Immune deficiency of cataract Shionogi (CTS) mouse. II. Impaired in vivo T cell-mediated immune response

Our previous study demonstrated that cataract Shionogi (CTS) mice, an inbred strain related to non-obese diabetic (NOD) mice, are T lymphocytopenic and that their T cell-mediated in vitro reactions, such as proliferative responses of spleen cells to T cell mitogens and alloantigens or production of IL 2 and IL 2 receptors after stimulation of spleen cells with Con A, are greatly reduced. To confirm these in vitro characteristics, in vivo immune responses of CTS mice to T-dependent and T-independent antigens were compared with those of some reference strains including NOD mice. Antibody responses of CTS mice after one injection of a high dose (10(8)) or one or two injections of a low dose (10(5)) of sheep red blood cells (SRBC) were markedly lower than those of the reference strains. The decrease was particularly striking in the IgM antibody production at primary response to both high and low doses, and the IgG antibody production at the secondary response to low dose. Similar lower antibody production was observed in CTS mice against bovine serum albumin (BSA). Little production of IgE antibody was observed from 1 through 3 weeks after an injection of BSA plus Bordetella pertussis. IgG1 response was observed at high incidence but lower in titer than those in the reference strains. Unexpectedly, in spite of the poor antibody production to BSA, potent systemic sensitization for anaphylactic shock was easily established; incidence of lethal shock being comparable with those in the reference strains. This suggests that CTS mice are highly susceptible to the effector phase of active anaphylactic shock. Cell-mediated immunity was also impaired. Delayed type of hypersensitivity to SRBC was low, and the rejection of the skin graft from NOD mouse did not occur. In contrast to the reduced T cell-mediated responses, no difference was found between CTS and reference strains with regard to the antibody production to LPS, a T-independent antigen. These in vivo findings are consistent with the previous in vitro study.     

Immune Deficiency of Cataract Shionogi (Cts) Mouse. H. Impaired in Vivo T Cell-Mediated Immune Response

Our previous study demonstrated that cataract Shionogi (CTS) mice, an inbred strain related to non-obese diabetic (NOD) mice, are T lymphocytopenic and that their T cell-mediated in vitro reactions, such as proliferative responses of spleen cells to T cell mitogens and alloantigens or production of IL 2 and IL 2 receptors after stimulation of spleen cells with Con A, are greatly reduced. To confirm these in vitro characteristics, in vivo immune responses of CTS mice to T-dependent and T-independent antigens were compared with those of some reference strains including NOD mice. Antibody responses of CTS mice after one injection of a high dose (10⁸) or one or two injections of a low dose (10⁵) of sheep red blood cells (SRBC) were markedly lower than those of the reference strains. The decrease was particularly striking in the IgM antibody production at primary response to both high and low doses, and the IgG antibody production at the secondary response to low dose. Similar lower antibody production was observed in CTS mice against bovine serum albumin (BSA). Little production of IgE antibody was observed from 1 through 3 weeks after an injection of BSA plus Bordetella pertussis. IgG₁ response was observed at high incidence but lower in titer than those in the reference strains. Unexpectedly, in spite of the poor antibody production to BSA, potent systemic sensitization for anaphylactic shock was easily established; incidence of lethal shock being comparable with those in the reference strains. This suggests that CTS mice are highly susceptible to the effector phase of active anaphylactic shock. Cell-mediated immunity was also impaired. Delayed type of hypersensitivity to SRBC was low, and the rejection of the skin graft from NOD mouse did not occur. In contrast to the reduced T cell-mediated responses, no difference was found between CTS and reference strains with regard to the antibody production to LPS, a T-independent antigen. These in vivo findings are consistent with the previous in vitro study.     

