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1.
J Microbiol Methods ; 67(2): 266-72, 2006 Nov.
Article in English | MEDLINE | ID: mdl-16704883

ABSTRACT

The resurgence of tuberculosis along with the increased resistance of Mycobacterium tuberculosis has emphasized the need for timely susceptibility testing for control of the disease. Previous studies have shown that rapid susceptibility testing can be accomplished for isoniazid, ethambutol, and rifampin using the flow cytometric assays. In this study we compared the flow cytometric susceptibility assay with the BACTEC TB 460 and BACTEC MGIT 960 for pyrazinamide (PZA). There was 93% agreement between the BACTEC MGIT 960 and the flow cytometric methods for 100 microg/mL of PZA. Additionally, there was a 95% and 86% agreement between the BACTEC TB 460 and flow cytometric methods for 50 microg/mL and 100 microg/mL of PZA, respectively. These findings show that susceptibility testing by the flow cytometric assay is accurate. Most importantly, susceptibility results by the flow cytometric assay were available 24 h after initiation of the testing procedure. The advantages of simplicity, speed and accuracy make the flow cytometric susceptibility assay an immediate impact technology to improve patient care.


Subject(s)
Antitubercular Agents/pharmacology , Flow Cytometry/methods , Microbial Sensitivity Tests/methods , Mycobacterium tuberculosis/drug effects , Pyrazinamide/pharmacology , Humans , Mycobacterium tuberculosis/isolation & purification , Reproducibility of Results , Tuberculosis/microbiology
2.
Clin Diagn Lab Immunol ; 11(1): 35-41, 2004 Jan.
Article in English | MEDLINE | ID: mdl-14715542

ABSTRACT

Development of a high level of sustained borreliacidal antibody is paramount for maintaining protection against infection with Borrelia burgdorferi. We show that production of borreliacidal antibody can be enhanced by preventing the effects of gamma interferon (IFN-gamma). When lymph node cells capable of producing borreliacidal antibody were cultured with anti-murine IFN-gamma, an eightfold increase in borreliacidal antibody production was obtained. However, anti-IFN-gamma treatment of these cells also enhanced their ability to adaptively induce arthritis. When anti-IFN-gamma-treated lymph node cells producing borreliacidal antibody were infused into C3H/HeJ mice and the mice were then challenged with B. burgdorferi, the mice developed severe destructive Lyme arthritis. Additional studies are needed to delineate the immune response responsible for the induction of arthritis and production of borreliacidal antibody. These studies are needed to ensure an effective and safe vaccine against infection with B. burgdorferi.


Subject(s)
Antibodies, Bacterial/biosynthesis , Borrelia burgdorferi/immunology , Interferon-gamma/antagonists & inhibitors , Lyme Disease/etiology , Animals , Lyme Disease/immunology , Lyme Disease/pathology , Lymph Nodes/immunology , Lymph Nodes/pathology , Mice , Mice, Inbred C3H , Neutralization Tests
3.
Curr Protoc Cytom ; Chapter 11: Unit 11.7, 2004 Feb.
Article in English | MEDLINE | ID: mdl-18770792

ABSTRACT

Here is another unit with clinical relevance. Tuberculosis remains a major health problem throughout the world, with approximately one-quarter of the population being infected. Rapid and accurate susceptibility testing for the tubercle bacillus is essential for control of the disease. Such testing can be accomplished by flow cytometry within twenty-four hours, instead of the days to weeks required by traditional methods. The use of flow cytometry both improves the quality of susceptibility testing and advances public health measures for the prevention and control of this ancient scourge.


Subject(s)
Flow Cytometry/methods , Microbial Sensitivity Tests/methods , Mycobacterium tuberculosis/cytology , Anti-Bacterial Agents/pharmacology , Drug Resistance , Mycobacterium tuberculosis/drug effects
4.
Infect Immun ; 71(6): 3437-42, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12761128

ABSTRACT

We showed that Borrelia burgdorferi-vaccinated interferon gamma-deficient (IFN-gamma(0)) mice challenged with the Lyme spirochete developed a prominent chronic severe destructive osteoarthropathy. The immune response underlying the development of the severe destructive arthritis involves interleukin-17 (IL-17). Treatment of vaccinated IFN-gamma(0) mice challenged with B. burgdorferi with anti-IL-17 antibody delayed the onset of swelling of the hind paws but, more importantly, inhibited the development of arthritis. Histopathologic examination confirmed that treatment with anti-IL-17 antibody prevented the destructive arthropathy seen in vaccinated and challenged IFN-gamma(0) mice. Similar preventive results were obtained when vaccinated and challenged IFN-gamma(0) mice were treated with anti-IL-17 receptor antibody or sequentially with anti-IL-17 antibody followed by anti-IL-17 receptor antibody. By contrast, treatment of vaccinated and challenged IFN-gamma(0) mice with recombinant IL-17 (rIL-17) did not alter the development and progression of arthritis found in vaccinated and challenged IFN-gamma(0) mice without treatment with rIL-17. Therapeutic intervention may be a realistic approach to prevent arthritis, especially if IL-17 is involved in the perpetuation of chronic or intermittent arthritis.


