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Background: Q fever fatigue syndrome (QFS) is a state of prolonged fatigue following around 20% of acute Q fever cases. It is thought that chronic inflammation plays a role in its etiology. To test this hypothesis we measured circulating cytokines and the ex-vivo cytokine production in patients with QFS and compared with various control groups., Materials/methods: Peripheral blood mononuclear cells (PBMCs), whole blood, and serum were collected from 20 QFS patients, 19 chronic fatigue syndrome (CFS) patients, 19 Q fever seropositive controls, and 25 age- and sex-matched healthy controls. Coxiella-specific ex-vivo production of tumor necrosis factor (TNF)α, interleukin (IL)-1β, IL-6, and interferon (IFN) was measured, together with a total of 92 circulating inflammatory proteins., Results: PBMCs of QFS patients produced more IL-6 (P = 0.0001), TNFα (P = 0.0002), and IL-1β (P = 0.0005) than the various control groups when stimulated with Coxiella antigen. QFS patients had distinct differences in circulating inflammatory markers compared to the other groups, including higher concentrations of circulating IL-6 and IFNγ., Conclusion: QFS patients showed signs of chronic inflammation compared to asymptomatic Q fever seropositive controls, CFS patients, and healthy controls, of which the monocyte-derived cytokines TNFα, IL-1β, and especially IL-6, are likely crucial components., (Copyright © 2019. Published by Elsevier Ltd.)

Genes involved in human immune response are well recognized to influence the clinical course of infection. The association of host genetics with susceptibility to and severity of clinical symptoms in acute Q fever was investigated. Single nucleotide polymorphisms (SNPs) in the IFNG (rs2430561/rs1861493), STAT1 (rs1914408), and VDR (rs2228570) genes were determined in 85 patients from the 2007 Dutch acute Q fever outbreak, and a symptom score was calculated. IFNG rs1861493 showed a significant association with the symptom score; IFNG rs2430561 showed a similar trend. These SNPs were then used to reproduce results in a 2009 outbreak population (n = 123). The median symptom score differed significantly in both populations: 2 versus 7. The significant association of IFNG rs1861493 with symptom score in the first population was not reproduced in the second population. We hypothesize that individuals in the second outbreak were exposed to a higher Coxiella burnetii dose compared to the first, which overruled the protection conferred by the A-allele of IFNG rs1861493 in the first population.

Dendritic cells (DCs) serve as the primers of adaptive immunity, which is indispensable for the control of the majority of infections. Interestingly, some pathogenic intracellular bacteria can subvert DC function and gain the advantage of an ineffective host immune reaction. This scenario appears to be the case particularly with so-called stealth pathogens, which are the causative agents of several under-diagnosed chronic diseases. However, there is no consensus how less explored stealth bacteria like Coxiella, Brucella and Francisella cross-talk with DCs. Therefore, the aim of this review was to explore the issue and to summarize the current knowledge regarding the interaction of above mentioned pathogens with DCs as crucial hosts from an infection strategy view. Evidence indicates that infected DCs are not sufficiently activated, do not undergo maturation and do not produce expected proinflammatory cytokines. In some cases, the infected DCs even display immunosuppressive behaviour that may be directly linked to the induction of tolerogenicity favouring pathogen survival and persistence., (© 2013 John Wiley & Sons Ltd.)

Lions (Panthera leo) are an endangered species threatened by illegal hunting, habitat loss, and infectious diseases. Little is known about the tick-borne pathogens that infect lions and could contribute to population declines. The objective of this study was to characterize Rickettsia spp., Anaplasma phagocytophilum, and Coxiella burnetii infections in 10 lions from the Fasano Safari Park in Italy by serology, polymerase chain reaction, and sequence analysis. Although animals did not show clinical signs of tick-borne diseases, evidence of infection with C. burnetii, spotted fever group Rickettsia sp., and A. phagocytophilum were found in 50%, 20%, and 10% of the lions, respectively. One of the lions tested positive for all three pathogens. This study is the first report of molecular evidence of infection with C. burnetii, Rickettsia sp., and A. phagocytophilum in lions and provides evidence that these felids become infected and serve as hosts for tick-transmitted bacteria.

Background: Q fever remains an important occupational zoonosis in rural Australia. Although Q fever vaccine is recommended in high-risk occupational groups, its availability has been limited in recent years., Method: A literature review of the efficacy of the human Q fever vaccine registered in Australia was conducted., Results: Seven relevant vaccine efficacy studies were identified but no large double-blind, randomised, placebo-controlled studies have been conducted. Vaccine efficacy has ranged from 83-100% but limitations of study designs hamper a precise estimate of vaccine efficacy., Conclusion: Despite the shortcomings of efficacy studies, the Q fever vaccine available in Australia has considerable protective benefit in established high-risk environments, particularly of an occupational nature.

