Production and Characterization of Guinea Pig Recombinant Gamma Interferon and Its Effect on Macrophage Activation

Jeevan, Amminikutty; McFarland, Christine T.; Yoshimura, Teizo; Skwor, Troy; Cho, H.; Lasco, Todd M.; McMurray, David N.

doi:10.1128/iai.74.1.213-224.2006

Cited by 45 publications

(39 citation statements)

References 51 publications

(55 reference statements)

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“…The inhibition of NO production chemically or genetically enhances bacillary proliferation in mice (5,20). On the other hand, the IFN-␥-mediated induction of NO in rabbit, guinea pig, and human monocytes/macrophages in vitro has been difficult to show (11,25). Both mechanisms have lethal effects on tubercle bacilli (4), and both mechanisms induce DosR and its regulon (33,34,39).…”

Section: Vol 77 2009mentioning

confidence: 99%

Role of the dosR - dosS Two-Component Regulatory System in Mycobacterium tuberculosis Virulence in Three Animal Models

et al. 2009

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The Mycobacterium tuberculosis dosR gene (Rv3133c) is part of an operon, Rv3134c-Rv3132c, and encodes a response regulator that has been shown to be upregulated by hypoxia and other in vitro stress conditions and may be important for bacterial survival within granulomatous lesions found in tuberculosis. DosR is activated in response to hypoxia and nitric oxide by DosS (Rv3132c) or DosT (Rv2027c). We compared the virulence levels of an M. tuberculosis dosR-dosS deletion mutant (⌬dosR-dosS [⌬dosR-S]), a dosR-complemented strain, and wild-type H37Rv in rabbits, guinea pigs, and mice infected by the aerosol route and in a mouse hollow-fiber model that may mimic in vivo granulomatous conditions. In the mouse and the guinea pig models, the ⌬dosR-S mutant exhibited a growth defect. In the rabbit, the ⌬dosR-S mutant did not replicate more than the wild type. In the hollow-fiber model, the mutant phenotype was not different from that of the wild-type strain. Our analyses reveal that the dosR and dosS genes are required for full virulence and that there may be differences in the patterns of attenuation of this mutant between the animal models studied.

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Section: Vol 77 2009mentioning

confidence: 99%

Role of the dosR - dosS Two-Component Regulatory System in Mycobacterium tuberculosis Virulence in Three Animal Models

et al. 2009

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“…To study the effect of TNF-␣ neutralization on the activation of AM by the soluble products of infected neutrophils, some of the wells containing AM with supernatants from infected neutrophils were also treated with rabbit anti-recombinant guinea pig TNF-␣ polyclonal antibody (1:600), obtained as previously described (10,23). Twenty-four hours later, all the medium from the wells was removed and the H 2 O 2 production by the alveolar macrophages was measured by using the horseradish peroxide-dependent oxidation of phenol red by H 2 O 2 as reported previously by us and others (19,33). Briefly, 100 l of the assay solution (28 mM phenol red [Sigma], 100 U/ml horseradish peroxidase type II [Sigma, St. Louis, Mo]) in Hanks balanced salt solution was added to the test wells.…”

Section: Vol 75 2007 M Tuberculosis-infected Neutrophils Produce Cmentioning

confidence: 99%

Guinea Pig Neutrophils Infected withMycobacterium tuberculosisProduce Cytokines Which Activate Alveolar Macrophages in Noncontact Cultures

Sawant

McMurray

2007

Infect Immun

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The early influx of neutrophils to the site of infection may be an important step in host resistance against Mycobacterium tuberculosis. In this study, we investigated the effect of M. tuberculosis infection on the ability of guinea pig neutrophils to produce interleukin-8 (IL-8; CXCL8) and tumor necrosis factor alpha (TNF-␣) and to activate alveolar macrophages. Neutrophils and alveolar macrophages were isolated from naïve guinea pigs, cultured together or alone, and infected with virulent M. tuberculosis for 3, 12, and 24 h. IL-8 protein production in cocultures, as measured by using an enzyme-linked immunosorbent assay, was found to be additive at 24 h and significantly greater in M. tuberculosis-infected cocultures than in uninfected cocultures and in cultures of the infected neutrophils or macrophages alone. The IL-8 mRNA levels, determined by real-time reverse transcription-PCR, were elevated at 24 h in infected cocultures and infected cells cultured alone. In order to elucidate the contributions of neutrophils and their soluble mediators to the activation of alveolar macrophages, neutrophils and alveolar macrophages were cultured in a contact-independent manner by using a Transwell insert system. Neutrophils were infected with virulent M. tuberculosis in the upper wells, and alveolar macrophages were cultured in the lower wells. The release of hydrogen peroxide from alveolar macrophages exposed to soluble products from infected neutrophils was significantly increased compared to that from unexposed alveolar macrophages. Significant up-regulation of IL-1␤ and TNF-␣ mRNA levels in alveolar macrophages was observed at 24 and 30 h, respectively, compared to those in cells not exposed to soluble neutrophil products. Treatment with anti-guinea pig TNF-␣ polyclonal antibody completely abolished the response of alveolar macrophages to neutrophil products. This finding suggests that TNF-␣ produced by infected neutrophils may be involved in the activation of alveolar macrophages and hence may contribute to the containment of M. tuberculosis infection during the early period of infection.