Subject(s)
Bacterial Vaccines/immunology , Borrelia burgdorferi/immunology , Interleukin-17/antagonists & inhibitors , Lyme Disease/prevention & control , Animals , Antibodies/therapeutic use , Interferon-gamma/physiology , Interleukin-17/physiology , Lyme Disease/pathology , Mice , Mice, Inbred C57BL , Receptors, Interleukin/antagonists & inhibitors , Receptors, Interleukin-17 , Recombinant Proteins/antagonists & inhibitors , Recombinant Proteins/toxicity , Vaccination
5.
Clin Diagn Lab Immunol ; 10(1): 44-52, 2003 Jan.
Article in English | MEDLINE | ID: mdl-12522038

ABSTRACT

We found that Borrelia burgdorferi-vaccinated gamma interferon-deficient (IFN-gamma(0)) mice challenged with B. burgdorferi developed prominent chronic destructive osteoarthropathy. When these mice were treated with anti-tumor necrosis factor alpha (TNF-alpha) antibody, the severity of the destructive osteoarthritis was enhanced and affected the mobility of the animals. In addition, extensive swelling of the hind paws occurred. In contrast, treatment of B. burgdorferi-vaccinated, challenged IFN-gamma(0) mice with recombinant TNF-alpha (rTNF-alpha) inhibited the development of arthritis, including swelling of the hind paws. Moreover, treatment of vaccinated, challenged IFN-gamma(0) mice with anti-TNF-alpha inhibited fourfold the production of an antibody that kills B. burgdorferi, while treatment of vaccinated, challenged IFN-gamma(0) mice with rTNF-alpha slightly elevated the level of the borreliacidal antibody. These results suggest that the level of TNF-alpha directly or indirectly regulates the production of borreliacidal antibody and the development of vaccine-induced destructive Lyme osteoarthritis. Studies are in progress to determine the mechanism by which TNF-alpha-dependent cytokines generate the destructive arthritis.


Subject(s)
Arthritis/chemically induced , Bacterial Vaccines/adverse effects , Interferon-gamma , Lyme Disease/complications , Tumor Necrosis Factor-alpha/pharmacology , Animals , Antibodies/pharmacology , Antibodies, Bacterial/biosynthesis , Antibodies, Bacterial/blood , Antibodies, Bacterial/drug effects , Arthritis/pathology , Borrelia burgdorferi/immunology , Drug Synergism , Mice , Mice, Knockout , Tumor Necrosis Factor-alpha/analysis , Tumor Necrosis Factor-alpha/immunology
6.
Clin Diagn Lab Immunol ; 9(5): 1095-101, 2002 Sep.
Article in English | MEDLINE | ID: mdl-12204965

ABSTRACT

The ability of a Lyme borreliosis vaccine to induce and maintain sustained levels of borreliacidal antibody is necessary for prolonged protection against infection with Borrelia burgdorferi. Vaccination against infection with B. burgdorferi could be improved by determining the mechanism(s) that influences the production of protective borreliacidal antibody. Borreliacidal antibody was inhibited in cultures of lymph node cells obtained from C3H/HeJ mice vaccinated with formalin-inactivated B. burgdorferi and cultured with macrophages and B. burgdorferi and treated with recombinant gamma interferon (rIFN-gamma). The suppression of production of outer surface protein A (OspA) borreliacidal antibody by rIFN-gamma was not affected by the time of treatment. In addition, treatment with rIFN-gamma inhibited the production of other anti-B. burgdorferi antibodies. By contrast, treatment of cultures of immune lymph node cells with anti-IFN-gamma marginally increased the production of borreliacidal antibody and enhanced the production of other antibodies directed against B. burgdorferi. These results show that IFN-gamma does not play a major role in the production of anti-OspA borreliacidal antibody. Additional studies are needed to determine which cytokine(s) will enhance production of borreliacidal antibody.


Subject(s)
Antibodies, Bacterial/biosynthesis , Antigens, Surface/immunology , Bacterial Outer Membrane Proteins/immunology , Borrelia burgdorferi/immunology , Interferon-gamma/pharmacology , Lipoproteins , Lyme Disease/immunology , Animals , Bacterial Vaccines , Cells, Cultured , Flow Cytometry , In Vitro Techniques , Interferon-gamma/immunology , Lyme Disease/prevention & control , Lyme Disease Vaccines , Lymph Nodes/cytology , Lymph Nodes/metabolism , Macrophages/cytology , Mice , Mice, Inbred C3H , Recombinant Proteins/immunology , Recombinant Proteins/pharmacology
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