Perinuclear ANCA are predominantly IgG antibodies directed against cytoplasmic antigens in neutrophils and other granulocytes and monocytes.1,2 For decades, pANCA have been used in humans as diagnostic and prognostic markers for several autoimmune diseases, such as autoimmune systemic vasculitis, glomerulonephritis,3–6 and IBD, including Crohn's disease and ulcerative colitis.1,7,8 Detection via IFA of pANCA on ethanol-fixed neutrophils is considered the most accurate assay for the detection of ANCA in dogs and humans with IBD.9–12
Only a few studies9,11,13,14 have been conducted to investigate the role of pANCA in diseases of dogs. Investigators in these studies have focused primarily on chronic enteropathies with or without protein-losing nephropathy. Similar to results for human patients, the presence of pANCA in the serum of dogs is associated with IBD.13–15 It has also been suggested that pANCA are associated with food-responsive disease in dogs and could become a useful noninvasive clinical marker for the diagnosis of chronic enteropathy in this subgroup of dogs.11
Various other immune-mediated diseases in humans, including a spectrum of inflammatory and infectious diseases, may be associated with positive results for pANCA testing.16 For example, pANCA have been identified in human patients with IMHA that developed concurrently with other autoimmune diseases.16–18 Although pANCA have been reported in humans with various infectious diseases, only a limited number of studies19–21 have been conducted to address pANCA status in human patients with specific infections, such as infections with Mycobacterium spp. In case reports,22–24 human patients with toxocariasis, visceral leishmaniasis, and endocarditis caused by Bartonella quintana and Bartonella henselae had positive results when tested for pANCA. There are limited data regarding the prevalence of pANCA in dogs with infectious diseases. In 1 study,14 investigators determined that the specificity of pANCA in dogs with IBD was only 76%, compared with the specificity for dogs with other inflammatory and infectious disorders. However, the dogs in that study14 were not infected with a specific pathogen. Infectious disorders caused by the vector-borne pathogens B henselae, Bartonella vinsonii subsp berkhoffii, Ehrlichia canis, and Leishmania infantum are associated with an immune-mediated pathogenesis.25–27 Investigators in a previous study25 found that dog sera reactive for B vinsonii subsp berkhoffii, L infantum, and E canis were likely to have positive results for ANA. Hence, pANCA might have a similar pattern. Thus, determining pANCA status in certain dog populations with defined exposure to various infectious organisms (such as vector-borne diseases), and in association with specific autoimmune diseases (such as IMHA), would provide clinically important diagnostic information regarding the usefulness of pANCA as a marker of disease for IBD as well as for infectious and immune-mediated inflammatory disease in dogs. Therefore, the objective of the study reported here was to investigate the prevalence of pANCA in dogs with IMHA and various vector-borne infectious diseases.
Materials and Methods
Dogs—To evaluate the diagnostic accuracy of pANCA for IMHA and vector-borne infectious diseases, serum samples were obtained from dogs in which IMHA was diagnosed or suspected clinically, dogs in which vector-borne infections were confirmed or suspected, and control dogs. Owner consent was not necessary because the samples were collected for diagnostic reasons in each case and the rest of the volume was stored and used retrospectively in the present study. The study was approved by the Royal Veterinary College's Ethics Committee.
Dogs in which IMHA was diagnosed or suspected clinically
Serum samplesa from 16 dogs with primary IMHA, as determined after examination at the University of London Royal Veterinary College Queen Mother Hospital, were used in the study. These dogs were all females, except for 1 male, and 13 of 16 were neutered. Mean ± SD age was 8.1 ± 2.66 years (range, 2.11 to 13.40 years). The diagnosis of IMHA required that the dogs were anemic (PCV < 37%), had positive results for a saline (0.9% NaCl) solution agglutination test, and had positive results for a direct Coombs' test or moderate to marked spherocytosis on a peripheral blood smear.28
Thirty-nine dogs were suspected clinically of having IMHA by the attending veterinarian. Serum samples from these dogs were obtained from the University of Bristol Clinical Immunology Diagnostic Laboratory at the School of Veterinary Sciences. Mean ± SD age of 36 dogs was 6.3 ± 3.39 years (range, 4 months to 15 years); the age of 3 dogs was not known. Twenty dogs were females, and 17 were males; sex of 2 dogs was not known. These 39 dogs were classified into dogs with positive (n = 19) and negative (20) results of a Coombs' test. Further clinical information was not available for these dogs.
