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PubMed2002

Positive polymerase chain reaction and histology with borderline serology in Parinaud's oculoglandular syndrome.

PURPOSE: To report a case of Parinaud's oculoglandular syndrome (POS) in which, despite a borderline serology, polymerase chain reaction (PCR) testing for a conjunctival biopsy was positive for Bartonella henselae, a source of cat-scratch disease. A Steiner silver stain demonstrated the organism. METHODS: Case Report. RESULTS: A 65-year-old man was seen for a foreign body in his left eye (OS) associated with chemosis and a preauricular node. CONCLUSION: B. henselae is a known cause of POS. This gram-negative pleomorphic rod has been more frequently discovered in connection with this syndrome due to improved diagnostic testing such as indirect immunofluorescence antibody and PCR testing. Frequently, serology is positive if the organism is present. This report describes a patient with clinical findings of POS who, despite borderline serology, had pleomorphic rods on Steiner silver stain and positive PCR testing compatible with Bartonella henselae.

Aged↗
PubMed2001

Survey of veterinary professionals for antibodies to Bartonella henselae in Japan.

The seroprevalence of Bartonella henselae among veterinary professionals in Japan was investigated by means of an immunoperoxidase (IP) test that used protein A-horseradish peroxidase conjugate. Sera were obtained from 233 veterinary professionals in the Tokyo and Chiba areas. As negative control group, sera from 155 healthy individuals (all medical students) were used. As positive control group, sera from 5 patients highly suspected of cat scratch disease (CSD) by clinical symptoms were tested. Serum antibody titers of > or = 200 to B. henselae were presumed seropositive, because the titer at which about 95.5% of all healthy individuals (148 of the 155) were negative, and 2 of the 5 suspected CSD patients' serum antibody titers were > or = 200. Of the individuals in the veterinary professionals group tested, 35 of the 233 (15.0%) were seropositive for B. henselae. Females were nearly twice as likely as males to have antibodies to B. henselae in the veterinary professionals group. Our data suggest that Japanese veterinary professionals, and in particular younger females who are veterinary assistants and animal beauticians are more often infected by B. henselae.

Adolescent↗
PubMed1993

Proteinase K-sensitive and filterable phagosome-lysosome fusion inhibiting factor in Afipia felis.

Afipia felis is one of the putative agents of cat scratch disease (CSD). Its intracellular location was previously demonstrated in tissue sections. It has been recently grown in human monocytes and in HeLa cells. We report that A. felis may infect human macrophages and the P 388 D1 murine macrophage cell line, yet is unable to grow in the cell-free culture medium. In both human macrophages and P 388 D1, the organisms multiply in a vacuole after active inhibition of phagosome-lysosome fusion (P-L) as demonstrated with cationized ferritin-labeled lysosomes. This P-L fusion inhibition is diffusable and is due to a filterable factor which is inactivated by protease digestion. The pathogenic role of A. felis in CSD may result from its ability to invade and multiply in macrophages.

Animals↗
PubMed1993

Antibiotic susceptibilities of Afipia felis in axenic medium and in cells.

Afipia felis, one of the putative agents of cat scratch disease (CSD), is a facultative intracellular bacterium. Although CSD is considered not to be susceptible to antibiotic therapy, sporadic case reports indicated that aminoglycosides may be effective. We determined the in vitro antibiotic susceptibilities of three A. felis strains in axenic medium and in a cell model. In axenic medium, A. felis was susceptible to imipenem, aminoglycosides, and rifampin when using either the broth dilution technique or the agar technique. When grown in HeLa cells, A. felis was susceptible to amikacin and tobramycin but was resistant to the other compounds tested. Despite its intracellular location, A. felis can apparently be reached by aminoglycosides. Thus, the in vitro data presented here are in accord with the clinical data obtained in patients suffering CSD.

Anti-Bacterial Agents↗
PubMed1997

Immunologic response to Bartonella henselae as determined by enzyme immunoassay and Western blot analysis.

Bartonella henselae is now regarded as the etiologic agent of cat-scratch disease and a cause of bacillary angiomatosis. We examined the human immune response to Bartonella henselae infection using a newly developed enzyme immunoassay (EIA) and a Western blot procedure using outer-membrane proteins. The EIA showed 98.6% and 91.4% agreement with an indirect fluorescence method (IFA) for detection of IgM and IgG antibodies, respectively. By using Western blot analysis, reactivity to an 8-kd band showed significant correlation with positive results by the IgM IFA and EIA. In contrast, reactivity to 209-, 208.5-, 208-, 116-, and 80-kd bands was identified only in positive IgG IFA serum samples. The EIA and Western blot should be useful tests in determining the antibody response to B. henselae infection and may also be important in determining the critical epitopes in the host-parasite interaction of this organism.

