Hypersensitivity Response In Mice Research Articles

A Monoclonal Antibody to cis-Urocanic Acid Prevents the Ultraviolet-Induced Changes in Langerhans Cells and Delayed Hypersensitivity Responses in Mice, Although Not Preventing Dendritic Cell Accumulation in Lymph Nodes Draining the Site of Irradiation and Contact Hypersensitivity Responses

Ultraviolet B (UVB) irradiation of C3H mice causes suppression of delayed hypersensitivity and contact hypersensitivity (CH) to antigens encountered following exposure, and is accompanied by a reduction in Langerhans cell (LC) numbers in the epidermis, loss of epidermal antigen-presenting cell function, and accumulation of dendritic cells in lymph nodes draining the site of irradiation. Various photoreceptors and mediators of these changes have been proposed, one of which is cis-urocanic acid (cis-UCA) formed from the naturally occurring trans-UCA in the epidermis on UV irradiation. A monoclonal antibody that reacts with cis-UCA has become available recently and has been used in this study to clarify the role of UCA. Pretreatment of C3H mice with the monoclonal antibody abrogated the UVB-induced and cis-UCA-induced reduction in epidermal LC numbers. It also prevented the UV-induced suppression of epidermal antigen-presenting cell ability as measured by the mixed skin lymphocyte response. However, it had no effect on the accumulation of dendritic cells in lymph nodes draining the site of UV exposure. With regard to hypersensitivity responses, it did not prevent UV-induced suppression of CH to oxazolone at a range of concentrations but it restored to normal the UV-suppressed delayed hypersensitivity to herpes simplex virus, if administered before exposure. Thus cis-UCA is involved in some UV-induced changes in murine skin but not in others, where alternative mediators, such as tumor necrosis factor-alpha, may be more important.

Functional evidence for epitope spreading in the relapsing pathology of experimental autoimmune encephalomyelitis.

The role of epitope spreading in the pathology of relapsing-remitting experimental autoimmune encephalomyelitis (R-EAE) was examined. Using peripherally induced immunologic tolerance as a probe to analyze the neuropathologic T cell repertoire, we show that the majority of the immunopathologic reactivity during the acute phase of R-EAE in SJL/J mice induced by active immunization with the intact proteolipid (PLP) molecule is directed at the PLP139-151 epitope and that responses to secondary encephalitogenic PLP epitopes may contribute to the later relapsing phases of disease. Intermolecular epitope spreading was demonstrated by showing the development of T cell responses to PLP139-151 after acute disease in mice in which R-EAE was initiated by the transfer of T cells specific for the non-cross-reactive MBP84-104 determinant. Intramolecular epitope spreading was demonstrated by showing that endogenous host T cells specific for a secondary encephalitogenic PLP epitope (PLP178-191) are demonstrable by both splenic T cell proliferative and in vivo delayed-type hypersensitivity responses in mice in which acute central nervous system damage was initiated by T cells reactive with the immunodominant, non-cross-reactive PLP139-151 sequence. The PLP178-191-specific responses are activated as a result of and correlate with the degree of acute tissue damage, since they do not develop in mice tolerized to the initiating epitope before expression of acute disease. Most importantly, we show that the PLP178-191-specific responses are capable of mediating R-EAE upon adoptive secondary transfer to naive recipient mice. Furthermore, induction of tolerance to intact PLP (which inhibits responses to both the initiating PLP139-151 epitope and to the PLP178-191 epitope) after the acute disease episode is sufficient to prevent relapsing disease. These results strongly support a contributory role of T cell responses to epitopes released as a result of acute tissue damage to the immunopathogenesis of relapsing clinical episodes and have important implications for the design of antigen-specific immunotherapies for the treatment of chronic autoimmune disorders in humans.