Dogs in which vector-borne disease was confirmed or suspected
Ninety-seven serum samples from dogs of various breeds were submitted to the Vector-Borne Diseases Diagnostic Laboratory at North Carolina State University by veterinarians who suspected infection with a tick-borne pathogen; these sera were found to be seroreactive for vector-borne pathogens. Information was not available regarding sex, age, and clinical status or final diagnosis. All samples were tested via IFA for seroreactivity for R rickettsii, B henselae, B vinsonii subsp berkhoffii, and E canis.25,29 An antibody titer ≥ 1:64 was considered indicative that the dog was seroreactive. Dogs with titers between 1:64 and 1:128 were classified as low seroreactors, and dogs with titers ≥ 1:256 were considered as medium to high seroreactors. Enzyme-linked immunosorbent assaysb,c were used to detect antibodies against B burgdorferi C6 peptide in the serum samples.30
Dogs seroreactive for L infantum included 43 dogs of various breeds with clinical signs and clinicopathologic abnormalities compatible with leishmaniasis and with high antibody titers against L infantum. Serum samples were submitted to the Laboratorio Veterinario Privato San Marco in Padova, Italy. An in-house quantitative ELISA was used to test all samples, as described elsewhere.31 Sera from these dogs were not tested for exposure to other vector-borne pathogens.
Control dogs
Serum samples were included from 20 dogs of unknown breeds that were suspected by the attending veterinarian of being infected with a tick-borne disease but in which antibodies against the defined vector-borne pathogens were not detected. Serum samples were tested by personnel at the Vector-Borne Diseases Diagnostic Laboratory. All of these dogs were seronegative for antigens of the tick-borne pathogens R rickettsii, B henselae, B vinsonii subsp berkhoffii, E canis, and B burgdorferi.
Sick dogs in which leishmaniasis was clinically suspected but antibodies against L infantum were not detected comprised 22 dogs of various breeds tested at the Laboratorio Privato Veterinario San Marco and found to be seronegative for L infantum. These dogs were not tested for exposure to other vector-borne pathogens, and no clinical information was available for these dogs.
Blood samples were collected from 20 dogs (13 males and 7 females) of various breeds during a general health screen at the Queen Mother Hospital. Mean ± SD age was 6.5 ± 4.37 years (range, 1.0 to 14.2 years). Residual serum samples were used as a healthy control group for the study. Nine dogs had orthopedic problems, 1 was admitted for castration, and 10 were healthy with no clinical signs. None of these dogs had clinical evidence of infectious or autoimmune diseases.
pANCA assay—Detection of pANCA via IFA and isolation of granulocytes was performed as described elsewhere.13 Briefly, prepared IFA slides were overlaid with thawed canine serum (dilutions 1:10 and 1:20), incubated for 1 hour, and then washed. Each slide contained a positive serum sample (1:10 dilution) and a negative control sample. Fluorescein isothiocyanate–labeled secondary antibodyd (1:100 dilution) was added, and the slides were incubated again (1 hour in a humid chamber at 22°C) and then washed. Fluorescence microscopy was used to evaluate the results. Slides were examined separately by 2 investigators (KA and AEK) who were not aware of the identity of dogs from which the serum samples were obtained. For samples with uncertain results, tests were repeated until samples were clearly identified as having positive or negative results. Samples with perinuclear immunofluorescence in the neutrophils were considered positive for pANCA, and samples with intranuclear or atypical labeling for pANCA were considered negative for pANCA (Figure 1).
Photomicrographs indicating the typical perinuclear staining pattern of canine granulocytes in a sample obtained from a dog with positive results (A) and a dog with negative results (B) for pANCA. Indirect immunofluorescence staining; bar = 100 μm.
Citation: American Journal of Veterinary Research 73, 9; 10.2460/ajvr.73.9.1403
Statistical analysis—The χ2 or Fisher exact test was used to identify significant differences in pANCA prevalence between the disease groups and the control groups. An OR was then calculated to measure the strength of the association. Significance was set at values of P < 0.05. Data analysis was performed with statistical software.e
Results
The percentage of pANCA-positive dogs in each group and subgroup was summarized (Table 1). Among the dogs in which IMHA was diagnosed or suspected clinically, the prevalence of pANCA-positive dogs (11/16) was highest in the subgroup of dogs in which IMHA was confirmed. Dogs with IMHA were 16.27 times (95% CI, 4.11 to 54.34; P < 0.001) as likely to have positive results for pANCA as were the healthy control dogs. There was no association between positive results for pANCA and positive or negative results for the Coombs' test for dogs suspected of having IMHA.