Adolescent↗
PubMed1999

Prevalence of Bartonella henselae immunoglobulin G antibodies in Singaporean cats.

BACKGROUND: Bartonella henselae causes several clinical diseases in humans. The most common infection is the classical cat-scratch disease (CSD) occurring in immunocompetent and immunocompromised children exposed to newly acquired kittens. This is the first study of B. henselae infection in Singaporean cat population. METHODS: Indirect fluorescent antibody tests were carried out on cat sera obtained from 80 cat blood samples. RESULTS: Of the cats studied in this project 47.5% tested seropositive, with high IgG titers (> or =1024) in 31 of 38 seropositive cats, 60.5% of which were males. CONCLUSIONS: Males cats had higher rates of infection than female cats. Serologic studies of cats in different parts of the United States have indicated that the prevalence of IgG antibody to B. henselae in cats is highest in regions with warm, humid climates and lowest in areas with cool, dry climates. The indirect fluorescent antibody test data for B. henselae infection in Singapore cats support this contention.

Animals↗
PubMed1994

Persistent generalized lymphadenopathy and non-Hodgkin's lymphoma in AIDS: association with Rochalimaea henselae infection.

Cat scratch disease, which is caused by infection with Rochalimaea henselae, is often manifested as lymphadenopathy. R. henselae has also been isolated from human immunodeficiency virus (HIV)-positive patients with bacillary angiomatosis. In order to determine the frequency of R. henselae-reactive antibodies in HIV-positive patients with persistent generalized lymphadenopathy (PGL) or non-Hodgkin's lymphoma (NHL), we tested a total of 124 HIV-positive patients for R. henselae-reactive immunoglobulin G (IgG), IgM, and IgA antibodies by an enzyme immunoassay procedure using whole R. henselae antigen. Of the patients, 7 had PGL, 17 had NHL, and 100 were HIV stage IV (Centers for Disease Control criteria). A total of 86% of PGL patients (6 of 7) were positive for R. henselae antibodies (three were positive for IgG, IgA, and IgM, one was positive for IgG and IgA only, and two were positive for IgG only). A total of 29% of NHL patients (5 of 17) were positive for R. henselae antibodies (two were positive for IgG, IgA, and IgM and three were positive for IgG only). Only 5% of HIV Stage IV patients without adenopathy (5 of 100) were positive for R. henselae-reactive IgG, IgA, and IgM. The high prevalence of R. henselae-reactive antibodies in HIV-positive PGL and NHL patients suggests that R. henselae is a potential etiologic agent or cofactor in these patients.

AIDS-Related Complex↗
PubMed1994

Downregulation of human polymorphonuclear cell activities exerted by microorganisms belonging to the alpha-2 subgroup of Proteobacteria (Afipia felis and Rochalimaea henselae).

Intracellular pathogens have evolved effective mechanisms in order to survive in an intracellular environment, thus avoiding destruction by phagocytic cells. In this regard, a correlation between resistance to phagocytic killing and expression of pathogenic potency has been established. In this report, we have studied the interaction between human polymorphonuclear cells (PMN) and two gram-negative microorganisms, Afipia felis and Rochalimaea henselae, which belong to the alpha-2 subgroup of the class Proteobacteria. A. falis has been previously proposed as the causative agent of Cat Scratch Disease (CSD), but several recent lines of evidence attribute a major role to R. henselae. Of note, CSD is a syndrome characterized by a chronic lymphoadenopathy, involving macrophages and endothelial cells with a progression towards a granulomatous process and/or angiogenesis. Since members of the alpha-2 subgroup of Proteobacteria have the property to survive intracellularly, we have evaluated the effects exerted by A. felis and R. henselae on human PMN in terms of chemotaxis locomotion, degranulation and oxidative metabolism. Results will show an impairment of PMN activities as a consequence of the challenge with both microrganisms. In particular, inhibition of PMN oxidative function occurred either as result of a direct exposure to both A. felis and R. henselae or when PMN were primed by bacteria for the N-formyl-methionyl-leucyl-phenylalanine enhancement of the oxidative burst. These findings may account for the ability of A. felis and R. henselae to survive within PMN as expression of a further mechanism of pathogenic potency, influencing also the nature and the evolution of inflammatory response in the lesion sites.

