Footpad Reaction Research Articles

Иммунопатогенные свойства SARS-CoV-2 как основа для выбора патогенетической терапии

Guinea-pigs were immunized via footpads with sheep red blood cells (SRBC) in saline. Histological examination of erythematous skin reaction was performed and effects of cyclophosphamide (CY) or BCG pre-treatment on the skin reaction were examined. Delayed-in-onset erythematous skin reaction accompanied by substantial basophil infiltration was elicited in guinea-pigs immunized with SRBC in saline. The erythema was augmented in size by CY which was injected 2 days before immunization. The reaction may be comparable to Jones-Mote type. In BCG pre-treated guinea-pigs, basophil infiltration at the skin reaction sites was reduced in number, but significant inhibition of macrophage migration was not detected in the presence of SRBC antigen. The reaction may be intermediate between Jones-Mote and the tuberculin type. Comparability of delayed skin reactions in guinea-pigs and delayed footpad reactions in mice or hamsters against SRBC is discussed.

Analysis of interferon-gamma production in killed BCG-pretreated mice after stimulation with staphylococcal enterotoxin A

Staphylococcal enterotoxin A (SEA), a T cell mitogen, was found to induce a high level of interferon-gamma (IFN-gamma) in mice which had been immunized with killed Bacillus Calmette-Guérin (BCG) in water-in-oil-in-water (W/O/W) emulsion. The phenomenon was analysed by in vivo and in vitro experiments, and the following results were obtained. 1. The SEA-induced IFN was inactivated by treatment with 0.2M glycine-HCl (pH 2.0) but not by heating at 56 degrees C for 30 min. nor by treatment with anti-IFN alpha/beta antibodies, and the fact suggest that the IFN belonged to the gamma type. 2. Treatment of the BCG-sensitized mice with silica and 2-chloroadenosine (2CA), specific lethal agents for macrophages, reduced the SEA-induced IFN production. 3. The SEA-induced IFN production occurred in mice immunized with BCG either intravenously or intraperitoneally, although they showed weak or no footpad reaction to purified protein derivatives (PPD). In contrast, mice sensitized subcutaneously with BCG showed strong foodpad reaction to PPD but not the SEA-induced IFN production. Thus, the mere presence of BCG-sensitized T cells does not appear to result in the SEA-induced IFN production. 4. In vitro experiments, in which SEA-induced IFN production was determined in the culture of BCG-sensitized spleen cells, showed that principal IFN-producing cells were Lyt-1+ T cells. 5. Deprivation of macrophages from BCG-sensitized spleen cell population by passing through Sephadex G-10 column reduced the SEA-induced IFN production in the culture. Addition of 2CA to the culture medium also reduced the SEA-induced IFN production by the BCG-sensitized spleen cells. 6. The SEA-induced IFN production in the culture of the BCG-sensitized spleen cells was suppressed in the presence of lipoxygenase inhibitor, i.e., caffeic acid or nordihydroguaiaretic acid. 7. The plastic adherent spleen cells (i.e. macrophages) from mice sensitized with BCG produced leukotriene C4 (LTC4). The suppression of the SEA-induced IFN production of BCG-sensitized spleen cells in the presence of the lipoxygenase inhibitor was prevented by addition of synthetic LTC4. These results suggest that LTC4 released from macrophages activated by BCG causes production of IFN-gamma by BCG-sensitized T cells responding to SEA.

Astilbin suppresses delayed-type hypersensitivity by inhibiting lymphocyte migration.

This study examined the effects of astilbin, a flavanone, on delayed-type hypersensitivity reactions and its mechanisms of action on cell migration. Astilbin significantly inhibited the sheep-red-blood-cell-induced footpad reaction and picryl-chloride-induced ear dermatitis without affecting the organ weights, when administered during the effector phase but not the induction phase. The flavanone also significantly inhibited the migration to gelatin of spleen cells isolated from mice with ear dermatitis in a transwell system. Furthermore, the isolated spleen cells from normal mice were incubated with astilbin in the presence of concanavalin A, or those from mice with ear dermatitis were cultured with astilbin. In the supernatants collected, the activity of matrix metalloproteinases (MMPs) MMP-2 and MMP-9 was remarkably inhibited by astilbin. These results suggest that astilbin may inhibit delayed-type hypersensitivity reactions through selectively suppressing the lymphocyte functions, including cell migration, via down-regulating MMP activity.