An Inhibition Elisa for the Quantification of Collagens Type I and Type II in Cyanogen Bromide-Digested Tissues Using Fragment-Directed Antibodies

The quantification of connective tissue components in small tissue samples is of great importance for the examination of drug-induced changes in the development of the mammalian embryo. An inhibition-ELISA for the quantification of collagens type I and type II in CNBr-digested tissue samples was developed. Fragments of type I collagen were produced by CNBr-cleavage of the pure collagen and partially purified by gel filtration chromatography. A mixture of fragments was used to immunize rabbits. Antisera with highest titres were absorbed with immobilized fibronectin and collagens type II, III, V and I. The eluted antibodies exhibited specificity for α₂(I)-CB4, exclusively. These antibodies, as well as the previously described antibodies with specificity for α₁(II)-CB8, were used for the development of an inhibition-ELISA. The sensitivity of the assay is 0.3ug/ml for collagen I and 3ug/ml for collagen II. To evaluate the value and practibility of the ELISA we have estimated the amounts of both collagens in biochemically well characterized tissues (skin, aorta, chondrosarcoma) and have performed an initial determination in mouse embryos.

Our previous study demonstrated that cataract Shionogi (CTS) mice, an inbred strain related to non-obese diabetic (NOD) mice, are T lymphocytopenic and that their T cell-mediated in vitro reactions, such as proliferative responses of spleen cells to T cell mitogens and alloantigens or production of IL 2 and IL 2 receptors after stimulation of spleen cells with Con A, are greatly reduced. To confirm these in vitro characteristics, in vivo immune responses of CTS mice to T-dependent and T-independent antigens were compared with those of some reference strains including NOD mice. Antibody responses of CTS mice after one injection of a high dose (10⁸) or one or two injections of a low dose (10⁵) of sheep red blood cells (SRBC) were markedly lower than those of the reference strains. The decrease was particularly striking in the IgM antibody production at primary response to both high and low doses, and the IgG antibody production at the secondary response to low dose. Similar lower antibody production was observed in CTS mice against bovine serum albumin (BSA). Little production of IgE antibody was observed from 1 through 3 weeks after an injection of BSA plus Bordetella pertussis. IgG₁ response was observed at high incidence but lower in titer than those in the reference strains. Unexpectedly, in spite of the poor antibody production to BSA, potent systemic sensitization for anaphylactic shock was easily established; incidence of lethal shock being comparable with those in the reference strains. This suggests that CTS mice are highly susceptible to the effector phase of active anaphylactic shock. Cell-mediated immunity was also impaired. Delayed type of hypersensitivity to SRBC was low, and the rejection of the skin graft from NOD mouse did not occur. In contrast to the reduced T cell-mediated responses, no difference was found between CTS and reference strains with regard to the antibody production to LPS, a T-independent antigen. These in vivo findings are consistent with the previous in vitro study.     

High Susceptibility of Cataract Shionogi (CTS) Mice to Passive Anaphylactic Shock Mediated by Allogeneic IgE and IgG1 Monoclonal Antibodies

The susceptibility of cataract Shionogi (GTS) mice as young as 8 to 10 weeks of age to passive anaphylactic shock mediated by anti-benzylpenicilloyl IgE and IgG₁ monoclonal antibodies was compared with those of other strains of the same age including sister strains such as nonobese-diabetic (NOD) and nonobese-nondiabetic (NON). When the animals had been treated with killed Bordetella pertussis organisms, potent sensitization, enough to cause lethal shock, was produced by either monoclonal antibody preparation in CTS, NOD, C57BL/6J and DS/Shi strains, but not at all in C3H/He, DBA/2 and BALB/c strains. In the NON strain, lethal shock was elicited in the animals sensitized with the IgG₁ antibody but not in those sensitized with the IgE antibody. Without the pertussis pretreatment, sensitization sufficient to cause lethal shock was produced at a high frequency by the IgG₁ antibody in CTS and NOD mice but not in the other strains. When the IgE antibody was used, lethal shock was not observed in any of the mouse strains tested except for one CTS mouse. These results indicate that CTS as well as NOD are highly susceptible strains, and that IgG₁ antibody is more effective than IgE antibody for producing systemic sensitization for anaphylactic shock. In addition to these findings, the results revealed an age-dependent potentiation of anaphylactic shock in CTS mice. The IgE antibody-mediated lethal shock was produced in all the aged animals of this mouse strain tested without the Bp treatment, but not in aged NOD and NON mice.     

