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1.
Infect Immun ; 88(11)2020 10 19.
Article En | MEDLINE | ID: mdl-32778610

The symptoms of Lyme disease are caused by inflammation induced by species of the Borrelia burgdorferisensu lato complex. The various presentations of Lyme disease in the population suggest that differences exist in the intensity and regulation of the host response to the spirochete. Previous work has described correlations between the presence of regulatory T cells and recovery from Lyme arthritis. However, the effects of Foxp3-expressing CD4+ T cells existing prior to, and during, B. burgdorferi infection have not been well characterized. Here, we used C57BL/6 "depletion of regulatory T cell" mice to assess the effects these cells have on the arthritis-resistant phenotype characteristic of this mouse strain. We showed that depletion of regulatory T cells prior to infection with B. burgdorferi resulted in sustained swelling, as well as histopathological changes, of the tibiotarsal joints that were not observed in infected control mice. Additionally, in vitro stimulation of splenocytes from these regulatory T cell-depleted mice resulted in increases in gamma interferon and interleukin-17 production and decreases in interleukin-10 production that were not evident among splenocytes of infected mice in which Treg cells were not depleted. Depletion of regulatory T cells at various times after infection also induced rapid joint swelling. Collectively, these findings provide evidence that regulatory T cells existing at the time of, and possibly after, B. burgdorferi infection may play an important role in limiting the development of arthritis.


Lyme Disease/immunology , T-Lymphocytes, Regulatory/immunology , Animals , Borrelia burgdorferi/immunology , Female , Male , Mice , Mice, Inbred C57BL
2.
Pathog Dis ; 75(6)2017 08 31.
Article En | MEDLINE | ID: mdl-28859307

The role that cytokines play in the induction of Lyme arthritis is gradually being delineated. We showed previously that severe arthritis developed in a T-cell-driven murine model, even in mice lacking interleukin-17A (IL-17A) and administered anti-gamma-interferon (IFN-γ) antibody. Increased levels of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6), two pro-inflammatory cytokines, were detected in cultures of popliteal lymph node cells obtained from these mice. We hypothesized that concomitantly administered anti-IL-6, anti-TNF-α and anti-IFN-γ antibodies would inhibit the development of arthritis in IL-17A-deficient mice. Our results showed that swelling of the hind paws and histopathological changes consistent with arthritis were significantly reduced in IL-17A-deficient mice that administered the three anti-cytokine antibodies. These results suggest that treatment with multiple anti-cytokine antibodies can abrogate the induction of Lyme arthritis in mice.


Anti-Inflammatory Agents/pharmacology , Antibodies/pharmacology , Interferon-gamma/antagonists & inhibitors , Interleukin-6/antagonists & inhibitors , Lyme Disease/drug therapy , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Animals , Borrelia burgdorferi/growth & development , Borrelia burgdorferi/pathogenicity , Disease Models, Animal , Gene Expression , Hindlimb/drug effects , Hindlimb/immunology , Hindlimb/microbiology , Hindlimb/pathology , Interferon-gamma/genetics , Interferon-gamma/immunology , Interleukin-17/deficiency , Interleukin-17/genetics , Interleukin-17/immunology , Interleukin-6/genetics , Interleukin-6/immunology , Lyme Disease/immunology , Lyme Disease/microbiology , Lyme Disease/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/immunology
3.
Pathog Dis ; 75(2)2017 03 01.
Article En | MEDLINE | ID: mdl-28175297

The immune mechanisms responsible for development of Lyme arthritis are partially understood with interleukin-17 (IL-17) and gamma-interferon (IFN-γ) playing a generally accepted role. Elevated levels of IL-17 and/or IFN-γ have been reported in samples from human Lyme arthritis patients and experimental mice. In addition, IL-17 and IFN-γ have been implicated in the onset of arthritis in Borrelia-primed and -infected C57BL/6 mice. Recently, we showed that IL-17-deficient mice developed swelling and histopathological changes consistent with arthritis in the presence of high levels of IFN-γ. We hypothesized that neutralization of IFN-γ in IL-17-deficient mice would inhibit Borrelia-induced arthritis. Our results, however, showed that swelling of the hind paws and histopathological changes of arthritis did not differ between Borrelia-primed and -infected IL-17-deficient and wild-type mice with or without neutralization of IFN-γ. We also found higher levels of tumor necrosis factor alpha (TNF-α) and IL-6 in the popliteal lymph node cells of Borrelia-primed and -infected IL-17-deficient mice after neutralization of IFN-γ. These results suggest that multiple cytokines interact in the development of Borrelia-induced arthritis.


