64 and its close analogues, increased the number of direct splenic hemolytic plaque forming cells (PFC). In addition, it was demonstrated that E-64 enhanced the PFC responses in the mice. These results suggested that some thiol proteases might be involved in the immune response process in mice.

Adjuvant and immunogenic properties of gonococcal R-type lipopolysaccharide in reaginic antibody formation in mice.

The ability of gonococcal R-type lipopolysaccharide (LPS) to function as an adjuvant and as an antigen in the formation of IgE and IgG1 antibody responses in mice was investigated. LPS failed to induce LPS-specific IgE or IgG1 under a variety of experimental conditions. LPS was capable of enhancing IgE and IgG1 antibody responses to gonococcal protein (ZAB), but its adjuvant effect was weaker than that of A1(OH)3. The LPS-induced anti-ZAB IgE antibody titers showed a cycling phenomenon with time, but the IgG1 response was delayed and peaked only once.

Specific antibody responses in rats and mice after daily immunization without adjuvant.

We have previously devised an immunization protocol to obtain IgE antibody formation in mice without the use of adjuvants. In this communication the observations were confirmed using several antigens and various routes of administration. The studies were also extended to rats. Subcutaneous (s.c.) administration to rats of 100-ng doses of trinitrophenylated human serum albumin (TNP-HSA) in 10-day periods resulted in production of IgE antibodies as determined with the RAST using antirat IgE. Higher antigen doses resulted in lower IgE antibody responses. Intranasal (i.n.) or peroral (p.o.) exposure of rats did not result in any IgE antibody responses. Exposure to antigen by the s.c. and i.n. route but not p.o. administration induced IgG, IgM and IgA antibody responses in rats as recorded with the enzyme-linked imunosorbent assay (ELISA). Several different antigens were given to CBA/Ca mice, i.e. penicilloylated bovine gammaglobulin (PO-BGG), FITC-conjugated dog serum albumin (FITC-DSA) or timothy pollen extract. All the antigens induced boosterable IgE antibody responses as determined with RAST utilizing antimouse IgE. Antibodies of IgG isotype were also recorded after such immunizations, whereas only low IgM and no IgA antibody titers were found in serum employing ELISA. After similar daily administration of antigen by the p.o. or i.n. route of antigen at doses from 10<sup>2</sup> to 10<sup>5</sup> ng, IgG antibody responses but no IgE antibodies were found. Intranasal administration of 10<sup>6</sup> ng induced some IgE and IgA antibodies. A single p.o. administration of 4 × 10<sup>7</sup> ng of antigen induced an antibody response of the IgG class. Pretreatment of animals with 4 × 10<sup>7</sup> ng of PO-BGG orally prior to the 10-day periods of s.c. injections suppressed the IgE antibody responses by 80% but enhanced the IgG antibody response. It was concluded that IgE antibody responses can be induced in rats and mice by daily s.c. administration of small doses of a variety of antigens without the use of any adjuvants. Antibody responses of the other isotypes may also be raised by such immunization. Intranasal and p.o. administration is less efficient in this respect, possibly requiring much higher antigen doses. The protocol for s.c. immunization appears to be suitable for studies of the regulation of an allergic immune response.

Kinetics and genetics of herpes simplex virus-induced antibody formation in mice.

The kinetics of antibody synthesis was investigated after intraperitoneal, subcutaneous, and footpad infection of various strains of mice with herpes simplex virus. Immunoglobulin M antibodies appeared 5 days after and immunoglobulin G antibodies appeared 10 to 12 days after intraperitoneal infection with herpes simplex virus type 1. The major histocompatibility complex and the background genome of inbred mice were not found to have a systematical influence on antibody synthesis. Female mice, however, consistently produced more antibodies than did male if the infection was done intraperitoneally, but not if it was done subcutaneously or into footpads. Castration considerably increased the amount of antibodies produced by male mice. The difference in antibody formation between females and males could be abolished by injection of silica; moreover, antibody titers were enhanced by this treatment. This has also been found by immunization with a Formalin-inactivated herpes simplex virus vaccine. The effect of silica in enhancing antibody formation could be observed up to 12 days after infection. Infectious virus could be detected up to 2 days after infection, and herpes simplex virus type 1 antibody-stimulating antigens could be detected up to 4 days in ultrasonicates of macrophages. The assumption is made that androgen-sensitive cell populations, including macrophages and their soluble products, are involved in antibody-depressing mechanisms.