High Susceptibility of Cataract Shionogi (CTS) Mice to Passive Anaphylactic Shock Mediated by Allogeneic IgE and IgG1 Monoclonal Antibodies

The susceptibility of cataract Shionogi (GTS) mice as young as 8 to 10 weeks of age to passive anaphylactic shock mediated by anti-benzylpenicilloyl IgE and IgG₁ monoclonal antibodies was compared with those of other strains of the same age including sister strains such as nonobese-diabetic (NOD) and nonobese-nondiabetic (NON). When the animals had been treated with killed Bordetella pertussis organisms, potent sensitization, enough to cause lethal shock, was produced by either monoclonal antibody preparation in CTS, NOD, C57BL/6J and DS/Shi strains, but not at all in C3H/He, DBA/2 and BALB/c strains. In the NON strain, lethal shock was elicited in the animals sensitized with the IgG₁ antibody but not in those sensitized with the IgE antibody. Without the pertussis pretreatment, sensitization sufficient to cause lethal shock was produced at a high frequency by the IgG₁ antibody in CTS and NOD mice but not in the other strains. When the IgE antibody was used, lethal shock was not observed in any of the mouse strains tested except for one CTS mouse. These results indicate that CTS as well as NOD are highly susceptible strains, and that IgG₁ antibody is more effective than IgE antibody for producing systemic sensitization for anaphylactic shock. In addition to these findings, the results revealed an age-dependent potentiation of anaphylactic shock in CTS mice. The IgE antibody-mediated lethal shock was produced in all the aged animals of this mouse strain tested without the Bp treatment, but not in aged NOD and NON mice.     

Immune deficiency of the cts mouse. I. Deficiency of in vitro t cell-mediated immune response

The cataract Shionogi (CTS) mouse characterized by cataracts and microphthalmia is a sister strain of the NOD mouse. We have made the immunological characterization of the CTS mouse by means of in vitro assays. Splenocytes of the CTS mouse were very low in the responsiveness to T cell mitogens such as Con A and PHA but not to a B cell mitogen, LPS. The production of IL 2 and expression of IL 2-receptor of spleen cells after in vitro stimulation with Con A decreased in the CTS mouse, when compared with those in the NOD and the other reference strains. In mixed lymphocyte culture, CTS splenocytes did not proliferate and did not generate cytotoxic T lymphocytes when cocultured with splenocytes of the C3H/He mouse. The NK activity against YAC-1 target cells was lower in the CTS mouse than in the C3H/He mouse, an NK high responder, but higher than in the NOD mouse, a low responder. These results suggest that the CTS mouse is deficient in T cells. Subset analysis of splenic lymphocytes of the CTS mouse using flow cytometry revealed that the percentage of T cells in the CTS mouse was significantly lower than those in the reference strains, which was consistent with the reduced responsiveness to T cell mitogens in the CTS mouse. The deficiency in the Ly-2⁺ T cell subset was particularly striking. However, the response to PHA of the splenocytes of the CTS mouse was normalized when T cells were enriched by nylon wool-passing and cell-sorting. Therefore, it seems that decreased T cell activity is due to a decrease in T cell number and not to dysfunction of individual T cells.     