Summary of pANCA results for serum samples obtained from several groups of dogs.
| Group | No. (%) of dogs with positive results for pANCA | 95% CI |
|---|---|---|
| Dogs in which IMHA was diagnosed or suspected clinically (n = 55) | ||
| IMHA diagnosed (n = 16) | 11 (69) | 46–92 |
| IMHA suspected; positive result for Coombs' test (n = 19) | 6 (32) | 11–53 |
| IMHA suspected; negative result for Coombs' test (n = 20) | 9 (45) | 23–67 |
| Dogs confirmed or suspected of having vector-borne infections (n = 140) | ||
| Clinically suspected and seroreactive for ≥ 1 tick-borne diseases including Rickettsia rickettsii, Bartonella henselae, Bartonella vinsonii subsp berkhoffii, Ehrlichia canis, and Borrelia burgdorferi infection (n = 97) | 39 (40) | 30–50 |
| Clinically suspected and seroreactive for Leishmania infantum (n = 43) | 28 (65) | 51–79 |
| Control dogs (n = 62) | ||
| Dogs clinically suspected but seronegative for tick-borne pathogens (n = 20) | 1 (5) | 0–25 |
| Dogs clinically suspected but seronegative for L infantum (n = 22) | 8 (36) | 20–57 |
| Healthy dogs (n = 20) | 1 (5) | 0–25 |
Of the 140 dogs seroreactive for ≥ 1 vector-borne disease pathogen, 67 (48%) had positive results for pANCA. Of the 97 dogs seroreactive for R rickettsii, B henselae, B vinsonii subsp berkhoffii, or E canis, 39 (40%) had positive results for pANCA. Of the 43 dogs seroreactive for L infantum, 28 (65%) had positive results for pANCA (Table 1).
A significant association was detected between a positive pANCA result and seroreactivity for any vector-borne pathogen evaluated in the study. Dogs seroreactive for any vector-borne pathogen were 17.4 times (95% CI, 2.27 to 133.85; P < 0.001) as likely to have positive results for pANCA as were healthy control dogs or dogs suspected by the attending veterinarian of being infected with a tick-borne pathogen (Table 2). The OR was 4.7 (95% CI, 1.72 to 12.96; P = 0.03) when the dogs seroreactive for a tick-borne pathogen were compared with the dogs suspected by the attending veterinarian of being infected with a tick-borne pathogen and healthy control dogs.
Summary of pANCA results for serum samples obtained from dogs confirmed or suspected of having vector-borne infections and comparison with results for healthy control dogs.
| Vector-borne pathogens | No. (%) of dogs with positive results for pANCA | OR* | 95% CI | P value† |
|---|---|---|---|---|
| Seroreactivity for at least 1 vector-borne pathogen (n = 140) | 67 (48) | 17.4 | 2.27–133.85 | < 0.001 |
| Seroreactivity for > 1 pathogen (n = 51) | 21 (41) | 13.3 | 1.65–107.18 | 0.003 |
| Seroreactivity for at least 1 tick-borne pathogen (n = 97) | 39 (40) | 4.7 | 1.72–12.96 | 0.003 |
| Rickettsia only (n = 15) | 5 (33) | 9.5 | 0.97–92.83 | 0.064 |
| Bartonella only (n = 29) | 13 (45) | 15.44 | 1.82–131.99 | 0.020 |
| Ehrlichia only (n = 12) | 5 (42) | 13.6 | 1.34–137.45 | 0.018 |
| Leishmania (n = 43) | 28 (65) | 35.5 | 4.32–291.49 | < 0.001 |
| Rickettsia, medium to high titers (n = 9)‡ | 3 (33) | 9.5 | 0.826–109.24 | 0.076 |
| Bartonella, medium to high titers (n = 17)‡ | 9 (53) | 21.4 | 2.31–197.79 | 0.001 |
| Ehrlichia, medium to high titers (n = 13)‡ | 8 (62) | 30.4 | 3.05–303.36 | < 0.001 |
Healthy control dogs were the referent group.
Values were considered significant at P < 0.05.
Medium to high titers were defined as titers ≥ 1:256.
To further investigate potential associations between pANCA and infection, dogs seroreactive for a tick-borne pathogen were allocated into subgroups on the basis of the antibody titer against vector-borne pathogens. For the 30 dogs with moderate to high IFA antibody titers against Bartonella or Ehrlichia spp antigens, there was a stronger association with positive results for pANCA (Table 2).
