[Clinical values of different vestibular function examinations to evaluate the curative effect of vestibular neuritis rehabilitation].

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.

[Influence of oxidative/antioxidative biomarkers and inflammatory cytokines on rats after sub-acute orally administration of titanium dioxide nanoparticles].

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.

[Early effects of low-level long-term occupational chromate exposure on workers'health].

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.

IL-17 rs2275913 genetic variation contributes to the development of gastric cancer in a Chinese population.

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.

Hemostatic/fibrinolytic protein changes in C3H/HeN mice infected with Rickettsia conorii--a model for Rocky Mountain spotted fever.

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.

Rickettsial phospholipase A2 as a pathogenic mechanism in a model of cell injury by typhus and spotted fever group rickettsiae.

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.

Analysis and ultrastructural localization of Ehrlichia chaffeensis proteins with monoclonal antibodies.

Ehrlichia chaffeensis, an obligately intracellular bacterium with tropism for monocytes, is the etiologic agent of human monocytic ehrlichiosis. To determine the nature and ultrastructural location of E. chaffeensis antigens, monoclonal antibodies (MAbs) to E. chaffeensis were developed. The MAbs were used for immunofluorescence and Western immunoblotting analysis of the antigens of density gradient-purified ehrlichiae. Monoclonal antibody 6A1 recognized an epitope of a 30-kD protein. This antibody reacted with a strain-specific epitope of E. chaffeensis, Arkansas strain, and did not cross-react with any other ehrlichia tested. Monoclonal antibodies 3C7 and 7C1-B recognized Arkansas strain proteins of 30 and 29 kD and reacted with E. chaffeensis (strain 91HE17) proteins of 31 and 29 kD and an E. canis protein of 30 kD. Lack of reactivity of these two MAbs with E. sennetsu and E. risticii suggests that the epitope is group-specific. Monoclonal antibody 5D11 recognized a 58-kD protein of both strains of E. chaffeensis as well as E. canis, apparently a group-specific, conformation-independent epitope. Monoclonal antibody 7C1-C reacted with 58- and 88-kD proteins of both the Arkansas and 91HE17 strains. Trypsin treatment destroyed the reactivity of E. chaffeensis antigens with all the MAbs when tested by Western immunoblotting, indicating that these antigens are proteins with trypsin-sensitive epitopes. Immunoelectron microscopy of negatively stained intact E. chaffeensis organisms showed that the 30- and 29-kD proteins are present on the surface of the ehrlichial cell wall along with the previously localized 28-kD protein.

Antigenic diversity of Rickettsia conorii.

Analysis of seven strains designated as Rickettsia conorii for reactivity with a panel of 12 monoclonal antibodies to surface-protein epitopes of spotted fever group rickettsiae and by Western immunoblotting with standard serotyping sera revealed remarkable antigenic diversity. Rickettsial strains from France, Morocco, Ethiopia, Kenya, South Africa, India, and the USSR differed from one another in reactivity with at least one and as many as five monoclonal antibodies. Simko and Indian strains were similar to one another and differed substantially from other R. conorii strains. All seven strains reacted with three R. conorii-specific monoclonal antibodies. Western immunoblotting demonstrated a major 120-kD protein and a major 135-kD protein in all strains. The principal differences were the presence of a major undenatured 130-kD protein in all strains except Indian and Simko, which had an analogous protein of 124 kD. Immunodominant antigenically related, heat-denatured protein bands of 170 kD (Malish 7 strain), 175 kD (Manuel strain), and 190 kD (Kenya tick typhus, Indian, and Simko strains) were not detected in the M-1 and Moroccan strains. This antigenic diversity is greater than that previously reported for other spotted fever group rickettsial species, suggesting that R. conorrii is an older species than R. rickettsii with a longer period of time for evolutionary divergence.

Rickettsial infection in murine models activates an early anti-rickettsial effect mediated by NK cells and associated with production of gamma interferon.

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.

Identification of the antigenic constituents of Ehrlichia chaffeensis.