Age-Dependent Difference in Susceptibility to IgE Antibody- and IgG1 Antibody-Mediated Passive Anaphylactic Shock in the Mouse

The age dependence of the susceptibility to passive anaphylactic shock was studied in the mouse. Anti-BPO IgE monoclonal antibody produced potent systemic sensitization sufficient for provocation of lethal shock in most aged (6 to 10 months) CTS, DS and C57BL/6J mice but only in a very few young (1.5 to 2.5 months) mice. A similar trend was found in the NOD strain, though it was not as definite as in the above three strains. Age-dependent potentiation of the IgE antibody-mediated anaphylactic shock was not found in both sexes of five other strains, C3H/He, DBA/2, NON, BALB/c and B6D2F₁. However, the potentiation became obvious even in these strains, when they were treated with Bordetella pertussis before the antigen challenge. Age-dependent potentiation was also clear with IgG₁ antibody-mediated anaphylactic shock in DS females and NON males. In contrast, no age-dependent difference was seen for the shock induced by PAF which is estimated to be the main mediator for anaphylactic shock in the mouse. This suggests that the age-dependent potentiation of anaphylactic shock does not seem to be due to elevated susceptibility to the mediator but to its increased release. The sex-dependent difference was also studied and found to be particularly clear in the case of IgG₁ antibody-mediated anaphylactic shock in young DS and aged NON mice.     

Age-dependent difference in susceptibility to IgE antibody- and IgG1 antibody-mediated passive anaphylactic shock in the mouse.

The age dependence of the susceptibility to passive anaphylactic shock was studied in the mouse. Anti-BPO IgE monoclonal antibody produced potent systemic sensitization sufficient for provocation of lethal shock in most aged (6 to 10 months) CTS, DS and C57BL/6J mice but only in a very few young (1.5 to 2.5 months) mice. A similar trend was found in the NOD strain, though it was not as definite as in the above three strains. Age-dependent potentiation of the IgE antibody-mediated anaphylactic shock was not found in both sexes of five other strains, C3H/He, DBA/2, NON, BALB/c and B6D2F1. However, the potentiation became obvious even in these strains, when they were treated with Bordetella pertussis before the antigen challenge. Age-dependent potentiation was also clear with IgG1 antibody-mediated anaphylactic shock in DS females and NON males. In contrast, no age-dependent difference was seen for the shock induced by PAF which is estimated to be the main mediator for anaphylactic shock in the mouse. This suggests that the age-dependent potentiation of anaphylactic shock does not seem to be due to elevated susceptibility to the mediator but to its increased release. The sex-dependent differences was also studied and found to be particularly clear in the case of IgG1 antibody-mediated anaphylactic shock in young DS and aged NON mice.     

Age-Dependent Difference in Susceptibility to IgE Antibody- and IgG1 Antibody-Mediated Passive Anaphylactic Shock in the Mouse

The age dependence of the susceptibility to passive anaphylactic shock was studied in the mouse. Anti-BPO IgE monoclonal antibody produced potent systemic sensitization sufficient for provocation of lethal shock in most aged (6 to 10 months) CTS, DS and C57BL/6J mice but only in a very few young (1.5 to 2.5 months) mice. A similar trend was found in the NOD strain, though it was not as definite as in the above three strains. Age-dependent potentiation of the IgE antibody-mediated anaphylactic shock was not found in both sexes of five other strains, C3H/He, DBA/2, NON, BALB/c and B6D2F₁. However, the potentiation became obvious even in these strains, when they were treated with Bordetella pertussis before the antigen challenge. Age-dependent potentiation was also clear with IgG₁ antibody-mediated anaphylactic shock in DS females and NON males. In contrast, no age-dependent difference was seen for the shock induced by PAF which is estimated to be the main mediator for anaphylactic shock in the mouse. This suggests that the age-dependent potentiation of anaphylactic shock does not seem to be due to elevated susceptibility to the mediator but to its increased release. The sex-dependent difference was also studied and found to be particularly clear in the case of IgG₁ antibody-mediated anaphylactic shock in young DS and aged NON mice.     

Immune reactivity during aging. I. T-helper dependent and independent antibody responses to different antigens, in vivo and in vitro.