Bartonella spp. have been identified as aetiological agents in culture-negative infective endocarditis (IE). Coxiella burnetii may cause chronic Q-fever with endocarditis, 334 blood samples collected from 329 patients (334 episodes) with IE diagnosed between 1984 and 1996 in Göteborg, Sweden, were investigated for antibodies to Bartonella spp. and C. burnetii. 71 of the episodes (21%) were blood culture negative. A microimmunofluorescence assay revealed immunoglobulin G (IgG) antibodies to Bartonella in 13 of the culture verified episodes and in 2 of the culture-negative episodes. Three of the patients had IgG antibodies to > or = 200 in the blood culture-verified group, but none had a titre > or = 800, the cut-off level for Bartonella endocarditis. One patient had elevated antibodies to C. burnetii, diagnosing chronic Q-fever endocarditis. In conclusion, serologically verified Bartonella endocarditis is not prevalent in western Sweden and Q-fever endocarditis is rare.

The treatment of infectious diseases caused by intracellular bacteria, such as Q fever, may benefit from cytokines acting on macrophages. Monocytic THP-1 cells were infected with Coxiella burnetii, the etiological agent of Q fever, and then treated with IFN-gamma. While C. burnetii multiplied in untreated monocytes, IFN-gamma reduced bacterial viability after 24 h of treatment and reached maximum inhibition after 96 h. IFN-gamma also affected the viability of infected cells. Cell death resulted from apoptosis; occurring 24 h after the addition of IFN-gamma, it reached a maximum after 48 h and was followed by necrosis. Reactive oxygen intermediates were not required for C. burnetii killing, since monocytes from patients with chronic granulomatous disease were microbicidal in response to IFN-gamma. The role of cytokines was also investigated. IFN-gamma elicited a moderate release of IL-1beta in infected monocytes. Moreover, the IL-1 receptor antagonist did not affect C. burnetii survival, suggesting that IL-1beta was not involved in the bacterial killing induced by IFN-gamma. TNF was involved in IFN-gamma-induced killing of C. burnetii and cell death. IFN-gamma induced mRNA expression and sustained secretion of TNF. Neutralizing Abs to TNF as well as Abs directed against TNF receptors I and II, significantly prevented IFN-gamma-dependent killing of C. burnetii and cell death. These results suggest that IFN-gamma promotes the killing of C. burnetii in monocytes through an apoptotic mechanism mediated in part by TNF.

Following biosynthesis, class II MHC molecules are transported through a lysosome-like compartment, where they acquire antigenic peptides for presentation to T cells at the cell surface. This compartment is characterized by the presence of HLA-DM, which catalyzes the peptide loading process. Here we report that the morphology and function of the class II loading compartment is affected in diseases with a phenotypic change in lysosome morphology. Swollen lysosomes are observed in cells from patients with the hereditary immunodeficiency Chediak-Higashi syndrome and in cells infected with Coxiella burnetii, the rickettsial organism that causes Q fever. In both disease states, we observed that HLA-DR and HLA-DM accumulate in enlarged intracellular compartments, which label with the lysosomal marker LAMP-1. The distribution of class I MHC molecules was not affected, localizing disease effects to the endocytic pathway. Thus, cellular mechanisms controlling lysosome biogenesis also affect formation of the class II loading compartment. Analysis of cell surface class II molecules revealed that their steady-state levels were not reduced on diseased cells. However, in both disease states, enhanced interaction between HLA-DR and HLA-DM was detected. In the Chediak-Higashi syndrome cells, this correlated with more efficient removal of the CLIP peptide. These findings suggest a mechanism for perturbation of Ag presentation by class II molecules and consequent immune deficiencies in both diseases.

An indirect fluorescent antibody test was performed on sera collected from dogs housed in the municipal kennel of Setúbal to assess the prevalence of antibodies to Ehrlichia canis, the causative agent of canine ehrlichiosis and to Rickettsia conorii, agent of boutonneuse fever in humans. Two other members of the family Rickettsiaceae, Coxiella burnetii and Rickettsia typhi, were included in the serosurvey. Of the 104 dogs tested, 85.6% had antibodies to R. conorii, 50% to E. canis, 26.9% to R. typhi, and 4.8% to C. burnetii. These high seroprevalence rates of dogs with antibodies all year around against Rickettsiaceae suggest that physicians, public health officers and veterinarians should more frequently consider the diagnosis of these infections in Portugal.

