Persistent interleukin-2 activity and molecular evidence for expression of lymphotoxin in the hapten-immune model for pulmonary interstitial fibrosis.

The hapten-immune model for pulmonary fibrosis shows that a specific T-cell-mediated immune response is essential for the induction of a nonresolving fibrosis. Here, we report results from studies that identify soluble factors released by activated T lymphocytes that might mediate long-lasting fibrosis. Pulmonary fibrosis was induced by priming hamsters for contact hypersensitivity responses with an epicutaneous application of 2,4,6-trinitro-1-chlorobenzene (TNCB) in carrier and challenging intratracheally (IT) 5 days later with a single dose of the soluble form of the immunizing hapten. Bronchoalveolar lavage fluid was harvested at various time points after IT challenge and assayed for tumor necrosis factor (TNF) and interleukin-2 (IL-2) bioactivity. After IT challenge with the sensitizing hapten, only the immune animals contained IL-2 activity in the bronchoalveolar lavage fluid. TNF activity was detected in lungs of both immune and nonimmune animals. Interestingly, the TNF activity was significantly higher (P less than 0.05) in nonimmune challenged than in immune challenged animals on day 5. Molecular hybridization studies showed that a similar amount of TNF-alpha mRNA was expressed in adherent cells from both groups. The nonadherent subpopulation of mononuclear cells harvested from challenged-immune animals expressed TNF-beta (lymphotoxin) mRNA. These data show, for the first time, an association of lymphotoxin with the appearance of pulmonary fibrotic disease in an animal model for pulmonary fibrosis. These observations are consistent with the postulates that lymphotoxin and IL-2 participate in the immunopathogenesis of hapten-immune induced pulmonary fibrosis and that TNF-alpha is associated with the healing of the fibrotic process initiated by toxic lung injury.     

Delayed type-hypersensitivity response of inbred strains of Syrian golden hamsters (Mesocricetus auratus) to lethal or non-lethal lymphocytic choriomeningitis virus (LCMV) infections

In adult Syrian golden hamsters (Mesocricetus auratus), intraperitoneal or footpad inoculation of the lymphocytic choriomeningitis virus (LCMV) strains, WE or Armstrong (ARM), caused systemic infection and induced serum LCMV-antibody. Hamster and virus strain-dependent lethal disease also occurred. With WE, MHA and PD4 inbred hamsters failed to eliminate infection and died of wasting disease. LSH and CB inbred hamsters resisted lethal WE-disease and cleared infection. LVG hamsters and inbred LHC hamsters were intermediate in WE-susceptibility; some died of wasting, while others survived with little illness. Resistance to lethal WE-disease directly correlated with a delayed-type hypersensitivity (DTH) response to live-virus footpad inoculation. In WE-resistant LSH and CB hamsters, DTH-responses were induced by intraplantar WE-inoculation; footpad edema began by 5 days, reached maximum thickness by 7 to 9 days, and subsided thereafter. In the other hamster strains, DTH to WE could not be elicited. Unlike WE, ARM was hamster-avirulent; infections were self-limited and did not induce DTH. All survivors of primary LCMV (WE or ARM)-infection resisted secondary WE-challenge, and did not develop DTH to LCMV. Immunosuppressive treatments, abrogating DTH and antibody responses to LCMV, rendered all hamsters susceptible to lethal WE-infection. Hamster DTH most likely mediated resistance to virulent LCMV-infection.     

A critical role for alveolar macrophages in elicitation of pulmonary immune fibrosis

Hapten immune pulmonary interstitial fibrosis (HIPIF) is induced by a recall cell-mediated immune response against the hapten 2,4, 6-trinitrobenzene sulphonic acid (TNBS) in the lung. Studies here dissect the role of the cellular components of the bronchoalveolar lavage (BAL) cells (alveolar macrophages [AMs] versus monocytes and immature dendritic cells) in the fibrogenic inflammatory response. BAL cells from HIPIF mice were generally more activated and produced a greater amount of tumour necrosis factor-alpha (TNF-alpha) than controls. Liposome-encapsulated dichloromethylene diphosphonate (Cl(2)MDP) that was inoculated intranasally (i.n.) into mice selectively depleted AMs. Following AM depletion, the number of TNF-alpha-containing cells was reduced, and both the number of immune inflammatory cells recruited into the alveolar space and the subsequent collagen deposition (hydroxyproline) were decreased in the sensitized and intratracheally (i.t.) challenged mice. In conclusion, AMs are required, in part, for the development of pulmonary fibrosis in HIPIF because AM-derived factors such as TNF-alpha are needed for initiation of chemokine and cytokine pathways and accumulation of immune inflammatory cells.     