Ehrlichia chaffeensis, the novel etiologic agent of human ehrlichiosis in the United States, was first isolated in 1990 and reported in 1991. To analyze the antigenic components of E. chaffeensis, we cultivated these obligate intracellular bacteria in DH82 cells, purified the ehrlichiae by renografin density gradient centrifugation, and examined the antigens by Western immunoblotting. Rabbit and human antisera to E. chaffeensis revealed more than 20 bands ranging from 20 to 200 kD. The distinct 22-kD protein was heat labile. The rest of the major immunoreactive components were heat stable. The immunoblots of E. chaffeensis were highly similar when probed with antisera to E. chaffeensis, E. canis, and E. ewingii, indicating the close antigenic relationships among the three species. The 22-kD protein cross-reacted only with anti-E. canis serum. The antibody against E. sennetsu reacted strongly with the 66-, 64-, 55-, and 44-kD antigens of E. chaffeensis. The E. risticii antisera reacted strongly with the 55- and 44-kD bands but only faintly with the 66-kD band. The major immunoreactive antigens of E. chaffeensis (66, 55, and 44 kD) showed cross-reactions with all the different antisera tested. The results indicated that E. chaffeensis is antigenically most closely related to E. canis, is less closely related to E. ewingii, and is only distantly related to E. sennetsu and E. risticii.

Antigen-capture enzyme immunoassay: a comparison with other methods for the detection of spotted fever group rickettsiae in ticks.

To evaluate the prevalence of spotted fever group rickettsiae along the Adriatic Coast of Croatia, 832 ticks were examined by hemolymph test, direct immunofluorescence, antigen-capture enzyme immunoassay, and polymerase chain reaction. Very good agreement was observed among direct immunofluorescence, polymerase chain reaction, and antigen-capture enzyme immunoassay. Twelve ticks that were positive by hemolymph test and negative by both direct immunofluorescence and polymerase chain reaction presumably do not represent spotted fever group rickettsiae. By direct immunofluorescence, spotted fever group rickettsiae were present in 12% of Rhipicephalus bursa, 10.6% of Rh. sanguineus, and 7.8% of Dermacentor marginatus. From the 98 ticks containing rickettsia-like organisms by hemolymph test, seven spotted fever group rickettsial isolates were established in cell culture. Four isolates were identified as Rickettsia conorii. The antigen-capture enzyme immunoassay, which utilizes a monoclonal antibody to antigens of the 135-kD surface protein shared among many members of the spotted fever group, is recommended for primary screening of tick samples because it is reliable and yet less labor-intensive than the hemolymph and direct immunofluorescence tests. Although the polymerase chain reaction is too expensive for use as a screening method, it is recommended for confirmation of positive screening results. In addition to the technologic advance of the antigen-capture enzyme immunoassay, this study documented by contemporary methods that R. conorii is present along the eastern Adriatic Coast not only in the classic vector, Rh. sanguineus, but also in Rh. bursa and D. marginatus.

Identification of protective components of two major outer membrane proteins of spotted fever group Rickettsiae.

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.

Comparative antigenic analysis of spotted fever group rickettsiae from Israel and other closely related organisms.

Spotted fever rickettsiosis in Israel has been considered as possibly somewhat more severe than boutonneuse fever, from which it also differs in having a very low proportion of cases with a tick-inoculation site eschar. This investigation was undertaken to determine whether the Israeli spotted fever group (SFG) rickettsiae differed sufficiently from Rickettsia conorii to be considered as a distinct species. Strains of Rickettsia conorii from Morocco and South Africa, four SGF rickettsial isolates from Israel, one from Russia, and one from Zimbabwe were compared by microimmunofluorescence serotyping, Western immunoblotting, monoclonal antibody reactivity, and polymerase chain reaction amplification of the repeat domain of the rickettsial outer membrane protein A (rOmpA). All are strains and isolates of R. conorii, yet there is considerable molecular and antigenic diversity of both rOmpA and rickettsial outer membrane protein B (rOmpB) among them. The rOmpA gene of the Israeli isolates and the Astrakhan strain from Russia is estimated to encode 15 rOmpA repeat units as compared with 10 for the South African strain and six for the strains from Morocco and Zimbabwe. The Israeli SFG rickettsial strains appear to be R. conorii, a species with substantial antigenic and genetic diversity. The Israeli strains appear to fall within the limit previously described for the genetic and antigenic diversity of R. conorii.

