Markers Of Autoimmune Disease Research Articles

Autolytic Mycobacterium leprae Hsp65 fragments may act as biological markers for autoimmune diseases

Investigating the proteolytic activity of the recombinant Mycobacterium leprae Heat Shock Protein of 65 kDa (rHsp65), chaperonin 2 (cpn2), we observed that it displays high instability. The fragmentation process starts at the C-terminus followed by progressive degradation of the N-terminus, which leads to a stable fragment comprising the middle region of the molecule. Urea was able to prevent autolysis, probably due to its denaturing action, while EDTA increased degradation levels indicating the need for metal ions. Peptides originated from autolysis were purified and analyzed by mass spectrometry, generating a continuous map. Since the bacteria and mammalian Hsp60 are known to be targets of the immune response and have been implicated in autoimmune diseases and chronic inflammation, the in vivo effect of rHsp65 peptides was evaluated in the spontaneous Systemic Lupus Erythematosus (SLE) model developed by the (NZB/NZW)F1 mouse hybrids, and their individual anti-rHsp65 IgG2a/IgG1 antibody titer ratio was determined. The results showed orientation toward a TH1 responsiveness, and the treatment with the rHsp65 peptides diminished the environmental variance of the survival time of treated animals. These results outline the fact that environmental factors may also act through the modified stability expression of Heat Shock Proteins intervening during autoimmune processes.

Differential diagnosis of plasma cell dyscrasias in rheumatological practice

Plasma cell dyscrasias (PCD) present certain difficulties in differentially diagnosing with autoimmune rheumatic diseases (RD) as they have some clinical and laboratory manifestations that are characteristic of the latter. Objective: to generalize the experience in diagnosing PCD at the Research Institute of Rheumatology (RIR), Russian Academy of Medical Sciences (RAMS). Subjects and methods. Nineteen patients admitted to the RIR, RAMS, for rheumatological referral diagnoses were diagnosed as having different types of PCD, both PCD in the presence of RD (n = 10) and primary PCD without RD (n = 9). Immunochemical studies of serum and urinary proteins and bone marrow trepanobiopsy were performed in all the patients. Histological studies were made of the minor salivary glands (n = 12), lymph nodes (n = 3), parotid salivary glands - PSG (n = 5), spleen (n = 1), cranial bones (n = 2) and humerus (n = 1). The immunophenotype of tumor lymphocytes in the biopsy and trepanobiopsy specimens were determined by an immunoflurenscence method with standard monoclonal antibody panels and the paraffin sections of biopsy specimens were examined by an immunoperoxidase technique. Biopsy and trepanobiopsy speciments and myelograms were assessed by the researchers of the N.N. Blokhin Russian Cancer Research Center, RAMS. Results. Over 10 years of follow-up, 19 patients (13 females and 6 males) were diagnosed as having different types of PCD: multiple myeloma (MM) (n = 9), extramedullary plasmacytoma of lymph nodes (n = 2), solitary plasmacytoma (SP) of bone (n = 2), Bence-Jones myeloma (BJM) (n = 2), primary amyloidosis (PA) (n = '), lymphoplasmacytic lymphoma (n = '), Waldenström's macroglobulinemia (WM) (n = 2). In the presence of RA, '0 patients with PCD developed Sjogren's disease (SD) (n = 7), rheumatoid arthritis (RA) + SD (n = 2), RA (n = '); and 9 patients had primary PCD (MM (n = 5), BJM (n = '), WM (n = '), SP (n = '), and PA (n = ')). These 9 patients with different rheumatological diagnoses were long followed up and referred to the RIR, RAMS, to specify these. One third of the patients had recurrent conjunctivitis, enlarged PSG and xerostomia (55%). Arthralgias, arthritis of minor and large joints, flexion contractures of hands, ossalgias, and pains in the lumbar spine and ribs were present in '0-45% of the patients. Vascular lesions, such as Reynaud's phenomenon, recurrent purpuras, lower extremity ulcerative lesions, and toe gangrenes, were observed in '0-35% of the patients with PCD. Lesions in the reticuloendothelial system, such as lymphadenopathy, splenomegaly ('5%), hepatomegaly (45%), and fever (25%), were also detected. Conclusion. Many clinical and laboratory manifestations of primary PCD and RD are similar and only the absence of classical laboratory markers of autoimmune disease, as well as the presence of serum monoclonal immunoglobulins and urinary Bence Jones protein suggest the presence of PCD, both primary PCDs and those with RD.

