Ambiguous Role of Interleukin-12 in Yersinia enterocolitica Infection in Susceptible and Resistant Mouse Strains

Endogenous interleukin-12 (IL-12) mediates protection against Yersinia enterocolitica in C57BL/6 mice by triggering gamma interferon (IFN-γ) production in NK and CD4⁺ T cells. Administration of exogenous IL-12 confers protection against yersiniae in Yersinia-susceptible BALB/c mice but exacerbates yersiniosis in resistant C57BL/6 mice. Therefore, we wanted to dissect the different mechanisms exerted by IL-12 during Yersinia infections by using different models of Yersinia-resistant and -susceptible mice, including resistant C57BL/6 mice, susceptible BALB/c mice, intermediate-susceptible wild-type 129/Sv mice, 129/Sv IFN-γ-receptor-deficient (IFN-γR^−/−) mice and C57BL/6 tumor necrosis factor (TNF) receptor p55 chain-deficient (TNFR p55^−/−) mice. IFN-γR^−/− mice turned out to be highly susceptible to infection by Y. enterocolitica compared with IFN-γR^+/+ mice. Administration of IL-12 was protective in IFN-γR^+/+ mice but not in IFN-γR^−/− mice, suggesting that IFN-γR-induced mechanisms are essential for IL-12-induced resistance against yersiniae. BALB/c mice could be rendered Yersinia resistant by administration of anti-CD4 antibodies or by administration of IL-12. In contrast, C57BL/6 mice could be rendered more resistant by administration of transforming growth factor β (TGF-β). Furthermore, IL-12-triggered toxic effects in C57BL/6 mice were abrogated by coadministration of TGF-β. While administration of IL-12 alone increased TNF-α levels, administration of TGF-β or TGF-β plus IL-12 decreased both TNF-α and IFN-γ levels in Yersinia-infected C57BL/6 mice. Moreover, IL-12 did not induce toxicity in Yersinia-infected TNFR p55^−/− mice, suggesting that TNF-α accounts for IL-12-induced toxicity. Taken together, IL-12 may induce different effector mechanisms in BALB/c and C57BL/6 mice resulting either in protection or exacerbation. These results are important for understanding the critical balance of proinflammatory and regulatory cytokines in bacterial infections which is decisive for beneficial effects of cytokine therapy.     

HK-ATPase Expression in the Susceptible BALB/c and the Resistant DBA/2 Strains of Mice to Autoimmune Gastritis

Neonatal thymectomy (NTx) in mice induces a group of alterations in the immune system homeostasis that results in the development of a variety of organ-specific autoimmune diseases such as gastritis, thyroiditis, oophoritis and orchitis. Given the importance of self-antigen expression in thymus for the control of autoreactive cells and generation of regulatory cells, we have compared the expression of parietal cell antigen in two strains of mice with the same H-2: BALB/c (susceptible to develop gastritis after NTx) and DBA/2 (resistant). We detected mRNA of HK-ATPase α and β chains in day 1 thymi of both strains. Fifty percent of BALB/c mice presented mRNA levels similar to DBA/2. However, lower mRNA levels were found in the remaining BALB/c mice that may correspond to those that would develop AIG after NTx. Since the presence of the antigen in periphery is also necessary for the induction of regulatory cells, we have compared both strains observing in day 1 stomachs from resistant DBA/2 strain, a significantly higher content of positive cells for HK-ATPase subunits than stomachs from susceptible BALB/c strain. Also, the presence of antinuclear Abs in NTx BALB/c mice makes this model a useful experimental system for analyzing the responsible mechanisms breaking the non-specific self-tolerance.     

Induction of Natural Killer Cells and Interferon during Mouse Hepatitis Virus Infection of Resistant and Susceptible Inbred Mouse Strains

Following infection by mouse hepatitis virus (JHM strain), an induction of natural killer (NK) cell activity was observed in C3H mice, which are considered to be sensitive to JHM virus infection. In contrast, mice of the resistant SJL strain did not show any increase of NK cell activity after JHM virus infection. However, infection of both SJL and C3H mice with mouse hepatitis virus type 3 (MHV3) resulted in an increase of NK level, comparable to that observed with the JHM virus infection in the C3H strain. No significant differences were observed in the NK cell activity of the peritoneal exudate or spleen cells of infected mice. Low levels of interferon were detected in serum or peritoneal exudate of C3H mice infected with JHM virus 18 or 24 hours before, but no detectable early interferon production was found. Also no interferon could be detected in the resistant SJL mice. After JHM virus infection, the number of peritoneal exudate cells (PEC) was increased significantly in C3H mice but not in SJL mice. Macrophages obtained from the C3H mice supported virus replication, whereas SJL macrophages did not. Our data suggest that NK cells do not play a role in the resistance of SJL mice against JHM virus infection but may participate in the defence mechanisms against this virus in C3H mice.     

