ABSTRACT
AIM: To construct three-dimensional (3D) and two-dimensional (2D) models to predict the malignancy probability of subsolid nodules (SSNs) and compare their effectiveness. MATERIALS AND METHODS: A total of 371 SSNs from 332 patients, collected between January 2020 and January 2024, were included in the study. The SSNs were divided into a training set for constructing the models and a test set for validating the models. Models were developed using binary logistic backward regression, based on factors that showed significant differences in univariate analyses. The performance of the models was assessed using the area under the curve (AUC) of the receiver operating characteristic (ROC). The AUCs of different models were compared using the DeLong test. RESULTS: The AUCs for the two 3D models, one 2D model, and the Brock model were 0.785 (0.733-0.836), 0.776 (0.723-0.829), 0.764 (0.710-0.818), and 0.738 (0.679-0.798) in the training set. In the test set, these AUCs were 0.817 (0.706-0.928), 0.796 (0.679-0.913), 0.771 (0.647-0.895), and 0.790 (0.678-0.903). The two 3D models demonstrated statistically significant differences from the Brock model in the training set (P=0.024 and P=0.046). None of the four models showed significant differences in the test set (all P>0.05). CONCLUSION: The 3D models outperform both the 2D model and the Brock model in predicting the malignancy probability of SSNs, and the 3D model incorporating volume, mean CT attenuation value, and lobulation as factors performed the best.
ABSTRACT
The clinical values of video head impulse test (vHIT), caloric test (CT) and sensory organization test (SOT) at different stages before and after rehabilitation of 30 patients with vestibular neuritis (VN) in Vertigo Center Ward of Air Force Special Medical Center from January 2019 to January 2020 were analyzed and compared. There were 19 males (63.3%) and 11 females (36.7%), respectively, aged 18-68 (44±14) years. After 1 week and 3 months of rehabilitation in VN patients, the results of the three examinations were detached, and the recovery rates among the three observed indicators of each examination were statistically different (P<0.001). After 1 week of rehabilitation, the total recovery rate of vHIT was 0, which was lower than that of CT (40.0%) and SOT (43.3%) (both P<0.001). After 3 months of rehabilitation, the total recovery rate of vHIT was 13.3%, which was also lower than CT (86.7%) and SOT (80.0%) (both P<0.001). The current study indicates that the results of observed indicators from vHIT, CT and SOT were detached at different stages of VN rehabilitation. Therefore, the clinical significance of different vestibular function examinations is different but complementary.
Subject(s)
Vestibular Neuronitis , Vestibule, Labyrinth , Caloric Tests , Female , Head Impulse Test , Humans , Male , Vertigo , Vestibular Neuronitis/diagnosisABSTRACT
OBJECTIVE: To evaluate the sub-acute oral effect of titanium dioxide (TiO2) nanoparticles on the oxidation/antioxidation biomarkers and inflammatory cytokines in blood, liver, intestine, and colon in rats. METHODS: Twenty four 4-week-old clean-grade Sprague Dawley (SD) rats were randomly devided into 4 groups by body weight (n=6, control, low, middle, and high), in which the rats were orally exposed to TiO2 nanoparticles at doses of 0, 2, 10 and 50 mg/kg body weight/day for 28 consecutive days separately. Food intake, body weight and abnormal behaviors during the experiment were recorded. The rats were euthanized on the 29th day. The blood was collected via abdominal aortic method and centrifuged to collect the serum. Tissues from liver, intestine and colon were collected and homogenated. Then enzyme-linked immunosorbent assay (ELISA) and microwell plate methods were used to detect oxidation/antioxidation biomarkers including superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px), total mercapto (T-SH), glutathione disulfide (GSSG), malomdialdehvde (MDA) and inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the serum, liver, intestine and colon in the rats. RESULTS: Compared with the control group, no significant differences in body weight, food intake and organ coefficients were observed in all the three groups after TiO2 gavage. No significant changes in GSH, GSH-Px, T-SH, and IL-6 were observed. Compared with the control group, significant increase of SOD activity in serum in high dose group, signi-ficant increase of GSSG concentration in intestine in middle and high dose group and significant increase of MDA concentration in liver in low and high dose group were observed. Compared with the control group, a significant increase of TNF-α in liver in middle and high dose group was observed. CONCLUSION: TiO2 nanoparticle can increase antioxidant enzymes activities in blood, increase oxidative biomarkers in liver and intestine, increase inflammatory cytokines in liver in rats after a 28-day sub-acute orally administration. Among blood, liver, intestine, and colon, liver is most sensitive to the toxicity induced by TiO2 nanoparticles, followed by intestine, blood, and colon in sequence.