Footpad reaction induced by Neospora caninum tachyzoite extract in infected BALB/c mice

Little information is available regarding a delayed type hypersensitivity (DTH) reaction in neosporosis. In this study, we examined the elicitation of a DTH reaction in mice infected with Neospora caninum by inoculation of the footpad with tachyzoite antigens. The footpads of BALB/c mice infected with N. caninum and those of non-infected were injected with either the tachyzoite extract, or paraformaldehyde-fixed tachyzoites. In mice inoculated with N. caninum antigens on day 7 p.i. swelling peaked at 6 h after injection of the tachyzoite extract. In mice inoculated on days 14, 28 and 56, swelling was observed between 6 and 72 h afterwards. Mice immunized with the tachyzoite extract plus adjuvant showed peak footpad swelling at 6 h post injection, and the swelling had decreased at 24 h or later. In contrast, mice injected before infection showed no specific swelling. In sections of footpads injected with the tachyzoite extract, exudate had accumulated at 6 h post injection and clusters of infiltrated lymphocytes were observed at 48 h post injection. In mice administered anti-CD4 + cell monoclonal antibodies swelling had decreased at 24 h post injection of the extract. These results indicate that mice infected with N. caninum produce a DTH reaction, which is a good indicator of the development of type 1 immune responses.

Nonmutated Self-Antigen-Derived Cancer Vaccine Peptides Elicit an IgE-Independent but Mast Cell-Dependent Immediate-Type Skin Reaction without Systemic Anaphylaxis

We previously reported an unexpected phenomenon, i.e., several cancer vaccine peptides, including a cyclophilin B-derived peptide (CypB-84), elicited an immediate-type skin reaction in prevaccination skin tests. These peptides were prohibited in the subsequent vaccinations because of a possible induction of systemic anaphylaxis. In this study, we investigated mechanisms involved in the peptide-elicited inflammatory reactions in BALB/c mice whose MHC class I molecule (Kd) shared similar binding motifs with the human HLA-A24 molecule. Among 11 peptides tested, all of which had been scheduled for use in clinical trials with HLA-A24+ cancer patients, three peptides (CypB-84, ART1-170, and ART4-13) elicited immediate footpad reactions in BALB/c mice similar to the skin reactions in humans. The footpad reaction was also observed in C57BL/6, athymic nu/nu, and CB17-SCID mice, but not in mast cell-deficient WBB6F1w/wv mice, indicating the reaction was not mediated by specific immunity, but was mast cell-dependent. Furthermore, the reactions were not correlated to in vivo antitumor effects of the peptides. An anaphylaxis was not elicited when the peptides were systemically injected due to a very rapid clearance of the peptides from the plasma by in vivo degradation. These results suggest that certain peptides of cancer vaccine candidates exhibit an IgE-independent but mast cell-dependent inflammatory response with no elicitation of systemic anaphylaxis, and may provide new insights for further development of peptide-based vaccinations for cancer patients.

Effect of Rolipram, a Phosphodiesterase IV Inhibitor, on Allergic Footpad Swelling using Various Adjuvants in Mice

We studied the effect of rolipram, a phosphodiesterase (PDE) IV inhibitor, on allergic footpad swelling in mice. For this study, varying adjuvants including complete Freund's adjuvant (CFA), incomplete Freund's adjuvant (IFA) and Imject Alum (Alum) were used because the extent of antigen-specifically induced T helper type 1 (Th1) and Th2 responses had been shown to depend on adjuvants used. To induce allergic footpad swelling, we immunized mice with ovalbumin (OVA) emulsified in either CFA or IFA, dissolved in Alum or in phosphate-buffered saline (PBS) as a control (day 0), followed by subcutaneous injection of the antigen into footpads on day 21. Rolipram was given orally to the animals daily from days 0-20. Results showed that treatment with rolipram was followed by an increase in early swelling at 0.5 h and a decrease in late swelling at 6 and 24 h in the CFA group. In the IFA group, rolipram significantly enhanced swelling at, but not after, 30 min. In the Alum and the PBS groups, the PDE inhibitor failed to affect the OVA-specific footpad reaction at all times examined. Treatment of the CFA and IFA groups with rolipram significantly inhibited the production of the Th1 antibody anti-OVA immunoglobulin G2a (IgG2a), and the drug enhanced Th2 cell-dependent anti-OVA IgE production. In both groups, rolipram also enhanced the secretion of Th2 cytokines including interleukin-4 (IL-4) and IL-10. These findings suggest that rolipram may facilitate early allergic footpad swelling mediated by Th2 immune responses, while the late phase of swelling associated with Th1 responses may be attenuated by the PDE IV inhibitor.

