Wangxue Chen

A Mutant of Francisella tularensis Strain SCHU S4 Lacking the Ability To Express a 58-Kilodalton Protein Is Attenuated for Virulence and Is an Effective Live Vaccine

ABSTRACT Francisella tularensis subsp. tularensis (type A) strain SCHU S4 is a prototypic strain of the pathogen that is highly virulent for humans and other mammals. Its intradermal (i.d.) 50% lethal dose (LD50) for mice is <10 CFU. We discovered a spontaneous mutant, designated FSC043, of SCHU S4 with an i.d. LD50 of >108 CFU. FSC043 effectively vaccinated mice against challenge with a highly virulent type A strain, and the protective efficacy was at least as good as that of F. tularensis LVS, an empirically attenuated strain which has been used as an efficacious human vaccine. Comparative proteomics was used to identify two proteins of unknown function that were identified as defective in LVS and FSC043, and deletion mutants of SCHU S4 were created for each of the two encoding genes. One mutant, the ΔFTT0918 strain, failed to express a 58-kDa protein, had an i.d. LD50 of ∼105 CFU, and was found to be less capable than SCHU S4 of growing in peritoneal mouse macrophages. Mice that recovered from sublethal infection with the ΔFTT0918 mutant survived when challenged 2 months later with >100 LD50s of the highly virulent type A strain FSC033. This is the first report of the generation of defined mutants of F. tularensis subsp. tularensis and their use as live vaccines.

Experimental tularemia in mice challenged by aerosol or intradermally with virulent strains of Francisella tularensis: bacteriologic and histopathologic studies.

BALB/c and C57BL/6 mice were challenged by aerosol or intradermally with low doses ( approximately 10-20 colony forming units) of virulent type A and type B strains of the facultative intracellular pathogen, Francisella tularensis, and the course of infection was monitored. Both mouse strains were equally susceptible to infection, but type A strains reached lethal numbers a few days earlier than type B strains regardless of challenge route. BALB/c mice showed overt signs of infection for several days, whereas C57BL/6 mice remained asymptomatic until a few hours before death. Histological changes were extensive and severe in the liver and spleen, but much more limited in the lungs, even in mice challenged by aerosol. Thus, it appears that regardless of the route of infection, systemic rather than pulmonary infection was the likely cause of death following low dose challenge with virulent F. tularensis.

Viral haemorrhagic disease in rabbits: A review

An acute haemorrhagic disease of rabbits was first reported in a southern province of China in 1984. It subsequently spread rapidly to South China and some parts of North China. The disease is characterized by an acute onset with fever, rapid respiration and sudden death. There is a high morbidity and mortality rate. The pathological changes are consistent with severe generalized circulatory dysfunction (hyperaemia, congestion and haemorrhage), marked degeneration of parenchymatous tissue, pronounced serous-haemorrhagic pneumonia and extensive disruption of reticulo-lymphoid tissue. The disease has been named rabbit viral haemorrhagic disease and it has been suggested that the aetiological agent is a picornavirus. A tissue-derived vaccine has been prepared by homogenizing the liver, lung, spleen and kidney of infected rabbits and inactivating with formaldehyde. This review summarizes the information on the aetiology, epidemiology and clinical and pathological aspects of this new rabbit disease.

Neutrophils Play an Important Role in Host Resistance to Respiratory Infection with Acinetobacter baumannii in Mice

ABSTRACT Acinetobacter baumannii has emerged as a major cause of both community-associated and nosocomial pneumonia, but little is known about the cellular and molecular mechanisms of host defense against respiratory infection with this bacterial pathogen. In this study, we examined the role of neutrophils in host resistance to pulmonary A. baumannii infection in a mouse model of intranasal (i.n.) infection. We found that neutrophils were rapidly recruited to the lungs following i.n. inoculation of the pathogen and declined to baseline level upon clearance of the infection. Depletion of neutrophils using monoclonal antibody RB6-8C5 prior to infection resulted in an acute lethal infection that was associated with enhanced bacterial burdens in the lung (P < 0.05) and extrapulmonary dissemination to the spleen. The increased susceptibility to A. baumannii in neutropenic mice was associated with a delay in the mRNA expression and production of early proinflammatory cytokines such as tumor necrosis factor alpha, interleukin-6, keratinocyte chemoattractant protein, monocyte chemoattractant protein 1, and macrophage inflammatory protein 2 (MIP-2) in the lungs and development of severe bronchopneumonia and lymphoid tissue destruction in the spleen. Moreover, i.n. administration of the neutrophil-inducing chemokine MIP-2 to normal mice induced a pulmonary influx of neutrophils and significantly enhanced the clearance of A. baumannii from the lungs (P < 0.01). These results imply that neutrophils play a critical role in host resistance to respiratory A. baumannii infection.

Tularemia in BALB/c and C57BL/6 mice vaccinated with Francisella tularensis LVS and challenged intradermally, or by aerosol with virulent isolates of the pathogen: protection varies depending on pathogen virulence, route of exposure, and host genetic background

In order to begin understanding the immunological basis for immunity to tularemia, and to establish a baseline for judging the efficacy of potential novel vaccines, the present study examined the ability of the live vaccine strain of Francisella tularensis (F. tularensis) LVS, to elicit immunity in mice against subsequent systemic and aerosol challenge with highly virulent strains of the pathogen. The results show, that infection with LVS protects BALB/c mice against systemic challenge with virulent Types A and B F. tularensis. In contrast, C57BL/6 mice vaccinated with LVS were only rendered immune to systemic challenge with Type B F. tularensis. Neither mouse strain immunized with LVS was able to resist aerosol challenge with Type A F. tularensis, and only immunized BALB/c mice withstood exposure to aerosols of Type B F. tularensis.