IgE antibody responses induced by repeated administration of antigens without adjuvant.

AbstractIt is generally considered that the induction of IgE antibody responses in laboratory animals to parenterally injected antigens requires the simultaneous administration of an adjuvant such as Al(OH)3 gel or Bordetella pertussis vaccine. In this study, we have been able to induce IgE antibody formation in mice to antigens such as heterologous whole serum or ovalbumin by giving multiple intraperitoneal injections of these substances without any added adjuvant. The IgE responses were measured by the passive cutaneous anaphylaxis (PCA) test performed in rats. The abrogation of PCA activity by heat treatment of undiluted serum for one hour at 56 ° was confirmed. We propose that this represents an ideal model to study induction of IgE antibody formation.

Effect of cadmium on japanese encephalitis virus infection in mice 1. Acute and single-dose exposure experiment

When mice were treated with 0.09 mg cadmium chloride (Cd) per mouse once and inoculated i.p. simultaneously with Japanese encephalitis virus (JEV) they showed a significant difference in the incubation time and mortality between the treated and untreated groups in repeated experiments. Cd treatment shortened the incubation time and the mortality increased > twice compared with the untreated control. This effect was not observed in the case of intracerebral inoculation of JEV. Effects of Cd on antibody formation in mice were also determined. Animals given a single s.c. dose of Cd were immunized with JE inactivated vaccine once simultaneously. When mice were treated with Cd, they did not show low neutralizing and hemagglutination inhibition activities compared with the control mice. Pretreatment of Cd did not affect any mortality or antibody formation.

Reaginic antibody formation in the mouse: XI. Participation of long-lived antibody-forming cells in persistent antibody formation

A persistent IgE anti-hapten antibody response was established by immunization of B 6D 2F1 mice with a minute dose of aluminum hydroxide-gel-absorbed dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH) or DNP-ovalbumin (OA). Irradiation of the immunized mice with a lethal dose of X-rays suppressed secondary anti-hapten antibody responses to DNP-homologous carrier conjugate, and depleted hapten-primed B memory cells in their spleen. However, persistent antibody formation of both IgE and IgG isotypes in the immunized mice was not affected by the lethal dose of X-rays. The results indicated that recruitment of B memory cells to antibody-forming cells is not responsible for the maintenance of antibody levels. The hapten-primed B cell population was expanded by adoptive transfer of DNP-KLH-primed spleen cells into irradiated mice followed by challenge with homologous antigen. Culture of T-cell-depleted spleen cells of the recipients, in the absence of exogenous antigen, resulted in the production of both IgE and IgG anti-DNP antibodies. The antibody titer in the culture supernatants continued to increase for 14 days, indicating that long-lasting antibody-forming cells are present in the spleen. The antibody-forming cells are radioresistant. The in vitro production of IgE anti-DNP antibodies by the spleen cells was not affected by the presence of IgG anti-DNP antibody, but was inhibited by exposure of the cells to multivalent DNP-heterologous carrier conjugates, such as DNP-bovine IgG (BGG) or DNP-mouse IgG (MGG). The persistent anti-hapten antibody formation of DNP-OA-primed and irradiated mice was depressed by an injection of DNP-MGG, which induced secondary anti-hapten antibody responses in the antigen-primed and nonirradiated mice. The results support the concept that radioresistant long-lived antibody-forming cells are involved in persistent antibody formation in the irradiated mice.

Immunosuppressive action of oral Streptococci.