The aim of this study was to investigate the antigenic structures of the morphologically distinct cells of the Coxiella burnetii developmental cycle. Postembedding immunoelectron microscopy with polyclonal antibodies produced in rabbits to (i) phase I cells, (ii) a chloroform-methanol residue fraction of cells, (iii) the cell walls (CW) of large and small cells and small dense cells (SDC), and (iv) the peptidoglycan-protein complexes of small cells and SDC labelled the continuum of morphologically distinct cells. But these antibodies did not distinguish between the antigenic structures of the various cells. Monoclonal antibodies to the phase I lipopolysaccharide labelled the CW of a majority of the smaller cells, but there was diminished reactivity to the larger cells. Although monoclonal antibodies to a 29.5-kDa outer membrane protein labelled the CW of the large mother cells, the large cells, and the small cells, a minority of the SDC with compact CW were not labelled. The endogenous spore within the mother cell was not labelled by the polyclonal or monoclonal antibodies to cellular components. A selected population of SDC was prepared by osmotic lysis of large cells, differential centrifugation, Renografin step-gradient fractionation, and breakage of the small cells in a French press at 20,000 lb/in2. The pressure-resistant SDC collected as fraction CL did not contain the 29.5-kDa protein, as evidenced by the lack of (i) Coomassie brilliant blue staining of protein in the 29.5-kDa region of sodium dodecyl sulfate-polyacrylamide gels and (ii) reactivity of the 29.5-kDa protein antigenic epitopes in immunoblotting with monoclonal antibodies to the protein. In contrast, CW of the pressure-sensitive small cells contained the 29.5-kDa protein. Therefore, the observed ultrastructural differences between large and small cells and SDC reflect differences in sensitivity to breakage by pressure treatment and in cell-associated antigens. Although the process of differentiation in C. burnetii remains an enigma, we have taken steps toward identifying cellular antigens as markers of differentiation. The pressure-resistant SDC in fraction CL that are devoid of the 29.5-kDa protein may be useful for answering questions about the physiological events required for triggering outgrowth and sequential regulation of the Coxiella developmental cycle.

Q fever is usually acquired by contact with aerosols generated during parturition of domestic ungulates (e.g., sheep, cows, goats). In the maritime provinces of Canada, parturient cats have also been implicated in its transmission. A 66-year-old woman from eastern Maine developed high fever, rigors, headache, myalgias, pulmonary infiltrates, and elevated hepatocellular enzymes, and the diagnosis of acute Q fever was confirmed serologically. She and 14 other family members had attended a family reunion in Maine 2 weeks earlier, when they were exposed to a parturient cat. All 11 adults and older children attending the reunion developed symptoms consistent with acute Q fever. Serum samples were obtained from 10 who attended the reunion and 8 who did not attend. Titers greater than or equal to 1:64 to Coxiella burnetii were present in all who attended the reunion but in none of those who did not. Cat-associated Q fever should be considered when sporadic cases of the disease occur in the United States.

Vaccination with an inactivated, whole cell, Q fever vaccine (Q-vax) induces lasting antibody conversion and a positive delayed-type hypersensitivity (DTH) skin reaction in about 60% of recipients but a long-lasting positive lymphoproliferative or mitogenic response to C. burnetii antigens with peripheral blood mononuclear cells (PBMC) in 85-95% of subjects. Analysis of the lymphoproliferative response to C. burnetii antigens has now been made by fractionation-reconstitution experiments with PBMC from vaccines, from past infections, and from healthy controls. The major contributor to the response in immune subjects proved to be the T lymphocyte. T cells were stimulated by both the phase I and phase II antigens of two prototype strains of C. burnetii and responses were greatly amplified by addition of IL-2. Similar T lymphocyte stimulation profiles were obtained with the 'Priscilla' strain of C. burnetii which represents a different biotype of Coxiella isolated from Q fever endocarditis; Q-vax is therefore likely to protect against endocarditis strains. Fractionation-reconstitution experiments with T and B cells from vaccines and subjects infected in the past, using various antigenic or haptenic fractions from C. burnetii indicate that protein, non-lipopolysaccharide components of the organism are responsible for the mitogenic response of immune T cells. However, the role of the lipopolysaccharide in the protective immunogen has still to be defined.