Degradation of Amyloid A Precursor Protein SAA by Macrophage Cell Lines Obtained from Amyloid Resistant and Susceptible Strains of Mice

Reactive AA amyloidosis can be induced in mice in a model of sustained inflammation following daily casein subcutaneous injections. However, the development of AA amyloidosis is known to vary in different strains of mice. The C57BL/6 strain is susceptible to the development of amyloidosis while the A/J strain is resistant. The degradation of purified serum amyloid A (SAA) protein by human monocytes as well as by mouse macrophages has been shown. The resistance/susceptibility of different mouse strains to the development of systemic amyloidosis may therefore be related to a difference in the ability of macrophages to degrade SAA. The authors have used bone marrow-derived macrophage cell lines obtained from susceptible C57BL/6 (ANA-1) and resistant A/J (A/J 10) mouse strains to compare their ability to degrade HDL-SAA in vitro . Cells were incubated with HDL-SAA for up to 72 h and the culture medium was analysed by SDS-PAGE to determine the rate of SAA degradation by the macrophages. The A/J 10 cells (resistant) were found to initiate a constant HDL-SAA degradation promptly whereas ANA-1 cells (susceptible) showed an intermittent block in the degradation of the precursor. Activation of macrophages by lipopolysaccharide (LPS) or interferon-γ (IFN-γ) hampered the precursor degradation suggesting that the activation process may favour extracellular accumulation of the precursor leading to a partial degradation and fibril formation.     

Methicillin-Resistant Staphylococcus aureus: Comparison of Susceptibility Testing Methods and Analysis of mecA-Positive Susceptible Strains

Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for an increasing number of serious nosocomial and community-acquired infections. Phenotypic heterogeneous drug resistance (heteroresistance) to antistaphylococcal beta-lactams affects the results of susceptibility testing. The present study compared the MRSA-Screen latex agglutination test (Denka Seiken Co., Ltd., Tokyo, Japan) for detection of PBP 2a with agar dilution, the VITEK-1 and VITEK-2 systems (bioMérieux, St. Louis, Mo.), and the oxacillin agar screen test for detection of MRSA, with PCR for the mecA gene used as the "gold standard" assay. Analysis of 107 methicillin-susceptible S. aureus (MSSA) isolates and 203 MRSA isolates revealed that the MRSA-Screen latex agglutination test is superior to any single phenotype-based susceptibility testing method, with a sensitivity of 100% and a specificity of 99.1%. Only one isolate that lacked mecA was weakly positive by the MRSA-Screen latex agglutination test. This isolate was phenotypically susceptible to oxacillin and did not contain the mecA gene by Southern blot hybridization. The oxacillin agar screen test, the VITEK-1 system, the VITEK-2 system, and agar dilution showed sensitivities of 99.0, 99.0, 99.5, and 99%, respectively, and specificities of 98.1, 100, 97.2, and 100%, respectively. The differences in sensitivity or specificity were not statistically significant. Oxacillin bactericidal assays showed that mecA- and PBP 2a-positive S. aureus isolates that are susceptible to antistaphylococcal beta-lactams by conventional methods are functionally resistant to oxacillin. We conclude that the accuracy of the MRSA-Screen latex agglutination method for detection of PBP 2a approaches the accuracy of PCR and is more accurate than any susceptibility testing method used alone for the detection of MRSA.     