Anti-CENP-B Antibodies Are Associated with Prolonged Survival in Breast Cancer

Presence of a relationship between autoimmunity and malignant diseases has been investigated for a long time. Anti-SS-B and anti-centromere protein (CENP)-B antibodies are used as serologic markers for autoimmune diseases. In this study, these autoantibodies were studied in breast cancer patients, and their effect on survival was evaluated. Breast cancer patients treated between January and June 2003 were prospectively included in this study. Antibodies were studied by ELISA, and serum values of greater than 10 U/ml were accepted as positive. In total, 55 patients with breast cancer were prospectively included in the study. Median follow-up time was 62 months. In univariate analysis, estrogen receptor status, anti-CENP-B antibody status and tumor size significantly affected disease-free survival, whereas overall survival was significantly dependent on tumor size and anti-CENP-B antibody status. In multivariate analysis, anti-CENP-B status was the independent prognostic factor for disease-free survival, whereas tumor size and anti-CENP-B status were independent prognostic factors for overall survival. Anti-CENP-B autoantibodies in breast cancer patients prolong disease-free and overall survival.

Tracking Early Autoimmune Disease by Bioluminescent Imaging of NF-κB Activation Reveals Pathology in Multiple Organ Systems

It is desirable to have an early and sensitive detection marker of autoimmune disease in intact animals. Nuclear factor (NF)-kappaB is a transcription factor that is associated with inflammatory responses and immune disorders. Previously, we demonstrated that so-called idiotypic-driven T-B cell collaboration in mice doubly transgenic for paired immunoglobulin and T cell receptor transgenes resulted in a systemic autoimmune disease with systemic lupus erythematosus-like features. Here, we investigated NF-kappaB activation by including an NF-kappaB-responsive luciferase reporter transgene in this animal model. Triply transgenic mice developed bioluminescence signals from diseased organs before onset of clinical symptoms and autoantibody production, and light emissions correlated with disease progression. Signals were obtained from secondary lymphoid organs, inflamed intestines, skin lesions, and arthritic joints. Moreover, bioluminescence imaging and immunohistochemistry demonstrated that a minority of mice suffered from an autoimmune disease of the small intestine, in which light emissions correlated with antibodies against tissue transglutaminase and gliadin. Detection of luciferase by immunohistochemistry revealed NF-kappaB activation in collaborating B and T cells, as well as in macrophages. These results demonstrate that bioluminescent in vivo imaging of NF-kappaB activation can be used for early and sensitive detection of autoimmune disease in an experimental mouse model, offering new possibilities for the evaluation of anti-inflammatory drugs.