The induction of carrier-specific helper cell tolerance in presensitized rats.

Lewis rats rendered tolerant to sheep IgG (SGG) show a markedly reduced antibody response to the 2,4,6-trinitrophenyl (TNP) hapten when later challenged with TNP-SGG. We have previously shown that this effect is due to functional unresponsiveness in the carrier SGG-specific helper T cell population. In this paper we demonstrate that induced helper cell tolerance is also maintained through a secondary immunogenic challenge. Furthermore, rats which are primed to the carrier SGG prior to tolerance induction also show a markedly reduced anti-TNP response upon secondary immunogenic challenge with TNP-SGG. The ability to specifically suppress a secondary response in this manner was found to be relatively long lasting, since rats showed reduced responsiveness when the secondary challenge was delayed for up to 4 weeks after tolerance induction. In addition, rats primed to the hapten (TNP) prior to carrier (SGG) tolerance induction also showed a marked reduction in anti-TNP antibody following challenge with TNP-SGG. These findings imply that helper cell tolerance can be induced in rats even after priming of carrier-specific (SGG) helper cells, or hapten-specific (TNP) B cells. These results parallel our other published findings that IgE responses in presensitized rats can be overcome by helper cell tolerance.     

Immunosuppression-induced susceptibility of inbred hamsters (Mesocricetus auratus) to lethal-disease by lymphocytic choriomeningitis virus infection

Summary The role of the immune response in the pathogenesis of lethal and non-lethal lymphocytic choriomeningitis virus (LCMV)-infections of young adult Syrian golden hamsters (Mesocricetus auratus) of different strains was examined using immunosuppressive treatment with cyclophosphamide or with whole-body gamma-irradiation. In all hamsters, the LCMV strains, WE and Armstrong (ARM), caused systemic infections and induced comparable serum LCMV-antibody titers. However, lethal wasting-disease occurred which was hamster-strain and virus-strain dependent. With WE-inocula, MHA and PD4 inbred hamsters were all susceptible to lethal-disease and failed to completely eliminate infection. All LSH and CB inbred hamsters resisted lethal-disease and totally cleared WE-infection. Random colony-bred LVG hamsters and inbred LHC hamsters were intermediate in WE-susceptibility; some died with wasting, while others survived with minimal to no illness. ARM was avirulent for all hamsters and infections were totally cleared. By immunosuppressive treatment, all hamsters were rendered completely susceptible to lethaldisease by WE, and had unresolved infections and diminished serum LCMV-antibody titers. Immunosuppression also rendered all hamster strains partially susceptible to lethal infection by ARM. The hamster immune response was thus shown to suppress LCMV-infection and protect against lethal illness.E.V.G. was funded by the Resident Research Associateship program of the National Research Council.     

Genetic analysis of resistance to lethal infections of vesicular stomatitis virus in Syrian hamsters.

Vesicular stomatitis virus (VSV), Indiana serotype, causes a lethal disease in adult Syrian hamsters. Susceptibility to low doses of VSV (10 to 100 plaque-forming units) was shown to be genetically determined by examining six inbred strains. Three strains, LSH, MHA, and CB, were found to be extremely susceptible, with more than 70% of the animals dying within 72 h after intraperitoneal injection. Two strains, MIT and UT2, were intermediately susceptible, with approximately 60% of VSV-infected animals surviving. One strain, UT1, was found to be highly resistant; however, resistance was not acquired until the 4th or 5th week of age, and 100% of VSV-infected neonatal hamsters died. Analyses of F1 hybrid and segregant backcross populations derived from the LSH and UT1 strains suggested that at least three independent genetic loci contribute to phenotypic resistance. One locus resides on the X chromosome; the others reside on autosomes. No obvious linkage to the hamster major histocompatibility complex was detected. F1 hybrids of two highly susceptible strains, CB and MHA, were more resistant than either parental strain, suggesting that alleles of unlinked genes in the susceptible strains interact to produce a partially resistant phenotype. These alleles probably are the cause of the resistance phenotype found in the random-bred LVG strain which shares a common heritage with the CB and MHA strains.     