Mycobacterial trehalose 6,6′-dimycolate preferentially induces type 1 helper T cell responses through signal transducer and activator of transcription 4 protein

Mycobacterium tuberculosis is an intracellular pathogen of tuberculosis and its pathogenicity is related to the ability to escape killing by ingested macrophages and induce delayed-type hypersensitivity (DTH). A major component of the cell wall of M. tuberculosis is trehalose 6,6′-dimycolate (TDM), which has been implicated as a pathogenetic factor. The expression of DTH and cell-mediated immunity is dependent on the macrophage-cytokine-type 1 helper T (Th1) lymphocyte axis. Cytokines, interleukin-12 (IL-12) and interferon-γ (IFN-γ), play a critical role in the process and IL-12-activated signal transducer and activator of transcription (STAT) 4 is required for the development of fully functional Th1 cells. To clarify host responses to mycobacterial TDM, we have analyzed footpad reaction, histopathology and cytokine profile of experimental granulomatous lesions using STAT4-deficient mice. In the present study, we have demonstrated that mycobacterial TDM selectively induces the Th1 response through the STAT4 signaling pathway, because mice lacking STAT4 protein significantly reduced to develop DTH, hypersensitivity granulomas, and Th1 cytokine responses, when compared to BALB/c mice. These results shed light on the molecular pathogenesis of mycobacterial disease. Taken together with previous studies, TDM is a pleiotropic molecule against the host and participates in the pathogenesis.

Further evidence regarding the effect of dietary protein on oral tolerance against beta-lactoglobulin through Th1-mediated immune response in mice.

Oral tolerance is a potential strategy for preventing or minimizing aberrant immune responses. Although, oral tolerance has been extensively studied, to date the effects of dietary protein on the induction of oral tolerance are poorly understood. We have previously shown that restricted dietary protein induces oral tolerance to ovalbumin. This study was designed to investigate whether or not such tolerance occurs with beta-lactoglobulin (BLG) instead of ovalbumin (OVA) and if the tolerance resulting from this feeding regimen involves Th1-mediated immune response. Female BALB/c mice fed either 20% or 5% dietary protein were given 5 mg BLG or water orally for four consecutive days and then immunized intraperitoneally (ip) twice with BLG at 3-wk intervals. Oral tolerance induction was compared in BLG-fed and water-fed mice by measuring total IgE, BLG-specific antibodies, footpad reactions, splenocyte proliferation, and cytokine production. When mice were given BLG orally before ip immunization, the Th1-mediated immune responses (production of IL-2, IFN-gamma, and IgG2a) were significantly reduced, whereas the Th2-mediated immune responses (production of IL-4 and IgG1) were unchanged. The Th1-mediated immune responses were markedly down-regulated in mice fed 5% protein as compared to those in mice fed 20% protein. Moreover, the production of total IgE, BLG-specific IgE, splenocyte proliferation, and footpad reactions were more reduced in mice fed 5% protein than those in mice fed 20% protein. The present study provides evidence that dietary protein plays an important role in the induction of oral tolerance against BLG as the result of, clear down-regulation of Th1 helper activity accompanied by a reduction in IgE.

Microanalysis of Tryptophan Metabolites and Suppressor Factor of Delayed‐Type Hypersensitivity in Mice

AbstractFor Abstract see ChemInform Abstract in Full Text.