Evidence-based laboratory medicine. Application in autoimmunity

Concepts of evidence based laboratory medicine are increasingly utilised in routine clinical and laboratory practice. Some basic concepts of evidence based laboratory medicine, such as likelihood ratios, are not fully comprehended by laboratory professionals and clinicians. This lecture aims to make laboratory professionals and clinicians more familiar with the basic concepts that deal with clinical performance characteristics of a laboratory test. Basic measures of the clinical performance characteristics of a laboratory test, such as sensitivity, specificity, likelihoods, likelihood ratio, post-test probability, odds, and Bayes theorem will be explained. The concepts will be illustrated with serological markers for autoimmune diseases (anti-tissue transglutaminase, anti-CCP antibodies, ASCA, anti-neutrophil cytoplasmic antibodies). The lecture seeks to provide the audience with a better understanding of the clinical performance characteristics of a laboratory test and of evidence-based laboratory medicine. Concepts of evidence based laboratory medicine are increasingly utilised in routine clinical and laboratory practice. Some basic concepts of evidence based laboratory medicine, such as likelihood ratios, are not fully comprehended by laboratory professionals and clinicians. This lecture aims to make laboratory professionals and clinicians more familiar with the basic concepts that deal with clinical performance characteristics of a laboratory test. Basic measures of the clinical performance characteristics of a laboratory test, such as sensitivity, specificity, likelihoods, likelihood ratio, post-test probability, odds, and Bayes theorem will be explained. The concepts will be illustrated with serological markers for autoimmune diseases (anti-tissue transglutaminase, anti-CCP antibodies, ASCA, anti-neutrophil cytoplasmic antibodies). The lecture seeks to provide the audience with a better understanding of the clinical performance characteristics of a laboratory test and of evidence-based laboratory medicine.

Lifetime exposure to trichloroethylene (TCE) does not accelerate autoimmune disease in MRL +/+ mice

A genetically-prone murine lupus model was used to assess the developmental effects of trichloroethylene (TCE) exposure on disease symptom onset (e.g., autoantibody production and proteinuria), lymphocyte proliferation, splenic B-cell populations, and thymic and splenic T-cell populations. MRL +/+ mice were exposed to TCE (0, 1,400 or 14,000 ppb) via drinking water beginning on gestation day (GD) 0 and continuing until 12 months of age. With the exception of splenic CD4-/CD8-cells in female mice only, no alterations were observed in splenic T-cell populations, numbers of splenic B220+ cells, or in lymphocyte proliferation at 12 months of age. Furthermore, populations of all thymic T-cell subpopulations were decreased in male but not female mice following exposure to 14,000 ppb TCE. Autoantibody levels (anti-dsDNA and anti-GA) were assessed periodically from 4 to 12 months of age. Over this period, no increase in autoantibody levels as compared to control was detected, suggesting that TCE did not contribute to or accelerate the development of autoimmune disease markers following lifetime exposure. Not only does this study offer encouraging results, but it is the first study to approach the development of autoimmunity in a novel lifetime exposure paradigm, using an autoimmune prone model, at environmentally relevant exposure levels.

Diagnostik der Autoimmunerkrankungen

Autoantibodies play a key role in diagnostic laboratories as markers of autoimmune diseases. In addition to their role as markers they mediate diverse effects in vivo. Autoantibodies with protective effect have been described. Natural protective IgM autoantibodies against tumour-antigens of malignant cells or their precursors may contribute to increased survival rates of carcinoma patients. In a mouse model of systemic lupus erythematosus it has been shown that anti-dsDNA IgM autoantibodies protect from glomerular damage. In contrast, a direct pathogenic role of autoantibodies has been well established e.g. in myasthenia gravis or in Goodpasture syndrome. Similarly autoantibodies against SSA Ro52 are detrimental in neonatal lupus erythematosus with congenital heart block. Moreover, putatively protective autoantibodies may become pathogenic during the course of the disease such as the onconeuronal autoantibodies whose pathogenicity depends on their compartmentalisation. In patients with paraneoplastic syndromes tumour cells express proteins that are also naturally present in the brain. Anti-tumour autoantibodies which temporarily suppress tumour growth can provoke an autoimmune attack on neurons once having crossed the blood-brain barrier and cause specific neurological symptoms. Only a restricted number of autoantibodies are useful follow-up markers for the effectiveness of treatment in autoimmune diseases. Certain autoantibodies hold prognostic value and appear years or even decades before the diagnosis of disease such as the antimitochondrial antibodies in primary biliary cirrhosis or anti-citrullinated protein (CCP)-antibodies in rheumatoid arthritis. It is crucial to know whether the autoantibodies in question recognise linear or conformational epitopes in order to choose the appropriate detection methods. Indirect immunofluorescence microscopy remains a very useful tool for confirmation of results of commercially available immunoassays and for detection of special and rare autoantibodies that otherwise often remain undetected. Standardisation of autoimmune diagnostics is still underway and requires joint efforts by laboratories, clinicians and industry.