Of the 43 dogs infected with L infantum, 28 (65%) had positive results for pANCA. There was a significant (P < 0.001) association between a positive pANCA status and L infantum seroreactivity (OR, 35.5; 95% CI, 4.32 to 291.49), compared with results for healthy dogs (Table 2). Compared with dogs in which leishmaniasis was suspected but antibodies against L infantum were not detected, seroreactive dogs were significantly (P = 0.027) more likely (OR, 3.28; 95% CI, 1.12 to 9.54) to have positive results for pANCA. Compared with healthy control dogs, dogs seronegative for L infantum were significantly (P = 0.022) more likely to be seroreactive for pANCA (OR, 6.16; P = 0.022; 95% CI, 1.43 to 26.49).
Discussion
Perinuclear ANCA have been described in association with various inflammatory and immune-mediated gastrointestinal diseases in dogs.9,13,14 To the authors' knowledge, the present study represents the first attempt to investigate pANCA status in dogs exposed to specific pathogens and in dogs with IMHA. A large proportion of dogs (67/140 [48%]) that were seroreactive for various pathogens in the present study also had positive results for pANCA. Moreover, 11 of 16 of dogs with confirmed IMHA also had positive results for pANCA. There was a significant association between positive results for pANCA and a diagnosis of IMHA or exposure to tick- or sandfly-transmitted infectious diseases.
Authors of several case reports32–34 have described pANCA seroreactivity in association with human autoimmune or immune-mediated diseases, including IMHA, glomerulonephritis, systemic lupus erythematosus, and IBD. The results of the study reported here suggested that pANCA production could also be associated with autoimmune disorders or vector-borne infectious diseases in dogs. There is a need for future studies with well-characterized serum samples from various dog populations with complete clinical and historical data to define the diagnostic specificity and sensitivity of pANCA in dogs.
Interestingly, the highest pANCA prevalence was found in dogs with stringent inclusion criteria for IMHA. In contrast, there was no significant association between pANCA reactivity and the outcome of a Coombs' test. The prevalence of pANCA in Coombs' test–positive and −negative dogs was 6 of 19 (32%) and 9 of 20 (45%), respectively. Because of a lack of sensitivity, the Coombs' test will yield negative results for dogs with IMHA and positive results for dogs with bartonellosis.25,35–37 Therefore, the Coombs' test–positive and −negative dogs included in the present study, although suspected of having primary IMHA, could have been assigned incorrectly to a group or could have had underlying infectious or neoplastic disease. No other clinicopathologic information was available on these dogs, and a positive result for a Coombs' test by itself is not diagnostic of IMHA, so it is not possible to draw any further conclusions regarding these dogs.
Mechanisms by which an infectious agent could induce autoimmune reactivity have been described.38–40 For example, pANCA cross-reactivity with colonic bacteria has been reported in humans with ulcerative colitis.39 In particular, pANCA appear to cross-react with both Bacteroides spp (Bacteroides caccae and Bacteroides thetaitaotaomicron) and Escherichia coli.41 In addition, bacterial permeability-increasing protein, an endogenous antimicrobial protein directed against gram-negative bacteria, appears to target pANCA antigens.42 Moreover, bacterial permeability-increasing protein has affinity for endotoxins of gram-negative bacteria, which might explain the reason that pANCA develop in humans and dogs infected with these enteric pathogens. Neutrophils are the first inflammatory cell types recruited in any infection, so it is possible that antibodies against neutrophils are produced in association with many acute infections that develop in human and other animal patients.43–45 Moreover, it is thought that superantigens are involved in the pathogenesis of pANCA-associated IBD in humans because superantigens stimulate the immune system greatly and may ultimately lead to autoantibody production, including pANCA.46,47
In the present study, serologic responses against several vector-borne gram-negative bacteria (R rickettsii, B henselae, B vinsonii subsp berkhoffii, E canis, and B burgdorferi) were investigated. Gram-negative bacteria can be a target antigen of pANCA.41,42 Although dogs were seroreactive, it is difficult to ascertain whether all of the dogs had an active infection at the time blood samples were collected because there was no clinicopathologic information available for these dogs. However, it is likely that these dogs had been exposed to the specified pathogens at some time point before collection of blood samples; therefore, we speculate that the formation of pANCA was likely associated with these vector-borne infectious diseases.