Subject(s)
Antioxidants , Nanoparticles , Animals , Biomarkers , Cytokines , Oxidative Stress , Rats , Rats, Sprague-Dawley , TitaniumABSTRACT
OBJECTIVE: To explore the effects of low-level long-term occupational exposure to chromate on the health of workers, and the potential biomarkers of early health effects in terms of lung function, immune toxicity and genetic damage. METHODS: A total of 22 chromate contact workers and 44 non-chromate contact workers from an electroplating enterprise with long-term occupational environment monitoring in line with the national standards in Inner Mongolia were investigated. The questionnaire survey was conducted to collect the basic situation, the history of smoking, drinking, diseases and so on. The portable lung function instrument, inductively coupled plasma mass spectrometry and cytokinesis-blocked micronucleus test were performed to measure the chromate contact workers'lung function, whole blood Cr (WB-Cr) and micronuclei frequency (MNF) of peripheral blood lymphocytes respectively. The cytometric bead array was used to detect the levels of IL-1ß, IL-6, IL-8, IL-10, IL-12P70 and TNFα in the serum among the two groups. The effects of chromate exposure on the above-mentioned indexes involved biological exposure, lung function, immune response and genetic damage, and their correlation were analyzed with different statistical methods. RESULTS: (1) the average length of service for chromate contact workers was 31 years, and their concentration of WB-Cr was 1.11-4.19 µg/L. They were divided into high and low exposure groups according to the median of 1.72 µg/L. The WB-Cr in the high exposure group (2.17 µg/L) was higher than that in the low exposure group (1.58 µg/L) as well as the reference value of the healthy population (1.74 µg/L, P<0.05); (2) the lung function test showed 10 (45.45%) chromate exposure workers had single or multiple abnormal lung function indexes, among which large airway injury index PEF, and small airway injury indexes MVV and FEF25%-75% were all negatively correlated with WB-Cr (r=-0.53, P<0.05; r=-0.52, P<0.05; r=-0.44, P<0.05); (3) IL-1ß, IL-6, IL-8 and TNFα in the serum of chromate contact workers were higher than those in the control group (P<0.05), and there was a positive correlation between TNFα and WB-Cr, and among these cytokines (P<0.05); (4) the average lymphocyte MNF in chromate contact workers was 1.341%, higher than the reference value of the general population (0.436%, P<0.01). Poisson regression analysis showed MNF in thehigh exposure group was higher than that in the low exposure group, OR (95%CI) =1.323 (1.049, 1.669); (5) multiple linear regression analysis showed that the lung function index FEF25%-75% decreased with the increase of TNFα (P<0.05), no significant correlation was found between other cytokines, MNF and lung function indexes. CONCLUSION: Long-term low-level occupational exposure to chromate can cause the decline of lung function, immune inflammatory reaction and genetic damage in workers, in which local or systemic inflammatory response is associated with decreased lung function. Lung function indexes PEF, FEF25%-75% and MVV, serum cytokines IL-1ß, IL-6, IL-8, and TNFα, and peripheral blood lymphocyte MNF may be used as early health effects biomarkers of chromate exposure.