The Immunology of Gluten Sensitivity beyond the Intestinal Tract

Celiac disease and gluten-sensitive enteropathy are terms that have been used to refer to a disease process affecting the small bowel. However, evidence has been accumulated in literature demonstrating that gluten sensitivity or celiac disease can exist even in the absence of enteropathy, but affecting many organs. Based on overwhelming evidence, immunological pathogenesis has been demonstrated in the joint, the heart, thyroid, bone, and, in particular, the brain cerebellum and neuronal synapsin I. When blood samples of patients with celiac disease are tested against gliadin and different tissue antigens, in addition to gliadin antibody, a significant percentage of them exhibit elevation in antibodies against transglutaminase, heat shock protein, collagen, thyroid, myosin, endothelial cell, bone antigen (transglutaminase), myelin basic protein, cerebellar and synapsin. This elevation of autoantibodies in patients with celiac disease may result in neuroimmune disorders. In fact, in comparison to the general population, the incidence of various autoimmune disorders, including gluten ataxia, is increased up to 30-fold in patients with celiac disease. Therefore, immune evaluation of patients with gluten sensitivity or celiac disease, in addition to gliadin and transglutaminase, should include antibody measurement against thyroglobulin, thyroid peroxidase, heat shock protein, bone transglutaminase, myelin basic protein, cerebellar peptide and synapsin. This novel laboratory approach to gluten sensitivity and autoimmunity may enable clinicians to detect markers of autoimmune diseases. Early identification of gluten sensitive and celiac disease patients and implementation of a gluten-free diet may result in significant improvement and control of associated diseases.

Generalized Nonspecific Pustular Lesions in Tietze's Syndrome

Tietze’s syndrome (TS) was defined as a nonsuppurative, benign, and self-limited arthropathy characterized by tenderness and painful swelling of one or more sternocostal, costochondral, or sternoclavicular joints in the absence of other possible causes. This clinical entity should not be confused with other chest wall pain in which there is no swelling. This disease can usually be seen in young people. Although the etiopathogenesis is not well known, symptoms generally follow microtraumas and/or respiratory tract infections. It is estimated that somewhere in the vicinity of 20%–25% of noncardiac chest pain has a musculoskeletal basis. Such a symptom generates anxiety in both patients and their medical attendants for fear that this symptom represents a life-threatening event, such as an acute coronary syndrome. Dermatologic comorbidities with TS have been reported previously, such as palmoplantar pustulosis (PPP) and psoriasis vulgaris. The majority of PPP associated with TS has skin eruptions not only confined to the palms and soles but also on the backs of hands and feet, the arms, the legs, and the trunk. Our patient with TS had pustular lesions that clinically resembled pustular psoriasis at a first glance. These should be recognized as a potential part of TS that as in our case may resolve with nonsteroidal anti-inflammatory drug therapy. A 21-year-old man, previously diagnosed with familial Mediterranean fever (FMF), was admitted to our clinic for the diagnosis and treatment of disseminated pustular lesions, especially localized to both legs and trunk, healing with atrophy. Medical history was consistent with disseminated and nonperiodic joint pain, 3 to 4 times per year, localized to the chest since he was 7 to 8 years old along with the truncal dermatologic lesions. Family history was negative for both FMF and TS. It was noticed that there was a localized swelling and an increased sensitivity at both sternoclavicular joints suggestive of TS during the chest wall examination. Dermatologic examination revealed wide macular plaques consisting of small atrophic scarred lesions surrounded with hyperpigmented area, which were found on both legs, especially on the pretibial areas. There were postinflammatory circular atrophic lesions, which were 1 to 2 mm in width and were scattered all over the body, especially on the trunk. Histopathologic examination of those lesions was consistent with subcorneal pustule formation containing polymorphonuclear leukocytes localized inside the epidermal layer, as well as pseudoepitheliomatous hyperplasia areas with granulation formation at the subepithelial area, and perivascular inflammatory infiltrate with neutrophils (perivascular dermatitis) (Fig. 1). Direct immune fluorescent investigation showed no deposit formation. Biochemical examination was normal except for microcytic anemia with an erythrocyte sedimentation rate of 40 mm/h. Serum immunoglobulin and complement levels, thyroid function tests, antithyroid peroxidase, antithyroglobulin, VDRL, antihuman immunodeficiency virus, hepatitis markers (hepatitis B surface antigen, antihepatitis C virus), autoimmune disease markers (antinuclear antibody, antidouble stranded DNA, anti-Sci-70, anticentromere antibody, anti-SSA-B, anti-SM), antistreptolysin O, rheumatoid factor, and tuberculin skin test were all normal. Culture of pustule contents was negative for bacteria. FMF gene (MDFV/pyrine gene) analysis was also negative. Conventional radiographic examination of the thorax was completely normal. However, bone scintigraphy of the whole body demonstrated an increased activity both at the right shoulder and both sternoclavicular joints consistent with arthritis (Fig. 2).

