Monocytogenes Infection In Mice Research Articles

The protective role of endogenous cytokines in host resistance against an intragastric infection with Listeria monocytogenes in mice.

It has been demonstrated that endogenous cytokines including gamma-interferon (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) play a protective role but that IL-4 plays a detrimental role in systemic Listeria monocytogenes infection in mice. The diverse roles of IL-10 have been reported in antilisterial resistance. In this paper, we studied the role of endogenous cytokines in host resistance against an intragastric infection with L. monocytogenes in mice. The expression of cytokine mRNAs including IFN-gamma, TNF-alpha, IL-4, IL-6, or IL-10, which were amplified by reverse transcription-PCR, was observed in intestinal intraepithelial lymphocytes irrespective of L. monocytogenes infection. Increased numbers of L. monocytogenes were detected in the ileal contents of infected mice which received monoclonal antibodies (mAbs) against IFN-gamma, TNF-alpha, IL-4, IL-6, or IL-10. By contrast, mAbs against IL-4 or IL-6 showed little effect on the growth of L. monocytogenes in the mesenteric lymph nodes (MLNs), spleen, and liver, but anti-IFN-gamma mAb and anti-TNF-alpha mAb suppressed the defense in these organs. Anti-IL-10 mAb enhanced bacterial elimination from the MLNs but not from the spleen or liver. These results suggest that the role of endogenous cytokines may differ between systemic and intestinal defenses.

Transforming growth factor beta is protective in host resistance against Listeria monocytogenes infection in mice.

The role of transforming growth factor beta (TGF-beta) in host resistance against Listeria monocytogenes infection was studied with mice. The constitutive expression of TGF-beta 1 mRNA was observed in the spleens and livers of mice before and after infection. Injecting the mice with anti-TGF-beta 1 peptide serum resulted in diminished antilisterial resistance, whereas the administration of human platelet-derived TGF-beta 1 enhanced the resistance. Moreover, mice were protected against lethal infection when treated with TGF-beta 1. These results suggest the TGF-beta 1 might be involved in antilisterial resistance. On the other hand, injecting the mice with TGF-beta 1 resulted in a decrease in the titers of endogenous gamma interferon, tumor necrosis factor alpha, and interleukin-6, which are crucial in antilisterial resistance, in sera and in extracts of spleen and liver. Thus, a complicated mechanism might be involved in the role of TGF-beta 1 in host resistance against L. monocytogenes infection.

Acquired resistance against a secondary infection with Listeria monocytogenes in mice is not dependent on reactive nitrogen intermediates.

During an infection, inflammatory mediators can induce the production of nitric oxide, a reactive nitrogen intermediate (RNI) which plays a role in antimicrobial activity against a wide variety of pathogens. In vitro experiments have shown that release of RNI by macrophages is mediated by tumor necrosis factor alpha (TNF). Since TNF is essential for acquired resistance during a secondary Listeria monocytogenes infection in mice, the aim of the present study was to determine whether RNI are also involved in the course of such an infection. Mice which had recovered from a sublethal primary infection with 0.1 50% lethal dose of (LD50) L. monocytogenes were infected intravenously with 10LD50 of L. monocytogenes. During a primary infection, the number of bacteria in the liver and spleen, as well as the concentration of RNI in plasma, increased. During a secondary infection, the number of bacteria in the liver and spleen decreased whereas no significant increase in the concentration of RNI in plasma was observed. Neutralization of endogenously produced TNF and gamma interferon by subcutaneous injection of alginate-encapsulated monoclonal antibody-forming cells during a secondary infection resulted in an increase in the number of bacteria in the liver and spleen an increase in the concentration of RNI in plasma. When the production of RNI was inhibited by treatment of mice with competitive NO-synthase inhibitor N omega-nitro-L-arginine methyl ester hydrochloride (L-Name) and an iota-arginine-deficient diet during a secondary infection, the proliferation of L. monocytogenes in the liver and spleen was not affected whereas the concentration of RNI in plasma of these mice was significantly reduced. Our findings that inhibition of RNI formation during a secondary infection does not affect the proliferation of L. monocytogenes in the liver and spleen and that enhanced elimination of bacteria from these organs is not accompanied by an increase in the concentration of RNI in plasma led to the conclusion that resistance against a secondary infection with L. monocytogenes is not dependent on RNI.