Bartonella henselae↗
PubMed1999

Identification of Bartonella species directly in clinical specimens by PCR-restriction fragment length polymorphism analysis of a 16S rRNA gene fragment.

It is now established that two species of Bartonella, namely, Bartonella henselae and B. quintana, cause bacillary angiomatosis in human immunodeficiency virus-infected patients. In addition, B. henselae causes cat scratch disease and B. quintana, B. henselae, and B. elizabethae can cause bacteremia and endocarditis in immunocompetent persons. We have developed a PCR-restriction fragment length polymorphism-based assay for direct detection and identification to species level of Bartonella in clinical specimens. This is accomplished by PCR amplification of Bartonella DNA using primers derived from conserved regions of the gene carrying the 16S ribosomal DNA, followed by restriction analysis using DdeI and MseI restriction endonucleases. We amplified a Bartonella genus-specific 296-bp fragment from 25 clinical samples obtained from 25 different individuals. Restriction analysis of amplicons showed that identical patterns were seen from digestion of B. henselae and B. quintana amplicons with DdeI, whereas a different unique pattern was seen by using the same enzyme with B. vinsonii and B. elizabethae. With MseI digestion, B. henselae and B. vinsonii gave nearly identical patterns while B. quintana and B. elizabethae gave a different pattern. By combining the restriction analysis data generated with MseI and DdeI, unique "signature" restriction patterns characteristic for each species were obtained. These patterns were useful in identifying the Bartonella species associated with each tissue specimen.

Angiomatosis, Bacillary↗
PubMed2001

[Serological cross-reaction among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody method].

We studied the serological cross-reactions among Bartonella henselae, Chlamydia pneumoniae and Coxiella burnetii by indirect fluorescence antibody (IFA) method, using sera from 8 patients with cat scratch disease (CSD), 13 patients with C. pneumoniae infection and 12 patients with acute Q fever. B. henselae IgG antibody was negative in 13 patients with C. pneumoniae infection, and was positive in 3 (titers being 1:64) of 12 patients with Q fever, whereas B. henselae IgM antibody was negative in all the patients with C. pneumoniae infection or Q fever. C. burnetii IgG antibody was removed by absorption of these 3 sera with C. burnetii antigens, whereas B. henselae IgG antibody did not change. C. pneumoniae IgG antibody was positive in 3 (titers being 1:125 in two, 1:32 in one) of 8 patients with CSD. Both C. pneumoniae and B. henselae IgG antibody titers were significantly reduced by absorption of these 3 sera with B. henselae antigens. C. burnetii IgG or IgM antibodies were negative in all patients with CSD. In conclusion, no serological cross-reaction between B. henselae and C. burnetii was observed. On the other hand. B. henselae IgG antibody cross-reacted to C. pneumoniae antigens, whereas C. pneumoniae IgG antibody did not cross-react to B. henselae antigens. Our findings suggest that determination of B. henselae IgG or IgM antibodies were not influenced by C. pneumoniae and C. burnetii antigens.

Bartonella henselae↗
PubMed1998

[Determination of anti-Bartonella henselae antibody by indirect fluorescence antibody test--comparison of two types of antigen: non-cocultivated B. henselae and cocultivated B. henselae with Vero cells].

Serum anti-Bartonella henselae IgG and IgM antibody titers for the diagnosis of cat scratch disease (CSD) were determined by indirect fluorescence antibody (IFA) tests. B. henselae as antigen were harvested either by cocultivating with Vero cells (cocultivated B. henselae) or by cultivating without them (non-cocultivated B. henselae). Based on the results on 110 healthy adults, cut off values were set at 1:32 for IgG, and < 1:20 for IgM antibodies. According to these criteria, IgG antibody was positive in 2.7% of the 110 adults, while nobody was positive for IgM antibody. The titers did not change depending on the types of antigen used. On the other hand, IgG antibody titers against cocultivated B. henselae tended to be higher than those against non-cocultivated B. henselae in 33 CSD suspected patients; 75.8% of the patients were anti-B. henselae IgG positive when tested with cocultivated B. henselae as antigen, while only 48.5% of the same patients gave positive results with non-cocultivated B. henselae. Anti-B. henselae IgM antibody was positive in 24.2% of the 33 CSD suspected patients against both types antigen. Vero cells themselves seemed to nonspecifically bind some IgM (but not IgG). We recommended cocultivated B. henselae as antigen for IgG IFA, and non-cocultivated B. henselae for IgM IFA in the serological tests of CSD.