Immunopharmacology and toxicology of the plant flavonoid baohuoside-1 in mice

The plant flavonoid baohuoside-1 (B-1), isolated from Epimedium davidii, was shown to suppress antibody and delayed-type hypersensitivity responses in mice a dose-dependent fashion. By contrast, similar dosage schedules of B-1 did not significantly prolong survival of cardiac grafts. Furthermore, B-1 did not potentiate the effects of the standard anti-rejection drug cyclosporine. Assessment of the mice for appearance, behaviour, biochemistry, haematology and histology revealed not toxic effects at the dosages used in the experiments. These results indicate that the immunosuppressive properties of B-1 are confined to the antibody-mediated system, and suggest that B-1 may be of value in the treatment of chronic inflammatoru and autoimmune diseases in which autoantibodies have a major role in pathogenesis.

Delayed type hypersensitivity responses in mice transplanted with rat hepatocytes.

The delayed type hypersensitivity (DTH) response to xenogeneic hepatocytes (HCs) was investigated in mice received subcutaneous (s.c.), intrasplenic (i.s.), or intravenous (i.v.) transplantation of rat HCs. The DTH response in mice preimmunized i.s. or i.v. with rat HCs (1 x 10(6) cells) was significantly lower than that in mice preimmunized s.c. with the same doses of rat HCs. Co-transfer of spleen cells from i.s. immunized mice with spleen cells from s.c. immunized mice to naive recipient mice did not suppress the DTH response induced by transfer of spleen cells from s.c. immunized mice. On the other hand, co-transfer of spleen cells from i.v. immunized mice with spleen cells from s.c. immunized mice suppressed the DTH response to rat HCs in recipients. Furthermore, the levels of DTH responses in recipients transferred with spleen cells from mice sensitized i.s. or i.v. with rat HCs, immunized s.c. with rat HCs 6 hr after transfer, and challenged with rat HCs 7 days later was almost similar to those in recipients transferred with spleen cells from s.c. immunized mice. These results suggest that antigen-specific suppression of DTH responses to rat HCs in mice is associated with the presence of suppressive spleen cells induced by i.s. or i.v. immunization with rat HCs.

Immunomodulating activities of orally administered SMANCS, a polymer-conjugated derivative of the proteinaceous antibiotic neocarzinostatin, in an oily formulation

Immunomodulatory effects of oily formulated SMANCS, a polymer conjugated derivative of the proteinsceous antibiotic neocarzinostatin, after oral administrations to mice was investigated. The oral administrations of SMANCS, dissolved in medium-chain triglyceride containing phosphatidylcholine and polyglycerine dioleate (oily SMANCS), resulted in: (1) augmentation of NK cell activity in naive mice, (2) activation of macrophage cytostasis in naive mice, (3) enhancement of the generation of cytotoxic T-lymphocytes in mice immunized with allogeneic lymphocytes, (4) production of circulating interferon in naive mice, and (5) increase of delayed-type hypersensitivity response in mice immunized with sheep red blood cells. The degree of immunomodulation orally stimulated with oily SMANCS was similar to that of the immunomodulation induced by i.v. or i.p. administrations of aqueous formulated SMANCS (aqueous SMANCS). Although SMANCS is a protein drug that may be digestable by various enzymes present in the stomach, in the present study immunomodulating activities of SMANCS were clearly demonstrated when an oily formulation of the compound was administered to mice orally. Since aqueous SMANCS administered parenterally and oily SMANCS administered orally exhibit the same immunomodulatory activities and the former has demonstrated antitumor activity in man and animals, the latter may possess this same antitumor activity.

The magnitude and kinetics of delayed-type hypersensitivity responses in mice vaccinated with irradiated cercariae ofSchistosoma mansoni

A footpad assay was used to measure the DTH of mice to soluble worm antigens (SWAP), and to living day 7 lung schistosomula, following vaccination and challenge infections with Schistosoma mansoni. DTH to SWAP was first observed on day 10, and reached its maximum on day 17 post-vaccination. Treatment of mice with anti-CD4 antibody on the 3 days prior to footpad challenge completely abrogated this response. Reactivity to living parasites was of a lower order than that to SWAP; it also peaked earlier, on day 10 post-vaccination. By day 35, responsiveness to both sets of antigens had declined almost to control levels. There was no correlation between the level of DTH to living schistosomula, at any time, and the degree of resistance subsequently developed. Percutaneous challenge of vaccinated mice was followed by a resurgence of reactivity to SWAP. This secondary response occurred more rapidly than the primary response, peaking on day 7 post-challenge, and was of a similar magnitude. We were unable to detect a similar recall of DTH to living schistosomula, possibly because the assay was insufficiently sensitive. We conclude that the intensity and kinetics of DTH responsiveness are crucial features of the irradiated vaccine model, and suggest that further investigation of cell-mediated immune reactions, particularly those occurring in the lungs, is vital to a better understanding of events underlying the development and expression of immunity.