Arthritis/etiology , Borrelia Infections/genetics , Borrelia Infections/immunology , Borrelia/immunology , Interferon-gamma/antagonists & inhibitors , Interleukin-17/deficiency , Animals , Antibodies, Monoclonal/pharmacology , Arthritis/pathology , Borrelia Infections/metabolism , Borrelia Infections/microbiology , Cytokines/metabolism , Disease Models, Animal , Interferon-gamma/metabolism , Lyme Disease/genetics , Lyme Disease/immunology , Lyme Disease/metabolism , Lyme Disease/microbiology , Lymphocytes/immunology , Lymphocytes/metabolism , Male , Mice , Mice, Knockout , Phenotype
4.
Expert Rev Anti Infect Ther ; 15(2): 133-145, 2017 02.
Article En | MEDLINE | ID: mdl-27894204

INTRODUCTION: Mycoplasma genitalium is an emerging agent of sexually-transmitted infection and is responsible for clinically-significant genital tract disease in both females and males. Similar to scenarios recently experienced with the urogenital flagellate Trichomonas vaginalis, an evolving molecular diagnostic reference standard based on transcription-mediated amplification allows for accurate detection of the organism, plus additional insight into disease epidemiology. Areas covered. The basis for this article includes primary peer-reviewed literature plus compilations of data derived from routine clinical laboratory screening of females and males for agents of sexually-transmitted infection. Introductory laboratory and epidemiologic data related to T. vaginalis provides not only a foreshadowing to the dichotomies inherent to M. genitalium prevalence but also advocacy of a common non-invasive specimen source that could be used to screen females for both agents. This review also documents increased prevalence rates of M. genitalium in both females and males by way of transcription-mediated amplification. Expert commentary. Molecular detection of M. genitalium should be a consideration in the development of comprehensive sexually-transmitted infection screening programs for both females and males. Transcription-mediated amplification has additionally identified novel facets of M. genitalium and T. vaginalis epidemiology that warrant further investigation.


Mycoplasma Infections/epidemiology , Mycoplasma genitalium/isolation & purification , Sexually Transmitted Diseases/epidemiology , Trichomonas Infections/epidemiology , Trichomonas vaginalis/isolation & purification , Female , Humans , Male , Mass Screening , Mycoplasma Infections/microbiology , Mycoplasma genitalium/genetics , Prevalence , Sexually Transmitted Diseases/microbiology , Trichomonas Infections/microbiology , Trichomonas vaginalis/genetics
5.
Pathog Dis ; 74(7)2016 10.
Article En | MEDLINE | ID: mdl-27549424

Lyme borreliosis, which is caused in the United States by the spirochete Borrelia burgdorferi, may manifest as different arrays of signs, symptoms and severities between infected individuals. Recent studies have indicated that particularly severe forms of Lyme borreliosis in humans are associated with an increased Th17 response. Here, we hypothesized that a murine model combining the dysregulated immune response of an environment lacking interleukin-10 (IL-10) with a robust T-cell-driven inflammatory response would reflect arthritis associated with the production of IL-17 by CD4+ cells. We demonstrate that IL-10 regulates the production of IL-17 by Borrelia-primed CD4+ cells early after interaction with Lyme spirochetes in vitro and that infection of Borrelia-primed mice with B. burgdorferi leads to significant production of IL-17 that contributes to the development of severe arthritis. These results extend our previous findings by demonstrating that a dysregulated adaptive immune response to Lyme spirochetes can contribute to severe, Th17-associated arthritis. These findings may lead to therapeutic measures for individuals with particularly severe symptoms of Lyme borreliosis.


Arthritis/immunology , Arthritis/metabolism , Borrelia/immunology , CD4-Positive T-Lymphocytes/immunology , CD4-Positive T-Lymphocytes/metabolism , Interleukin-17/metabolism , Lyme Disease/immunology , Lyme Disease/metabolism , Animals , Arthritis/microbiology , Arthritis/pathology , Disease Models, Animal , Edema/pathology , Female , Interleukin-10/metabolism , Lyme Disease/microbiology , Lyme Disease/pathology , Male , Mice , Mice, Knockout , Phenotype , Spleen/cytology , Spleen/immunology , Spleen/metabolism , Time Factors
6.
Pathog Dis ; 74(7)2016 10.
Article En | MEDLINE | ID: mdl-27493062