Susceptibility of LSH hamsters to intraperitoneal inoculation with Legionella pneumophila

The guinea pig is the most widely used animal model in the study of Legionellosis. The hamster should also be considered, since it acquires virtually no spontaneous epidemic lung infection, possesses similar cellular immune components observed in other mammals, has a normal body temperature identical to that of man, and is readily available for laboratory investigation. We studied the pathologic findings of four 12 week old inbred London School of Hygiene (LSH) hamsters inoculated intraperitoneally with 0.2 ml of 10(9) organisms per ml suspension of a viable Philadelphia 1 strain of Legionella pneumophila. Four LSH hamsters (control group) received 0.2 ml of sterile phosphate buffered saline, intraperitoneally. All animals of the test group became clinically ill and two of the four spontaneously expired on days 1 and 2 after inoculation. The remainder were sacrificed on day 3. In three out of four animals of the test group, a suppurative peritonitis and an interstitial pneumonitis were observed. It was characterized by infiltrates of neutrophils and macrophages. The test group also exhibited acute splenitis, including microabscesses, and two of four test animals showed hepatic congestion, vacuolization of hepatocytes, and microabscesses. None of the controls appeared sick or died after three days, and neither gross nor microscopic lesions were found at autopsy. Culture results documented L pneumophila in lung and spleen of all test animals and the absence of organisms in the control group. Hence, the LSH hamster is rapidly infected with the Philadelphia 1 strain of L pneumophila given intraperitoneally, and pathological changes can be readily observed. The findings of our study add hamsters to the list of animals susceptible to intraperitoneal infection by L pneumophila.     

Radiation-induced pulmonary fibrosis resolves spontaneously if dense scars are not formed

The relation of static compliance of excised lungs to collagen accumulation and histologic fibrosis was examined in Syrian hamsters inhaling sufficient 238PuO2 particles to achieve initial lung burdens of 50 or 100 nCi. Control animals were exposed to nonradioactive aerosols. Irradiated lungs from hamsters at both dose levels had compliance reduced to the same extent at point of maximal reduction. However, collagen accumulation was more closely related to 238Pu exposure level than the compliance measurements. Histologic examination revealed both diffuse alveolar thickening and some dense fibrous scars, the former predominating at lower dose levels. Hamsters exposed to 50 nCi 238PuO2 showed normal collagen content and static lung compliance with minimal histologic fibrosis 288 days after exposure. In contrast, hamsters exposed to 100 nCi had significant pulmonary fibrosis at that time and the highest incidence of dense scars at any time period. Such findings are consistent with a stiffening of lung parenchyma. They suggest that the diffuse interstitial fibrosis developed by this injury resolves spontaneously; dense fibrous scars, however, do not.     

Allergic hepatitis in guinea pigs induced by 2,4,6-trinitrophenyl liver protein conjugate

Experimental drug-induced allergic hepatitis was induced in guinea pigs which had been immunized to the 2,4,6-trinitrophenyl (TNP)-conjugated liver protein first peak (TNP-LP1) emulsified in complete Freund's adjuvant (CFA). Delayed-type hypersensitivity (DTH) was elicited in the immunized animals by intradermal challenge with TNP-LP1. When TNP-LP1 was introduced into the liver via the mesenteric vein, allergic hepatitis was provoked. Histological examination of the liver revealed massive monocytic infiltration and focal hepatic necrosis in the periportal areas. Blood biochemical analysis showed increased levels of glutamate oxalacetate transaminase (GOT) and total bilirubin. TNP-modified hepatocytes were found to be effective as a challenge elicitor to the immunized animals to induce both DTH skin reaction and allergic hepatitis. In the latter case, the severity of the lesion was stronger than that observed by the challenge with TNP-LP1.     