The immunological status during aging was assessed by measuring the antibody response of the long-lived (C3H/eb X C57Bl/6J)F1 mice to various antigens in vivo and in vitro. In vivo, a decrease in antibody production to DNP and NIP haptenic determinants coupled on to BGG, as well as the response to SRBC, was observed. The decline was more pronounced in the IgG as compared to IgM antibodies. The results were recorded when various parameters such as antigen dose and kinetics of the response, were considered. Reduction of the antibody response was also noted when PVP was employed as immunogen. Similar results were noted when the responses to SRBC and DNP--polylysine were induced and followed in spleen organ cultures. In all of these experimental systems, the peak response was observed in mice 6--12 months old. From then on a gradual decrease was manifested, mice 30--36 months old producing significantly low responses. The results demonstrate that decrease in antibody production is expressed in the isolated spleen tissue in the same manner as in the intact animal. Furthermore, they were interpreted as indicating that the lesion may be at the T helper and the B cell compartments.     

Cellular and humoral immune responses in mice. II. Effect of intraperitoneal or subcutaneous injection of carrier of anti-hapten antibody and delayed hypersensitivity responses.

The subcutaneous (s.c.) injection of sheep red blood cells (SRBC) without adjuvant into mice preferentially induced delayed hypersensitivity (DH) reaction, as measured by footpad swelling, while intraperitoneal (i.p.) measured by the haemagglutinin test. Under these conditions, the properties of the helper activities of thymus-derived (T) cells for humoral responses were examined, in association with the features of the DH response, by measuring the anti-hapten and anticarrier antibody responses after a booster injection of trinitrophenylated (TNP) SRBC and by changing the combination of doses and injections routes of the carrier and the hapten-carrier conjugates. When mice were presensitized with i.p. injections of SRBC and boosted with i.p. injections of TNP-SRBC, the anti-TNP antibody production was maximally enhanced by presensitization with a low dose of SRBC, and gradually abolished with higher doses of SRBC for pre-sensitization. In the latter case, anti-SRBC antibody production was increases with increasing doses of SRBC..     

Peritoneal exudate T lymphocytes with specificity to sheep red blood cells. II. Inflammatory helper T cells and effector T cells in mice with delayed-type hypersensitivity and in suppressed mice.

Peritoneal exudate cells were induced in mice 4 days after immunization with SRBC. A low dose of SRBC (10(6) i.v.) caused T lymphocytes to appear in inflammatory exudates. These cells, not only transferred DTH reactions, but also functioned as helper T cells in antibody production after transfer to syngeneic nu/nu recipient mice. After a high dose of SRBC (10(9) i.v.), very few helper T cells and no DTH transferring T cells were found in inflammatory exudates, although they were present in the spleen. It is postulated that T cells mediating DTH reactions and helper T cells behave similarly as far as those dose dependency of appearance in inflammatory exudates is concerned. A high dose of sensitizing antigen causes retention of helper and effector T cells in the spleen, in this way favouring antibody formation; low doses of antigen allow them to leave the spleen, thus favouring mediation of DTH reactions in the periphery.     

The effect of disodium 4-chloro-2-iminodibenzoate (CCA) on IgE levels and anaphylactic shock

The effect of disodium 4-chloro-2,2-iminodibenzoate (CCA) on IgE antibody response was examined in C₃H/A and (BALB/c×C57BL/6J) F1 hybrid mice immunized with low doses of ovalbumin (OA) adsorbed on aluminium hydroxide gel. CCA administered orally at the doses of 5 and 50 mg/kg/day reduced IgE antibody production in these mice as determined by PCA test. High doses of CCA (100 mg/kg/day) given from day 7 before immunization of C57BL mice and during 1 week after immunization of mice with OA and Bordetella Pertussis Vaccine reduced the mortality of these mice subjected to anaphylactic shock on day 7 of immunization. CCA treatment was ineffective in anaphylactic shock of C57BL mice immunized with very high dose of OA, known to elicit little or no IgE antibody production but high IgG antibody response.The treatment of OA-immunized Guinea pigs with one oral dose of CCA (100 mg/kg) did not reduce mortality in protracted anaphylactic shock. Our results demonstrate that CCA inhibits IgE production as well as IgE mediated hypersensitivity reactions in mice.     