Interleukin-17 (IL-17) has been shown to participate in the development of Lyme arthritis in experimental mice. For example, neutralization of IL-17 with antibodies inhibits induction of arthritis in Borrelia-primed and -infected C57BL/6 wild-type mice. We hypothesized that mice lacking IL-17 would fail to develop Borrelia-induced arthritis. IL-17-deficient and wild-type C57BL/6 mice were primed with heat-inactivated Borrelia and then infected with viable spirochetes 3 weeks later. No swelling or major histopathological changes of the hind paws were detected in IL-17-deficient or wild-type mice that were primed with Borrelia or infected with viable spirochetes. By contrast, IL-17-deficient and wild-type mice that were primed and subsequently infected with heterologous Borrelia developed severe swelling and histopathological changes of the hind paws. In addition, Borrelia-primed and -infected IL-17-deficient mice exhibited elevated gamma-interferon (IFN-γ) levels in sera and increased frequencies of IFN-γ-expressing lymphocytes in popliteal lymph nodes compared to Borrelia-primed and -infected wild-type mice. These results demonstrate that IL-17 is not required for development of severe pathology in response to infection with Borrelia burgdorferi, but may contribute to disease through an interaction with IFN-γ.


Arthritis/genetics , Arthritis/microbiology , Borrelia , Interleukin-17/deficiency , Lyme Disease/genetics , Lyme Disease/microbiology , Animals , Arthritis/pathology , Disease Models, Animal , Edema/pathology , Gene Expression , Interferon-gamma/blood , Interferon-gamma/metabolism , Interleukin-17/genetics , Interleukin-17/metabolism , Lyme Disease/pathology , Lymph Nodes/immunology , Lymph Nodes/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Knockout
7.
J Clin Microbiol ; 54(2): 432-8, 2016 Feb.
Article En | MEDLINE | ID: mdl-26659209

Following analysis of primary cervix, vagina, and first-void female urine specimens for Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis via commercial transcription-mediated amplification (TMA), residual material was subjected to Mycoplasma genitalium research-use-only TMA. Representation within a 2,478-specimen retrospective study set was established by comparison to a 6-month audit of clinical C. trachomatis TMA (12,999 specimens) on the basis of the C. trachomatis detection rate, specimen source distribution, clinic location, and age. M. genitalium was detected in 282 (11.4%) patients. This rate was higher than those seen with T. vaginalis (9.0%; P = 0.005), C. trachomatis (6.2%), and N. gonorrhoeae (1.4%). Positive M. genitalium results were confirmed by repeat testing or alternative-target TMA at a rate of 98.7%. The mean age of the M. genitalium-infected females (24.7 years) was lower than that of the T. vaginalis-infected females (mean, 30.1 years; P < 0.0001) and higher than that of the C. trachomatis-infected females (mean, 23.8 years; P = 0.003). Of 566 patient encounters positive for at least one sexually transmitted infection (STI), 35.9% exhibited sole detection of M. genitalium (P ≤ 0.0004 versus sole detection of other STI agents) and 26.1% were solely positive for T. vaginalis (P < 0.0002 versus C. trachomatis). The M. genitalium and T. vaginalis detection rates among 755 patients at urban emergency departments were 14.6% and 13.0%, respectively (P = 0.37). A 10.0% M. genitalium detection rate from other facilities exceeded that of T. vaginalis (7.2%; P = 0.004). Incorporation of M. genitalium TMA into comprehensive testing programs would detect M. genitalium in a significant proportion of females, particularly those in outpatient obstetrics and gynecology (OB/GYN) settings.


Mycoplasma Infections/diagnosis , Mycoplasma Infections/microbiology , Mycoplasma genitalium/genetics , Nucleic Acid Amplification Techniques , Sexually Transmitted Diseases/diagnosis , Sexually Transmitted Diseases/microbiology , Adolescent , Adult , Aged , Female , Humans , Middle Aged , Mycoplasma genitalium/isolation & purification , Phenotype , RNA, Bacterial , Retrospective Studies , Young Adult
8.
Diagn Microbiol Infect Dis ; 82(3): 194-8, 2015 Jul.
Article En | MEDLINE | ID: mdl-25934156

A total of 2750 male urines subjected to a transcription-mediated amplification (TMA)-based Mycoplasma genitalium assay yielded 188 positive results (6.84%). This rate was similar to Chlamydia trachomatis (6.87%; P = 0.96) and greater than Neisseria gonorrhoeae (4.0%) and Trichomonas vaginalis (2.3%; P < 0.0002). Mean age of M. genitalium-infected males (30.8) was similar to N. gonorrhoeae (P = 0.78) but less than T. vaginalis (mean, 41.6; P < 0.0001). A total of 266 STI clinic encounters had at least 1 sexually transmitted infection (STI); 36.5% of these encounters had sole detection of M. genitalium (P ≤ 0.009 versus sole detection of other STI agents). In 209 community encounters with at least 1 STI, 22.0% exhibited sole detection of M. genitalium (P = 0.0007 versus sole M. genitalium detection in STI clinic males), while 18.7% had sole detection of T. vaginalis (P < 0.0002 versus detection in STI clinic males). TMA-based M. genitalium screening identifies additional cases of nongonococcal urethritis.