Another theory supports defective apoptosis as a cause of pANCA development in patients with autoimmune and inflammatory disorders. Ineffective apoptosis or defective removal of cell debris by neutrophils and macrophages might lead to production of pANCA because substances that are normally intracellular are released and detected as antigens by the immune system.48,49 A better understanding of the immunopathogenesis of pANCA in a spectrum of inflammatory and infectious diseases would benefit both canine and human medicine. Dogs and humans share the same environment and similar or identical genetic diseases, so it is conceivable that dogs could be used to investigate environmental and genetic factors that influence pANCA induction, as described in human patients.48,50
Similar to the results of the present study, it was reported by other investigators25 that dogs seroreactive for Bartonella spp, E canis, and L infantum develop another type of autoantibody (ie, ANA). It is speculated that dogs infected with these vector-transmitted pathogens may develop persistent infection within the systemic circulation and therefore may be more prone to develop ANA. On the basis of results of that study,25 this likelihood may also be evident in the production of pANCA. Interestingly, dogs that were seroreactive for > 1 organism were more likely to develop ANA, compared with results for dogs that were seroreactive for only 1 pathogen.25 Similar results were found in the present study, in which there was a higher prevalence of pANCA in samples with medium to high antibody titers and coinfections, compared with results for samples with reactivity for single pathogens. Interestingly, the highest pANCA prevalence among all vector pathogen groups was found in the dogs seroreactive for L infantum. This may be explained by the fact that most dogs in this group had strong antibody responses against L infantum; alternatively, some of these dogs may have been coinfected with other vector-borne pathogens.26,51,52
Investigators in several studies29,53–55 have reported a spectrum of clinical signs similar to those seen for IMHA, immune-mediated thrombocytopenia, endocarditis, vasculitis, and nonseptic neutrophilic polyarthritis in dogs infected with various Bartonella spp. In Bartonella-infected dogs, it is also possible that the pathogen triggered a secondary autoimmune response that can include the production of pANCA, as has been described in humans with endocarditis attributable to Bartonella infection.24,56 Although the common pathway or pathways have yet to be definitively characterized, it is likely that many intravascular infections induce an immune response that results in the production of a spectrum of autoantibodies (ie, antierythrocyte, antiplatelet, ANA, ANCA, and others). For example, several autoantibodies (antiendothelial, antierythrocyte, antiplatelet, and antinuclear) have been found in sera of dogs with ehrlichiosis and leishmaniosis.25,26,57–60 Analysis of results of the present study suggested that pANCA are an additional autoantibody that can be found in a high percentage of dogs with these infections. Because of deposition of immune complexes, glomerulonephritis, uveitis, polyarthritis, and neutrophilic vasculitis are common diagnoses in dogs persistently infected with vector-borne organisms.55,61–64 The extent to which deposition of immune complexes is linked to the development of ANA and pANCA needs further investigation.25
In the present study, we investigated the prevalence of pANCA in healthy dogs, dogs with IMHA, and dogs that were seroreactive for selected vector-borne pathogens. We detected pANCA in a high percentage of dogs with IMHA and vector-borne infectious diseases; therefore, pANCA may be a relatively nonspecific marker for dogs with IBD. The low prevalence of pANCA in healthy dogs is similar to results reported in a previous study.13 This may imply that although pANCA have poor diagnostic specificity the development of pANCA in a dog may represent a biomarker for systemic illness. Consequently positive results for pANCA in an individual dog should be interpreted with caution because pANCA are associated with chronic enteropathies as well as with immune-mediated and vector-borne infectious diseases.9,13 Further prospective studies are needed to define the diagnostic utility of pANCA in dogs with immune-mediated and infectious diseases.
ABBREVIATIONS
| ANA | Antinuclear antibodies |
| ANCA | Antineutrophil cytoplasmic autoantibodies |
| CI | Confidence interval |
| IBD | Inflammatory bowel disease |
| IFA | Indirect immunofluorescence assay |
| IMHA | Immune-mediated hemolytic anemia |
| pANCA | Perinuclear antineutrophil cytoplasmic autoantibodies |
Provided by Robert Goggs, Queen Mother Hospital, Royal Veterinary College, University of London, Hatfield, Hertfordshire, AL9 7TA, England.
SNAP3Dx, IDEXX Laboratories Inc, Westbrook, Me.
SNAP4Dx, IDEXX Laboratories Inc, Westbrook, Me.
Sheep anti-canine IgG:fluorescein isothiocyanate, Serotec AA132, Kidlington, Oxford, England.
SPSS, version 18.0, SPSS Inc, Chicago, Ill.
References
- 1.
Rump JAScholmerich JGross V, et al. A new type of perinuclear anti-neutrophil cytoplasmic antibody (p-ANCA) in active ulcerative colitis but not in Crohn's disease. Immunobiology 1990; 181:406–413.
- 2.
Radice ASinico RA. Antineutrophilic cytoplasmic antibodies (ANCA). Autoimmunity 2005; 38:93–103.
- 3.
Hagen ECDaha MRHermans J, et al. Diagnostic value of standardized assays for anti-neutrophil cytoplasmic antibodies in idiopathic systemic vasculitis. EC/BCR Project for ANCA Assay Standardization. Kidney Int 1998; 53:743–753.