Administration of antigranulocyte monoclonal antibody RB6-8C5 prevents expression of acquired resistance to Listeria monocytogenes infection in previously immunized mice

Recent studies have established the importance of neutrophils in innate resistance to Listeria monocytogenes infection in mice. The purpose of this study was to determine the importance of neutrophils in acquired resistance to L. monocytogenes infection. Previously immunized mice that were depleted of neutrophils by administration of the antigranulocyte monoclonal antibody RB6-8C5 demonstrated less resistance to L. monocytogenes challenge than did nonimmunized control mice. In contrast, immunized control mice exhibited a heightened resistance to rechallenge, as expected. These results suggest that neutrophils make previously unrecognized contributions to acquired immunity to a facultative intracellular pathogen.

Resistance to infection and activation of the monocyto-macrophage system caused by Bacillus firmus and its fractions.

Crude lipids isolated from Bacillus firmus, but not from other bacilli, were previously found to induce significant resistance against Listeria monocytogenes infection in mice. In this study, formaldehyde- and heat-killed bacterins of eight Bacillus species and some cellular fractions of B. firmus were prepared and tested for further immunomodulatory activities. Crude lipids, their aqueous extract, LTA, Protodyne and Pex-residue preparations exhibited a strong anti-infection activity, whereas Pextract, P40 and all bacterins tested had no effect. Formaldehyde-killed bacterins, live bacteria and the P40 preparation of both B. firmus strains, as well as bacterins of both B. subtilis strains, induced pronounced splenomegaly in mice. Peptidoglycan and Pex-residue induced significant depression of cytochrome P-450 in mouse liver microsomes after application of 0.1 mg per mouse. Optimal conditions for obtaining a bacterial suspension exhibiting these immunomodulatory properties were elaborated.

Treatment with anti-interleukin-10 monoclonal antibody enhances early resistance to but impairs complete clearance of Listeria monocytogenes infection in mice.

Mice that received an anti-interleukin-10 (anti-IL-10) neutralizing monoclonal antibody (MAb) (SXC-1) prior to infection with Listeria monocytogenes initially demonstrated resistance to the infection, as indicated by reduced recovery of L. monocytogenes from their spleens and livers during the first 5 days after challenge. Anti-IL-10 MAb-treated mice then demonstrated reduced resistance during the later stage of infection, as indicated by persistent infection with L. monocytogenes in their livers 11 days after challenge. Aspartate aminotransferase (AST) levels (a measure of liver damage) in the sera of control mice increased between 1 and 5 days after challenge, while anti-IL-10 MAb-treated mice maintained lower AST levels. At 7 days after challenge, AST levels in the sera of control mice decreased as the numbers of organisms declined. In contrast, AST levels increased as the infections persisted in anti-IL-10 MAb-treated mice. The AST levels in serum reflected liver histopathology as anti-IL-10 MAb-treated mice exhibited fewer granulomatous lesions and less necrosis of liver tissue than the control mice during the first 5 days after challenge. Anti-IL-10 MAb treatment altered the expression of inflammatory cytokine mRNAs during L. monocytogenes infection. Control MAb-treated mice exhibited increased expression of tumor necrosis factor alpha and granulocyte-macrophage colony-stimulating factor mRNA in their lives during L. monocytogenes infection, but this increase did not occur in anti-IL-10 MAb-treated mice. Gamma interferon mRNA expression in the livers of the control MAb-treated mice was increased between 1 and 5 days after L. monocytogenes challenge and then decreased at 7 days after challenge. In contrast, gamma interferon mRNA expression in the livers of anti-IL-10 MAb-treated mice was not decreased until 7 days after challenge. These results indicate that endogenous IL-10 has both beneficial and detrimental effects on the host response to L. monocytogenes infection in mice.