Variation Between Strains of Hamsters in the Lethality of Pichinde Virus Infections

Infection by Pichinde virus, a member of the arenavirus group, was studied in Golden Syrian hamsters (Mesocricetus auratus) with regard to possible mechanisms of resistance to virus infection in adult hamsters. Two hamster strains were found to differ in their susceptibility to lethal Pichinde virus infection. LVG/Lak randomly bred hamsters were found to be 100% susceptible to low doses of Pichinde virus during the first 6 days of life, but after 8 days of life, mortality was uncommon. Peak virus titers in the serum of animals infected at 3 days of life were 4 logs greater than in animals infected at 12 days. MHA/Lak inbred hamsters, in contrast, were found to be susceptible to lethal virus infection both as newborns and as adults. Peak virus titers of greater than 10(8) plaque-forming units/ml were observed in serum 8 days after infection of adult MHA hamsters as compared with less than 10(3) plaque-forming units/ml in the serum of adult LVG hamsters. Cultured primary kidney cells and peritoneal macrophages from either hamster strain supported Pichinde virus replication equally well in vitro. Antibodies to the complement-fixing antigens and to antigens at the surface of virus-infected cells were produced by both strains of hamsters. Cyclophosphamide immunosuppression rendered adult LVG animals susceptible to lethal infections, and virus grew to high titers in the treated animals. These findings suggest that immunological factors that appear early in life in LVG hamsters and are deficient in MHA hamsters limit Pichinde virus infection. Unlike previously reported arenavirus diseases, the observations suggest that death is produced by a direct viral effect and not through immunopathological mechanisms.     

Abatement of bleomycin-induced increases in vascular permeability, inflammatory cell infiltration, and fibrotic lesions in hamster lungs by combined treatment with taurine and niacin.

BACKGROUND: The bleomycin (BL) hamster model of interstitial pulmonary fibrosis has been widely used to study the pathogenesis of interstitial pulmonary fibrosis and to screen potentially desirable antifibrotic agents. We have recently shown that taurine and niacin in combination, diminished BL-induced increases in lung lipid peroxidation and hydroxyproline content in hamsters. In the present study, we have evaluated the effects of taurine and niacin on the bronchoalveolar lavage (BAL) cells, and morphologic and morphometric features of the lung in the same model of pulmonary fibrosis. EXPERIMENTAL DESIGN: The hamsters were divided into 4 groups: saline; taurine + niacin + saline; BL; and taurine + niacin + BL. Treatment of taurine and niacin began 2 days before the first intratracheal instillation of saline or BL and thereafter daily throughout the study for taurine + niacin + saline and taurine + niacin + BL groups. Hamsters received BL or saline in three consecutive doses at weekly intervals by intratracheal route. Twenty days after the last intratracheal instillation, the hamsters were sacrificed for various studies. RESULTS: Combined treatment with taurine and niacin suppressed BL-induced inflammation and almost completely abrogated pulmonary fibrosis in hamsters. Two independent studies showed that taurine and niacin in combination significantly reduced BL-induced increases in bronchoalveolar inflammatory cell counts, protein content, and acid phosphatase activity. By both light and electron microscopy, the lungs of hamsters treated with BL and taurine and niacin had much fewer inflammatory cells, less epithelial necrosis and collagen deposition than hamsters treated with BL alone. CONCLUSIONS: The results of this investigation suggest that combined treatment with taurine and niacin is effective against the development of lung fibrosis in the BL-hamster model and offers a novel therapeutic modality in the prevention of the fibrotic processes.     

Hapten-Specific Unresponsiveness in Mice

The features of B-cell tolerance induced in mice with the chemically reactive hapten 2,4,6-trinitrobenzenesulphonic acid were investigated after various antigenic challenges. A complete abolition of the IgG response was observed after challenge with trinitrophenol (TNP)-coupled haemocyanin (TNP-KLH) associated or not with Escherichia coli lipopolysaccharide. Carrier immunization with KLH (3 micrograms) followed by challenge with TNP-KLH led to a normal IgG anti-TNP response. However, tolerant mice given 3 micrograms TNP-KLH (instead of KLH) and a repeat injection of TNP-KLH exhibited a depressed IgG anti-TNP response. These results are discussed within the framework of tolerance induction in adult mature B cells.     