- 4.
Choi HKLiu SMerkel PA, et al. Diagnostic performance of antineutrophil cytoplasmic antibody tests for idiopathic vasculitides: metaanalysis with a focus on antimyeloperoxidase antibodies. J Rheumatol 2001; 28:1584–1590.
- 5.
Gross WL. Aniteutrophil cytoplasmic autoantibody testing in vasculitides. Rheum Dis Clin North Am 1995; 21:987–1011.
- 6.
Falk RJHogan SCarey TS, et al. Clinical course of anti-neutrophil cytoplasmic autoantibody–associated glomerulonephritis and systemic vasculitis. Ann Intern Med 1990; 113:656–663.
- 7.
Homsak EMicetic-Turk DBozic B. Autoantibodies pANCA, GAB and PAB in inflammatory bowel disease: prevalence, characteristics and diagnostic value. Wien Klin Wochenschr 2010; 122(suppl 2):19–25.
- 8.
Anand VRussell ASTsuyuki R, et al. Perinuclear antineutrophil cytoplasmic autoantibodies and anti–Saccharomyces cerevisiae antibodies as serological markers are not specific in the identification of Crohn's disease and ulcerative colitis. Can J Gastroenterol 2008; 22:33–36.
- 9.
Allenspach KLomas BWieland B, et al. Evaluation of perinuclear anti-neutrophilic cytoplasmic autoantibodies as an early marker of protein-losing enteropathy and protein-losing nephropathy in Soft Coated Wheaten Terriers. Am J Vet Res 2008; 69:1301–1304.
- 10.
Jaskowski TDLitwin CAHill HR. Analysis of serum antibodies in patients suspected of having inflammatory bowel disease. Clin Vaccine Immunol 2006; 13:655–660.
- 11.↑
Luckschander NAllenspach KHall J, et al. Perinuclear anti-neutrophilic cytoplasmic antibody and response to treatment in diarrheic dogs with food responsive disease or inflammatory bowel disease. J Vet Intern Med 2006; 20:221–227.
- 12.
Mokrowiecka AGasiorowska AMalecka-Panas E. pANCA and ASCA in the diagnosis of different subtypes of inflammatory bowel disease. Hepatogastroenterology 2007; 54:1443–1448.
- 13.↑
Allenspach KLuckschander NStyner M, et al. Evaluation of assays for perinuclear antineutrophilic cytoplasmic antibodies and antibodies to Saccharomyces cerevisiae in dogs with inflammatory bowel disease. Am J Vet Res 2004; 65:1279–1283.
- 14.↑
Mancho CRodriguez-Franco FGarcia-Sancho M, et al. Detection of anti-neutrophil cytoplasmic antibodies (ANCA) in serum from dogs with inflammatory bowel disease. Clin Vet Pequenos Anim 2007; 27:121–126.
- 15.
Nakamura RMBarry M. Serologic markers in inflammatory bowel disease (IBD). MLO Med Lab Obs 2001; 33:8–19.
- 16.↑
Hirai YIyoda MShibata T, et al. Lupus nephritis associated with positive MPO-ANCA in a patient with underlying autoimmune hemolytic anemia. Clin Exp Nephrol 2008; 12:393–397.
- 17.
Ohhashi JMiyamoto MIshikawa S, et al. Crescentic glomerulonephritis with positive antineutrophil cytoplasmic autoantibody specific for myeloperoxidase associated with autoimmune hemolytic anemia and thrombocytopenic purpura. Intern Med 2000; 39:650–654.
- 18.
Hirokawa M. Paraneoplastic autoimmune disorders. Gan To Kagaku Ryoho 2010; 37:980–983.
- 19.
Faruqi SKastelik JAMcGivem DV. Diagnostic pitfall: Mycobacterium avium complex pulmonary infection and positive ANCA. Eur J Intern Med 2008; 19:216–218.
- 20.
Teixeira LMahr AJaureguy F, et al. Low seroprevalence and poor specificity of antineutrophil cytoplasmic antibodies in tuberculosis. Rheumatology 2005; 44:247–250.
- 21.
Flores-Suarez LFCabiedes JVilla AR, et al. Prevalence of anti-neutrophil cytoplasmic autoantibodies in patients with tuberculosis. Rheumatology 2003; 42:223–229.
- 22.
Holmes AHGreenough TCBalady GJ, et al. Bartonella henselae endocarditis in an immunecompetent adult. Clin Infect Dis 1995; 21:1004–1007.
- 23.