Cytokine mRNA expression in livers of mice infected with Listeria monocytogenes

Temporally distinct groups of cytokine expression was observed by reverse transcription-polymerase chain reaction assay, in situ hybridization, and immunohistochemistry in the livers of Listeria monocytogenes-infected mice. One group consisted of interferon-gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and interleukin-10 (IL-10), for which mRNAs were induced within 1 day after challenge. A second group consisted of IL-2 and IL-4, for which mRNA was strongly expressed at 1 day but then suppressed at 3 days into the infection. Granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-1 alpha, and IL-6 mRNA constituted a third group, which was increased at 3 days after challenge. Distributions of cytokine mRNA-expressing cells in the liver was observed by in situ hybridization. Cells expressing TNF-alpha and IL-1 alpha mRNA were present throughout liver granulomas, whereas cells that expressed IFN-gamma mRNA were observed mostly along the periphery of granulomas. Cells expressing IL-2, IL-4, IL-6, IL-10, and GM-CSF mRNA were distributed principally in the hepatic sinuses. Cells expressing IL-10 mRNA increased in number early in the infection when L. monocytogenes was multiplying in the liver. We conclude that cytokine mRNA expression during the early phases of L. monocytogenes infection in mice is temporally regulated and that IFN-gamma, TNF-alpha, and IL-1 alpha are expressed by cells associated with hepatic granulomas.

The protective activity of immunostimulants against Listeria monocytogenes infection in mice

The function of peritoneal macrophages induced by intraperitoneal (i.p.) injection of attenuated Streptococcus pyogenes (OK-432), Bacillus Calmette Guérin (BCG), protein-bound polysaccharide preparation isolated from Coriolus vesicolor (PSK) or Lactobacillus casei was examined. The PMA-triggered respiratory burst (production of O2- and H2O2) of macrophages induced by OK-432, BCG or Lac. casei was greater than that of resident or thioglycollate-stimulated macrophages and was similar to that of BCG-activated macrophages. PSK failed to enhance the production of O2- or H2O2 by macrophages. Alkaline phosphodiesterase (APD) activity was reduced in macrophages induced by OK-432, BCG or Lac. casei injection and in BCG-activated macrophages. The APD activity of macrophages obtained 7 and 13 days after i.p. injection of PSK was elevated, as with thioglycollate-stimulated macrophages. Listericidal activity in vitro was enhanced in macrophages obtained in 13 and 7 days, but suppressed in macrophages obtained 2 days after OK-432, BCG or Lac. casei injection. Lac. casei administered either 2 or 13 days before infection with Listeria monocytogenes was protective but OK-432, BCG (0.1 mg) and PSK were not, even though they were able to stimulate macrophage function.

Endogenous tumor necrosis factor, interleukin-6, and gamma interferon levels during Listeria monocytogenes infection in mice

Mice were infected intravenously with a sublethal dose of Listeria monocytogenes cells and then levels of tumor necrosis factor (TNF), interleukin-6 (IL-6), and gamma interferon (IFN-gamma) in the bloodstreams, spleens, and livers were monitored. The maximum level of TNF was detected at 72 h in the spleens and livers, but TNF was never detected in the bloodstreams. IL-6 appeared in the bloodstreams and spleens and peaked at 48 h. The maximum level of IFN-gamma could be detected in all three specimens, and the highest titer was shown in the spleens. Endogenous TNF production was suppressed by in vivo administration of anti-CD4 monoclonal antibody (MAb) or anti-asialo GM1 antibody but not by anti-CD8 MAb, whereas none of these antibodies suppressed endogenous IL-6 production. Endogenous production of neither IL-6 nor IFN-gamma was inhibited in rabbit anti-recombinant mouse TNF-alpha antibody-treated mice. Similarly, production of TNF and IL-6 did not decrease in anti-mouse IFN-gamma MAb-treated animals, but TNF production was augmented in these animals. These results suggest that the these endogenous cytokines are produced by different mechanisms in L. monocytogenes infection.