A 38-year-old man was admitted to the hospital with complaints of persistent fever up to 40 degrees C, arthralgias, headache and a nonproductive cough. The white-cell count was within the normal range but was markedly shifted to the left and demonstrated toxic granulations. Sonographic examination of the abdomen revealed a slight enlargement of the spleen. A nonspecific reactive hepatitis which was clinically asymptomatic was detected by laboratory evaluation. Diagnosis of an acute Q fever was made by demonstration of antibodies against C. burnetii. Following therapy with doxycycline, the patient became afebrile within 48 hours.

A serological survey for Coxiella burnetii was undertaken on a randomly selected population of 103 Ontario sheep flocks. Twenty-two flocks had at least one positive ewe; seven flocks had two or more reactors. The positive flocks were geographically clustered northwest of Guelph. Crutch-clipping of the ewe's wool prior to lambing, and total confinement housing at lambing in winter and spring seemed to lower the probability of seroreactivity of the flock (p less than 0.05). The study suggests that sheep are not a major reservoir for Coxiella burnetii in Ontario.

Materials on the development of an enzyme immunoassay (EIA) system for the detection of the antigens of C. burnetii, the causative agent of Q rickettsiosis, are presented. The system is highly specific and effective with respect to both corpuscular antigens of phases 1 and 2 and soluble antigen (lipopolysaccharide). The sensitivity of this method varies within the range 5-100 ng/ml. The effectiveness of EIA as a quantitative (semiquantitative) control test used in the process of the production of Coxiella preparations has been demonstrated.

Q fever is known to be a worldwide disease, with Sweden supposed to be one of a few exceptions. The purpose of this pilot study was to elucidate whether or not a potential risk group for obtaining Q fever in Sweden was seropositive to the causative agent Coxiella burnetii. Blood samples were collected from sheep farmers on the island of Gotland, and from members of their families. Serum samples were examined by ELISA for the presence of antibodies against C. burnetii, phases I and II. Positive reactions were confirmed with Western blot analysis. It was found that 30% of the study group were seropositive to C. burnetii, thus indicating that Q fever is endemic in this area of Sweden.

The comparative study of the dynamics of morphological changes in tissues of guinea pigs after the subcutaneous injection of chemical, live and combined vaccines against Q fever during the period from 12 hours to 90 days was made. All vaccines under study were shown to produce a pronounced local damaging effect. Two periods were tentatively discriminated in the dynamics of changes: the early phase (till 48 hours) and the late phase (days 2-90). At the early stage the most pronounced changes were registered after the injection of the combined vaccine. At the late phase the use of the chemical and combined vaccines was accompanied by the appearance of secondary hemorrhages into newly formed connective tissue. Starting from day 30, practically no deviation from the normal state of tissues were registered at the site of injection.

Evidence of past zoonotic infection was investigated serologically in randomly selected Northern Ireland farmers. The percentage of farmers with antibody was: Brucella abortus (0.7), Leptospira interrogans serovars (8.1), Borrelia burgdorferi (14.3), Toxoplasma gondii (73.5), Coxiella burnetii (28.0), Chlamydia psittaci (11.1) and Hantavirus (1.2). The results show that Northern Ireland farmers have been exposed in the past to zoonotic infections. It is not known if these infections contributed to ill health in farmers but it is now time for the health of farm workers and their medical services to be reassessed.

Complement binding antibodies against Coxiella burnetii were measured in 699 seric samples in order to ascertain the possible relationship between seric prevalence and the following factors: age, sex, living in rural or urban areas and socio-economical level. All patients were healthy: 180 were from a rural area 86 from Guijo (Cáceres); 36 from Chiloeches (Guadalajara) and 58 from Bustarviejo (Madrid); 519 were from an urban area (City of Madrid). The samples from Madrid city were classified by socio-economical level: 386 from medium-low level (MLL) and 133 from medium-high socio-economical (MHL) level. The global seroprevalence was 32.3%, being 46.1% from rural areas; --59.3 from Guijo, 33.3% from Chiloeches and 34.5% from Bustarviejo. In urban areas the seroprevalence was 27.5% this being 30.8% MLL and 18% MHL. We found a statistically significant relationship between both socio-economical levels and the area of living.