Ultrastructural variation of cultured Ehrlichia chaffeensis.

The ultrastructure of Ehrlichia chaffeensis (Arkansas strain) was studied in non-irradiated and irradiated monolayers of mouse embryo, Vero, BGM and L929 cells, and in non-irradiated DH82 cells. Within the intracellular parasitophorous vacuoles (morulae), two types of ehrlichial cells were found regularly--those with uniformly dispersed nucleoid filaments and ribosomes (dense-cored cells), which represent the normal life cycle of ehrlichiae. In addition, large reticulate cells were observed, forming long projections of the cell wall, protrusions of cytoplasmic membrane into the periplasmic space, or budding of protoplast fragments (minute forms) into the periplasmic space. Ehrlichiae with abnormalities of protoplast fission were found, apparently leading to formation of giant, multilobular or elongated rod-like ehrlichiae. Morulae were usually surrounded by cisterns of granular endoplasmic reticulum and mitochondria and often contained vesicles, long tubules 25nm in diameter, probably originating from the ehrlichial cell wall, and fibrillar ehrlichial antigen apparently shed from the surface of the cell wall. Some cells contained, in addition to normal morulae, a whole morula that had become dense and contained degenerating ehrlichiae. These results indicate that as well as normal growth and reproduction, ehrlichiae exhibit pathological events: they can be remarkably damaged inside the host cell vacuoles, presumably phagolysosomes, or enter a process morphologically similar to bacterial L-transformation.

Ultrastructural differentiation of the genogroups in the genus Ehrlichia.

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.

Molecular cloning of a new variant of human papillomavirus type 6 isolated from a Chinese woman and expression of its L1 gene in E. coli--molecular cloning & expression of HPV6 gene.

A human papillomavirus genome DNA of 7.9 kb from a Chinese woman with genital condyloma acuminata was cloned in BamHI site of pAT153. According to the results obtained from Southern blotting, restriction mapping as well as partial DNA sequencing, the isolated genome (HPV6BV) had obvious variance and was referred to as a new variant of HPV6 found in China the first time. HPV6BV L1 gene was successfully expressed in E. coli as a fusion protein with pUR288. The beta-galactosidase/L1 fusion protein reacted with both beta-galactosidase antiserum and HPV antibody using Western blot technique. The E. coli-produced fusion protein, possessing HPV antigenicity, may provide a reagent for clinical diagnosis and epidemiological survey.

Interferon-gamma and tumor necrosis factor-alpha exert their antirickettsial effect via induction of synthesis of nitric oxide.

How the host defenses control rickettsiae in the cytosol of nonphagocytic host cells, where they are not exposed to antibodies or phagocytes, has posed a difficult question. Rickettsia conorii infection of a mouse fibroblast cell line was inhibited in a dose-dependent manner by nitrogen oxide synthesized by eukaryotic host cells stimulated by interferon-gamma or tumor necrosis factor-alpha. L-arginine was the source of the nitric oxide as demonstrated by competitive inhibition by NG-monomethyl-L-arginine. Nitric oxide synthesis required host cell protein synthesis and had an approximately 48-hour lag phase following cytokine stimulation. At low doses of interferon-gamma and tumor necrosis factor-alpha, which had no detectable response as single agents, dramatic synergistic nitric oxide synthesis and antirickettsial effects were observed.

Mechanisms of intracellular killing of Rickettsia conorii in infected human endothelial cells, hepatocytes, and macrophages.

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.

Establishment of a novel endothelial target mouse model of a typhus group rickettsiosis: evidence for critical roles for gamma interferon and CD8 T lymphocytes.

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.