Effect of a Traditional Chinese Medicine, Bu-Zhong-Yi-Qi-Tang (Japanese Name: Hochu-Ekki-To) on the Protection Against Listeria Monocytogenes Infection in Mice

Effects of Bu-Zhong-Yi-Qi-Tang (Japanese name: Hochu-ekki-to) on the resistance against Listeria monocytogenes were observed in ICR mice orally administered this medicine daily for 10 days. Survival rates were increased by the pretreatment in mice inoculated i.v. with bacteria 1 day after the last administration and in mice inoculated i.p. 4 days after the last administration. After an i.v. inoculation of L. monocytogenes, the numbers of bacteria in the spleen and liver increased gradually to kill mice by day 5 in untreated group but the bacterial numbers increased slightly by day 3 and decreased from day 3 to day 8 in Hochu-ekki-to pretreated group. After an i.p. inoculation, the number of bacteria in the peritoneal cavity decreased very rapidly within 6h in Hochu-ekki-to treated group compared to that in untreated group. After the administration, number of polymorphonuclear cells increased in the peripheral blood, peritoneal cavity and spleen. In treated mice, macrophages increased in number in the peritoneal cavity and the spleen but decreased in the peripheral blood. Peritoneal macrophages from treated mice showed an enhanced activity to kill L. monocytogenes in vitro within 60 min after ingestion of bacteria. Hochu-ekki-to may augment the host defense against L. monocytogenes through the activation of macrophage series during an early phase of infection.

RIL-1α enhances adoptive transfer of resistance to Listeria monocytogenes infection

We have previously demonstrated that administration of recombinant rIL-1α enhances resistance against Listeria monocytogenes infection in mice. In this study we considered the possibility that this cytokine might also augment adoptive immunity conferred by the transfer of listeria-immune spleen cells. Concomitant administration of rIL-1α with large numbers (2 × 10 7 or 10 8) of listeria-immune spleen cells reduced the protection mediated by the transferred cells. Conversely, rIL-1α co-administered with suboptimal numbers (1 − 5 × 10 6) of immune splenocytes augmented anti-listeria resistance in an additive fashion. Although transfer of 10 6 listeria-immune spleen cells alone did not result in significant protection, when 10 6 immune cells were incubated with rIL-1α prior to transfer they conferred significant protection to naive recipients. Time course experiments indicated that the greatest protection was achieved when listeria-immune spleen cells were pretreated with rIL-1α for 2 h prior to adoptive transfer. The protection transferred by 10 6 rIL-1α-pretreated immune spleen cells was not inhibited by TGF β. This study is the first to use riL-1α to potentiate the adoptive transfer of resistance to an infectious agent by immune cells.

Validation of a role for endogenously produced IFN gamma in resolution of Listeria monocytogenes infection in mice.

It is now well recognized that activated macrophages can express enhanced levels of cytocidal activity toward a variety of microbial targets and neoplastic cells (1–6). Three to five years ago, work from several laboratories indicated that IFNγ was one of the major classes of lymphokines capable of activating macrophages under well-defined in vitro conditions (7–14). Recently, there has been an explosive growth of experiments directed at defining IFNγ’s mechanism of action at the molecular level. These investigations have led to the identification and characterization of a specific IFNγ receptor on the macrophage plasma membrane (15–17), have provided insights into the mechanism of transmembrane signal transduction, (18–20), and have elucidated the ultimate fate of receptor bound ligand (16,21).

Changes in serum colony-stimulating factor and monocytic progenitor cells during Listeria monocytogenes infection in mice.

The capacity of a host to produce and mobilize monocytes is an essential component of host defenses during the early phases of infection caused by Listeria monocytogenes. In this study, the concentrations of colony stimulating factor (CSF) and the numbers of monocyte progenitor cells (CFUm) were measured in mice during infection with L. monocytogenes. The concentration of CSF in serum increased sharply during the first 24 h of infection and remained elevated for the next 7 days. The number of CFUm in the bone marrow, however, decreased during the first 4 days after injection of L. monocytogenes. Thereafter, the number increased slowly, returning to normal on day 14. The decrease in marrow progenitor cells did not appear to result from a reduced sensitivity to CSF. In contrast to bone marrow changes, spleen progenitor cells increased greater than 400%, reaching a peak 7 days after bacterial challenge. These data indicate that monocyte production during L. monocytogenes infection is correlated with a rise in serum CSF concentration, depletion of bone marrow CFUm, and an increase in the number of spleen CFUm.