Fibrin-mediated Protection Against Infection-stimulated Immunopathology

Fibrin, a product of the blood coagulation cascade, accompanies many type 1 immune responses, including delayed-type hypersensitivity, autoimmunity, and graft rejection. In those settings, fibrin is thought to exacerbate inflammation and disease. Here, we evaluate roles for coagulation during infection with Toxoplasma gondii, a pathogen whose control requires robust type 1 immunity. We establish that fibrin prevents infection-stimulated blood loss, thereby performing a protective function that is essential for survival. Remarkably, fibrin does not simply protect against vascular damage caused directly by the infectious agent, but rather, protects against hemorrhage evoked by interferon-γ, a critical mediator of type 1 immunity. This finding, to our knowledge, is the first to document a beneficial role for coagulation during type 1 immunity, and suggests that fibrin deposition protects host tissue from collateral damage caused by the immune system as it combats infection.

Distinct roles of reactive nitrogen and oxygen species to control infection with the facultative intracellular bacterium Francisella tularensis.

ABSTRACT Reactive nitrogen species (RNS) and reactive oxygen species (ROS) are important mediators of the bactericidal host response. We investigated the contribution of these two mediators to the control of infection with the facultative intracellular bacterium Francisella tularensis. When intradermally infected with the live vaccine strain F. tularensis LVS, mice deficient in production of RNS (iNOS−/− mice) or in production of ROS by the phagocyte oxidase (p47phox−/− mice) showed compromised resistance to infection. The 50% lethal dose (LD50) for iNOS−/− mice was <20 CFU, and the LD50 for p47phox−/− mice was 4,400 CFU, compared to an LD50 of >500,000 CFU for wild-type mice. The iNOS−/− mice survived for 26.4 ± 1.8 days, and the p47phox−/− mice survived for 10.1 ± 1.3 days. During the course of infection, the serum levels of gamma interferon (IFN-γ) and interleukin-6 were higher in iNOS−/− and p47phox−/− mice than in wild-type mice. Histological examination of livers of iNOS−/− mice revealed severe liver pathology. Splenocytes obtained 5 weeks after primary infection from antibiotic-treated iNOS−/− mice showed an in vitro recall response that was similar in magnitude and greater secretion of IFN-γ compared to cells obtained from wild-type mice. In summary, mice lacking expression of RNS or ROS showed extreme susceptibility to infection with F. tularensis LVS. The roles of RNS and ROS seemed to be distinct since mice deficient in production of ROS showed dissemination of infection and died during the early phase of infection, whereas RNS deficiency led to severe liver pathology and a contracted course of infection.

Reduced colonization of gastric mucosa by Helicobacter pylori in mice deficient in interleukin-101

Background and Aims: Interleukin‐10 (IL‐10) is a potent anti‐inflammatory and immunoregulatory cytokine. Mice deficient in IL‐10 production (IL‐10−/−mice) develop a spontaneous chronic enterocolitis, suggesting that IL‐10 is an important regulator of the mucosal immune response in vivo. The objective of this study was to determine the role of endogenous IL‐10 in the host defense against gastric colonization by Helicobacter pylori by using IL‐10‐deficient mice.

Anti-CD25 antibody-mediated depletion of effector T cell populations enhances susceptibility of mice to acute but not chronic Toxoplasma gondii infection

Natural regulatory T cells (Tregs) constitutively express the IL-2R α-chain (CD25) on their surface. Consequently, administration of anti-CD25 Abs is a commonly used technique to deplete Treg populations in vivo. However, activated effector T cells may also transiently express CD25, and are thus also potential targets for anti-CD25 Abs. In this study using Toxoplasma gondii as a model proinflammatory infection, we have examined the capacity of anti-CD25 Abs to target effector T cell populations during an inflammatory episode, to determine to what extent that this action may modulate the outcome of disease. Anti-CD25 Ab-treated C57BL/6 mice displayed significantly reduced CD4+ T cell IFN-γ production during acute T. gondii infection and exhibited reduced weight loss and liver pathology during early acute infection; aspects of infection previously associated with effector CD4+ T cell responses. In agreement, anti-CD25 Ab administration impaired parasite control and caused mice to succumb to infection during late acute/early chronic stages of infection with elevated tissue parasite burdens. In contrast, anti-CD25 Ab treatment of mice with established chronic infections did not markedly affect brain parasite burdens, suggesting that protective T cell populations do not express CD25 during chronic stages of T. gondii infection. In summary, we have demonstrated that anti-CD25 Abs may directly abrogate effector T cell responses during an inflammatory episode, highlighting important limitations of the use of anti-CD25 Ab administration to examine Treg function during inflammatory settings.

Detection of Listeria monocytogenes by polymerase chain reaction in intestinal mucosal biopsies from patients with inflammatory bowel disease and controls.

Abstract Background and Aims: Components of the intestinal microflora are believed to play an important role in the pathogenesis of inflammatory bowel disease (IBD) in genetically susceptible hosts acting either as a non‐specific antigenic stimulus or as a specific pathogen. Listeria monocytogenes has been suggested as an organism with the potential to cause IBD. The objective of the present study was to investigate the prevalence of L. monocytogenes DNA in intestinal biopsies from patients with IBD and from non‐IBD controls by using nested polymerase chain reaction (PCR).