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“…In contrast to ROS, RNS, produced by the nitric oxide synthase (iNOS) that is induced by IFN-␥, inhibit the growth of mycobacteria and are critical to maintenance of the persistent state in mice (7,8,22,27,29,42). Although the role of RNS in the control of M. tuberculosis growth in mice seems well proven, the involvement of RNS in human (58) and in other animal, including guinea pig (36,68), tuberculosis models is less clear, possibly because the presence of NO has been difficult to demonstrate (9,24,46,54,67). These observations suggest that M. tuberculosis genes involved in resistance to either ROS or RNS will play a role in persistence.…”

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Protection ofMycobacterium tuberculosisfrom Reactive Oxygen Species Conferred by themel2Locus Impacts Persistence and Dissemination

et al. 2009

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Persistence of Mycobacterium tuberculosis in humans represents a major roadblock to elimination of tuberculosis. We describe identification of a locus in M. tuberculosis, mel2, that displays similarity to bacterial bioluminescent loci and plays an important role during persistence in mice. We constructed a deletion of the mel2 locus and found that the mutant displays increased susceptibility to reactive oxygen species (ROS). Upon infection of mice by aerosol the mutant grows normally until the persistent stage, where it does not persist as well as wild type. Histopathological analyses show that infection with the mel2 mutant results in reduced pathology and both CFU and histopathology indicate that dissemination of the mel2 mutant to the spleen is delayed. These data along with growth in activated macrophages and infection of Phox ؊/؊ and iNOS ؊/؊ mice and bone marrow-derived macrophages suggest that the primary mechanism by which mel2 affects pathogenesis is through its ability to confer resistance to ROS. These studies provide the first insight into the mechanism of action for this novel class of genes that are related to bioluminescence genes. The role of mel2 in resistance to ROS is important for persistence and dissemination of M. tuberculosis and suggests that homologues in other bacterial species are likely to play a role in pathogenesis.Despite extensive efforts to eradicate tuberculosis, caused by Mycobacterium tuberculosis, worldwide and prevent the spread of antibiotic-resistant strains, tuberculosis remains one of the most frequent causes of death in humans. Currently, one-third of the world's population is thought to be persistently infected with tuberculosis (6,19,20). A better understanding of the mechanisms that lead to persistence in humans is needed before it will be possible to develop rational strategies to prevent establishment of latency and block reactivation from it. Although the mouse model, even when infected by the natural low-dose aerosol route, does not replicate all aspects of pathogenesis by tuberculosis, its cost-effectiveness and the presence of numerous reagents make it an important tool for examination of the acute and persistent stages of infection (26,50,51). As a result, much of our knowledge regarding the role of the host in controlling infections as well as the bacterial factors involved has been obtained using the mouse model (26,31,34).Initially, tubercle bacilli encounter naïve alveolar macrophages in the mouse lung that produce low levels of reactive oxygen species (ROS) and undergo an oxidative burst in response to infection (8, 52). In the absence of ROS production, there is a modest transient advantage for tuberculosis in the lung, suggesting that ROS initially assist in the control of bacterial growth (16). This innate immune response does little to prevent the growth of the bacteria in mouse lungs, since for the first month after infection, bacterial numbers increase to a million or more. This time point corresponds to the peaks in the numbers of CD4 and CD8 T cel...

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Production and Characterization of Guinea Pig Recombinant Gamma Interferon and Its Effect on Macrophage Activation

Abstract: Gamma interferon (IFN-␥

Cited by 45 publications

References 51 publications

Role of the dosR - dosS Two-Component Regulatory System in Mycobacterium tuberculosis Virulence in Three Animal Models

Role of the dosR - dosS Two-Component Regulatory System in Mycobacterium tuberculosis Virulence in Three Animal Models

Guinea Pig Neutrophils Infected withMycobacterium tuberculosisProduce Cytokines Which Activate Alveolar Macrophages in Noncontact Cultures

Protection ofMycobacterium tuberculosisfrom Reactive Oxygen Species Conferred by themel2Locus Impacts Persistence and Dissemination

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