Host Glycosaminoglycan Confers Susceptibility to Bacterial Infection in Drosophila melanogaster

Many pathogens engage host cell surface glycosaminoglycans, but redundancy in pathogen adhesins and host glycosaminoglycan-anchoring proteins (heparan sulfate proteoglycans) has limited the understanding of the importance of glycosaminoglycan binding during infection. The alpha C protein of group B streptococcus, a virulence determinant for this neonatal human pathogen, binds to host glycosaminoglycan and mediates the entry of bacteria into human cells. We studied alpha C protein-glycosaminoglycan binding in Drosophila melanogaster, whose glycosaminoglycan repertoire resembles that of humans but whose genome includes only three characterized membrane heparan sulfate proteoglycan genes. The knockdown of glycosaminoglycan polymerases or of heparan sulfate proteoglycans reduced the cellular binding of alpha C protein. The interruption of alpha C protein-glycosaminoglycan binding was associated with longer host survival and a lower bacterial burden. These data indicate that the glycosaminoglycan-alpha C protein interaction involves multiple heparan sulfate proteoglycans and impairs bacterial killing. Host glycosaminoglycans, anchored by multiple proteoglycans, thereby determine susceptibility to infection. Because there is homology between Drosophila and human glycosaminoglycan/proteoglycan structures and many pathogens express glycosaminoglycan-binding structures, our data suggest that interfering with glycosaminoglycan binding may protect against infections in humans.     

Decreased Susceptibility to Listeria monocytogenes in Mice After Infection with Trichinella spiralis

Mice challenged intravenously with Listeria monocytogenes at 7 or 21 days after Trichinella spiralis infection demonstrated a higher mean lethal dose and longer survival time than normal mice. This effect of helminthic infection was abrogated after 49 days.     

Enhancement of the Humoral Immune Response and Resistance to Bacterial Infection in Mice by the Oral Administration of a Bacterial Immunomodulator (OM-89)

Abstract

We investigated the effects of an Escherichia coli-derived product (OM-89) in mice. The oral administration of OM-89 led to a significant (p < 0.05. Student's t test) increase in the levels of IgA in intestinal secretions, which was at maximum 25 days after the end of the treatment, when a two-fold increase in IgA levels was observed. The i.p. inoculation of OM-89 induced the stimulation of anti-SRBC plaque-forming cells (PFC) in the spleen. The effect of OM-89 was dose-dependent and produced up to a 9-fold increase in PFC in the treated mice when compared to untreated controls. The oral administration of OM-89 proved to be effective in the enhancement of resistance to challenge i.p. inoculation with E. coli. 32% of OM-89-treated mice showed resistance to this experimental infection at minimal LD₁₀₀. The combined effects of low environmental temperature and cyclophosphamide (CY) immunosuppression enabled us to enhance differences in survival rates in experiments on the modulation of resistance towards Pseudomonas aeruginosa infection. The oral treatment with the immunoraodulator induced a significant (p < 0.05, Student's t test) level of protection in CY-immunosuppressed mice to the intranasal infection with P. aeruginosa, when mice were kept at low environmental temperature right after the bacterial challenge. The protective effect of OM-89 treatment was dependent on both the environmental temperature and the timing of the experiment.     

Fowl Immunoglobulins: Quantitation in Birds Genetically Resistant and Susceptible to Marek's Disease

The development of serum immunoglobulins was observed in chicks genetically resistant (N line) and susceptible (P line) to Marek's disease (MD), in conditions free from infection with MD. IgG was present at hatching at about 5.0 mg/ml and decreased to about 0.5 mg/ml by 12 to 15 days. Active production of IgG was apparent at about 20 days of age and reached levels of 2.0 to 3.0 mg/ml by 67 days. Low levels of IgM were present at hatching, and a gradual increase was seen to 1.0 to 2.0 mg/ml by 67 days. IgA was not detectable at hatching; it appeared in N-line birds at 5 days and in P-line birds at 13 days, and by 67 days was about 0.10 to 0.13 mg/ml. After 30 to 40 days, immunoglobulin levels were generally higher in N-line than in P-line birds. However, it was concluded that no primary immunoglobin deficiency existed sufficient to explain the susceptibility of the P-line birds to MD     

Circadian Rhythm of the Resistance of Mice to Acute Pneumococcal Infection

The resistance of both male and female mice to an acute infection by type I pneumococci varied rhythmically in an approximately 24-hr cycle. Mice inoculated intraperitoneally with 10(5.4+/-0.1) organisms during the dark period of the daily cycle survived significantly longer than mice similarly inoculated during the light period. The variation in resistance was correlated with the rate of increase of bacteremia, the rate being greatest after inoculation during the light period. Neither the capacity of the blood to support growth of pneumococci in vitro nor clearance of pneumococci from the blood in vivo varied between phases of the daily cycle. Adrenalectomized mice were equally and highly susceptible to infection during both light and dark periods. Stress of mice during the light period, normally a time of relative inactivity of mice, disturbed the rhythm; resistance to infection initiated during the light period increased and equalled that during the dark period.     