Mycoplasma genitalium/isolation & purification , Urethritis/diagnosis , Urethritis/epidemiology , Urine/microbiology , Adolescent , Adult , Aged , Chlamydia trachomatis/isolation & purification , Humans , Male , Middle Aged , Molecular Diagnostic Techniques/methods , Neisseria gonorrhoeae/isolation & purification , Prevalence , Sexually Transmitted Diseases/diagnosis , Sexually Transmitted Diseases/epidemiology , Sexually Transmitted Diseases/microbiology , Sexually Transmitted Diseases/parasitology , Trichomonas vaginalis/isolation & purification , Urethritis/microbiology , Urethritis/parasitology , Young Adult
9.
Infect Immun ; 81(12): 4421-30, 2013 Dec.
Article En | MEDLINE | ID: mdl-24042116

Previous studies have shown that cells and cytokines associated with interleukin-17 (IL-17)-driven inflammation are involved in the arthritic response to Borrelia burgdorferi infection. Here, we report that IL-17 is a contributing factor in the development of Lyme arthritis and show that its production and histopathological effects are regulated by interleukin-10 (IL-10). Spleen cells obtained from B. burgdorferi-infected, "arthritis-resistant" wild-type C57BL/6 mice produced low levels of IL-17 following stimulation with the spirochete. In contrast, spleen cells obtained from infected, IL-10-deficient C57BL/6 mice produced a significant amount of IL-17 following stimulation with B. burgdorferi. These mice developed significant arthritis, including erosion of the bones in the ankle joints. We further show that treatment with antibody to IL-17 partially inhibited the significant hind paw swelling and histopathological changes observed in B. burgdorferi-infected, IL-10-deficient mice. Taken together, these findings provide additional evidence of a role for IL-17 in Lyme arthritis and reveal an additional regulatory target of IL-10 following borrelial infection.


Arthritis/immunology , Borrelia burgdorferi/immunology , Interleukin-10/immunology , Interleukin-17/metabolism , Lyme Disease/pathology , Animals , Antibodies/immunology , Cells, Cultured , Inflammation/immunology , Interleukin-10/deficiency , Interleukin-10/genetics , Interleukin-17/biosynthesis , Interleukin-17/immunology , Lyme Disease/immunology , Lyme Disease/microbiology , Mice , Mice, Inbred C57BL , Mice, Knockout , Th17 Cells/immunology , Th17 Cells/metabolism
10.
Expert Rev Anti Infect Ther ; 11(8): 845-63, 2013 Aug.
Article En | MEDLINE | ID: mdl-23944153

We briefly examine the clinical significance and pathogenesis of Trichomonas vaginalis and provide a comprehensive summary of non-molecular and molecular diagnostics for the organism. Transcription-mediated amplification (TMA) identifies more cases of trichomoniasis than other detection modalities. In our high-prevalence sexually transmitted infection community, TMA has allowed us to investigate female and male trichomoniasis epidemiology. Distribution of the organism is community-wide and similar between Caucasian-majority geographical areas and African-American-majority locales. First-void urine provides an excellent means for laboratory diagnosis of T. vaginalis in both genders. While trichomoniasis affects older age demographics, urine screening in younger females demonstrates elevated T. vaginalis prevalence. These data promote widespread adoption of TMA for diagnosis of T. vaginalis and further epidemiological assessments. Since trichomoniasis is the most-prevalent non-viral etiology of sexually transmitted infection worldwide, utilization of TMA would enhance accurate treatment with oral nitroimidazole agents that have cure rates of 90-100%.


Trichomonas Vaginitis/diagnosis , Trichomonas vaginalis/genetics , Female , HIV Infections/complications , Humans , Male , Nucleic Acid Hybridization , Polymerase Chain Reaction , Prevalence , Risk Factors , Sexually Transmitted Diseases/complications , Socioeconomic Factors , Trichomonas Vaginitis/complications , Trichomonas Vaginitis/epidemiology , United States/epidemiology , Urine/microbiology
11.
J Clin Microbiol ; 51(6): 1855-60, 2013 Jun.
Article En | MEDLINE | ID: mdl-23554208