The Salamanca Province of Spain is an endemic zone for Mediterranean spotted fever. In this area, only one case of Q fever has been reported and typhus group rickettsial diseases have never been diagnosed. To obtain a panoramic view of the presence of antibodies to the most ubiquitous rickettsial agents, 400 sera specimens from a statistically representative sample of the human population of Salamanca City and its surrounding province were subjected to indirect microimmunofluorescent test for antibodies against Rickettsia conorii, Coxiella burnetti, phase II, and Rickettsia typhi antigens. Titers greater than or equal to 1:40 to R. conorii were found in 73.5% of the sera. Positivity was more common in subjects who reported closer contact with the natural environment and/or with domestic animals. Seropositivity to C. burnetti, phase II, was detected in 50.2% of sera. These positive cases were related to rural environmental factors and to previous contact with animals. The frequency of antibodies increased with age showing a progressive exposure to the rickettsial antigen. The prevalence of antibodies to R. conorii and C. burnetii in the human population of Salamanca Province is higher than that reported from any other geographic zone. The study of antibodies to R. typhi showed that 12.5% of the sera had titers greater than or equal to 1:40. According to our results, seropositivity to this rickettsial antigen cannot be related to any particular group of population nor interpreted totally as cross reactivity with R. conorii. Our data show a wide distribution of R. conorii and C. burnetii antigens in Salamanca Province, and also indicate the presence of R. typhi antigens in this area.

Sheep erythrocytes sensibilized with the lipopolysaccharide (LPS) extracted from Coxiella burnetii phase 1 cells were used in indirect haemolysis test (IHT). The test was highly sensitive and specific. Its use seemed reasonable when following the chronic course of Q-fever by serologic methods.

Coxiella burnetii antigens stimulate the defence against growth of hepatoma 22a cells. The antigen-stimulated mice survived longer, they considerably later developed palpable tumours and showed a retarded tumour growth. The enhanced resistance to tumour growth may be explained by at least 2 interrelated phenomena; namely by the induction of interferon-like activity and an increased NK cell activity.

A 51 year old woman with pneumonia developed Miller Fisher syndrome. Indirect immunofluorescence tests showed antibodies against Coxiella burnetti. Miller Fisher syndrome associated with Q fever has not been described previously.

Five cases of clinical Q fever were identified amongst students and staff of a Somerset secondary school between 23 October 1987 and 21 December 1987. Five goats which were kept at the school were found to have antibodies to Coxiella burnetii phase II. A cross-sectional study was conducted at the school in July 1988. A single CF test was used to identify evidence of prior infection, and a self-administered questionnaire was used to collect data on exposure variables and illness during 1987. Four hundred and ninety-nine eligible subjects took part in the study, and serological information was obtained from 461 of these. Eighty-seven subjects (18.9%) had CF titres of 20 or greater. It was estimated that only 1 in every 30 individuals with evidence of past C. burnetii infection had been recognized as a clinical case of Q fever. Antibody positive subjects were more likely to have been off school sick and to report having suffered chest pain than negative subjects. Contact with school animals, specifically cleaning the school poultry, collecting their eggs and visiting a school goat on the day of kidding, was associated with the presence of antibodies to C. burnetii. However, a large proportion of the antibody positives (24.2%) had no known contact with the school animals. Spread of organisms, either wind-borne or in straw or manure, may have been responsible for the high prevalence of unexplained infection.

R. prowazekii antigens have been tested with the use of monoclonal antibodies (McAb) to different epitopes of the microorganism. As revealed in these tests, McAb B4/4 and A-3/D, active against species-specific thermolabile antigen, interact with protein having a molecular weight of 90-120 KD. McAb C5/2, active against thermostable group antigen common with that of Rickettsia typhi, interact with LPS-like antigen having a molecular weight of 30 KD. Ultrastructural immunochemical studies have revealed that both R. prowazekii antigens are located on surface structures of rickettsiae, such as the microcapsule and cell wall.

A limited, randomized, blind, placebo-controlled trial of Q fever and influenza vaccines has been conducted in three Queensland abattoirs on a sequential analysis design. Ninety-eight subjects were given Q fever vaccine and 102 influenza vaccine. Q fever cases were observed in unvaccinated workers in all three abattoirs during the period of observation. A total of seven Q fever cases in one group, one more than the number required to achieve statistical significance between the two vaccine groups, was reached after 15 months with the cases coming from two of the abattoirs. These Q fever cases were in the group which had been given influenza vaccine and none in that given Q fever vaccine. Symptomless seroconversion rates of 24% were found in the remaining influenza virus vaccinees, and those without immunity were given Q fever vaccine.