Microanalysis of tryptophan metabolites and suppressor factor of delayed-type hypersensitivity in mice

We developed methods for the fluorometric assay of 3-hydroxykynurenine and 3-hydroxy-anthranilic acid, which are suspected as carcinogens in bladder cancer. It was shown that the urinary excretion of 3-hydroxyanthranilic acid increased in patients with bladder cancer. We also developed methods for the fluorometric assay of glucuronide and sulfate of 3-hydroxyanthranilic acid and showed that the excretion of these conjugated forms was minor in humans. The distribution of 3-hydroxykynurenine was studied and data obtained suggested that it has an affinity for the pancreas. We then developed methods for determination of the related compounds of tryptophan. Fluorescence reaction with UV radiation was applied to the determinations of kynurenic acid, kynurenine, quinolinic acid, nicotinic acid, nicotinamide, N1-methyl-nicotinamide, isatin, xanthrenic acid, and melatonin in the serum or urine. Furthermore, the fluorescence reaction with UV radiation was applied to some drugs, e.g., indomethacin, isoniazid, naldixic acid, nicorandil, and disodium cromoglycate. The relationship was investigated between the tumor promoter, 12-tetradecanoyl-phobol-13-acetate (TPA), and delayed hypersensitivity in mice. The foot pad reaction (FPR) in mice was suppressed by the application of TPA following the application of 7,12-dimethylbenz[alpha]-anthracene (DMBA), a tumor initiator, in BALB/c mice, while the FPR was suppressed by the application of TPA alone in C3H/He mice. CD8+ and CD4+ T cells, which suppress the FPR, were induced in BALB/c and C3H/He mice, respectively. These T cells produced soluble factors that inhibited the FPR in mice.

Th1 adjuvant N-acetyl-D-glucosamine polymer up-regulates Th1 immunity but down-regulates Th2 immunity against a mycobacterial protein (MPB-59) in interleukin-10-knockout and wild-type mice.

Treatment of mice with heat-killed (HK) Mycobacterium bovis BCG or 1- to 10-microm chitin particles (nonantigenic N-acetyl-D-glucosamine polymers) is known to induce innate immune responses, including gamma interferon (IFN-gamma) production, which plays a Th1 adjuvant role. However, HK BCG further induces prostaglandin E2-releasing spleen macrophages (Mphi) (PGE2-Mphi), which potentially inhibit Th1 adjuvant activities. We found that chitin particles did not induce PGE2-Mphi formation. To further assess whether chitin has Th1 adjuvant effects, interleukin-10 (IL-10)-knockout (KO) mice and their wild-type (WT, C57BL/6) controls were immunized with a 30-kDa MPB-59 mycobacterial protein mixed with chitin. Immunization with MPB-59 alone induced Th2 responses, characterized by increases in total serum immunoglobulin E (IgE) and specific serum IgG1 levels and spleen Th2 cells producing IL-4, IL-5, and IL-10. No IFN-gamma-producing spleen Th1 cells, specific serum IgG2a, or delayed-type hypersensitivity (DTH) footpad reactions were detected. On the other hand, chitin-MPB-59 immunization significantly increased spleen Th1 responses, DTH reaction, and serum IgG2a levels along with decreases of Th2 responses. The magnitude of these Th1 adjuvant effects was greater in IL-10-KO mice than in WT mice. In contrast, immunization with HK BCG-MPB-59 showed little or no Th1 adjuvant effect. These data indicate that chitin has a unique Th1 adjuvant effect on the development of Th1 immunity against a mycobacterial antigen. IL-10 down-regulates the adjuvant effect of chitin.

Trehalose 6,6'-dimycolate (cord factor) of Mycobacterium tuberculosis induces foreign-body- and hypersensitivity-type granulomas in mice.

Granulomatous inflammation is characterized morphologically by a compact organized collection of macrophages and their derivatives. It is classified as either a hypersensitivity type or a foreign-body type. Lipid components of the Mycobacterium tuberculosis cell wall participate in the pathogenesis of infection. Strains of M. tuberculosis have cord factor (trehalose 6,6'-dimycolate [TDM]) on their surface. To clarify host responses to TDM, including immunogenicity and pathogenicity, we have analyzed the footpad reaction, histopathology, and cytokine profiles of experimental granulomatous lesions in immunized and unimmunized mice challenged with TDM. In the present study, we have demonstrated for the first time that TDM can induce both foreign-body-type (nonimmune) and hypersensitivity-type (immune) granulomas by acting as a nonspecific irritant and T-cell-dependent antigen. Immunized mice challenged with TDM developed more severe lesions than unimmunized mice. At the active lesion, we found monocyte chemotactic, proinflammatory, and immunoregulatory cytokines. The level was enhanced in immunized mice challenged with TDM. This result implies that both nonimmune and immune mechanisms participate in granulomatous inflammation induced by mycobacterial infection. Taken together with a previous report, this study shows that TDM is a pleiotropic molecule against the host and plays an important role in the pathogenesis of tuberculosis.