The study on the immunosuppressive activity of the cytoplasmic component (SF) from Streptococcus mitis on the immune response of mice to the sheep red blood cells (SRBC) was carried out. The bacterial cell suspension was disrupted in a Braun shaker. The disrupted cell suspension was diluted and centrifuged at 10,000 x g for 30 minutes to remove the cell walls. The supernatant was centrifuged at 31,000 x g for 45 minutes and the final supernatant was dialyzed against distilled water, frozen and lyophilized. These products were employed as the cytoplasmic component (SF). The SF in the extracts of Streptococcus mitis suppresses the antibody formation in mice against SRBC. It is effective to inject it just one day before the inoculation with SRBC.

Antibody response to bovine serum albumin in mice: the efffects of psychosocial environmental change.

AbstractThe effects of psychosocial environmental change upon circulating antibody response to antigenic challenge was investigated in CBA/USC mice. Mice were reared in isolation and selected groups were subsequently exposed to psychosocial stimulation. Antibody titers of mice that remained in isolation were significantly higher than the titers of mice exposed to psychosocial stimulation. One group of mice exposed to psychosocial stimulation and then returned to isolation showed titers significantly below those of mice exposed to psychosocial stimulation only. These data indicate that psychosocial environmental changes can be productive of significant suppression of antibody formation in mice.

Administration of antigen to digestive tract and formation of antibodies in mice (author's transl)

Administration of antigen to digestive tract and formation of antibodies in mice (author's transl)

Suppression of IgE Antibody Formation in Mice with Nonspecific Stimulation

Effects of administration of various adjuvants were examined for the induction of IgE antibody response against a hapten-carrier conjugate in the mouse. It was confirmed that alum was an excellent adjuvant for induction of IgE antibody response, while poly A:U, CFA and LPS had much less activity. However, when these adjuvants were administered before immunization with antigen and alum, they suppressed the forthcoming antibody response to various degrees. Even alum exerted a profound suppressive effect when it was given before immunization. Such suppressive effect of alum and poly A:U was partially overcome by incorporating in the adjuvant the carrier protein. In contrast the suppressive effect of CFA was overcome by incorporating in the CFA an unrelated carrier conjugated to the same hapten (DNP). The results indicate that the target of the adjuvants is different from one to the other and further give a clue to study the possible application for the suppression of IgE antibody formation.

Experimental evidence of a decreased incidence of penicillin allergy through use of pure penicillins.

The results revealed the presence of high molecular weight impurities in commercially available penicillins, measured with a radioimmuno assay. The impurities had penicilloyl specificity and induced antibody formation in mice when the contaminated penicillin was administered in 50 mg/kg body weight daily for ten day periods with a 20 to 30 day interval. Penicillin of high purity similarly administered produced very few antibodies. Furthermore, experimentally contaminated penicillin given according to the same schedule caused IgE antibody formation against the penicilloyl moiety, while pure penicillin did not. These findings were explained by the weak immunogenicity of isologous penicilloylated serum albumin in rabbits both regarding the IgE and the IgG/IgM antibody formation compared to the immunogenicity of heterologous bovine serum albumin similarly penicilloylated.

Synergism of methylmercury and selenium producing enhanced antibody formation in mice.

Mice fed 1, 5, and 10 ppm methylmercury plus 6 ppm selenium for 10 wk had a significant increase in antibody synthesis. Since methylmercury singly depresses antibody synthesis and the response was greater than that produced by selenium alone, synergism between methylmercury and selenium occurred. In this case, the synergism is considered to be advantageous to a host, while exposure by other combinations of environmental contaminants may be detrimental. Mercury concentrations in the kidney were markedly elevated when methylmercury and selenium were administered simultaneously compared to when methylmercury was given without selenium supplement. These results indicate that data collected from individual pollutants may not be of value in predicting responses to multiple exposure.