CTLA4 (CD152) modulates the Th subset response and alters the course of experimental Leishmania major infection

Since both the nature and the amplitude of an antigen-specific T cell response are dependent on co-stimulatory signals, we have invesigated the role of CD28 / CD152-mediated T cell co-stimulation in the regulation of experimental cutaneous leishmaniasis. CD28-deficient mice and their wild-type littermates are equally susceptible to Leishmania major infection. Whole anti-CD152 antibody significantly exacerbates the disease while anti-CD152 Fab ameliorates the disease in genetically susceptible BALB / c mice but not in C57BL / 6, a resistant strain. The anti-CD152-induced exacerbation of the disease is accompanied by increased IL-4-secreting cell number, diminished parasite-specific delayed-type hypersensitivity (DTH) response and augmented anti-2,4,6-trinitrophenyl (TNP) IgG1 in response to TNP-leishmanial antigen crude soluble antigen (CSA), suggesting an exaggerated Th2 type of response. Anti-CD152 Fab-mediated amelioration of the disease is associated with increased IFN-γ-secreting cell number, increased parasite-specific DTH response and enhanced IgG2a isotype in response to TNP-CSA suggesting a Th1 type of response. Unlike TNP-CSA, TNP-keyhole limpet hemocyanin does not induce the change in Ig isotype, indicating that the immunomodulatory effect of anti-CD152 is antigen specific. Anti-CD152 antibody-induced early change in Th subsets suggests an important role for CD152 in determining the course of L. major infection, perhaps by alteration of Th subset differentiation.     

B-cell and T-cell lymphomas and other associated diseases induced by an infectious DNA viroid-like agent in hamsters (Mesocricetus auratus).

Five epidemics of diffuse, poorly differentiated lymphocytic, immunoblastic, and plasmacytoid lymphoma induced by an infectious, horizontally transmitting viroidlike agent have occurred in two hamster facilities. Incidence summaries and pathologic characteristics of the lymphomas induced in LSH and LVG hamsters are presented. An elevated leukocyte count with a marked increase in neutrophils and a significant decrease in small mononuclear lymphocytes was detected in 5-week-old but not in 10- or 25-week-old LVG hamsters born in the facility contaminated with the lymphoma-inducing agent. Three-week-old LVG hamsters exposed to the contaminated facility showed no similar hematologic change at 5 weeks of age or 5 weeks of exposure. Several associated syndromes, including an intussusception disease, pyelonephritis, inflammatory bowel disease, and body warts associated with the presence of the causative viroidlike agent in the contaminated colonies are described. Details of the epidemiology of the disease, karyology, viral studies, and correlation with several epidemics in other laboratories are presented.     

Differing responses of hamsters to infection by vesicular stomatitis virus Indiana and New Jersey serotypes

Intraperitoneal injection of vesicular stomatitis virus, New Jersey serotype (VSV-NJ), into inbred LSH hamsters resulted in an inapparent infection and survival of the majority of the animals. Infectivity titrations of tissues from VSV-NJ-infected hamsters showed that little or no virus was present following infection. The few animals that died from VSV-NJ succumbed to neurological disease. This is in contrast to our previous work where we found that LSH hamsters are exquisitely sensitive to i.p. infection by VSV, Indiana serotype (VSV-IND), and that large amounts of VSV-IND could be detected in tissues. The 50% lethal doses of VSV-NJ and VSV-IND for LSH hamsters are approximately 10(7) pfu and 1 pfu, respectively. When peritoneal cells from LSH hamsters were infected in vitro with both VSV serotypes, the yields of VSV-NJ consistently were lower than yields of VSV-IND. The growth of the two serotypes in fibroblast and epithelial cell lines of hamster origin was similar. VSV-NJ was not more efficient than VSV-IND in inducing interferon in vitro or in vivo, and there appeared to be no difference in the sensitivities of the two serotypes to the antiviral activity of hamster interferon. Thus, i.p. infection with less than 10(7) pfu of VSV-NJ is avirulent for LSH hamsters and this avirulence may be due, in part, to partial intrinsic resistance of peritoneal macrophages to infection by VSV-NJ. When four LSH hamsters that had been immunized with VSV-NJ were challenged with 10(6) LD50 of VSV-IND, three of the four animals survived. Despite the fact that neutralizing antibodies to VSV-NJ did not cross-neutralize VSV-IND, five out of five LSH hamsters were protected by passive transfer of 1 ml of immune hamster anti-VSV-NJ antiserum prior to challenge with VSV-IND. This suggests an important role for non-neutralizing antibodies.     