Brahn EPegues DAYao QP, et al. Mucocutaneous leishmaniasis masquerading as Wegener granulomatosis. J Clin Rheumatol 2010; 16:125–128.
- 24.
Sugiyama HSahara MImai Y, et al. Infective endocarditis by Bartonella quintana masquerading as antineutrophil cytoplasmic antibody-associated small vessel vasculitis. Cardiology 2009; 114:208–211.
- 25.↑
Smith BETompkins MBBreitschwerdt EB. Antinuclear antibodies can be detected in dog sera reactive to Bartonella vinsonii subsp berkhoffii, Ehrlichia canis, or Leishmania infantum antigens. J Vet Intern Med 2004; 18:47–51.
- 26.
Ciaramella POliva GDeLuna R, et al. A retrospective clinical study of canine leishmaniasis in 150 dogs naturally infected by Leishmania infantum. Vet Rec 1997; 141:539–543.
- 27.
Henn JBLiu CHKasten RW, et al. Seroprevalence of antibodies against Bartonella species and evaluation of risk factors and clinical signs associated with seropositivity in dogs. Am J Vet Res 2005; 66:688–694.
- 28.↑
Overmann JASharkey LCWeiss DJ, et al. Performance of 2 microtiter canine Coombs' tests. Vet Clin Pathol 2007; 36:179–183.
- 29.
Solano-Gallego LLlull JOsso M, et al. A serological study of exposure to arthropod-borne pathogens in dogs from northeastern Spain. Vet Res 2006; 37:231–244.
- 30.↑
Gerber BHaug KEichenberger S, et al. Comparison of a rapid immunoassay for antibodies to the C6 antigen with conventional tests for antibodies to Borrelia burgdorferi in dogs in Europe. Vet Rec 2009; 165:594–597.
- 31.↑
Riera CValladares JEGallego M, et al. Serological and parasitological follow-up in dogs experimentally infected with Leishmania infantum and treated with meglumine antimoniate. Vet Parasitol 1999; 84:33–47.
- 32.
Katrib ASturgess ABertouch JV. Systemic sclerosis and antineutrophil cytoplasmic autoantibady-associated renal failure. Rheumatol Int 1999; 19:61–63.
- 33.
Spornk PEBootsma HHorst G, et al. Antineutrophil cytoplasmic antibodies in systemic lupus erythematosus. Rheumatology 1996; 35:625–631.
- 34.
Hochman JA. Autoimmune hemolytic anemia associated with Crohn's disease. Inflamm Bowel Dis 2002; 8:98–100.
- 35.
Carr APPanciera DLKidd L. Prognostic factors for mortality and thromboembolism in canine immune-mediated hemolytic anemia: a retrospective study of 72 dogs. J Vet Intern Med 2002; 16:504–509.
- 36.
Breitschwerdt EBBlann KRStebbins ME, et al. Clinicopathological abnormalities and treatment response in 24 dogs seroreactive to Bartonella vinsonii (berkhoffii) antigens. J Am Anim Hosp Assoc 2004; 40:92–101.
- 37.
Goodman RABreitschwerdt EB. Clinicopathologic findings in dogs seroreactive to Bartonella henselae antigens. Am J Vet Res 2005; 66:2060–2064.
- 38.
Gionchetti PVecchi MRizzello F, et al. Lack of effect of anti-neutrophil cytoplasmic antibodies associated with ulcerative colitis on superoxide anion production from neutrophils. Gut 1997; 40:102–104.
- 39.↑
Schultz HSchinke SMosier K, et al. BPI-ANCA of pediatric cystic fibrosis patients can impair BPI-mediated killing of E. coli DH5 alpha in vitro. Pediatr Pulmonol 2004; 37:158–164.
- 40.
Schinke SFellermann KHerlyn K, et al. Autoantibodies against the bactericidal/permeability-increasing protein from inflammatory bowel disease patients can impair the antibiotic activity of bactericidal/permeability-increasing protein. Inflamm Bowel Dis 2004; 10:763–770.
- 41.↑
Cohavy OBruckner DGordon LK, et al. Colonic bacteria express an ulcerative colitis pANCA-related protein epitope. Infect Immun 2000; 68:1542–1548.
- 42.↑
Zhao MHJones SJLockwood CM. Bacterial permeability-increasing protein (PBI) is an important antigen of antineutrophil cytoplasmic autoantibodies (ANCA) in vasculitis. Clin Exp Immunol 1995; 99:49–56.
- 43.
Vonandrian UHChambers JDMcEvoy LM, et al. 2-step model of leukocyte endothelial-cell interaction in inflammation-distinct roles for lECAM-1 and the leukocyte beta 2 intergrins in vivo. Proc Natl Acad Sci U S A 1991; 88:7538–7542.