Endogenous Immune Response to Glutamic Acid Decarboxylase (GAD67) in NOD Mice is Modulated by Adjuvant Immunotherapy

We have shown that immunization of non-obese diabetic (NOD) mice with adjuvants (CFA or BCG) prevents the onset of diabetes by induction of regulatory cells. Since autoimmune responses to glutamic acid decarboxylase (GAD) are up-regulated in insulin-dependent diabetes mellitus (IDDM), in this study GAD67-specific antibody, T cell proliferation and lymphokine production patterns were analysed in the adjuvant-treated mice to characterize the regulatory mechanisms underlying the protection. We used both spontaneous diabetes and syngeneic islet transplantation models in NOD mice. Protection against spontaneous diabetes and prevention of syngeneic islet graft rejection by CFA or BCG treatment was found to be accompanied by the production of long lasting and high titre anti-GAD67 antibody of IgG1 isotype in the sera. Uponin vitrostimulation with GAD67, draining lymph node and spleen cells from CFA-immunized NOD mice or syngeneic islet-grafted and BCG-protected NOD mice produced much more IL-4, whereas there was no significant change in IFN-γ production. The strong early T cell proliferative response to GAD67 in CFA or BCG-immunized NOD mice was followed by a low or unresponsiveness state. Taken together, these results suggest a shift in Th1/Th2 balance in the GAD67-specific endogenous immune response to a change in Th2 levels after adjuvant treatment. We postulate that the protective effect of CFA or BCG is due to the diversion of GAD-specific endogenous cellular immune response to a non-pathogenic humoral response.     

Influence of Arecoline on Immune System: III. Suppression of B Cell-Mediated Immune Response in Mice After Short-Term Exposure

Abstract

Arecoline, a major alkaloid of arecanut was examined to explore its modulatory influence on B cell-mediated immune response in a murine model system. The in vivo and in vitro effects were evaluated at sub-toxic concentrations of arecoline. The number of primary antibody forming cells (AFC) and hemagglutinating and hemolysis antibody titers to Sheep Red Blood Cells (SRBC) were evaluated in male mice. Arecoline exposure for a week invoked dose-dependent effect on primary antibody forming cells to SRBC with a maximum reduction at the dosage of 20 mg/kg bw, a moderate reduction at 10 mg/kg bw and no effect at 5 mg/kg bw dose level. HA and HL antibody titers to SRBC were suppressed markedly at arecoline dosage of 20 mg/kg bw and moderately at a dose of 10 mg/kg bw, given daily for 1, 2 or 3 weeks. The inhibitory effect of arecoline was not dependent on the duration of treatment. Like the primary antibody response, the secondary HA and HL antibody titers were also decreased after arecoline exposure. The administration of arecoline dosages 10 and 20 mg/kg bw daily for 4 days following SRBC immunization also, exerted dose-dependent suppression of primary antibody response. Similarly, when treated after 12 h following immunization, significant reduction in response was observed with arecoline dosage of 20 mg/kg bw. While moderate suppression of antibody response was noticed at the dose level of 10 mg/kg bw, there was no alteration in response at a dosage of 5 mg/kg bw. In contrast, arecoline dosages 5, 10 or 20 mg/kg bw given after 1, 2 or 4 days following immunization did not alter the HA and HL antibody titers to SRBC. Recovery experiments in mice revealed that arecoline-mediated suppression of antibody response is of a reversible nature. Concomitant exposure of arecoline at the concentrations of 10^?5 – 10^?4 M with PWM suppressed ³H-thymidine incorporation of splenic cells in vitro. Taken together, the findings reported in this paper suggest that the intensity of arecoline-mediated suppression of antibody response to SRBC and PWM-induced splenic cell proliferation is dependent upon the dosage and the mode of treatment.     