Transcription-mediated amplification (TMA) enhances detection of Neisseria gonorrhoeae and Chlamydia trachomatis from rectal and pharyngeal sources. The utility of TMA for detection of Trichomonas vaginalis has recently been described. We report on the performance of TMA for detection of sexually transmitted infection (STI) agents from extraurogenital sources, with a focus on T. vaginalis. Within a 21-month interval, 1,314 consecutive male patient encounters at an STI clinic resulted in collection of 2,408 specimens for C. trachomatis, N. gonorrhoeae, and T. vaginalis TMA screening. A total of 471 encounters were managed with a single specimen collection (94.9% urine), with 12.7% positive for at least one STI agent. This detection percentage increased to 14.4% with collection of specimens from two sources and to 20.3% with collection from three sources (P = 0.03 versus single-source sampling). A total of 44.4% of encounters were managed by collection of urine and pharyngeal specimens and 19.1% by the addition of a third (rectal) collection. While procurement of urine and rectal specimens resulted in greater detection of C. trachomatis (6.1% and 11.3% rates, respectively) than of other STI agents, 858 pharyngeal specimens yielded a 2.9% T. vaginalis detection rate compared with 2.1% for N. gonorrhoeae and 1.6% for C. trachomatis. All T. vaginalis pharyngeal detections were confirmed by TMA-based alternative target testing. A total of 38.1% of T. vaginalis-positive pharyngeal specimens were derived from symptomatic patient encounters. A total of 85.7% of males with T. vaginalis-positive pharyngeal collections indicated strictly heterosexual preference. Additional specimen source sampling is necessary to make STI screening comprehensive. Incorporation of extraurogenital sources into assessment for T. vaginalis detection may identify additional symptomatic and asymptomatic male STI carriers.


Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , Parasitology/methods , Sexually Transmitted Diseases/diagnosis , Trichomonas Infections/diagnosis , Trichomonas vaginalis/isolation & purification , Adult , Ambulatory Care Facilities , Humans , Male , Mass Screening/methods , Middle Aged , Pharynx/parasitology , Prevalence , Sexually Transmitted Diseases/epidemiology , Trichomonas Infections/epidemiology , Young Adult
12.
J Clin Microbiol ; 51(1): 101-4, 2013 Jan.
Article En | MEDLINE | ID: mdl-23100348

Trichomonas vaginalis infection in males has been largely uncharacterized. Past reports indicated increased susceptibility to other sexually transmitted infection (STI) agents such as human immunodeficiency virus and Neisseria gonorrhoeae with concurrent T. vaginalis infection. This warrants a more thorough review of male T. vaginalis incidence. A retrospective 3-year investigation of transcription-mediated amplification (TMA)-based urethral swab and first-void urine screening for T. vaginalis within a regional health care system was performed to address T. vaginalis prevalence in males. Of 622 total samples tested, 6.6% were positive for T. vaginalis. Delineation of all specimens by ZIP code of patient residence revealed 11 predominant ZIP codes with respect to testing volume and detection rates. Within these 11 ZIP codes, representing 78.3% of total testing volume, urine was the preferred specimen source compared to urethral swabs. Seven of these 11 ZIP codes contained majority African American populations. The aggregate T. vaginalis detection rate trended higher than that of the remaining four ZIP codes, which were comprised primarily of Caucasian populations (8.9% versus 5.0%, respectively; P = 0.15). The average age of a T. vaginalis-infected male (39.9 years) was significantly greater than those for Chlamydia trachomatis or N. gonorrhoeae (27.6 and 25.9 years, respectively; P < 0.001). Given the significant rate of T. vaginalis detection, with age distribution analogous to that reported in females, TMA-based detection of T. vaginalis can be a routine constituent within a comprehensive STI screening panel for males in high-prevalence STI communities.


Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , Parasitology/methods , Sexually Transmitted Diseases/diagnosis , Trichomonas Infections/diagnosis , Trichomonas vaginalis/isolation & purification , Adolescent , Adult , Aged , Aged, 80 and over , Humans , Male , Mass Screening/methods , Middle Aged , Prevalence , Retrospective Studies , Sexually Transmitted Diseases/epidemiology , Sexually Transmitted Diseases/parasitology , Trichomonas Infections/epidemiology , Trichomonas Infections/parasitology , Trichomonas vaginalis/genetics , Urethra/parasitology , Urine/parasitology , Young Adult
13.
WMJ ; 111(5): 233-6, 2012 Oct.
Article En | MEDLINE | ID: mdl-23189457

OBJECTIVE: Trichomonas vaginalis analyte-specific reagent is a highly sensitive assay for T vaginalis detection. We report how this diagnostic innovation influenced the sexually transmitted infection ordering practice patterns of 20 subacute-care clinicians. METHODS: T vaginalis, Neisseria gonorrhoeae, and/or Chlamydia trachomatis screening data were audited on female swab submissions when only wet mount testing was available for detection of T vaginalis (2004-2007) and when T vaginalis detection options included analyte-specific reagent and wet mount (2008-2010). RESULTS: Analyte-specific reagent availability resulted in more screening and detection of T vaginalis, prompted less utilization of wet mount microscopy, and increased overall RNA-based screening for N gonorrhoeae and C trachomatis (P < 0.0002). CONCLUSION: Clinician familiarity with T vaginalis analyte-specific reagent can benefit both clinical practice and public health.