Subject(s)
Occupational Exposure , Biomarkers , China , Chromates , Humans , SmokingABSTRACT
The purpose of this hospital-based case-control study was to assess whether the interleukin (IL)-17 rs2275913 genetic variation can influence susceptibility to gastric cancer. Samples from a total of 202 gastric cancer patients and 237 controls were collected from the Linyi People's Hospital between March 2013 and March 2015. The IL-17 rs2275913 gene polymorphism was identified by polymerase chain reaction and restriction fragment length polymorphism. When compared with control subjects, gastric cancer patients were older in age (OR = 3.89, 95%CI = 2.55-5.95), male (OR = 2.08, 95%CI = 1.39-3.10), had a habit of alcohol consumption (OR = 1.71, 95%CI = 1.15-2.55), and were more likely to be infected with Helicobacter pylori (OR = 2.76, 95%CI = 1.83-4.16). We observed that the AA genotype of the IL-17 rs2275913 polymorphism resulted in a 2.32-fold risk of gastric cancer compared to the GG genotype (OR = 2.32, 95%CI = 1.20-4.54; P = 0.01). The AG combined with AA genotype of the IL-17 rs2275913 polymorphism had more risk of developing gastric cancer than the GG genotype (OR = 1.50, 95%CI = 1.01-2.23; P = 0.04). Moreover, the AA genotype of the IL-17 rs2275913 polymorphism was correlated with a higher risk of developing gastric cancer than the GG and AG genotypes combined (OR = 2.01, 95%CI = 1.08-3.79; P = 0.02). In conclusion, the results of our study suggest that the IL-17 rs2275913 polymorphism could contribute to the risk of gastric cancer.
Subject(s)
Genetic Association Studies , Helicobacter Infections/genetics , Interleukin-17/genetics , Stomach Neoplasms/genetics , Adult , Aged , Asian People , Female , Genetic Predisposition to Disease , Helicobacter Infections/microbiology , Helicobacter Infections/pathology , Helicobacter pylori/pathogenicity , Humans , Male , Middle Aged , Polymorphism, Single Nucleotide , Risk Factors , Stomach Neoplasms/microbiology , Stomach Neoplasms/pathologyABSTRACT
Two approaches for presentation of a part of the rickettsial outer membrane protein A (OmpA) of Rickettsia rickettsii, namely (1) recombinant Mycobacterium vaccae (rMV) or (2) recombinant DNA vaccine, stimulated protective immunity against a lethal challenge with the closely related bacterium, R. conorii, in mice. After primary immunization with rMV and booster immunization with homologous recombinant protein, 67 and 55% of mice were protected against challenge in two experiments. DNA vaccination with booster recombinant protein immunization protected six out of eight animals from a lethal challenge. Production of IFN-gamma by antigen-exposed T-lymphocytes of DNA vaccine recipients indicated that cellular immunity had been stimulated.
Subject(s)
Bacterial Outer Membrane Proteins/immunology , Rickettsia rickettsii/immunology , Rickettsial Vaccines/pharmacology , Rocky Mountain Spotted Fever/prevention & control , Animals , Bacterial Outer Membrane Proteins/genetics , Base Sequence , DNA, Bacterial/genetics , Escherichia coli/genetics , Immunization, Secondary , Interferon-gamma/biosynthesis , Male , Mice , Mice, Inbred C3H , Mycobacterium/genetics , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Rickettsia rickettsii/genetics , Rickettsial Vaccines/genetics , Rocky Mountain Spotted Fever/immunology , T-Lymphocytes/immunology , Transformation, Genetic , Vaccines, DNA/genetics , Vaccines, DNA/pharmacologyABSTRACT
Fragments representing the genes of the two major outer membrane proteins of spotted fever group rickettsiae (rOmpA and rOmpB) were tested as DNA vaccines. Immunizations with each of three fragments (rompA4999-6710, rompB1550-2738, and rompB2459-4123) conferred a degree of protection on vaccinated mice against virulent rickettsial challenge. Protection was achieved when DNA immunizations were followed by booster immunizations with the homologous recombinant protein. Proliferation and gamma-interferon secretion were detected after in vitro stimulation of lymphocytes from immunized animals with whole Rickettsia conorii antigen. The data validate particular segments of rOmpA and rOmpB as potent immunogens and hence as sources of immunostimulatory elements with specificity for T lymphocytes, which are the key effectors of protective immunity against rickettsial infections.