Adult↗
PubMed2002

Comparison of in-house and commercial slides for detection by immunofluorescence of immunoglobulins G and M against Bartonella henselae and Bartonella quintana.

We compared the sensitivities and specificities of indirect fluorescent antibody tests developed in our laboratory and commercially available from Focus Technologies (FT; formerly MRL Diagnostic) for detection of serum antibodies to Bartonella spp. Serum samples tested were from patients with culture- or PCR-confirmed Bartonella quintana or B. henselae infections causing cat scratch disease (CSD), chronic bacteremia, or endocarditis. At a cutoff titer of 64, the FT test had higher sensitivity than our in-house test in detecting anti-B. henselae immunoglobulin G (IgG) antibodies in CSD patients (91.2 versus 52.9%; P < 0.001). The specificity in serum samples from 85 control patients was, however, lower with the FT test (87%) than with the in-house test (98.8%) (P = 0.002). A cutoff titer of 128 improves the specificity for the FT test but lowers the sensitivity to 85%. For patients infected with B. henselae, our in-house test, but not the FT test, enabled endocarditis to be detected more reliably. With the in-house test, titers of IgG against B. henselae of >/=1,024 were found only in endocarditis patients and not in CSD patients. With the FT test, 19.1% of CSD patients had titers of IgG against B. henselae of >/=1,024 (P < 0.001). Our in-house technique also improved detection of anti-B. quintana antibodies in homeless patients with endocarditis. IgG titers of >/=1,024 were present in 75% of serum samples, but only in 16.7% of serum samples with the FT test (P = 0.004). Since each test has advantages over the other, the serological diagnosis of Bartonella infections would benefit if both tests were used concurrently.

Antibodies, Bacterial↗
PubMed1998

Use of the cell division protein FtsZ as a means of differentiating among Bartonella species.

Genes coding for homologs of the highly conserved cell division protein FtsZ were isolated from Bartonella henselae and Bartonella quintana, the causative agents of cat scratch disease and trench fever, respectively. DNA fragments coding for the ftsZ open reading frames (ORFs) were cloned into Escherichia coli following PCR amplification with primers based on the ftsZ sequence of the closely related species Bartonella bacilliformis. The amino acid sequences predicted from the cloned B. henselae and B. quintana ftsZ ORFs are 81 to 83% identical to the corresponding protein in B. bacilliformis. Like the FtsZ protein of B. bacilliformis, the B. henselae and B. quintana homologs are about twice as large as the FtsZ proteins reported in most other organisms. Localized sequence differences within the C-terminal coding regions of the Bartonella ftsZ genes were used as the basis for species-specific identification of these organisms at both the DNA and protein levels. Oligonucleotide primers which permit the amplification of an ftsZ fragment from each of the Bartonella species without amplifying DNA from the other two species were designed. Anti-FtsZ antisera raised in rabbits against synthetic peptides corresponding to the relatively divergent C-terminal regions were shown via Western blot analysis to react only with the FtsZ protein from the cognate Bartonella species. These observations raise the possibility that the differences in ftsZ sequences can be used as the basis for diagnostic tests to differentiate among these closely related pathogens.

Amino Acid Sequence↗
PubMed1998

Characterization of human immunoglobulin (Ig) isotype and IgG subclass response to Bartonella henselae infection.

Serologic parameters of cat scratch disease (CSD) were evaluated by Western blot analysis. Sera from patients with serologically confirmed CSD antigen were screened for immunoglobulin (Ig) isotype-specific as well as IgG subclass-specific reactivity against Bartonella henselae whole-cell antigen. Bartonella-negative control sera were used to determine baseline antibody activity. Heterogeneous B. henselae-specific IgG reactivity with numerous protein bands, ranging from >150 to <17 kDa, was observed. Though individual banding patterns were variable, one approximately 83-kDa B. henselae protein (Bh83) was immunoreactive with all CSD sera tested, suggesting it is a conserved antigen during infection. Bh83 was not recognized by reference human antisera against Rickettsia rickettsii, Chlamydia group positive, Treponema pallidum, Orientia tsutsugamushi, Fransciscella tularensis, Ehrlichia chaffeensis, Mycoplasma pneumoniae, and Escherichia coli, although other cross-reactive proteins were evident. Significantly, CSD sera failed to recognize the 83-kDa protein when tested against Bartonella quintana antigen, though sera from B. quintana-infected patients did react to Bh83. This cross-reactivity suggests epitope conservation during infection with B. henselae or B. quintana. Western blot analysis further revealed similar banding patterns when B. henselae was reacted against the Ig isotypes IgG and IgG1 and both secretory and alpha chains of IgA. Neither IgM nor IgE reacted significantly to Bartonella antigen by our Western blot analysis. Dissection of the antibody response at the IgG subclass level indicated that prominent antigen recognition was limited to IgG1. These observations provide insight into induced immunity during CSD and provide evidence for conserved epitope expression during infection with B. henselae or B. quintana.