The isotypic immune response of 16 individuals who developed Q fever pneumonia following exposure to an infected parturient cat was studied. The enzyme-linked immunosorbent (ELISA) test was used to detect IgM, IgA, and IgG antibodies to phase I and phase II Coxiella burnetii whole-cell antigens and to the phase I lipopolysaccharide. The indirect immunofluorescent antibody (IFA) test was also used to detect antibodies to phase I and phase II whole cells. None of the 16 subjects developed antibodies to the phase I lipopolysaccharide. The ELISA was more sensitive than the IFA test. IgM antibodies to phase II antigen were detectable by ELISA in 80% of the subjects at the time of onset of symptoms and were still present in 7 of the 8 tested at 32 weeks following the onset of symptoms. In all instances (ELISA: IgG, IgM; IFA: IgG, IgM) phase II antibodies developed earlier and reached higher levels than did phase I antibodies. The absence of antibodies to phase I lipopolysaccharide in acute Q fever combined with our unpublished findings of antibodies to phase I lipopolysaccharide in chronic Q fever suggests that this test may be used to distinguish acute from chronic Q fever.

During the period 1981-8 a clinical trial of a Q fever vaccine (Q-vax; Commonwealth Serum Laboratories, Melbourne) has been conducted in abattoir workers and other at-risk groups in South Australia. Volunteers in four abattoirs and visitors to the abattoirs were given one subcutaneous dose of 30 micrograms of a formalin-inactivated, highly-purified Coxiella burnetii cells, Henzerling strain, Phase 1 antigenic state, in a volume of 0.5 ml. During the period, over 4000 subjects have been vaccinated and the programme continues in the abattoirs and related groups. 'Common' reactions to the vaccine comprised tenderness and erythema, rarely oedema at the inoculation site and sometimes transient headache. Two more serious 'uncommon' reactions, immune abscess at the inoculation site, were observed in two subjects, and two others developed small subcutaneous lumps which gradually dispersed without intervention. Protective efficacy of the vaccine appeared to be absolute and to last for 5 years at least. Eight Q fever cases were observed in vaccinees, but all were in persons vaccinated during the incubation period of a natural attack of Q fever before vaccine-induced immunity had had time (greater than or equal to 13 days after vaccination) to develop. On the other hand, 97 Q fever cases were detected in persons working in, or visiting the same abattoir environments. Assays for antibody and cellular immunity showed an 80-82% seroconversion after vaccination, mostly IgM antibody to Phase 2 antigen, in the 3 months after vaccination. This fell to about 60%, mostly IgG antibody to Phase 1 antigen, after 20 months. On the other hand, 85-95% of vaccinees developed markers of cell mediated immunity as judged by lymphoproliferative responses with C. burnetii antigens; these rates remained elevated for at least 5 years. The Q fever vaccine, unlike other killed rickettsial vaccines, has the property of stimulating long-lasting T lymphocyte memory and this may account for its unusual protective efficacy as a killed vaccine.

This study investigated the release of superoxide anion (O2-) as an indicator of the oxidative metabolism of human neutrophils during the phagocytosis of Phase I Coxiella burnetti. Human neutrophils were incubated for 1 hr at 37 degrees C with opsonized or unopsonized viable Phase I Coxiella burnetii (MOI was 100 : 1) and superoxide anion formation was measured by the reduction of ferricytochrome C. The data revealed that during its phagocytosis by human neutrophils, C. burnetii (opsonized or unopsonized) fails to stimulate superoxide anion production. In contrast, the uptake of Staphylococcus aureus or zymosan was accompanied by the release of measurable O2-. This release of O2- was abrogated by the addition of 100 micrograms/ml of superoxide dismutase (SOD). These results suggest that the establishment of C. burnetii within neutrophils, as occurs during persistent infection, may be due to the failure to stimulate the metabolic burst during phagocytosis.

The indirect microimmunofluorescence test was used to determine the presence of antibodies to phase I and phase II Coxiella burnetii antigens in New Brunswick and Prince Edward Island cats. Twenty of 104 (19.2%) New Brunswick cats tested had antibodies to phase II antigen; five of these (4.8%) also had antibodies to phase I antigen. Six of 97 (6.2%) Prince Edward Island cats tested had antibodies to phase I and phase II antigens. Our data suggest that cats may be important in the epidemiology of Q fever in these provinces.