Cell-mediated immune responses (CMIR) to Rhinosporidium seeberi in mice.

There is no published data on Cell Mediated Immune Responses in experimental animals to Rhinosporidium seeberi the causative agent of human and animal rhinosporidiosis. The quantitative mouse foot-pad model was used to assay the Delayed-Type Hypersensitivity (DTH) cell-mediated immune response to extracts of purified endospores and sporangia of R. seeberi. Histological examination was used to confirm that the foot-pad reactions were compatible with DTH reactions in the mouse. We report that sonically disintegrated rhinosporidial endospores/sporangia induced DTH responses in the foot-pads of sensitized mice which were comparable in intensity and histological profile to that induced by sheep red blood cells in SRBC sensitized mice. Anti-rhinosporidial antibody was also induced. Filtrates of the soluble antigens in sonicated suspensions failed to evoke a DTH-foot-pad (DTH-FP) response in sensitized mice although an anti-rhinosporidial antibody response to this preparation was detected. Prolonged pre-treatment with sonicated suspensions of endospores and sporangia resulted in a decrease of DTH reactivity as compared with reactions following pre-treatment of a shorter duration.

Delayed-type hypersensitivity reaction induced in broilers by killed Staphylococcus aureus

A trial was conducted to determine whether the delayed footpad reaction (DFR) induced by killed Staphylococcus aureus in chickens is a delayed-type hypersensitivity (DTH) reaction. Five criteria were used to assess DTH: 1) DFR with a peak response at 24 to 48 h postchallenge, 2) inhibition of monocyte/macrophage migration, 3) lymphocyte blastogenic response, 4) mononuclear cell infiltration at the challenge site, and 5) passive transfer of DFR by splenic lymphocytes. Broilers were sensitized twice with a s.c. injection in the neck of S. aureus antigen (150 microg/bird) diluted in polyethylene glycol at 3 and 4 wk of age. Controls were s.c. injected with polyethylene glycol. At 6 wk of age, a migration inhibition test was conducted before the birds were challenged intradermally with S. aureus antigen (75 microg/bird) in PBS in the right footpad. The left footpad was injected with PBS. The thickness of the footpad was measured at 0, 4, 24, and 48 h postchallenge to evaluate the DFR. After challenge, blood was collected for the lymphocyte blastogenesis assay. Birds were euthanatized, and both footpads were removed for histology. The spleens were collected aseptically; splenic lymphocytes were injected i.v. into recipient birds. Sensitized birds showed an increase in the DFR (P < 0.02) and blastogenic response (P < 0.01) compared with nonsensitized birds. Delayed footpad reaction reached a maximum response at 24 h postchallenge. The in vitro migration of monocytes/macrophages from sensitized birds was significantly inhibited (P < 0.01). The histological appearance of S. aureus-injected footpads was characterized by dermal edema and perivascular infiltrates of small lymphocytes and macrophages. Birds that received sensitized splenic lymphocytes had a significantly pronounced DFR following challenge with S. aureus when compared with birds that received nonsensitized lymphocytes (P < 0.0001). These results indicated that the DFR can be used as a standard in vivo test for cell-mediated DTH reaction induced by killed S. aureus antigen in chickens.