Rickettsioses in Australia

Australia, an island continent in the southern hemisphere, has a range of rickettsial diseases that include typhus group rickettsiae (Rickettsia typhi), spotted fever group rickettsiae (R. australis, R. honei), scrub typhus group rickettsiae (R. tsutsugamushi), and Q fever (C. burnetii). Our knowledge of Australian rickettsiae is expanding with the recognition of an expanded range of R. honei (Flinders Island spotted fever) to Tasmania and southeastern mainland Australia (not just on Flinders Island), and the detection of a new SFG species (or subspecies), tentatively named "R. marmionii" in the eastern half of Australia. This rickettsia causes both acute disease (7 cases, recognized so far) and is also associated (as a "R. marmionii" bacteriaemia) with patients having a chronic illness. The significance of the latter is under investigation. It may be a marker of autoimmune disease or chronic fatigue in some patients.

A Rare Tricuspid Anomaly

54-year-old Mexican American woman was admitted to the Erie County Medical Center, Buffalo, NY, with the general symptoms of tiredness, weight loss with loss of appetite, leg weakness, and low blood pressure (80/60 mm Hg). The patient denied any paroxysmal nocturnal dyspnea, orthopnea, or leg edema. She did have some facial swelling, a skin rash on both hands, and joint swelling, which was consistent with a previous diagnosis of polymyositis and dermatomyositis. The patient also had evidence of septic arthritis of the left elbow. Physical examination revealed extensive wheezing and a grade 2/3 systolic ejection murmur, heard predominantly over the tricuspid valve area. Laboratory workup revealed pancytopenia and an elevation in liver function tests. All the markers for autoimmune disease were negative. Biopsies and computerized tomographic scans performed to search for visceral malignancy were negative. Blood cultures grew methicillin-sensitive Staphylococcus aureus. Electrocardiography revealed normal sinus rhythm with evidence of right atrial enlarge

A rare tricuspid anomaly.

54-year-old Mexican American woman was admitted to the Erie County Medical Center, Buffalo, NY, with the general symptoms of tiredness, weight loss with loss of appetite, leg weakness, and low blood pressure (80/60 mm Hg). The patient denied any paroxysmal nocturnal dyspnea, orthopnea, or leg edema. She did have some facial swelling, a skin rash on both hands, and joint swelling, which was consistent with a previous diagnosis of polymyositis and dermatomyositis. The patient also had evidence of septic arthritis of the left elbow. Physical examination revealed extensive wheezing and a grade 2/3 systolic ejection murmur, heard predominantly over the tricuspid valve area. Laboratory workup revealed pancytopenia and an elevation in liver function tests. All the markers for autoimmune disease were negative. Biopsies and computerized tomographic scans performed to search for visceral malignancy were negative. Blood cultures grew methicillin-sensitive Staphylococcus aureus. Electrocardiography revealed normal sinus rhythm with evidence of right atrial enlarge

Dysimmune disorders and migraine: is there a possible common denominator?