- 44.
Fiebig ELey KArfors KE. Rapid leukocyte accumulation by spontaneous rolling and adhesion in the exteriorised rabbit mesentery. Int J Microcirc Clin Exp 1991; 10:127–144.
- 45.
Hickey MJKubes P. Intravascular immunity: the host-pathogen encounter in blood vessels. Nat Rev Immunol 2009; 9:364–375.
- 46.
McKay DM. Bacterial superantigens: provocateurs of gut dysfunction and inflammation? Trends Immunol 2001; 22:497–501.
- 47.
Dalwadi HWei BKronenberg M, et al. The Crohn's disease-associated bacterial protein I2 is a novel enteric T cell superantigen. Immunity 2001; 15:149–158.
- 48.
Reumaux DDuthilleul PRoos D. Pathogenesis of diseases associated with antineutrophil cytoplasm autoantibodies. Hum Immunol 2004; 65:1–12.
- 49.
Utz PJAnderson P. Posttranslational protein modifications, apoptosis, and the bypass of tolerance to autoantigens. Arthritis Rheum 1998; 41:1152–1160.
- 50.
Bell EKChugh SSCook WJ. A case of infection-associated antiproteinase-3-negative cytoplasmic antineutrophil cytoplasmic antibody pauci-immune focal necrotizing glomerulonephritis. Nephrol Dial Transplant 2010; 25:3119–3123.
- 51.
Alvar JCanavate CMolina R, et al. Canine leishmaniasis. Adv Parasitol 2004; 57:1–88.
- 52.
Otranto DTestini GDantas-Torres F, et al. Diagnosis of canine vector-borne diseases in young dogs: a longitudinal study. J Clin Microbiol 2010; 48:3316–3324.
- 53.
Pappalardo BLCorrea MTYork CC, et al. Epidemiologic evaluation of the risk factors associated with exposure and seroreactivity to Bartonella vinsonii in dogs. Am J Vet Res 1997; 58:467–471.
- 54.
Henn JBVanhorn BAKasten RW, et al. Short report: antibodies to Bartonella vinsonii subsp berkhoffii in Moroccan dogs. Am J Trop Med Hyg 2006; 74:222–223.
- 55.
Breitschwerdt EBAtkins CEBrown TT, et al. Bartonella vinsonii subsp berkhoffi and related members of the alpha subdivision of the Proteobacteria in dogs with cardiac arrhythmias, endocarditis, or myocarditis. J Clin Microbiol 1999; 37:3618–3626.
- 56.
Riding AMD'Cruz DP. A case of mistaken identity: subacute bacterial endocarditis associated with p-antineutrophil cytoplasmic antibody [published online ahead of print Dec 15, 2010]. BMJ Case Rep doi:10.1136/bcr.09.2010.3299.
- 57.
Frank JRBreitschwerdt EB. A retrospective study of ehrlichiosis in 62 dogs from North Carolina and Virginia. J Vet Intern Med 1999; 13:194–201.
- 58.
Waner THarrus SWeiss DJ, et al. Demonstration of serum antiplatelet antibodies in experimental acute canine ehrlichiois. Vet Immunol Immunopathol 1995; 48:177–182.
- 59.
Harrus SWaner TWeiss DJ, et al. Kinetics of serum antiplatelet antibodies in experimental acute canine ehrlichiosis. Vet Immunol Immunopathol 1996; 51:13–20.
- 60.
Cortese LTerrazzano GPiantedosi D, et al. Prevalence of antiplatelet antibodies in dogs naturally co-infected by Leishmania infantum and Ehrlichia canis. Vet J 2011; 188:118–121.
- 61.
Agut ACorzo NMurciano J, et al. Clinical and radiographic study of bone and joint lesions in 26 dogs with leishmaniasis. Vet Rec 2003; 153:648–652.
- 62.
Vamvakidis CDKoutinas AFKanakoudis G, et al. Masticatory and skeletal muscle myositis in canine leishmaniasis (Leishmania infantum). Vet Rec 2000; 146:698–703.
- 63.
Koutinas AFPolizopoulou ZSSaridomichelakis MN, et al. Clinical considerations on canine visceral leishmaniasis in Greece: a retrospective study of 158 cases (1989–1996). J Am Anim Hosp Assoc 1999; 35:376–383.
- 64.
Breitschwerdt EBKordick DLMalarkey DE, et al. Endocarditis in a dog due to infection with a novel Bartonella subspecies. J Clin Microbiol 1995; 33:154–160.