Involvement of cells of hematopoietic origin in genetically determined resistance of Syrian hamsters to vesicular stomatitis virus.

Susceptibility of Syrian hamsters of the inbred LSH and MHA strains to injection of as few as 10 plaque-forming units of vesicular stomatitis virus (VSV) was shown to occur only after intraperitoneal and intrapleural injection and not after injection of VSV intravenously, intranasally, or in the footpads. Despite the fact that fewer LSH hamsters died when VSV was injected via the latter routes, the histopathology of the VSV-induced disease at early times after infection was identical irrespective of the route of virus administration. Histological examination of tissues at various times after administration of VSV by the various routes revealed that VSV exhibited tropism for lymphoreticular tissue, with the greatest amount of necrosis in the splenic periarteriolar lymphoid sheath. A similar pattern also was observed in VSV-infected tissues from genetically resistant UT1 hamsters. Infectivity titrations of various tissues at different times after intraperitoneal injection of VSV revealed that resistant UT1 hamsters began to clear virus from tissues between 40 and 48 h postinfection, whereas virus titers remained high in susceptible animals. Resistance of UT1 hamsters appeared to require an intact spleen since survival of splenectomized animals was less than that of sham-splenectomized UT1 controls. Sublethal whole-body irradiation was also able to reduce resistance of UT1 hamsters (survival was reduced from 100 to 50%). Bone marrow cells from resistant (UT1 X LSH) F1 females were transferred into lethally irradiated susceptible LSH hamsters, and hematopoietic chimeras were produced. After intraperitoneal injection of 100 plaque-forming units of VSV, all of the female chimeras survived, but only 33% of male chimeras survived. These data indicate that resistance to VSV in Syrian hamsters is mediated, at least partially, by cells of hematopoietic origin.     

The effect of alpha-difluoromethylornithine on the development of bleomycin-induced pulmonary fibrosis in hamsters.

The development of bleomycin-induced lung fibrosis was studied in hamsters drinking tap water or 2% alpha-difluoromethylornithine (DFMO) dissolved in tap water for 14 days. The fibrotic lesions in the lung were evaluated by biochemical measurements of total neutral salt soluble (NSS) and insoluble (NSI) collagens and by morphometric histopathologic techniques. Daily ingestion of DFMO failed to offer any protection against bleomycin-induced lung fibrosis; instead, it increased the deposition of total lung NSI collagen to 396% of control, as compared with 145% of control caused by bleomycin treatment alone. Daily intake of DFMO by itself increased the accumulation of total lung NSI collagen to 250% of control, as opposed to a 145% increase caused by bleomycin treatment alone. Histopathologically, the lung lesions in hamsters treated with bleomycin and DFMO were qualitatively similar to those of hamsters treated with bleomycin alone. However, morphometric estimates revealed that of lung lesions were more diffuse and severe in the former than in the latter group.     

A new assay system detecting antibody production and delayed-type hypersensitivity responses to trinitrophenyl hapten in an individual mouse