To determine the utility of Western immunoblotting in distinguishing chronic Q fever from acute Q fever, we first examined serum samples from individuals who had no antibodies to Coxiella burnetii by either the indirect immunofluorescence antibody test or by the enzyme-linked immunosorbent assay. In these subjects, the IgG fraction in low dilutions of serum (1:8, 1:16) reacted with as many as 10 proteins in phase I and phase II antigens. This number of reacting bands seen in Western blots was reduced by using serum in a dilution of 1:1024. In contrast, IgA antibodies were uncommon even at low dilutions. Likewise, IgA antibodies were infrequently observed in patients with acute Q fever. However, in chronic Q fever there were many IgA antibodies to phase I and phase II proteins. Antibodies to phase I proteins were more common than those to phase II proteins. Several antigenic protein bands were recognized only by serum from chronic Q fever patients. Three of these antigens had molecular masses of, respectively, 50 kDa, 80 kDa, and 160 kDa. Serial serum samples from patients with chronic Q fever revealed that the number of antigens recognized by the IgA fraction decreased after the initiation of antibiotic therapy. The decline was faster for antibodies to phase II proteins. We conclude that immunoblotting is useful in the diagnosis of chronic Q fever.

There is very little information available about the prevalence of Coxiella burnetii in Africa. We obtained blood from 75 Nigerians hospitalized in Sokoto for a variety of acute medical conditions. Their age range was 7 months to 50 years; there were 39 males and 36 females. Antibody titers were determined to phase I and phase II C. burnetii antigens using a microimmunofluorescence test. Thirty-three (44%) had an antibody titer of greater than or equal to 1:8 to phase II C. burnetii. We conclude on the basis of this limited survey that Q fever is common in Nigeria.

The antigenic structure of Coxiella burnetii is being investigated by identifying both external and internal cellular epitopes of the morphologic cell types. Both the phase I lipopolysaccharide (LPSI) and several surface proteins are candidates for the development of subunit multivalent vaccines. The protective efficacy of purified LPSI was demonstrated in A/J mice. The purified LPSI preparations contained residual peptides detected by amino acid analysis. Therefore, the protection afforded by LPSI may be, in part, due to the presence of peptides. The purification of proteins free of LPSI must be accomplished before the protective efficacy of proteins or peptides can be established. We have identified three proteins that are both antigenic and immunogenic, as indicated by either enzyme immunoassay, radioimmunoprecipitation, immunoblot assay, or lymphocyte transformation. A 62-kDa protein antigen encoded by the htpB gene of C. burnetii was analyzed for immunogenicity. The purified protein antigen was immunogenic, as it elicited specific antibodies and performed as recall antigen in lymphocyte stimulation assays. The antigen was not detected on the surface of phase I cells but was highly represented on the surface of phase II cells. Therefore, the protein may not be a good candidate for vaccine development. The diagnostic utility of the 62-kDa protein antigen lies in the fact that convalescent and chronic Q fever sera from human patients reacted with the antigen, whereas acute sera did not. Although the 62-kDa protein is a "common antigen," specific peptide-based diagnostic reagents may be useful in the detection of Q fever disease progression. A major surface protein (P1) of roughly 29.5 kDa was purified from the phase I Nine Mile (clone 7) strain. No LPSI was detected in the P1 preparation by three different LPSI monoclonal antibodies. Monoclonal antibodies prepared against P1 were effective in localizing the protein on the cell surface, in the cell wall, and associated with the peptidoglycan of large cells of C. burnetii. Small, pressure-resistant cells did not contain P1. Mice immunized with two 25-micrograms injections of LPSI produced antibodies against LPSI and phase I whole cells. No antibody was detected against phase II whole cells. Immunization with P1 induced antibody against the LPSI fraction and phase I and phase II whole cells. P1 was more effective than LPSI in reducing the number of infectious C. burnetii in the spleens of challenged mice. The gene encoding another protein (P2) recognized by P1 monoclonal antibodies was cloned and sequenced.(ABSTRACT TRUNCATED AT 400 WORDS)

A survey was carried out on 23 sheep flocks to estimate the frequency of abortion as well as the prevalence of antibodies against abortive infections. During the visit of each farm, a questionnaire was completed with the collaboration of the owner and blood samples were collected from all aborted ewes and some of those with normal lambing. A rate of 7% abortion was reached in both aborted and normally lambed ewes. Anti-Chlamydia psittaci antibodies were the most frequently detected (14 flocks). Anti-Coxiella burnetii and anti-Toxoplasma gondii antibodies were found in 9 flocks, whereas anti-Brucella and anti-Salmonella abortus ovis were present in only 1 flock each. None of the 5 infections was detected in 2 flocks. Mixed infections were prevalent: 13 flocks were simultaneously infected by at least 2 abortive pathogens. The procedure used does not allow the cause of abortion to be identified in all cases.