Increased Susceptibility of Melanin-Concentrating Hormone-Deficient Mice to Infection with Salmonella enterica Serovar Typhimurium

Melanin-concentrating hormone (MCH) was initially identified in mammals as a hypothalamic neuropeptide regulating appetite and energy balance. However, the wide distribution of MCH receptors in peripheral tissues suggests additional functions for MCH which remain largely unknown. We have previously reported that mice lacking MCH develop attenuated intestinal inflammation when exposed to Clostridium difficile toxin A. To further characterize the role of MCH in host defense mechanisms against intestinal pathogens, Salmonella enterocolitis (using Salmonella enterica serovar Typhimurium) was induced in MCH-deficient mice and their wild-type littermates. In the absence of MCH, infected mice had increased mortality associated with higher bacterial loads in blood, liver, and spleen. Moreover, the knockout mice developed more-severe intestinal inflammation, based on epithelial damage, immune cell infiltrates, and local and systemic cytokine levels. Paradoxically, these enhanced inflammatory responses in the MCH knockout mice were associated with disproportionally lower levels of macrophages infiltrating the intestine. Hence, we investigated potential direct effects of MCH on monocyte/macrophage functions critical for defense against intestinal pathogens. Using RAW 264.7 mouse monocytic cells, which express endogenous MCH receptor, we found that treatment with MCH enhanced the phagocytic capacity of these cells. Taken together, these findings reveal a previously unappreciated role for MCH in host-bacterial interactions.     

Interferon Induction and Resistance to Virus Infection in Mice Infected with Brucella abortus

Induction of circulating interferon and protection against vaccinia virus infection in mice by injection of Brucella abortus were studied. It was demonstrated that morphologically intact brucellae (either live or killed by heat or exposure to NaOH) induce high and prolonged levels of circulating interferon in mice. In each instance, the inducing principle remained associated with the bacterial particle. Disruption of brucellae by mechanical means destroyed the interferon-inducing capacity. However, by alkalinization of the water extract of disrupted bacilli, an interferon inducer could be rescued. On intravenous injection, this inducer caused a typical endotoxin type of interferon response with a peak value at 2 hr. Mice pretreated with cycloheximide showed an enhanced interferon response to the brucella extract, but a reduced reaction to live brucellae. The significance of these data, in relation to the triggering of de novo interferon synthesis by brucella, is discussed. It was also observed that small doses of brucellae protected mice for at least 1 month against vaccinia virus infection. High doses of heat-or alkali-killed brucellae protected the animals for only a short time, and disrupted brucellae did not afford any protection. Thus, there was a good correlation between interferon-inducing capacity and short-term protective activity. Long-term protection, on the other hand, seemed to be related to multiplication of brucellae in the body.     

Decreased Resistance to Primary Intravenous Cryptococcus neoformans Infection in Aged Mice despite Adequate Resistance to Intravenous Rechallenge

It is often stated that impaired immune functions in the aged underlie their greater susceptibility to infections. Indeed, in many experimental settings, T-cell responses in aged mice have been shown to be deficient compared with those from young adults. Nonetheless, there are very few examples where a greater susceptibility to infection in aged mice has been demonstrated to result from impaired T-cell function. The clinical importance of understanding the basis for increased susceptibility to infection that accompanies advanced age dictates a need for experimental models with which to study the effect that aging has on immunological resistance to infection. This study was undertaken to investigate whether aged mice were less resistant than young adult control mice to infection with the fungus Cryptococcus neoformans. After a primary intravenous challenge with yeast, aged mice died sooner and developed higher organ burdens of yeast than did young adults. Deficient in vitro responses were observed in T cells from aged mice; however, greater susceptibility to intravenous infection appeared not to result from less effective T-cell-dependent resistance in vivo. In fact, T-cell-replete aged mice were more susceptible to intravenous cryptococcal infection than were T-cell-depleted young adults. Furthermore, aged mice were as resistant to primary pulmonary challenge with Cryptococcus as were young adults. Similarly, vaccinated aged mice were as resistant to rechallenge as were young adult counterparts. Therefore, despite demonstrably deficient in vitro responses of T cells from aged mice, their T-cell-dependent resistance to C. neoformans is as effective as that of young adults.