Delayed-type hypersensitivity reaction induced in broilers via trachea inoculation of killed Staphylococcus aureus

A study was conducted to determine whether the delayed-type hypersensitivity (DTH) reaction to killed Staphylococcus aureus antigen in chickens could be induced through multiple intratracheal inoculations. Three criteria were used to assess DTH: 1) delayed footpad reaction (DFR) with a peak response at 24 to 48 h postchallenge, 2) inhibition of monocyte/macrophage migration, and 3) mononuclear cell infiltration at the challenge site. Broilers were sensitized three times with a s.c. injection in the neck or intratracheal inoculation of killed S. aureus in polyethylene glycol at 2, 3, and 4 wk of age. Controls were given polyethylene glycol with a s.c. injection in the neck or intratracheal inoculation. Migration inhibition tests were conducted at 6 wk of age. At 7 wk of age, all birds were challenged intradermally with S. aureus antigen in PBS in the right footpad. The left footpad was injected with PBS. The thickness of the footpad was measured at 0, 4, 24, and 48 h postchallenge to evaluate the DFR. Birds were euthanatized, and both footpads were removed for histopathological examination. Subcutaneously or intratracheally sensitized birds showed significant DFR compared with nonsensitized birds (P < 0.0001), which reached maximum response at 24 h postchallenge. The s.c. sensitization resulted in an inhibition of the in vitro migration of monocytes/macrophages (P < 0.0001), whereas intratracheally sensitized birds did not show migration inhibition of monocytes/macrophages. Histological examination showed typical perivascular infiltration of small lymphocytes in S. aureus-injected footpads from s.c. and intratracheally sensitized birds. These results indicate that multiple intratracheal inoculation, as well as s.c. injection of killed S. aureus antigen, can be used to induce a cell-mediated DTH reaction in chickens.

Induction of the delayed footpad and wattle reaction to killed Staphylococcus aureus in chickens

Two experiments were conducted to induce the delayed footpad reaction (DFR) to killed Staphylococcus aureus antigen. In Experiment 1, tracheal, cloacal, and choanal swabs were collected from chickens prior to sensitization with S. aureus to determine the carrier status of S. aureus. The second experiment compared the DFR to the delayed wattle reaction (DWR). Chickens were subjected to single or multiple sensitizations in the neck with S. aureus antigen between 4 and 6 wk of age. One week later, birds were challenged with S. aureus either in the right footpad or wattle. The left footpad or wattle was injected with PBS. The thicknesses of the footpad or the wattle were measured up to 96 h postchallenge. The recoveries of S. aureus from the choanal slit and trachea were significantly higher than that of the cloaca (P < 0.001). Birds of Experiment 1 showed a significant DFR (P < 0.0001) following intradermal challenge with killed S. aureus that was sustained through 48 h postchallenge with no difference in the DFR between carrier and noncarrier birds. In Experiment 2, the thicknesses of the footpad and wattle were significantly increased following challenge with S. aureus (P < 0.0001), with the footpad showing a greater response than the wattle (P < 0.001). Three sensitizing dosages, as compared to two dosages, resulted in a less pronounced DFR and DWR (P < 0.02). These results indicate that the DFR can be used as a delayed reaction model in the study of staphylococcosis in poultry.

Effects of erythromycin on experimental extrinsic allergic alveolitis.

Recent clinical studies have demonstrated the efficacy of erythromycin for treating patients with chronic lower respiratory tract inflammation. Mechanisms related to the anti-inflammatory action are yet to be determined. The therapeutic efficacy of erythromycin in experimental extrinsic allergic alveolitis (EAA) was evaluated. A murine model of EAA was developed by intratracheal inoculations with particulate Trichosporon mucoides followed by erythromycin or josamycin treatment. Cell populations, specific antibodies, chemotactic activities, TNF-alpha, IL-1beta, MIP-2 and KC of bronchoalveolar lavage fluid (BALF); histopathology of the lung and footpad reaction; myeloperoxidase of the whole lung; and immunohistochemistry of intercellular adhesion molecule- (ICAM-1), at 6 and 96 h after the challenge, were examined. There was a marked neutrophilic alveolitis and bronchiolitis at 6 h, and lymphocytic alveolitis and perivenule cuffing at 96 h after the challenge. Increase in total inflammatory cells and neutrophils in BALF at 6h was significantly suppressed by pretreatment with 5 mg/kg/day of erythromycin intraperitoneally for 5 days (P<0.01), with no apparent effect on specific antibodies, chemotactic activity or cytokines. Erythromycin also suppressed the Arthus-type reaction in the footpad (P<0.01). Histopathological studies revealed that erythromycin markedly decreased neutrophils in the lung and skin lesions and myeloperoxidase in the lung, simultaneously with inhibiting ICAM-1 expression. The therapy has no remarkable effects on lymphocytes or 96 h response. Josamycin had no effects on the model. The therapeutic dosage of erythromycin significantly suppressed acute neutrophil influx into the lung, intradermal Arthus reaction and the expression of ICAM-1 in the lesions of experimental EAA. Erythromycin may be effective for treating subjects with acute EAA.