The encounter of phase I C. burnetii with the host results in seemingly disparate consequences. On the one hand, in vitro lymphocyte responses to mitogens and homologous recall antigen are suppressed. On the other, host resistance to a variety of infectious agents and to a tumor is increased. An explanation for this augmented immune response surely involves the ability of C. burnetii to stimulate cytokines, such as interferon and TNF, which enhance host immune function.

A retrospective serological survey was carried out using sera obtained from women at childbirth in the southern desert region of Israel to determine exposure experience to three rickettsial agents: Coxiella burnetii, Rickettsia typhi, and spotted fever group rickettsiae. Using the indirect fluorescent antibody method for determining IgG antibodies, it was found that about 40% of all sera examined demonstrated antibodies to one or more rickettsiae. Bedouin women appeared to be at greater risk of having antibodies to C. burnetii and spotted fever group rickettsiae than did Jewish residents of Beersheba, agricultural settlements, and development towns. The residents of development towns appeared to be at lower risk of developing antibodies to spotted fever group rickettsiae than did other populations sampled. Possible reasons for these differences are discussed.

There are very few reports in the literature on mass seroepidemiological studies of the prevalence of Q fever antibodies. Those to be found are mostly concerned with focal epidemics of Q fever. Out of more than 2000 serum specimens from blood donors analyzed at random in Switzerland in 1982 for Q fever antibodies, 3.5% gave seropositive results (CBR titer at least 1:10). In selected population samples, e.g. forestry workers or vets, the prevalence was distinctly higher (7.6% and 25.7%). Contact with sheep and the consumption of unboiled and not commercially processed milk proved to be mutually independent risk factors. The foregoing findings indicate that the incidence of Q fever in Switzerland is higher than would appear from the number of cases reported to the Federal Health Department. They are discussed in view of the recent renewed interest in Q fever endocarditis.

A lipopolysaccharide obtained in a dialyzed phenol extract from the rickettsia Coxiella burneti produced the following effects in guinea pigs after intraperitoneal injection: hyperthermia, loss of body weight, increased liver weight and concomitant lipid infiltration, elevated levels of hepatic and plasma cortisol, increased incorporation of [(3)H]orotic acid into hepatic 28S and 18S ribosomal ribonucleic acid, increased incorporation of (14)C-labeled amino acids into liver and plasma protein, and leukocytosis. Most of these events also occur during infection of guinea pigs with C. burneti, and a causal relationship between the rickettsial lipopolysaccharide and the biochemical changes that occur during Q fever is suggested.

21 sera of 13 patients with erythema chronicum migrans (ECM) and related diseases were examined for the presence of antibodies against 14 rickettsial antigens by using 3 serological techniques in 2 laboratories. Most tests were negative. This indicates that ECM and related diseases are not due to an hitherto known rickettsia. 5 out of 158 tests showed, however, positive reactions in low titers against Rickettsia akari and Coxiella burnetii. Thus, an antigenic relationship may exist between one or more rickettsia(e) and the etiologic agent of ECM.

The combined use of the complement fixation test, the indirect immunofluorescent test and the ring precipitation test with C. burnetii antigen greatly enhances the effectiveness or serological study, as it allows not only to find out the spread of infection among population more completely, but also to differentiate, to a certain extent, "fresh" infectious process from immunological trace reaction. The arguments are presented in favor of introducing the antigen of C. burnetii, phase I, into practice, especially in surveying the sera of farm animals.

Mild acid hydrolysis of phase I Coxiella burnetii cells gradually cleaved off polysaccharide chains, which contain determinants of antigen 1, thus unmasking determinants of the diagnostically important antigen 2. This artificial phase I to phase II conversion was reflected by changed serological activity in the complement-fixation (CF) test and by changed nonspecific phagocytosis. The product of mild acid hydrolysis can serve as a complex diagnostic preparation, because it reacts in the CF test with antibodies directed to both C. burnetii antigens.

In France Q fever is a notifiable disease, but information could only be obtained from fragmental inquiries. Routine serologic examination of 75 901 sera during the years 1982-85 showed 430 Q fever positive sera (mean rate 0.57%). Our laboratory could confirm 135 cases of clinical Q fever. In the region of Dijon, 4.4% of the country population was seropositive, but among breeders 25% were positive. 36 persons (8%) among 245 employees of the departmental veterinary services were seropositive, some with very high titers, but without obvious clinical symptoms. It seems that infection by Coxiella burnetii is still largely underestimated.

A case of Legionnaires' disease associated with Q fever is presented; this is probably the first reported case of concomitant infections with Legionella pneumophila and Coxiella burnetii. The pulmonary features of the illness were not typical of either disease. The difficulties of making a diagnosis in such cases are discussed.