Nucleic Acid Amplification Techniques/methods , Sexually Transmitted Diseases, Bacterial/diagnosis , Trichomonas Vaginitis/diagnosis , Chlamydia Infections/diagnosis , Female , Gonorrhea/diagnosis , Humans , Mass Screening/methods , Sensitivity and Specificity
14.
Clin Dev Immunol ; 2012: 504215, 2012.
Article En | MEDLINE | ID: mdl-22461836

Arthritis is a frequent complication of infection in humans with Borrelia burgdorferi. Weeks to months following the onset of Lyme borreliosis, a histopathological reaction characteristic of synovitis including bone, joint, muscle, or tendon pain may occur. A subpopulation of patients may progress to a chronic, debilitating arthritis months to years after infection which has been classified as severe destructive Lyme arthritis. This arthritis involves focal bone erosion and destruction of articular cartilage. Hamsters and mice are animal models that have been utilized to study articular manifestations of Lyme borreliosis. Infection of immunocompetent LSH hamsters or C3H mice results in a transient synovitis. However, severe destructive Lyme arthritis can be induced by infecting irradiated hamsters or mice and immunocompetent Borrelia-vaccinated hamsters, mice, and interferon-gamma- (IFN-γ-) deficient mice with viable B. burgdorferi. The hamster model of severe destructive Lyme arthritis facilitates easy assessment of Lyme borreliosis vaccine preparations for deleterious effects while murine models of severe destructive Lyme arthritis allow for investigation of mechanisms of immunopathology.


Borrelia burgdorferi/immunology , Disease Models, Animal , Immunocompromised Host , Joints/immunology , Lyme Disease/immunology , Animals , Cricetinae , Humans , Immunocompetence , Interferon-gamma/deficiency , Interferon-gamma/immunology , Joints/pathology , Lyme Disease/microbiology , Lyme Disease/pathology , Lyme Disease Vaccines/biosynthesis , Lyme Disease Vaccines/immunology , Mice , Mice, Knockout , Whole-Body Irradiation
15.
Clin Vaccine Immunol ; 18(7): 1125-32, 2011 Jul.
Article En | MEDLINE | ID: mdl-21613457

Interleukin-35 (IL-35) has been reported to inhibit the production of interleukin-17 (IL-17) as a means of preventing arthritis and other inflammatory diseases. We previously showed that treatment of Borrelia-vaccinated and -infected mice with anti-IL-17 antibody at the time of infection prevented the development of arthritis. The anti-IL-17 antibody-treated mice lacked the extensive tissue damage, such as bone and cartilage erosion, that occurred in the tibiotarsal joints of untreated Borrelia-vaccinated and -infected control mice. We hypothesized that IL-35 would reduce the severity of arthritis by suppressing the production of IL-17 in Borrelia-vaccinated and -infected mice. Here, we show that administration of recombinant IL-35 (rIL-35) to Borrelia-vaccinated and -infected mice augments the development of severe arthritis compared to the results seen with untreated control mice. Borrelia-vaccinated and -infected mice treated with rIL-35 had significantly (P < 0.05) greater hind paw swelling and histopathological changes from day 4 through day 10 than non-rIL-35-treated Borrelia-vaccinated and -infected mice. In addition, the treatment with IL-35 only slightly decreased the production of IL-17 in Borrelia-primed immune cells and did not prevent the development of borreliacidal antibody. Our data do not support a role for IL-35 as a potential therapeutic agent to reduce inflammation in Lyme arthritis.


Interleukins/adverse effects , Lyme Disease Vaccines/pharmacology , Lyme Disease/drug therapy , Animals , Borrelia burgdorferi , Humans , Inflammation , Interleukin-17/biosynthesis , Interleukins/administration & dosage , Mice , Recombinant Proteins/administration & dosage , Recombinant Proteins/pharmacology , Treatment Failure , Vaccination
16.
J Food Prot ; 73(10): 1864-74, 2010 Oct.
Article En | MEDLINE | ID: mdl-21067675