Hymenolepis nana: Adoptive transfer of protective immunity and delayed type hypersensitivity response with mesenteric lymph node cells in mice

A marked degree of footpad swelling was observed in BALB/c mice infected with Hymenolepis nana eggs, when soluble egg antigen was injected into their footpads 4 to 21 days after the egg infection, indicating delayed type hypersensitivity responses in infected mice. Adoptive transfer with mesenteric lymph node cells from donor mice ( BALB c strain; +/+) infected with eggs 4 days before cell collection could confer this hypersensitivity to recipient nude mice (BALB/c strain; nu/nu). These mesenteric lymph node cells were then divided into two fractions, blastenriched and blast-depleted cells, by density gradient centrifugation with Percoll. The recipients intravenously injected with the blast-depleted cell fraction showed a marked increase in footpad thickness, whereas the intravenous transfer of the blast-enriched cell fraction resulted in an insignificant increase in footpad thickness. The transfer of the blastenriched cell fraction, but not of the blast-depleted cell fraction, conferred a strong adoptive immunity on syngeneic recipient nude mice, when the immunity transferred was assessed by examining cysticercoids developed in the intestinal villi on Day 4 of challenge infection. The lack of delayed type hypersensitivity response in mice that received the blast-enriched cell population was not due to a lack of the capacity of the cells to induce the response, because the cells were capable of inducing a significant increase in thickness of footpads of normal mice when these cells were locally injected into the footpad together with soluble egg antigen. Furthermore, experiments involving passive transfer and preferential localization of radiolabeled cell populations suggested that footpad swelling could be induced by a very small proportion of sensitized migratory lymphocytes, while at least a significant number of sensitized blast lymphocytes is required for the rejection of cysticercoids in the intestine.

776 IL-6 mediated regulation of IgE dependent immediate hypersensitivity responses in mice. Lack of effect of interferon alpha and interferon gamma

776 IL-6 mediated regulation of IgE dependent immediate hypersensitivity responses in mice. Lack of effect of interferon alpha and interferon gamma

Contact Hypersensitivity Response to O-Benzyl-P-Chlorophenol in Mice

o-Benzyl-p-chlorophenol was evaluated for its potential as a sensitizing agent for allergic contact hypersensitivity in mice. Female B6C3F1 mice were sensitized with 1.0, 3.0, and 10.0% o-benzyl-p-chlorophenol and challenged with 20.0% o-benzyl-p-chlorophenol. Doses of o-benzyl-p-chlorophenol were selected from assays for primary irritancy. Mice received 20 microliters by direct dermal application, for 5 days, to sites prepared by shaving, dermabrading and, in some mice, with intra dermal injection of Freund's complete adjuvant. The rest period was 7 days. Measurement of the contact hypersensitivity response in mice was by radioisotopic assay two days after challenge and mouse ear swelling test one and two days after challenge. Mice demonstrated statistically significant dose-dependent contact hypersensitivity response to o-benzyl-p-chlorophenol with or without adjuvant pretreatment.

Suppression of Urushiol-Induced Delayed-Type Hypersensitivity Responses in Mice with Serum IgG Immunoglobulin form Human Hyposensitized Donors

Serum IgG immunoglobulin fractions from human subjects hyposensitized to poison ivy/oak by oral administration of urushiol suppressed the induction of delayed-type hypersensitivity (DTH) responses in mice to this hapten. This suppressive activity was hapten specific because it did not modify DTH responses to dinitrofluorobenzene (DNFB). Absorption of human serum with lymph node cells from urushiolsensitized but not DNFB-sensitized mice removed the suppressive activity, suggesting that anti-idiotypic antibodies reacting with T-cell receptors are involved.