Enhancement of host resistance against Listeria infection by Lactobacillus casei: role of macrophages

Among the 10 species of the genus Lactobacillus, L. casei showed the strongest protective action against Listeria monocytogenes infection in mice. The activity of L. casei differed with regard to the dose of administration. The anti-L. monocytogenes resistance in mice intravenously administered 5.5 X 10(7), 2.8 X 10(8), or 1.1 X 10(9) L. casei cells was most manifest at ca. 2, 2 and 13, and 3 to 21 days after its administration, respectively. The growth of L. monocytogenes in the liver of mice injected with L. casei (10(7), 10(8), or 10(9) cells) 48 h after infection was suppressed, particularly when 10(8) or 10(9) L. casei cells were given 2 or 13 days before the induced infection, respectively. This suppression of L. monocytogenes growth was overcome by carrageenan treatment or X-ray irradiation. [3H]thymidine incorporation into the liver DNA increased 13 days after administration of L. casei, and augmentation of [3H]thymidine incorporation during 6 to 48 h after infection was dependent on the dose of L. casei. Peritoneal macrophage accumulation observed 1 to 5 days after intraperitoneal injection of UV-killed L. monocytogenes was markedly enhanced when the mice were treated with L. casei cells 13 days before macrophage elicitation. Therefore, the enhanced host resistance by L. casei to L. monocytogenes infection may be mediated by macrophages migrating from the blood stream to the reticuloendothelial system in response to L. casei injection before or after L. monocytogenes infection.

Decreased resistance to Listeria monocytogenes in mice injected with killed corynebacterium parvum: association with suppression of cell-mediated immunity.

To investigate the therapeutic potential of killed Corynebacterium parvum, its effects on the course of Listeria monocytogenes infection in mice was studied. Mortality in mice given C. parvum after L. monocytogenes infection was greater than in mice given C. parvum before infection or infected with only L. monocytogenes. C. parvum alone resulted in no mortality. Spleens from infected mice given C. parvum had increased numbers of L. monocytogenes. Peritoneal macrophages from mice infected with only L. monocytogenes were activated by four days after infection, whereas those from mice given L. monocytogenes plus C. parvum were not. By seven days macrophages from both were activated. Delayed hypersensitivity and in vitro lymphocyte transformation in response to L. monocytogenes antigen were strikingly suppressed in infected mice given C. parvum. The increased susceptibility of L. monocytogenes-infected mice given C. parvum may have been due to delayed macrophage activation resulting from suppression of thymus-derived lymphocyte responses.

Immunologic Events during Listeria Monocytogenes Infection in Mice: Adjuvanticity and Immunogenicity of Macrophage-Bound Antigens

Abstract There was a marked enhancement of the immune response to keyhole limpet hemocyanin administered at the time of inoculation with Listeria monocytogenes. Such enhancement was best seen if the antigen was given close to the time of inoculation with Listeria monocytogenes. Such enhancement was best seen if the antigen was given close to the time of inoculation with the bacteria. Bacterial filtrates also produced an adjuvant effect. Peritoneal exudate cells harvested a few days after Listeria infection contained large activated macrophages. Such macrophages had a greater uptake of 125I-labeled hemocyanin. Metabolic studies indicated that the percentage of hemocyanin retained on the membrane or released from macrophages was increased in activated cells. Hemocyanin bound to activated macrophages was slightly more immunogenic. Activated macrophages (without antigen) did not transfer adjuvant effects. We conclude that the adjuvant effect was not associated with the presence of activated macrophages.

Synthetic polyanions protect mice against intracellular bacterial infection.

A NUMBER of synthetic polyanions stimulate interferon production1–7 and these materials are active against infection with viruses and also with parasites of a more complex nature8–12. Mice inoculated intraperitoneally with pyran, a random copolymer of divinyl ether and maleic acid anhydride, are resistant to an ordinarily lethal challenge of mengo virus7. Such antiviral resistance persists for as long as 60 days in spite of the absence of measurable levels of interferon in the sera of the mice after 4 to 6 days7,13. These polyanions modify a number of host defence mechanisms including delayed hypersensitivity14, antibody response15 and the number of circulating white blood cells16. In view of the suggestion that a common mechanism of host resistance to intracellular pathogens may exist17, we were interested to determine if the long term resistance against viral challenge observed following administration of such polymers may also be extended to intracellular bacterial infection. In these studies, three anionic polymers*, pyran copolymer, polyacrylic acid and the double stranded synthetic ribonucleic acid poly I·poly C, each conferred resistance to Listeria monocytogenes infection in mice.