Subject(s)
Antigens, Surface/immunology , Bacterial Outer Membrane Proteins/immunology , Boutonneuse Fever/prevention & control , Rickettsia conorii/immunology , T-Lymphocytes/immunology , Vaccines, DNA , Animals , Antigens, Bacterial/immunology , Antigens, Surface/analysis , Antigens, Surface/genetics , Bacterial Outer Membrane Proteins/analysis , Bacterial Outer Membrane Proteins/genetics , Base Sequence , Humans , Immunization, Secondary , Interferon-gamma/metabolism , Lymphocyte Activation , Male , Mice , Mice, Inbred C3H , Molecular Sequence Data , Molecular Weight , Polymerase Chain Reaction , Rickettsia conorii/genetics , VirulenceABSTRACT
Changes in plasma hemostatic and fibrinolytic proteins were determined during courses of a murine model of fatal and non-fatal Rocky Mountain spotted fever. C3H/HeN mice were infected with Rickettsia conorii and coagulation and histopathologic studies were performed at prescribed periods of time. A significant decrease in plasma factor VIII activity and rise in plasma factor V procoagulant activity correlated with a fatal infection. Factor VII levels were unchanged; factor XI levels dropped early in the course in the lethally infected animals, but returned to normal. Factor XII, high molecular weight kininogen, and prekallikrein levels were unchanged by the sublethal infection. Prekallikrein levels fell during the lethal infection. Antithrombin concentrations were decreased significantly in all animals, but plasma plasminogen levels did not change in either group of animals. Nonocclusive thrombi were microscopically observed rarely and only in animals surviving a sublethal infection. A fall in tissue plasminogen activator activity and a rise in plasminogen activator inhibitor activity highly correlated with a lethal outcome. Lethal infection with R. conorii is associated with primary endothelial cell injury resulting in decreased tissue plasminogen activator and increased plasminogen activator inhibitor.
Subject(s)
Endothelium, Vascular/pathology , Hemostasis , Rickettsia conorii/physiology , Rocky Mountain Spotted Fever/blood , Animals , Blood Coagulation Factors/metabolism , Chick Embryo , Endothelium, Vascular/metabolism , Endothelium, Vascular/microbiology , Factor V/analysis , Fibrinolysis , Lethal Dose 50 , Mice , Mice, Inbred C3H , Models, Animal , Partial Thromboplastin Time , Plasminogen Activator Inhibitor 1/analysis , Prothrombin Time , Rocky Mountain Spotted Fever/complications , Specific Pathogen-Free Organisms , Thrombophilia/etiology , Thrombophilia/pathology , Tissue Plasminogen Activator/analysisABSTRACT
Natural killer (NK) cell activity was significantly increased on days 2-6 of infection in the Rickettsia conorii-infected C3H/HeN mice and on day 2 in the Rickettsia typhi-infected C57BL/6 mice. Depletion of NK cell activity utilizing anti-NK1.1 monoclonal antibody enhanced the susceptibility of normally resistant C57BL/6 mice to infection with R. typhi, and depletion of NK cell activity with antibody to asialo GM1 enhanced the susceptibility of C3H/HeN mice to infection with R. conorii. Serum gamma interferon was increased in R. conorii-infected C3H/HeN mice compared with NK cell-depleted, infected mice during the early course of infection. Additionally, the NK cell activating cytokine IL-12 was elevated in the sera of infected mice during the time period representing enhanced NK cell activity compared with uninfected mice. Thus, it appears that NK cells contribute to the early anti-rickettsial immune response, likely via a mechanism involving gamma interferon.
Subject(s)
Interferon-gamma/biosynthesis , Killer Cells, Natural/immunology , Rickettsia Infections/immunology , Rickettsia conorii/immunology , Rickettsia typhi/immunology , Animals , Antibodies, Monoclonal , Boutonneuse Fever/blood , Boutonneuse Fever/immunology , Chick Embryo , Chlorocebus aethiops , Disease Models, Animal , Flow Cytometry , Interferon-gamma/blood , Interferon-gamma/immunology , Interleukin-12/blood , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , Mice, Inbred C57BL , Rickettsia Infections/blood , Spleen/immunology , Typhus, Endemic Flea-Borne/blood , Typhus, Endemic Flea-Borne/immunology , Vero CellsABSTRACT
Orientia tsutsugamushi is the etiologic agent of scrub typhus, a chigger-borne zoonosis that is a highly prevalent, life-threatening illness of greatest public health importance in tropical Asia and the islands of the western Pacific Ocean. The target cell of this bacterium is poorly defined in humans. In this study, O. tsutsugamushi were identified by immunohistochemistry using a rabbit polyclonal antibody raised against O. tsutsugamushi Karp strain in paraffin-embedded archived autopsy tissues of three patients with clinical suspicion of scrub typhus who died during World War II and the Vietnam War. Rickettsiae were located in endothelial cells in all of the organs evaluated, namely heart, lung, brain, kidney, pancreas, and skin, and within cardiac muscle cells and in macrophages located in liver and spleen. Electron microscopy confirmed the location of rickettsiae in endothelium and cardiac myocytes.