Antibodies, Bacterial↗
PubMed1994

Rochalimaea henselae infection. A new zoonosis with the domestic cat as reservoir.

OBJECTIVE: To determine the reservoir and vector(s) for Rochalimaea henselae, a causative agent of bacillary angiomatosis (BA) and cat scratch disease, and to estimate the percentage of domestic cats with R henselae bacteremia in the Greater San Francisco Bay Region of Northern California. DESIGN: Hospital-based survey of patients diagnosed with BA who also had significant exposure to at least one pet cat, as well as a convenience sampling of pet or impounded cats for prevalence of Rochalimaea bacteremia. SETTING: Community and university hospitals and clinics; veterinary clinics treating privately owned or impounded cats. PATIENTS: Patients with or without human immunodeficiency virus infection, with biopsy-confirmed BA, who had prolonged exposure to pet cats prior to developing BA. MAIN OUTCOME MEASURES: Cultures and laboratory studies were performed on blood drawn from pet cats associated with patients with BA. The Rochalimaea species infecting pet cats and fleas and causing the BA lesions in human contacts of these cats was identified by culture, polymerase chain reaction-restriction fragment length polymorphism analysis, and DNA sequencing. The presence of R henselae bacteremia in pet cats was documented, and predictor variables for culture positivity were evaluated. RESULTS: Four patients diagnosed with BA who had prolonged contact with seven pet cats were identified. The Rochalimaea species causing BA lesions in these patients was determined to be R henselae. The seven pet cats were found to be bacteremic with R henselae; this bacterium was also detected in fleas taken from an infected cat by both direct culture and polymerase chain reaction. Blood samples were cultured from pet and impounded cats (N = 61) in the Greater San Francisco Bay Region, and R henselae was isolated from 41% (25/61) of these cats. CONCLUSION: We have documented that the domestic cat serves as a major persistent reservoir for R henselae, with prolonged, asymptomatic bacteremia from which humans, especially the immunocompromised, may acquire potentially serious infections. Antibiotic treatment of infected cats and control of flea infestation are potential strategies for decreasing human exposure to R henselae.

Adult↗
PubMed1995

Bartonella henselae prevalence in domestic cats in California: risk factors and association between bacteremia and antibody titers.

The isolation of Bartonella henselae, the agent of cat scratch disease, from the blood of naturally infected domestic cats and the demonstration that cats remain bacteremic for several months suggest that cats play a major role as a reservoir for this bacterium. A convenience sample of 205 cats from northern California was selected between 1992 and 1994 to evaluate the B. henselae antibody and bacteremia prevalences and to determine the risk factors and associations between bacteremia and antibody titers. B. henselae was isolated from the blood of 81 cats (39.5%). Forty-two (52%) of these bacteremic cats were found to be infected with > or = 1,000 CFU/ml of blood. Impounded or former stray cats were 2.86 (95% confidence interval [CI] = 1.94, 4.22) times more likely to be bacteremic than the pet cats. Young cats ( < 1 year old) were more likely than adult cats to be bacteremic (relative risk = 1.64; (95% CI = 1.19, 2.28). Bacteremic cats were more likely than nonbacteremic cats to be infested with fleas (relative risk = 1.64; 95% CI = 1.38, 1.96). No association between B. henselae infection and feline immunodeficiency virus antibody prevalence was observed. Eighty-one percent of the cats (166 of 205) tested positive for B. henselae antibodies, and titers were higher in bacteremic than in nonbacteremic cats. Multiple logistic regression analysis indicated that younger age and seropositivity for B. henselae antibodies were associated with bacteremia. Serological screening for Bartonella antibodies may not be useful for the identification of bacteremic cats (positive predictive value = 46.4%), but the lack of antibodies to B. henselae was highly predictive of the absence of bacteremia (negative predictive value = 89.7%). Seronegative cats may be more appropriate pets for immunocompromised individuals who are at increased risk for developing severe B. henselae disease.

Aging↗