A new assay system detecting antibody production and delayed-type hypersensitivity (DTH) responses to trinitrophenyl hapten in an individual mouse (AS-DAD) was established. BALB/c mice were immunized intraperitoneally with varying amounts of 2,4,6-trinitrophenylated sheep red blood cells (TNP-SRBC) on day 0. Venous blood was collected on days 2, 4, 6, 8 and 10. Levels of anti-TNP IgM and IgG in serum were assayed by enzyme-linked immunosorbent assay (ELISA). After series of bleeding the mice were challenged with 2,4,6-trinitrobenzene sulfonic acid (TNBS) solution in the footpad on day 14. Footpad swelling was measured 24 or 48 h after the challenge. Peak responses of the anti-TNP IgM and IgG production were detected 4 or 6 days after the immunization with 10⁹ TNP-SRBC. Maximum DTH response was also observed with 10⁹ TNP-SRBC 24 h after the challenge on day 14. The antibody and DTH responses were also induced in other normal inbred strains such as C3H/He and DBA/1 but not BALB/c nu/nu mice. To evaluate AS-DAD in immunopharmacological studies, various immunomodulating agents were examined in BALB/c mice by subcutaneous administration on days 0, 1, 2 and 3. Cyclosporin or cyclophosphamide at 100 mg/kg/day completely inhibited not only the anti-TNP IgM and IgG production but also the TNP-specific DTH response. Prednisolone at 0.5 mg/kg/day had no significant effect on the IgM and IgG production, whereas it inhibited the TNP-specific DTH response. Interestingly, histamine-added mouse γ-globulin at 150 mg/kg/day clearly enhanced the anti-TNP IgM and IgG production, while it showed a suppressive effect on the TNP-specific DTH response. Levamisole at 5.0 mg/kg/day showed suppressive effects on the anti-TNP IgG production without affecting the IgM production and the DTH response. These results suggest that AS-DAD is useful for evaluating the immunopharmacological action of various agents.     

Effect of rifampin and its cyclopentyl analogue, MDL 473, on the expression of delayed type hypersensitivity to oxazolone

The purpose of this study was to determine the effect of rifampin and its cyclopentyl analogue, MDL 473, on delayed type hypersensitivity (DTH) using a murine model. Neither compound has any significant effect on DTH (as measured by ear swelling) to oxazolone when administered i.p. to BALB/c mice at doses of 1, 10, 50 or 100 mg/kg beginning 3 days before oxazolone sensitization and continuing until hapten challenge 5 days later. In contrast, sex and age matched controls receiving 200 mg hydrocortisone/kg i.p. beginning on the day of oxazolone sensitization and continuing to hapten challenge demonstrate a significant (P less than 0.001) abrogation of DTH.     

Auto-Delayed-Type Hypersensitivity Induced in Immunodeficient Mice with Syngeneic Modified Self-Antigens

The control of the autoimmune response to modified self-antigens was explored, using immunodeficient mice injected with syngeneic trinitrophenylated spleen cells (TNP-SC) as an experimental model system. X-irradiated (250 rad) A mice injected with TNP-SC and footpad-challenged 7 to 14 days later with syngeneic lymphoblasts generated a delayed-type hypersensitivity (DTH) response that was expressed by footpad swelling measured 24 h, 48 h and 72 h later. Histopathological examination showed massive inflammatory infiltration in the soft tissues of the limbs with extensive necrosis. This was not observed in X-irradiated mice that received the lymphoblast challenge only. The immunological activity was transferred from the X-irradiated TNP-SC-immunized mice to naive recipients by T cells (Lyt-1+) and not by serum, thus excluding the possibility that the inflammatory reaction is mediated by antibodies. We have previously presented evidence that the differentiation status of the lymphoblasts, and not contaminants from the incubation media, was the determinant factor eliciting the DTH response of immunodeficient mice injected with TNP-SC. Since only syngeneic lymphoblasts were able to elicit the DTH response of immunodeficient mice injected with syngeneic TNP-SC, we suggested that immunological activity was directed against self-antigens, thus expressing an autoimmune reactivity. The ability of immunodeficient mice to generate syngeneic DTH was not restricted to the TNP hapten or to inbred A-strain mice. X-irradiated BALB/c mice injected with syngeneic penicillinated spleen cells and challenged with syngeneic lymphoblasts generated a significant DTH response, in contrast to X-irradiated BALB/c mice exposed to the challenge dose only. X-irradiated A mice injected with syngeneic TNP-SC and simultaneously reconstituted with syngeneic splenocytes failed to generate a DTH response after the lymphoblast challenge, indicating that the syngeneic DTH response is controlled by normal suppressor cells. The suppressor cells were characterized as T cells carrying I-Jk, Lyt-1+, Lyt-2+ and Lyt-3+ antigenic markers. The suppressor cells abrogated the syngeneic DTH response of immunodeficient mice injected with TNP-SC, even when transferred a few days after the induction of immunological activity, but not when transferred 1 h before the lymphoblast challenge, indicating that even the established immunological activity can be restrained. Various immunological aspects of these observations and the significance of the findings in illuminating human autoimmune disorders are considered.