Increased contact hypersensitivity response in mice by topical application of 1?,25-dihydroxyvitamin D3 to elicitation site

Recent evidence indicates that the biologically active metabolite of vitamin D3, 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], has an effect on the regulation of the immune response. We investigated whether topical treatment of mice with 1 alpha,25(OH)2D3 influences the contact hypersensitivity (CHS) response to trinitrochlorobenzene (TNCB). 1 alpha,25(OH)2D3 was applied to the dorsal trunk of A/J mice on days 0-3, and on day 4 topical application of 5% TNCB on the 1 alpha,25(OH)2D3-treated site was performed. The mice were tested for CHS on day 10 by applying 1% TNCB to the ears. No effect on induction of CHS response to TNCB was observed in 1 alpha,25(OH)2D3-treated mice compared with 24,25-dihydroxyvitamin D3[24,25(OH)2D3]-treated mice as control. In a second experiment, the dorsal trunk of A/J mice was treated with 5% TNCB on day 0. The topical application of 1 alpha,25(OH)2D3 on the ears was performed from days 2 to 5. On day 6, the mice were tested for CHS by applying 1% TNCB to the 1 alpha,25(OH)2D3-treated ears. When 1 alpha,25(OH)2D3 was administered to the elicitation site prior to the challenge, pretreatment of mice with 1 alpha,25(OH)2D3 increased their response to TNCB by 40% compared with 24,25(OH)2D3-treated mice as control (P less than 0.01). There were no findings suggesting that the pretreatment of the challenge site with 1 alpha,25(OH)2D3 induced an irritant dermatitis that was superimposed on a subsequent CHS reaction. The 1 alpha,25(OH)2D3 modulation of CHS response to TNCB in mice suggests that the hormone may play a role in the regulation of the immune response in vivo.

Contact hypersensitivity response to isophorone diisocyanate in mice.

Isophorone diisocyanate was evaluated for its potential as a sensitizing agent for allergic contact hypersensitivity in mice. Female B6C3F1 mice were sensitized with 0.1, 0.3, and 1.0% isophorone diisocyanate and challenged with 3.0% isophorone diisocyanate. Doses of isophorone diisocyanate were selected from assays for primary irritancy. Mice received 20 microliters by direct dermal application, for 5 days, to sites prepared by shaving, dermabrading and, in some mice, with intra dermal injection of complete Freund's adjuvant. The rest period was 7 days. Measurement of the contact hypersensitivity response in mice was by radioisotopic assay two days after challenge and mouse ear swelling one and two days after challenge. Mice demonstrated statistically significant dose-dependent contact hypersensitivity responses to isophorone diisocyanate with or without adjuvant pretreatment.

Parallel Recovery of Epidermal Antigen-Presenting Cell Activity and Contact Hypersensitivity Responses in Mice Exposed to Ultraviolet Irradiation: The Role of a Prostaglandin-Dependent Mechanism

Contact hypersensitivity (CH) responsiveness to 2-4-dinitro-1-fluorobenzene (DNFB) is depressed in mice that are sensitized through skin sites exposed to ultraviolet radiation (UVR). This is partially due to a reduction in antigen-presenting cell (APC) activity within UVR-exposed skin, a condition marked by a decrease in the density of ATPase/Ia-positive epidermal cells. The purpose of this study was to correlate the histological and functional recovery of APC activity in the skin of C3H mice exposed to low-dose (4 X 450 J/m2) or high-dose (1 X 15 kJ/m2) UVR with the normalization of CH responsiveness. Skin biopsy specimens taken at various intervals after UVR exposure revealed a rapid recovery in the density of ATPase/Ia positive cells: about 70% of normal by 3 days, and normal after 5 days. Functional analyses showed that lymph node cells obtained from donors that were sensitized with DNFB 3 days after UVR treatment transferred normal ear-swelling responsiveness to non-primed recipients, thus indicating that APC activity in UVR-exposed skin paralleled the recovery of ATPase/Ia-positive epidermal cells. This suggested that an alternative mechanism causes the persistent depression of CH in mice exposed to UVR. Mice pretreated with indomethacin prior to UVR exposure demonstrated a capacity to elicit CH responses to DNFB, which paralleled the histological and functional recovery of APC in the skin (i.e., normal CH responses were elicited 3 days after exposure to UVR). We conclude from this study that APC activity in the skin recovers rapidly after exposure to UVR, and that a PG-dependent mechanism is responsible for many of the persistent and systemic effects that cause a depression in the CH responsiveness of mice treated with UVR.