CELL-MEDIATED AUTOIMMUNE RESPONSE TO TESTIS INDUCED BY BILATERAL TESTICULAR INJURY CAN BE SUPPRESSED BY CYCLOSPORIN A

Purpose Since the delayed type hypersensitivity (DTH) response to testis antigens plays a key role in the induction and/or maintenance of experimental autoimmune orchitis (EAO), an animal model for human immunological male infertility or aspermatogenesis, we have investigated the immunosuppressive effect of cyclosporin A (CsA) on the DTH response to autologous testicular cells (TC). Materials and Methods A DTH response to autologous TC was induced in C3H/HeN mice by bilateral testicular injury (trauma). CsA was administered intraperitoneally for 10 consecutive days before and after injury. A DTH response was assessed by measuring delayed footpad reaction (DFR) to autologous TC 9 days after injury. Results When the mice were traumatized alone, 10 mg./kg. or more of CsA suppressed the DTH response to autologous TC significantly. In mice traumatized with 100 mg./kg. of cyclosphosphamide (CY)-pretreatment, 30 mg./kg. or more of CsA was needed to suppress the DTH response. In mice traumatized with 200 mg./kg. of CY-pretreatment, 50 mg./kg. of CsA was needed to suppress the autoimmune response. Conclusions The DTH response to autologous TC was suppressed significantly by administration of CsA in a dose-dependent manner. We have also shown the direct suppressive effect of CsA on effector cells for DTH by means of local passive transfer of DTH. Administration of CsA had no augmenting or suppressive effect on suppressor cells for DTH. CsA might be a significant drug for the immunosuppression of EAO and possibly for immunological male infertility.

The induction of renal autoantigen-specific T cells by a local Listeria monocytogenes infection.

In order to examine the possibility that a local chronic infection can induce organ-specific autoimmune disease, we inoculated unilateral kidneys with viable Listeria monocytogenes (intrarenal infection). The delayed footpad reaction against syngeneic kidney homogenate (KH) became positive from 1 week after initiating the intrarenal infection. A proliferative response of the spleen T cells from the infected mice was also observed against KH from 1 week after initiating the intrarenal infection, but no such response was seen against liver homogenate (LH). In contrast, an intravenous Listeria infection did not induce a delayed footpad reaction or proliferative response against KH, suggesting that these autoimmune responses were not caused by molecular mimicry between renal antigens and Listeria antigens. Furthermore, the ability to transfer the autoimmune response of spleen T cells from intrarenally infected mice was examined. The transferred mice showed a positive delayed footpad reaction against KH and an interstitial infiltration of mononuclear cells in their kidneys. These results demonstrate that the intrarenal Listeria infection induced renal autoantigen-specific T cells, which subsequently induced an autoimmune interstitial nephritis (AIN). The autoreactive T cells were all induced without immunization by autoantigens mixed with complete Freund's adjuvant. Based on these findings, we propose that a local bacterial infection may induce an autoimmune response against autoantigens in the infected organ and subsequently trigger organ-specific autoimmune disease.

Schistosome eggs in the portal vein can induce tolerance

Schistosoma japonicum lives in the portal vein and/or mesenteric vein of patients, producing numerous eggs which eventually induce multiple granulomas in the liver and the intestime. The experimental administration of Schistosoma japonicum eggs via the portal vein in the mouse induces telerance both in the footpad reaction and granuloma formation in the lung. Thus, the presence of Schistosoma japonicum in the portal vein is beneficial to the parasites, not only for receiving enriched nutrition from the blood of the portal vein, but also for inducing immunological tolerance in the hosts. However, prior intravenous injection of gadolinium chloride abolishes such tolerance, indicating the role of Kupffer cells in the induction of tolerance via the portal vein.