Suppressive effects of antihistaminic and/or anti-PAF agents on passive anaphylactic shock in mice sensitized with allogeneic monoclonal IgE and IgG1 antibodies and hyperimmune serum

For the immunopharmacological characterization of murine passive anaphylactic shock, the effects of antihistaminics and/or anti-platelet-activating factor (anti-PAF) agents were studied on the shock mediated by allogeneic monoclonal IgE and IgG₁ antibodies and hyperimmune serum. IgE antibody-mediated shock was strongly suppressed by cyproheptadine (10 mg/kg, ip) in every strain regardless of the age and sex of the mice and the presence or absence of a shock potentiator. As far as tested with CTS, DS, and B6D2F1 mice, IgE antibody-mediated shock was also suppressed by the other two antihistamines, triprolidine (10 mg/kg, ip) and oxatomide (100 mg/kg, Po). This type of shock was not suppressed by an anti-PAF agent, CV-6209 (3.3 mg/kg, iv), when tested on aged CTS mice given no shock potentiator and young DS mice given a potentiator such as Bordetella pertussis organisms or DL-propranolol. IgG₁ antibody-mediated shock was also suppressed by cyproheptadine in general except for CTS mice. Suppression in the DL-propranolol-treated DS and C3H/He mice was not very marked on sensitization with undiluted or slightly diluted IgG₁ ascites but quite striking on sensitization with properly diluted ascites. In contrast with the effect of cyproheptadine, suppression by CV-6209 was obvious in aged CTS mice but not in young DL-propranolol-treated DS mice. The shock in DL-propranolol-treated DS mice sensitized with undiluted or slightly diluted ascites was completely abolished by the combined use of these two agents. These results suggest that histamine and/or PAF play a major role in IgE antibody- and IgG₁ antibody-mediated shock. However, so far as tested in young DS mice, the shock mediated by serum differed in drug susceptibility from that mediated by the monoclonal antibodies. In the absence of shock potentiators, prevention was produced by cyproheptadine in the males which had been sensitized with the 1:4 or 1:8 dilution of the immune serum. In the presence of DL-propranolol, prevention was not produced even by the combined treatment with cyproheptadine and CV-6209. Therefore, it is likely that some mediators other than histamine and PAF, whose release is triggered by antibody isotypes other than IgE and IgG₁, play a greater role for the shock mediated by hyperimmune serum than for the shock mediated by IgE or IgG₁ antibody, especially in the presence of shock potentiators.     

Intraperitoneal systemic anaphylaxis in the mouse. I. Age-dependence of fatal anaphylactic shock

Following the i.p. challenge of a shocking dose of BSA from 9 days up to 133 days after the s.c. injection of BSA in CFA, fatal anaphylaxis was induced regularly in female ICR mice that had been given the immunizing antigen when 8 weeks old. These immunized mice provided an antiserum to BSA that had the capacity to transfer fatal shock to normal recipient mice at a minimum Ab-N dose of 8 microgram when the i.p. route for challenge was employed. The optimal dose-range of antigen and antibody in order to elicit fatal shock following the i.p. challenge was much broader than that obtained by i.v. injection. Age is critical in producing fatal shock in mice; a 100% fatal anaphylaxis never occurred in groups of 6- and 7-week-old recipient mice although those at 8 weeks and older were sufficiently sensitized by the amounts of antibody given.     

Dextran sulphate: an adjuvant for cell-mediated immune responses.