Subject(s)
Endothelium, Vascular/microbiology , Orientia tsutsugamushi , Scrub Typhus/microbiology , Adult , Animals , Brain/microbiology , Endothelium, Vascular/cytology , Endothelium, Vascular/ultrastructure , Fatal Outcome , Heart/microbiology , Humans , Immunohistochemistry , Kidney/microbiology , Liver/microbiology , Lung/microbiology , Male , Mice , Mice, Inbred BALB C , Microscopy, Electron , Orientia tsutsugamushi/immunology , Orientia tsutsugamushi/ultrastructure , Scrub Typhus/pathology , Spleen/microbiologyABSTRACT
Cytotoxic T-lymphocyte (CTL) activity developed against the major infected target cells of rickettsial infections, endothelial cells and macrophages. Spleen cells from mice immune to Rickettsia conorii exerted specific major histocompatibility complex (MHC) class I-matched CTL activity against R. conorii-infected SVEC-10 endothelial cells, with peak activity on day 10. Similarly, spleen cells from Rickettsia australis-immune mice exerted specific CTL activity against an R. australis-infected macrophage-like cell line. Gamma interferon (IFN-gamma) gene knockout mice were more than 100-fold more susceptible to R. australis infection than wild-type C57BL/6 mice. MHC class I gene knockout mice were the most susceptible, more than 50,000-fold more susceptible to a lethal outcome of R. australis infection than wild-type C57BL/6 mice. These results indicate that CTL activity was more critical to recovery from rickettsial infection than were the effects of IFN-gamma. The observation that perforin gene knockout mice were more than 100-fold more susceptible than wild-type C57BL/6 mice indicates that perforin-mediated activity accounts for a large component, but not all, of the CTL-mediated antirickettsial effect. CTL activity was expressed by immune CD8 T lymphocytes. Adoptive transfer of immune CD8 T lymphocytes from IFN-gamma gene knockout mice into R. australis-infected IFN-gamma gene knockout mice dramatically reduced the infectious rickettsial content in the organs, confirming that CD8 T lymphocytes provide immunity against rickettsiae besides that provided by the secretion of IFN-gamma. CTLs appear to be crucial to recovery from rickettsial infection.
Subject(s)
Boutonneuse Fever/immunology , Rickettsia Infections/immunology , T-Lymphocytes, Cytotoxic , Adoptive Transfer , Animals , Apoptosis , Boutonneuse Fever/therapy , Endothelium, Vascular/microbiology , Genes, MHC Class I , Immunity, Innate , Interferon-gamma/genetics , Macrophages/microbiology , Membrane Glycoproteins , Mice , Mice, Knockout , Perforin , Pore Forming Cytotoxic Proteins , Rickettsia Infections/therapyABSTRACT
Human monocytotropic ehrlichiosis caused by Ehrlichia chaffeensis is a life-threatening, tick-borne, emerging infectious disease for which no satisfactory animal model has been developed. Strain HF565, an ehrlichial organism closely related to E. chaffeensis isolated from Ixodes ovatus ticks in Japan, causes fatal infection of mice. C57BL/6 mice became ill on day 7 after inoculation and died on day 9. The liver revealed confluent necrosis, ballooning cell injury, apoptosis, poorly formed granulomas, Kupffer cell hyperplasia, erythrophagocytosis, and microvesicular fatty metamorphosis. The other significant histological findings consisted of marked expansion of the marginal zone and infiltration of the red pulp of the spleen by macrophages, interstitial pneumonitis, and increased numbers of immature myeloid cells and areas of necrosis in the bone marrow. Ehrlichiae were detected by immunohistology and electron microscopy in the liver, lungs, and spleen. The main target cells were macrophages, including Kupffer cells, hepatocytes, and endothelial cells. Apoptosis was detected in Kupffer cells, hepatocytes, and macrophages in the lungs and spleen. This tropism for macrophages and the pathological lesions closely resemble those of human monocytotropic ehrlichiosis for which it is a promising model for investigation of immunity and pathogenesis.