Immunomodulatory Effect of Neurotropin on Delayed Type Hypersensitivity Response in Mice

Effect of Neurotropin on the delayed type hypersensitivity (DTH) response to sheep red blood cells was examined in mice. Neurotropin suppressed the high DTH responses in ddY and BALB/c mice, while it enhanced the low DTH response in C57BL/6 mice when administered after sensitization. In addition, Neurotropin exerted a suppressive effect on the enhanced DTH response by cyclophosphamide pretreatment in C57BL/6 mice, but had a restorative effect on the suppressed DTH response in ddY mice loaded with restraint stress.

Immunomodulating effect of neurotropin on delayed type hypersensitivity response in mice.

Effect of Neurotropin on the delayed type hypersensitivity (DTH) response to sheep red blood cells was examined in mice. Neurotropin suppressed the high DTH responses in ddY and BALB/c mice, while it enhanced the low DTH response in C57BL/6 mice when administered after sensitization. In addition, Neurotropin exerted a suppressive effect on the enhanced DTH response by cyclophosphamide pretreatment in C57BL/6 mice, but had a restorative effect on the suppressed DTH response in ddY mice loaded with restraint stress.

Pharmacodynamics of cyclosporine-ketoconazole interaction in mice. Combined therapy potentiates cyclosporine immunosuppression and toxicity.

We have previously studied the pharmacokinetics of the cyclosporine-ketoconazole interaction and shown that ketoconazole inhibits the metabolism of cyclosporine in mice. In the present study we investigated the pharmacodynamics of the interaction and found that ketoconazole increased the immunosuppressive activity and toxicity of cyclosporine in vivo. Mice were treated orally with cyclosporine (25-200 mg/kg/day), ketoconazole (100 mg/kg/day), drug vehicle, or drug combination for 14 days. Immunosuppression was studied by assaying radiometrically the delayed hypersensitivity response in mice that were sensitized and challenged to keyhole limpet hemocyanin. The dose-response curve for cyclosporine was shifted to the left when cyclosporine and ketoconazole were coadministered compared with treatment with cyclosporine alone. The dose that produced 50% of the maximal immunosuppression (ED50) for cyclosporine alone was 89 mg/kg/day, whereas the ED50 for cyclosporine, when combined with ketoconazole, was 33 mg/kg/day. Ketoconazole alone was not immunosuppressive. Toxicity was measured by the length of survival during a ten-day period of treatment. Animals receiving both drugs had a significantly lower survival: the ten-day survival of animals treated with 200 mg/kg/day of cyclosporine was 100% in the absence of ketoconazole treatment and 62.5% with 100 mg/kg/day of ketoconazole. The results of our study have two important implications. First, the effect of ketoconazole on cyclosporine immunosuppression is quantitatively similar to its effect on cyclosporine kinetics. Because of this two- to three-fold increase in both unchanged cyclosporine blood levels and immunosuppression, our results suggest that unchanged cyclosporine--not its metabolites--is the primary pharmacological agent of cyclosporine in vivo immunosuppression. Second, to avoid cyclosporine-induced nephrotoxicity, it has been recommended that the dosage of cyclosporine be decreased when this drug combination is used clinically. The results of this study suggest that a decrease in cyclosporine dosage will not lower the level of immunosuppression in these patients.

Correlation between keratinocyte expression of Ia and the intensity and duration of contact hypersensitivity responses in mice.