The effect of high mol. wt dextran sulphate (DS) on cell-mediated immune responses was studied. Three criteria were used to assess cell-mediated delayed-type hypersensitivity: footpad swelling, i.d. skin tests and macrophage migration inhibitory factor (MIF) production. Guinea-pigs sensitized with egg albumin (EA) and treated with DS showed strong positive delayed skin tests. Control animals given only EA showed negative skin tests. Lymphocytes from mice sensitized s.c. with EA and treated with DS showed an increase in MIF production. Delayed footpad swelling responses in mice sensitized s.c. with sheep red blood cells (SRBC) and treated with DS were increased when the mice were challenged 8 days after sensitization. Doses of DS which were effective in increasing delayed footpad swelling ranged from 50-200 mg DS/kg body weight. DS was only capable of increasing delayed footpad swelling responses when both SRBC and DS were injected s.c. at the same site. Intraperitoneal injection of both SRBC and DS obeebction of both s.c. but at different sites did not result in increased delayed footpad swelling. DS was capable of augmenting footpad swelling responses when given s.c. as much as 6 days before SRBC. The optimal time for administration of DS was 2 days before SRBC. Injection of DS 2 days or more after SRBC resulted in no increase in delayed footpad swelling responses. The results of this study indicate that dextran sulphate is a potent adjuvant for cell-mediated delayed-type hypersensitivity immune responses in both mice and guinea-pigs.     

Modulation of Antibody-Mediated Immune Response by Probiotics in Chickens

Probiotic bacteria, including Lactobacillus acidophilus and Bifidobacterium bifidum, have been shown to enhance antibody responses in mammals. The objective of this study was to examine the effects of a probiotic product containing the above bacteria in addition to Streptococcus faecalis on the induction of the chicken antibody response to various antigens, both systemically and in the gut. The birds received probiotics via oral gavage and subsequently were immunized with sheep red blood cells (SRBC) and bovine serum albumin (BSA) to evaluate antibody responses in serum or with tetanus toxoid (TT) to measure the mucosal antibody response in gut contents. Control groups received phosphate-buffered saline. Overall, BSA and SRBC induced a detectable antibody response as early as week 1 postimmunization (p.i.), which lasted until week 3 p.i. Probiotic-treated birds had significantly (P ≤ 0.001) more serum antibody (predominantly immunoglobulin M [IgM]) to SRBC than the birds that were not treated with probiotics. However, treatment with probiotics did not enhance the serum IgM and IgG antibody responses to BSA. Immunization with TT resulted in the presence of specific IgA and IgG antibody responses in the gut. Again, treatment with probiotics did not change the level or duration of the antibody response in the gut. In conclusion, probiotics enhance the systemic antibody response to some antigens in chickens, but it remains to be seen whether probiotics have an effect on the generation of the mucosal antibody response.     

Non-specific stimulation of immunoglobulin synthesis in mice by capsular polysaccharide of Klebsiella pneumoniae

Intramuscular injection of a non-immunogenic dose (1 mg) of the capsular polysaccharide of Klebsiella pneumoniae (CPS-K) into mice caused a marked and prolonged increase in the amount of the serum IgM and IgG. The increase of IgM after this injection was over 100 times greater than that of antigen-specific IgM antibodies produced by injection of an immunogenic dose of CPS-K (5 μg) or sheep red blood cells (SRBC) (2 X 10⁸). The increase in the amount of immunoglobulins was not related to induction of immunological paralysis to the major subcomponent of CPS-K (acidic CPS-K) by a large dose of antigen, but was induced by non-antigenic stimulation by the minor subcomponent of CPS-K (neutral CPS-K), which was non-immunogenic at any dose. In mice injected with a non-immunogenic dose of CPS-K, antibody levels in the serum to three kinds of non-cross-reacting antigens, SRBC, bovine serum albumin (BSA) and a possibly autochthonous IgG (IgM—IgG mixed type cryoglobulin) were also increased. The number of plaque-forming cells (PFC) for SRBC in the spleens of mice was also markedly increased after CPS-K injection. These antibodies to SRBC and BSA and PFC for SRBC were exclusively of IgM type and increased to as high levels as those found after specific antigenic stimulation. This marked increase in IgM antibody was neither followed by an increase in IgG antibody nor by immunological memory for a secondary IgG response.