Subject(s)
Ehrlichia , Ehrlichiosis/pathology , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/metabolism , Apoptosis , Bone Marrow/microbiology , Bone Marrow/pathology , Disease Models, Animal , Ehrlichia/immunology , Ehrlichia chaffeensis/immunology , Ehrlichiosis/metabolism , Ehrlichiosis/microbiology , Humans , Immunohistochemistry , In Situ Nick-End Labeling , Leukocyte Count , Liver/microbiology , Liver/pathology , Liver/ultrastructure , Lung/microbiology , Lung/pathology , Lung/ultrastructure , Mice , Mice, Inbred C57BL , Platelet Count , Spleen/microbiology , Spleen/pathology , Spleen/ultrastructure , Tissue DistributionABSTRACT
Phospholipase A2 activity by typhus group rickettsiae causes hemolysis in vitro. Rickettsial phospholipase A2 has been proposed to mediate entry into the host cell, escape from the phagosome, and cause injury to host cells by both typhus and spotted fever group rickettsiae. In a rickettsial contact-associated cytotoxicity model, the interaction of Rickettsia prowazekii or R. conorii with Vero cells caused temperature-dependent release of 51Cr from the cells. Treatment of rickettsiae, but not the cells, with a phospholipase A2 inhibitor (bromophenacyl bromide) or with antibody to king cobra venom inhibited cell injury. Rickettsial treatment with bromophenacyl bromide inhibited the release of free fatty acids from the host cell. Neither the inhibitor nor antivenom impaired rickettsial active transport of L-lysine. Thus, host cell injury was mediated by a rickettsial phospholipase A2-dependent mechanism.
Subject(s)
Acetophenones/pharmacology , Antibodies/pharmacology , Enzyme Inhibitors/pharmacology , Phospholipases A/immunology , Rickettsia conorii/pathogenicity , Rickettsia prowazekii/pathogenicity , Vero Cells/ultrastructure , Acetophenones/therapeutic use , Animals , Antibodies/therapeutic use , Antivenins/pharmacology , Antivenins/therapeutic use , Boutonneuse Fever/drug therapy , Chlorocebus aethiops , Cytotoxicity Tests, Immunologic , Elapid Venoms/enzymology , Elapid Venoms/immunology , Enzyme Inhibitors/therapeutic use , Humans , Models, Biological , Phospholipases A/antagonists & inhibitors , Phospholipases A2 , Rickettsia conorii/drug effects , Rickettsia conorii/enzymology , Rickettsia prowazekii/drug effects , Rickettsia prowazekii/enzymology , Typhus, Epidemic Louse-Borne/drug therapy , Vero Cells/drug effects , Vero Cells/microbiologyABSTRACT
The mechanism of killing of obligately intracellular Rickettsia conorii within human target cells, mainly endothelium and, to a lesser extent, macrophages and hepatocytes, has not been determined. It has been a controversial issue as to whether or not human cells produce nitric oxide. AKN-1 cells (human hepatocytes) stimulated by gamma interferon, tumor necrosis factor alpha, interleukin 1beta, and RANTES (regulated by activation, normal T-cell-expressed and -secreted chemokine) killed intracellular rickettsiae by a nitric oxide-dependent mechanism. Human umbilical vein endothelial cells (HUVECs), when stimulated with the same concentrations of cytokines and RANTES, differed in their capacity to kill rickettsiae by a nitric oxide-dependent mechanism and in the quantity of nitric oxide synthesized. Hydrogen peroxide-dependent intracellular killing of R. conorii was demonstrated in HUVECs, THP-1 cells (human macrophages), and human peripheral blood monocytes activated with the cytokines. Rickettsial killing in the human macrophage cell line was also mediated by a limitation of the availability of tryptophan in association with the expression of the tryptophan-degrading enzyme indoleamine-2,3-dioxygenase. The rates of survival of all of the cell types investigated under the conditions of activation and infection in these experiments indicated that death of the host cells was not the explanation for the control of rickettsial infection. This finding represents the first demonstration that activated human hepatocytes and, in some cases, endothelium can kill intracellular pathogens via nitric oxide and that RANTES plays a role in immunity to rickettsiae. Human cells are capable of controlling rickettsial infections intracellularly, the most relevant location in these infections, by one or a combination of three mechanisms involving nitric oxide synthesis, hydrogen peroxide production, and tryptophan degradation.