The association between headache, especially migraine, and some immunomediated diseases, in particular autoimmune diseases, is well known. The evidence in favor of this association comes prevailingly from epidemiological studies, as well as research aimed at assessing the onset and the course of migraine in relation with the onset and activity and the biological markers of autoimmune diseases. The most relevant findings in this regard concern celiac disease, systemic lupus erythematosus, and multiple sclerosis.

ENDOSONOGRAPHIC FEATURES OF LYMPHOPLASMACYTIC SCLEROSING PANCREATITIS

The role of endoscopic ultrasound (EUS) in the evaluation of pancreatic disorders is well established. However, little information on the endosonographic features of uncommon pancreatic diseases is available. We describe the case of a rare form of autoimmune chronic pancreatitis evaluated with EUS. Clinical Vignette: A 31 year-old Mexican male was referred to our institution with jaundice and a common bile duct (CBD) stricture on endoscopic retrograde cholangiopancreatography (ERCP). The patient described a three-month history of malaise, abdominal pain, a forty-pound weight loss, and the recent onset of jaundice; on ERCP he had a 6 cm distal biliary stricture with proximal dilation of the biliary tree. EUS was performed revealing severe changes of chronic pancreatitis including marked lobularity, prominent interlobular septae and a diffusely inhomogeneous echo pattern, features suggestive of diffuse fibrosis. The pancreatic duct was hardly visible. At the level of the pancreatic head, these changes coalesced into a mass-like lesion which involved the distal CBD and produced near-complete biliary obstruction. No lymphadenopathy was present. Concerned for the possibility of an underlying malignancy, no EUS-guided fine needle aspiration (FNA) was performed and the patient was taken for pancreaticoduodenectomy. Histopathologic examination of the surgical specimen demonstrated diffuse lymphoplasmacytic infiltration of the pancreas with marked acinar atrophy, obliterated phlebitis, and extensive fibrosis. The CBD and gallbladder showed marked inflammatory wall thickness. These features were compatible with lymphoplasmacytic sclerosing pancreatitis (LPSP). Serologic markers for autoimmune disease were negative. The patient recovered satisfactorily from the procedure and was discharged home. His follow-up at 21 months has included the development of insulin-requiring diabetes, an episode of acute pancreatitis of the remaining gland, and the need for chronic immunosuppressive therapy. Conclusion: Lymphoplasmacytic sclerosing pancreatitis is a rare form of autoimmune pancreatitis that can only be accurately diagnosed with histopathology. However, the clinical and endosonographic features (including the information obtained with EUS-guided FNA) can provide important clues to consider this disease in the differential diagnosis of patients with chronic pancreatic disorders.

Drinking water exposure to cadmium, an environmental contaminant, results in the exacerbation of autoimmune disease in the murine model