We compared the survival of potential pathogen surrogates-meat-hygiene indicators (non-Escherichia coli coliforms), biotype I E. coli, and lactic acid bacteria starter cultures-with survival of an E. coli O157:H7 (ECO157:H7) inoculum in beef carcass intervention trials. Survival of one lactic acid bacteria starter culture (Bactoferm LHP Dry [Pediococcus acidilactici and Pediococcus pentosaceus]), a five-isolate biotype I inoculum, and a five-isolate non-E. coli coliform inoculum, was compared with survival of a 12-isolate ECO157:H7 inoculum in interventions by using beef brisket (adipose and lean), cod fat membrane, or neck tissue. Treatments were grouped by abattoir size: small (6-day dry aging; 22°C acid treatment [2.5% acetic acid, 2% lactic acid, or Fresh Bloom], followed by 1-day dry aging; hot water) and large (warm acid treatment [5% acetic acid or 2% lactic acid] with or without a preceding hot water treatment). Reductions in pathogen and surrogate inocula were determined with excision sampling. A surrogate was considered a suitable replacement for ECO157:H7 if the intervention produced a reduction in surrogate levels that was not significantly greater (P≥0.05) than that observed for ECO157:H7. All three surrogate inocula were suitable as ECO157 surrogates for dry aging and acid spray plus dry-aging treatments used by small abattoirs. No one inoculum was suitable as an ECO157 surrogate across all intervention treatments used by large abattoirs. Effects seen on neck tissue were representative of other tissues, and the low value of the neck supports its use as the location for evaluating treatment efficacy in in-plant trials. Results support using nonpathogenic surrogate organisms to validate beef carcass intervention efficacy.


Abattoirs/standards , Anti-Infective Agents/pharmacology , Cattle/microbiology , Decontamination/methods , Escherichia coli O157 , Food Handling/methods , Animals , Colony Count, Microbial , Consumer Product Safety , Escherichia coli O157/drug effects , Escherichia coli O157/growth & development , Food Contamination/prevention & control , Humans , Hydrogen-Ion Concentration , Lactic Acid/pharmacology , Meat/microbiology , Temperature
17.
Diagn Microbiol Infect Dis ; 68(1): 66-72, 2010 Sep.
Article En | MEDLINE | ID: mdl-20727473

Following wet mount analysis, 255 vaginal saline suspensions were aliquoted to lysis medium for transcription-mediated amplification (TMA)-based Trichomonas vaginalis analyte-specific reagent testing (ASR) (Gen-Probe, San Diego, CA). Specimens with visible T. vaginalis were then refrigerated, with additional aliquoting at later intervals. Twenty-four wet mount-positive specimens (9.4%) yielded a median luminescent value (x1000, relative light unit [RLU]) of 4736. In contrast, RLU ranged from 1 to 21 following ASR of 204 wet mount-negative specimens. Twenty-seven wet mount-negative specimens (10.5%) were positive by ASR and subsequently positive via T. vaginalis alternative target TMA (Gen-Probe). Discrepancies were additionally resolved by demonstration of T. vaginalis nucleic acid from a separate endocervical collection. T. vaginalis nucleic acid was detectable following prolonged storage, following minimal incubation in lysis medium, and from low-volume aliquots of sparsely populated specimens. T. vaginalis ASR adequately detects T. vaginalis from vaginal saline suspension aliquots, providing a simple specimen alternative for a highly sensitive laboratory diagnosis of trichomoniasis.


Nucleic Acid Amplification Techniques/methods , Sodium Chloride , Specimen Handling/methods , Transcription, Genetic , Trichomonas Vaginitis/parasitology , Trichomonas vaginalis/metabolism , Vagina/parasitology , Animals , Female , Humans , Trichomonas Vaginitis/diagnosis , Trichomonas vaginalis/genetics , Trichomonas vaginalis/isolation & purification
18.
FEMS Immunol Med Microbiol ; 60(1): 78-89, 2010 Oct.
Article En | MEDLINE | ID: mdl-20662925

The immunological events leading to the development of Lyme arthritis in humans are partially understood. Much of this information has been gained by studying the course of infection of naïve or vaccinated mice with Borrelia burgdorferi. However, the Borrelia-vaccination and -infection model has not been described using the organismal parameters commonly used in the widely accepted Borrelia-infection model. This is the first comparison between the Borrelia-infection and the Borrelia-vaccination and -infection models of arthritis. Borrelia-vaccinated and -infected C3H/HeN mice develop acute inflammation comparable to that of nonvaccinated, Borrelia-infected C3H/HeN mice. The duration and severity of arthritis in Borrelia-vaccinated and -infected mice was slightly increased compared with Borrelia-infected mice. Significantly, Borrelia-vaccinated and -infected C3H/HeN mice produce interleukin-17 (IL-17), while Borrelia-infected mice that had not been previously vaccinated do not. Neutralization of IL-17 in Borrelia-vaccinated and -infected C3H/HeN mice decreased the severity of arthritis, although not to the degree we observed previously in C57BL/6 mice. Collectively, these findings show that the Borrelia-vaccination and -infection model of Lyme arthritis incorporates elements of adaptive immunity that likely have relevance to human disease, but may not be observed in Borrelia-infected C3H/HeN mice.