Subject(s)
Endothelium, Vascular/microbiology , Hepatocytes/microbiology , Macrophages/microbiology , Rickettsia conorii/immunology , Cells, Cultured , Endothelium, Vascular/cytology , Humans , Hydrogen Peroxide/metabolism , Nitric Oxide/physiology , Tryptophan/physiology , Tryptophan Oxygenase/physiology , Tumor Necrosis Factor-alpha/physiologyABSTRACT
A mouse model of typhus rickettsiosis that reproduces the hematogenous dissemination to the critical target organs, including brain, lungs, heart, and kidneys, primary endothelial and, to a lesser degree, macrophage intracellular rickettsial infection, and typical vascular-based lesions of louse-borne typhus and murine typhus was established. Intravenous inoculation of C3H/HeN mice with Rickettsia typhi caused disease with a duration of the incubation period and mortality rate that were dependent on the infective dose of rickettsiae. Lethal infection was associated with high concentrations of R. typhi in the lungs and brain, despite a brisker humoral immune response to the rickettsiae than in the sublethal infection. Gamma interferon and CD8 T lymphocytes were demonstrated to be crucial to clearance of the rickettsiae and recovery from infection in experiments in which specific monoclonal antibodies were administered to deplete these components. Death of animals depleted of gamma interferon or CD8 T lymphocytes was associated with overwhelming rickettsial infection demonstrated by titers of infectious rickettsiae and by immunohistochemistry. An effective antirickettsial immune response was associated with elevated serum concentrations of IL-12 on Day 5 and increased secretion of IL-12 by concanavalin-A-stimulated spleen cells on Day 5. Evidence for transient suppression of the immune response consisted of marked reduction in the secretion of IL-2 and IL-12 by concanavalin-A-stimulated spleen cells on Days 10 and 15. This model offers excellent opportunities for study of attenuation and pathogenetic mechanisms of typhus rickettsiae, which are established biologic weapons of potential use in bioterrorism.
Subject(s)
CD8-Positive T-Lymphocytes/physiology , Disease Models, Animal , Endothelium, Vascular/microbiology , Interferon-gamma/physiology , Typhus, Epidemic Louse-Borne/etiology , Animals , Immunohistochemistry , Male , Mice , Mice, Inbred C3H , Rickettsia typhi/isolation & purification , Typhus, Epidemic Louse-Borne/immunology , Typhus, Epidemic Louse-Borne/pathologyABSTRACT
Cutaneous biopsies of five eschars and two rash lesions from five patients from New York City with documented rickettsialpox were examined by immunohistochemical methods with a monoclonal antibody directed against spotted fever group rickettsial lipopolysaccharide for the presence and cellular location of Rickettsia akari Rickettsiae were identified in all of the five patients, with good concordance of results for the same biopsy tissues with previously reported results by the direct immunofluorescence method. In contrast with immunofluorescence, which did not reveal the location of the organisms, immunohistochemical examination demonstrated R. akari to be in perivascular cells, morphologically resembling macrophages. Evaluation with double staining for rickettsiae and either CD68 or Factor VIII-related antigen revealed that the predominant infected cell type was CD68-positive macrophages, and only a rare rickettsia was detected in vascular endothelium, the major target cell for other rickettsioses. These results provide a diagnostic method for rickettsialpox and other spotted fever group rickettsioses and indicate that the elucidation of the pathogenesis of rickettsialpox must take into account that its target cell differs from that of Rocky Mountain spotted fever, boutonneuse fever, louse-borne typhus fever, and murine typhus.