Cadmium is a pervasive environmental contaminant. The primary route of exposure to the general population occurs via contaminated drinking water or food supplies. Our hypothesis was that cadmium could be a trigger for inducing autoimmune disease (AD) in genetically predisposed populations. Therefore, New Zealand Black/White F1 (NZBW) mice were exposed to cadmium via drinking water. Mice were exposed to: 0, 3, 30, 3000 or 10 000 parts per billion (ppb) of cadmium in tap water for 2, 4, 28, or 31 weeks. After 4 weeks of exposure, in the group of mice exposed to 10 000 ppb cadmium, there was an increased incidence of antinuclear antibodies (ANA). There was also deposition of immune complexes in all groups after 4 weeks of exposure. After 31 weeks, there were increases in IgG2a in mice exposed to low doses of cadmium. In an attempt to establish the progression from an autoimmune reaction to the development of AD, the biological marker for AD, proteinuria, was assessed. Onset of proteinuria was exacerbated by 11 weeks in mice exposed to cadmium. This data suggests that short-term exposure may result in a type of autoimmune reaction since the mice are beginning to produce ANA after only 4 weeks of exposure and there is immune-complex deposition in the kidney. Long-term exposure to cadmium appears to result in the exacerbation of AD as indicated by the development of proteinuria and continued presence of immune complexes in the kidney. The mechanism may involve the increased production of IgG2a, which is capable of forming immune complexes and causing autoimmune glomerulonephritis.

Hepatitis autoinmune y anticuerpos anti-Ro positivos. ¿Alguna relación?

Antinuclear antibodies (ANA) are considered to be markers of autoimmune disease. Several specific reactivities of ANA are known, among them the Ro (SS-a) complex. Although the presence of anti-Ro antibodies is widely known in systemic connective tissue diseases such as lupus erythematous or Sjögren's syndrome, few studies have attempted to link this finding with autoimmune hepatitis (AIH). We present a case of acute AIH in a 55-year-old woman who tested positive for anti-Ro 60 kD antibodies. The Ro (SS-A) antigen is a complex ribonucleoprotein whose structure is still the subject of debate. Two fractions, of 50 and 60 kD, have been identified. A review of the literature revealed frequencies of 21-34% for anti-Ro 52 kD and of 9-13-6% for anti-Ro 60 kD in AIH. Although no relationship has been demonstrated between this complex and the etiopathogenic mechanisms or clinical patterns of AIH, we propose that multicenter investigations with large series should be performed before the possible involvement of anti-Ro antibodies in these diseases is definitively ruled out.

Ovarian autoimmune disease and ovarian autoantibodies.

Detection of specific autoantibodies remains the most practical clinical and research marker of autoimmune disease. The lack of consensus on ovary specific antibodies as a marker for ovarian autoimmunity has clinical and research consequences. The objective of this review is to summarize the evidence for ovarian autoimmunity and the detection of ovary specific autoantibodies in humans. Evidence favors the presence of an autoimmune disease of the ovary. Ovarian autoantibodies are associated primarily with premature ovarian failure (POF) and unexplained infertility. Variations in detection of ovarian autoantibodies are likely to be due to study design elements such as antibody test format, antigen preparation, and criteria for study and comparison groups. In addition, multiple targets appear to be involved in ovarian autoimmunity including ovarian cellular elements and oocyte related antigens. Many studies only assess one target antigen, leaving individuals with ovarian autoimmunity unidentified. The next most significant advance in characterizing ovarian autoimmunity will be definitive identification of the specific antigens and development of standardized tests based on use of specific antigens.

Autoantibodies associated with acute rejection after liver transplantation for type-2 autoimmune hepatitis

Autoantibodies, markers of autoimmune diseases, can also be detected in chronic allograft rejection. However, the appearance of these autoantibodies in acute rejection after orthotopic liver tranplantation has not yet been reported. Liver-kidney-microsome type-1 (LKM-1) antibodies directed against the autoantigen cytochrome CYP2D6 define a group of patients with autoimmune hepatitis type-2 (AIH-2), distinct from autoimmune hepatitis type-1 (AIH-1) in which antinuclear antibodies and anti-smooth muscle antibodies (SMA) with actin specificity are present in patient sera. Autoantibodies were studied by the quantitative CYP2D6 radioligand assay (RLA) that uses a radiolabeled CYP2D6 as antigen, immunoblotting using recombinant CYP2D6 protein and human liver microsomal and cytosolic fractions, and indirect immunofluorescence (IIF) using rat kidney-stomach-liver cryostat sections. In addition, the specificity of anti-SMA was detected by IIF on HEp2 cell line harvested with colchicin. This report describes the time course of CYP2D6 antibodies and the appearance of anti-SMA (without anti-actin, cytokeratin and vimentin reactivity) associated with acute rejection during a 2-year follow-up, in a patient who underwent transplantation at end-stage type 2 autoimmune hepatitis. In addition, we report a new reactivity against an unknown 40-kDa protein using a rat cytosolic fraction. The detection of autoantibodies in sequential samples may be important to better predict rejection or relapse, and to establish adequate therapy.