Suppression of IgE Antibody Formation in Mice: Requirement for T-T Lymphocyte Interaction

Norman SWR mice injected with syngeneic spleen cells from ovalbumin (Ov)-primed mice were unable to make IgE anti-Ov antibodies when challenged with alum-pertussis-Ov. Immune T lymphocytes were shown to be responsible for the inhibitory effects of adoptively transferred spleen cells. Treatment of recipient mice with mild x-irradiation or with cyclophosphamide 2 or 3 days before cell transfer resulted in abrogation of the suppressor effect of immune cells. The injection of T lymphocytes into x-irradiated milce restored the suppressive effect of immune cells. It thus appears that immune T cells provide the stimulus for activation of suppressor T cells of the host. Although the generation of suppression is antigen-specific, the expression of suppression appears to be nonspecific.

Antigens in Penicillin Allergy

Using a radioimmunoassay, it was shown that commercially available ampicillin preparations often contain penicilloylated high molecular weight impurities. These possess immunological activities and stimulate penicilloyl-specific antibody formation in mice treated according to a therapeutic schedule. Using purified and experimentally contaminated preparations it was also found that exposure of the animals to <i>Escherichia coli </i>and <i>Bordetella pertussis </i>bacteria could increase the antibody formation to small amounts of impurities. In addition, penicilloylated antigen could be recorded in serum from treated animals. The antigen formed by penicilloylation <i>in vivo, </i>however, was very weak and did not induce much antibody formation when injected together with Freund’s adjuvant in mice or rabbits.

21-Methyl-20,21-diketocorticosteroid類の抗炎症作用(第2報)11β, 17α-Dihydroxy-21-methylpregna-1,4-diene-3,20,21-trione 17-Acetate(TSC-5)の抗炎症作用

A new steroid, 11β, 17α-dihydroxy-21-methylpregna-1, 4-diene-3, 20, 21-trione 17-acetate (TSC-5) was tested for its inhibitory effect on acute, subacute and chronic inflammation by systemic administration (p.o. and s.c.) or local application, as well as its glucocorticoid effects in experimental animals. TSC-5 showed potent anti-inflammatory activities comparable to those of prednisolone in acute and subacute inflammation models, but its inhibitory effect on chronic inflammation was less than that of prednisolone in rats. Unlike prednisolone, TSC-5 was found to have local anti-inflammatory activity in rats, where TSC-5 was one half as active as prednisolone 21-tert-butylacetate. TSC-5 was 1/12 as active as prednisolone in glycogendeposition activity by s.c. administration. The inhibitory effect of TSC-5 on antibody formation in mice by s.c. and p.o. administrations was less potent than that of prednisolone. TSC-5 was less potent than prednisolone in thymus-involution effect, but the adrenalinvolution effects of both compounds were almost the same in rats. Thus, TSC-5 showed a significant gap between anti-inflammatory and glucocorticoid activities.

Effect of colchicine on the antibody response. I. Enhancement of antibody formation in mice.

Colchicine (CC) enhances the antibody response in mice to protein antigens, like diphtheria toxoid and human gamma globulin, as well as to the 2,4,6-trinitrophenyl hapten. Maximal enhancement was observed when CC was administered to animals on the same day as the injection of antigen. The optimal dose of CC was in the range of 1.0-1.5 mg/kg body weight. The enhanced antibody formation was evident from elevated circulating antibody titers and from an increased number of antibody plaque-forming cells (PFC) of the spleen. The circulating antibody titer of CC-treated animals was higher than that of control animals by a factor of about 3-7 in the primary response, and by a factor of at least 15 in the secondary response. In terms of the number of antibody forming cells, CC enhanced the primary PFC response by approximately equal to 100%, and the secondary PFC response by as high as fivefold. The enhancing effect of CC seemed to be related to its mitosis-blocking capacity since (a) vinblastine another antimitotic drug, was found to be as effective as CC and (b) lumicolchicine, the non-anti-mitotic structural isomer of CC, was ineffective in potentiating antibody responses. The critical timing in the administration of CC on the same day as antigen suggests that most likely, the mitotic poison was acting on antigen-stimulated early dividing suppressor cells.