Subject(s)
Macrophages/microbiology , Rickettsiaceae Infections/diagnosis , Rickettsiaceae Infections/immunology , Animals , Antibodies, Monoclonal , Antigens, CD/metabolism , Antigens, Differentiation, Myelomonocytic/metabolism , Biopsy , Humans , Immunohistochemistry , Lipopolysaccharides/metabolism , Macrophages/metabolism , Mice , Rickettsiaceae/isolation & purification , Rickettsiaceae Infections/metabolism , Rickettsiaceae Infections/microbiology , Skin Diseases/immunology , Skin Diseases/metabolism , Skin Diseases/microbiologyABSTRACT
Well-documented cases of simultaneous human infection with more than one tick-borne pathogen are rare. To our knowledge only two dual infections have been reported: simultaneous human infection with the agent of human granulocytic ehrlichiosis and Borrelia burgdorferi and simultaneous human infection with B. burgdorferi and Babesia microti (1-2). Rocky Mountain spotted fever has long been known to be endemic in North Carolina; cases of human ehrlichial infection were recognized there soon after Ehrlichia chaffeensis was recognized as an important cause of tick-borne disease in the southeastern United States. Because both Rocky Mountain spotted fever and ehrlichiosis are prevalent in North Carolina, occasional cases of simultaneous human infection by rickettsial and ehrlichial agents would not be surprising; however, no such cases seem to have been reported.
Subject(s)
Ehrlichia chaffeensis , Ehrlichiosis/complications , Rocky Mountain Spotted Fever/complications , Adult , Ehrlichia chaffeensis/genetics , Ehrlichia chaffeensis/isolation & purification , Ehrlichiosis/diagnosis , Fluorescent Antibody Technique , Genes, Bacterial , Humans , Male , Polymerase Chain Reaction , Rickettsia rickettsii/isolation & purification , Rocky Mountain Spotted Fever/blood , Rocky Mountain Spotted Fever/diagnosis , Sequence Analysis, DNA , Skin/microbiologyABSTRACT
Ultrastructural characteristics of 15 strains and isolates of ehrlichiae belonging to three genogroups, or clades of genetically related organisms united in the genera Ehrlichia, Cowdria, Anaplasma, Neorickettsia and a strain of Wolbachia pipientis which represents a fourth genogroup in this cluster of species, were studied in continuous cell culture or in vivo: E. canis (Oklahoma strain and VHE isolate), E. muris (AS 145), E. chaffeensis (Arkansas, 91HE17 and Sapulpa), human granulocytic ehrlichiae (HGE)(BDS, 96HE27, 96HE37, #54, #55 and #72), E. equi (MRK), E. sennetsu (Miyayama), E. risticii (HRC-IL). Wolbachia pipientis was studied in the naturally infected Aedes albopictus mosquito cell line Aa23. All organisms were similar in the normal ultrastructure of individual cells and in the ability to form abnormal, pathological ehrlichial cells of the same type irrespective of the species. Normally all ehrlichiae studied in cell culture existed in two morphological forms - reticulate and dense-cored cells, both of which could divide by binary fission. Most alterations were related to their membranes, especially the cell wall. Differences in the structure of intravacuolar microcolonies (morulae) of ehrlichiae and their inter-relations with the host cells allowed differentiation of the genogroups: the E. canis-E. chaffeensis-E. muris genogroup formed large morulae, with many ehrlichiae, often suspended in a fibrillar matrix, and the host cell mitochondria and endoplasmic reticulum usually aggregated near the morulae and were in contact with the morula membrane; the E. phagocytophila-E. equi-HGE group morulae had no fibrillar matrix, no contacts with host cell mitochodria, and they did not aggregate around the morulae; E. sennetsu-E. risticii group usually developed in small individual vacuoles that did not fuse with each other and divided along with the ehrlichiae.
Subject(s)
Ehrlichia/ultrastructure , Animals , Cells, Cultured , Ehrlichia/classification , Ehrlichia/genetics , Ehrlichia chaffeensis/ultrastructure , Horses , Humans , Macrophages/microbiology , Mice , Mice, Inbred BALB C , RNA, Ribosomal, 16S/analysisABSTRACT
A monoclonal antibody directed against an epitope on the lipopolysaccharide of typhus-group rickettsiae was developed for the purpose of detecting this heat-stable, proteinase-resistant antigen in formalin-fixed, paraffin-embedded tissues. Rickettsia prowazekii organisms were identified in endothelium and macrophages in sections of the brains of three Egyptian men who died of epidemic louse-borne typhus in Cairo during World War II and in the brain from a recent case of typhus fever acquired in Burundi. R. typhi organisms were identified in endothelial cells from a fatal case of murine typhus and in experimentally infected mice. This approach is applicable not only to the study of archival tissues and experimental animal models but also could be used to establish a timely diagnosis of typhus-group rickettsiosis by immunohistochemical examination of cutaneous biopsies of rash lesions during the acute stage of illness.