Borrelia burgdorferi/immunology , Borrelia burgdorferi/pathogenicity , Lyme Disease Vaccines/immunology , Lyme Disease/immunology , Lyme Disease/pathology , Animals , Disease Models, Animal , Interleukin-17/antagonists & inhibitors , Interleukin-17/metabolism , Male , Mice , Mice, Inbred C3H
19.
Clin Vaccine Immunol ; 17(5): 870-4, 2010 May.
Article En | MEDLINE | ID: mdl-20237200

Laboratory-reared beagles were vaccinated with a placebo or a bacterin comprised of Borrelia burgdorferi S-1-10 and ospA-negative/ospB-negative B. burgdorferi 50772 and challenged after 1 year with B. burgdorferi-infected Ixodes scapularis ticks. For the placebo recipients, spirochetes were recovered from 9 (60%) skin biopsy specimens collected after 1 month, and the organisms persisted in the skin thereafter. Ten (67%) dogs also developed joint infection (3 dogs), lameness or synovitis (7 dogs), or B. burgdorferi-specific antibodies (8 dogs). For the vaccine recipients, spirochetes were recovered from 6 (40%) skin biopsy specimens collected after 1 month. However, subsequent biopsy specimens were negative, and the dogs failed to develop joint infection (P = 0.224), lameness/synovitis (P = 0.006), or Lyme disease-specific antibody responses (P = 0.002). The bacterin provided a high level of protection for 1 year after immunization, and the addition of the OspC-producing B. burgdorferi 50772 provided enhanced protection.


Borrelia burgdorferi/immunology , Dog Diseases/prevention & control , Lyme Disease Vaccines/immunology , Lyme Disease/veterinary , Animals , Antibodies, Bacterial/blood , Arthritis, Infectious/microbiology , Arthritis, Infectious/prevention & control , Biopsy , Borrelia burgdorferi/isolation & purification , Borrelia burgdorferi/pathogenicity , Dogs , Ixodes/microbiology , Lyme Disease/prevention & control , Lyme Disease Vaccines/administration & dosage , Placebos/administration & dosage , Skin Diseases, Bacterial/microbiology , Skin Diseases, Bacterial/prevention & control , Time Factors
20.
J Clin Microbiol ; 48(4): 1408-12, 2010 Apr.
Article En | MEDLINE | ID: mdl-20147649

Real-time detection of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) in cases of clinical bacteremia may promote appropriate antimicrobial therapy and infection control. Expense inherent to molecular diagnostics may prevent laboratories from utilizing real-time PCR for this purpose. BD GeneOhm StaphSR assay master mix was reconstituted and aliquoted into SmartCycler tubes in 25-mul volumes (freshly reconstituted master mix), with a portion being frozen at -70 degrees C (frozen master mix). Incubation of 40 previously analyzed lysates from positive BacT/Alert SA and SN blood culture bottles (identified as 10 MRSA strains, 10 MSSA strains, 12 coagulase-negative Staphylococcus strains, and 8 Micrococcus strains) in freshly reconstituted master mix and master mix frozen between 1 week and 6 months generated the expected results in all PCRs. Similarly, positive- and negative-control reagents stored frozen at -70 degrees C for up to 18 weeks yielded the expected reactions. Prospective analysis of 244 positive blood culture samples utilizing 1-week-frozen master mix and freshly reconstituted master mix yielded a 1.2% discordant rate upon initial testing due to three unresolved results (two unresolved results for freshly reconstituted master mix and one unresolved result for frozen master mix). Repeat testing produced a final 100% concordance rate between the two master mix preparations. Use of master mix that was frozen up to 6 months did not compromise performance of the BD GeneOhm StaphSR assay. This modification, resulting in less reagent waste, may allow a greater number of laboratories to consider real-time PCR methodology for detection of bacteremia caused by MRSA and MSSA.


Bacteremia/diagnosis , Bacteriological Techniques/methods , Methicillin Resistance , Polymerase Chain Reaction/methods , Staphylococcal Infections/diagnosis , Staphylococcus aureus/isolation & purification , Bacteremia/microbiology , Bacteriological Techniques/economics , Blood/microbiology , Humans , Polymerase Chain Reaction/economics , Prospective Studies , Sensitivity and Specificity , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/genetics
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