Soluble FcϵRII/CD23 in Patients with Autoimmune Diseases and Epstein-Barr Virus-Related Disorders: Analysis by ELISA for Soluble FcϵRII/CD23

The low-affinity Fc receptor for IgE (FcϵRII/CD23) and its soluble form (sCD23, IgE-binding factor) have multiple functions, and enhanced levels of these are associated with various immunological diseases. We established two sensitive ELISA systems using enzyme-conjugated mAb and biotinylated mAb. The detection limits of the ELISA systems were 0.03 and 1.0 ng/ml, which showed good correlation in the range 1.0-10 ng/ml. In the ELISA system using enzyme-conjugated mAb, the average sCD23 concentration in 303 normal healthy volunteers was 1.4 ± 0.3 ng/ml. In the ELISA system using biotinylated mAb, sCD23 levels in normal healthy volunteers showed almost the same values. In patients with autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Sjögren syndrome, progressive systemic sclerosis, and mixed connective tissue disease, the sCD23 levels were significantly higher than those in normal individuals. Furthermore, in Epstein-Barr virus-related disorders after liver transplantation with immunosuppression, plasma levels of sCD23 rapidly Increased to more than 12 ng/ml when clinical symptoms were evident. In addition, the sCD23 values remained high, although elevated GOT levels gradually decreased to standard values and EBV hepatitis improved. These data suggest that sCD23 levels are a sensitive marker of autoimmune diseases and EBV-related disorders in addition to allergic disorders. The ELISA system for sCD23 may be an additional diagnostic tool in estimating the clinical courses of these diseases.

Inappropriate major histocompatibility complex expression on cardiac tissue in dilated cardiomyopathy. Relevance for autoimmunity?

The inappropriate expression of major histocompatibility complex (MHC) molecules on epithelial and endothelial cells is a recognized marker of autoimmune disease. An autoimmune pathogenesis has been suspected in dilated cardiomyopathy (DCM). In the normal heart, MHC products are usually not detectable on myocytes using immunochemical techniques. MHC molecule expression has not, however, been assessed on cardiac endothelial cells. The aim of this study was to investigate possible autoimmune phenomena and MHC molecule expression in fresh endomyocardial biopsies from 29 patients with DCM. These were compared with those observed in surgical specimens from 63 patients with other acquired cardiac disease and from 22 with congenital heart disease (CHD) as normal controls. Conventional immunofluorescence (IFL) with monoclonal antibodies (MoAbs) to lymphocyte and macrophage markers and to MHC molecules was employed, and double IFL with antiserum to human Factor VIII was used for the identification of endothelial cells. Myocytes did not express MHC molecules in either DCM or controls. In normal hearts, Class II molecules were detected on endothelial and endocardial cells in only a few cases ( 3 22 and 2 22 respectively). By contrast, endothelial and endocardial cells inappropriately expressed Class II in a high proportion of DCM patients ( 28 29 and 22 29 ) but less frequently in other acquired cardiac diseases ( 19 63 , P < 0.001 and 11 63 , P < 0.001 respectively). In all the DCM biopsies examined there was a hierarchy of Class II subloci product expression (DR > DP > DQ); lymphocytic infiltration was a rare finding and macrophages/dendritic cells were not prominent. The finding of inappropriate MHC Class II molecule expression on cardiac endothelial and on endocardial cells suggests a possible pathogenic role for these cells in the initiation and/or perpetuation of DCM.