Langerhans cells are the only cells within the epidermis that normally express immune response-associated antigens (referred to as Ia in mice and HLA-D in humans). However, in the epidermis of patients with allergic contact dermatitis or individuals undergoing a delayed-type hypersensitivity response, the keratinocytes at the reaction site are induced to express HLA-DR. In this study the inducible expression of Ia by the keratinocytes of mice was found to be directly correlated with the intensity and duration of experimentally induced contact hypersensitivity (CH) responses. During a CH response in animals that were sensitized on the belly and challenged on the ear with the contact-sensitizing (CS) agent oxazolone, the keratinocytes in the challenged, but not the unchallenged, ear were induced to express Ia. In comparison with animals that were sensitized and challenged at different sites, an intensified expression of Ia by the keratinocytes was associated with a twofold increase in ear swelling in mice that were sensitized and challenged with oxazolone at the same site. Curiously, the challenge of oxazolone-sensitized ears with dinitrofluorobenzene (an unrelated CS agent), croton oil (a nonspecific inflammatory agent), or acetone/olive oil (a noninflammatory agent) also induced both a marked keratinocyte expression of Ia and an enhanced CH response. These results suggest that residual antigen at the original sensitization site may be mobilized to function as the challenge stimulus to elicit a CH response, in association with keratinocyte expression of Ia, when CS-sensitized skin is perturbed with a nonspecific agent. Further evidence of an association between CH responsiveness and keratinocyte expression of Ia came from the following observations. First, the magnitude and duration of a CH response was markedly increased in pertussis toxin (PT)-treated mice. These enhanced responses were associated with intense Ia expression by the keratinocytes in the epidermis at the reaction site. Because PT is known to have an adjuvant effect on delayed-type hypersensitivity reactions, as well as to alter the normal regulatory mechanisms associated with this type of response, it is possible that Ia+ keratinocytes play a synergistic role in the enhanced CH responses that are observed in PT-treated animals. Second, a direct correlation between keratinocyte expression of Ia and CH responsiveness was observed in athymic nude mice that were challenged with oxazolone after receiving an adoptive transfer of lymphoid cells from oxazolone-primed normal syngeneic donors.(ABSTRACT TRUNCATED AT 400 WORDS)

Different effects of psychotropic drugs on delayed hypersensitivity responses in mice

The influence of certain psychotropic drugs on the delayed-type hypersensitivity (DTH) response to sheep red blood cells in BALB/c mice was studied. The effects on the overall response and the induction and elicitation phases were evaluated, using two alternative dosage schedules for each agent. It was found that diazepam had no effect on the DTH reaction but the administration of imipramine, haloperidol, chlorpromazine or meprobramate all resulted in depression of the response, impairing both the induction or elicitation phases. The results indicate that psychotropic drugs may produce in vivo depression of cell-mediated immunity by different mechanisms.

Phenolic glycolipid 1 of Mycobacterium leprae causes nonspecific inflammation but has no effect on cell-mediated responses in mice.

The involvement of the phenolic glycolipid from Mycobacterium leprae in cell-mediated immunity has been investigated in this study. The phenolic glycolipid itself does not appear to stimulate cell-mediated immunity directly, as shown by its failure to elicit a classical delayed-type hypersensitivity response in mice immunized with M. leprae or to stimulate M. leprae-immune lymph node cells in a lymphoproliferative assay. Intradermal vaccination with the phenolic glycolipid failed to influence the growth of M. leprae in mouse footpads. A nonspecific inflammatory response to the sonicated glycolipid was observed in mice vaccinated with whole M. leprae and in control animals. No evidence was obtained for any adjuvant or suppressive effect on cell-mediated immunity by the phenolic glycolipid either to M. leprae or an unrelated antigen (sheep erythrocytes); neither sensitization nor elicitation to either antigen was affected.

Specific cutaneous hypersensitivity responses in mice infected or immunized with Trypanosoma cruzi.

The appearance and development of cutaneous hypersensitivity to epimastigote antigen was followed during the early phase of Trypanosoma cruzi infection in mice. The effect of living BCG on the kinetics of skin reactivity was studied in animals infected with T. cruzi blood trypomastigotes or artificially immunized with disrupted epimastigotes. BCG stimulated the immediate response in infected animals while preferentially stimulating delayed responses in immunized animals. Infection with living blood trypomastigotes depressed already existing delayed-type responses. The development of delayed-type responses was also impaired in animals immunized in the course of the infection.