Antinuclear antibodies and histocompatibility antigens in patients on long-term lithium therapy

A survey of antinuclear factor, histocompatibility antigens, red blood cell groups and red blood cell isoenzyme variants is reported in 54 patients on long-term lithium therapy. Eleven patients with detectable antinuclear factor could not be distinguished from 43 patients without antinuclear factor using age, sex, diagnosis, previous medication, time on lithium or usual dose of lithium. The presence of antinuclear factor was not associated with any particular genetic marker.     

Antinuclear antibodies

The presence of abnormal levels of autoantibodies to intracellular antigens is a hallmark of systemic connective tissue disease. The indirect immunofluorescence assay is the most commonly used routine test for the detection of antinuclear antibodies. In this text, several representative patterns of fluorescence are reviewed and some pitfalls for application of the results to the clinical field are mentioned.     

Antinuclear antibodies

The presence of abnormal levels of autoantibodies to intracellular antigens is a hallmark of systemic connective tissue disease. The indirect immunofluorescence assay is the most commonly used routine test for the detection of antinuclear antibodies. In this text, several representative patterns of fluorescence are reviewed and some pitfalls for application of the results to the clinical field are mentioned.     

Antinuclear antibodies and survival in cardiac transplant patients

Serum from 48 patients with cardiac transplants was tested for the presence of antinuclear antibodies (ANA). Thirty-two of the sera were negative for ANA (ANA-), and 16 were positive for ANA (ANA+). Six of the ANA+ sera had a diffuse pattern, six had a peripheral pattern, three had a combined diffuse and rim pattern, and one had a diffuse and nucleolar pattern. Seven of the ANA+ sera were positive at a 1:40 titer, one at a 1:80 titer, four at a 1:160 titer, two at a 1:320 titer, and two at a 1:640 titer. Thirteen of the 32 ANA- patients have died, at intervals of one month to four years after transplantation. Fourteen of the 16 ANA+ patients have died, at intervals ranging from less than one day to four years after transplantation. The mean time period from transplantation to death was 13 +/- 15 months for the ANA+ patients: these were not statistically significantly different. However, the mean interval from transplantation to death of the ANA+ patients with an ANA titer greater than 1:40 was 0.8 +/- 0.8 months, which was significantly different from the mean survival period of the ANA+ group as a whole (P less than or equal to 0.05) and the mean survival of the ANA- group (P less than or equal to 0.005). Although the mechanism is not clear, there appears to be an association between higher titer serum ANA positivity and increased mortality in patients with cardiac transplants with significantly decreased duration of survival after transplantation.     

Antinuclear antibodies in Thai patients with connective tissue diseases

A study of antinuclear antibodies (ANA) among Thai patients with various connective tissue diseases revealed that the prevalence of ANA was similar to that in other countries, but that the ANA patterns showed interesting contrasts in most diseases. Rather than the predominant homogeneous pattern seen elsewhere in systemic lupus erythematosus and rheumatoid arthritis, the speckled pattern was commonest among Thai patients with these two diseases (67.9% and 76.9% respectively). Patients with scleroderma exhibited a much lower percentage of the nucleolar pattern (17%) than reported elsewhere. The discrepancy between our findings and those from other studies may reflect differences in genetics, the environment or the severity of disease.     

Antinuclear antibodies in patients with endometriosis: A cross-sectional study in 94 patients

Markers of autoimmunity, such as autoantibodies, have been found in patients with endometriosis. These include the antinuclear antibodies (ANA). We aimed to evaluate the prevalence of ANA in a sample of patients with endometriosis and its possible clinical associations. Ninety-four patients with endometriosis and 91 controls were studied for ANA and extractable nuclear antigen (ENA; anti-Ro, anti-La, anti-Sm, and anti-RNP) profiles and anti-dsDNA. Epidemiological, clinical, and staging data in endometriosis were obtained. Patients with autoimmune disorders were excluded. Patients with endometriosis had a 21.2% prevalence of positive ANA vs. 5.4% in controls (P = 0.001). The ENA profile and anti-dsDNA were negative. Patients with positive ANA were more asymptomatic (P = 0.03) and had less dysmenorrhea. No associations with disease duration, patient age, or endometriosis stage were found. We found a high prevalence of positive ANA in patients with endometriosis. The presence of this autoantibody may be linked to a milder clinical expression of the disease.     

Antinuclear antibodies and elevated anti-Epstein-Barr virus titers in cancer patients.

One and one-half percent of human sera from patients seen at a clinic for treatment of cancer contained antibodies to the nuclei of chick kidney cells by indirect immunofluorescence tests. In the group of sera containing antinuclear antibodies, the geometric mean titer to Epstein-Barr virus (EBV) capsid antigen was significantly elevated. Sera obtained from normal adults or from patients with similar histological types of tumors that possessed no antinuclear antibodies contained lower levels of anti-EBV antibodies. The elevated titers to EBV were correlated with the presence or absence of antinuclear antibodies and not with a particular type or site of neoplastic disease.     

Antinuclear antibodies: clinical applications

One of the common serological hallmarks of autoimmune disorders is the presence of various autoantibodies in the sera of patients affected by these disorders. Antinuclear antibodies (ANA) detection is often needed to aid the diagnosis in several autoimmune disorders. In view of the different methodologies available for their detection, it becomes essential to understand the advantages and pitfalls of each procedure. This brief review discusses some methodological aspects of ANA detection and the clinical relevance of the presence of some of the autoantibodies found in the sera of patients with autoimmune disorders.     

Antinuclear antibodies: two-step detection strategy

In order to evaluate the performance of the chemiluminescent immunoassay (CLIA) in antinuclear antibodies (ANA) testing, using indirect fluorescent antibody (IFA) on HEp-2 cells as a standard, 209 samples were studied from September to October/2010. The tests were conducted according to the procedures recommended by the manufacturers of the reagents. The interpretation of the IFA results was done according to the Brazilian standards. The charts of patients showing different results between the two techniques were analyzed. The CLIA efficiency was 89%, with a sensitivity of 65%, and a specificity of 94.7%. Nine (4.3%) false-positive and 14 (6.7%) false-negative results were detected. Of these, 13 (93%) represented no risk for the diagnosis and therapeutic management of the patients. The CLIA methodology reduced the need for the IFA manual technique by 77% (160/209). The ANA screening test proved to be a fast and acceptable methodology in the studied population. We established the following criteria for the introduction of an automated ANA screening: (1) Positive results must be confirmed by IFA to characterize the pattern and titer of the antibody. (2) Negative results are issued with a notice to request a new test by IFA when the clinical suspicion of autoimmune disease persists.     

Antinuclear antibodies in the oldest-old women and men

The aim of this study was to compare the prevalence of antinuclear antibodies (ANA) in very old individuals (>or=86 years of age) with healthy younger (18-68 years) blood donors (n=200) regarding gender, health status, ratio of circulating CD4/CD8 cells and cytomegalovirus (CMV) serology. Frozen plasma was used for ANA detection in two study groups, i.e. 'OCTO' (97 persons aged 86-92 years, 65% women) and 'NONA' (136 persons aged 86-95 years, 70% women). OCTO participants were recruited on the basis that they were healthy or moderately healthy according to a selection protocol. No exclusion criteria regarding health status were applied in the NONA sample. The prevalence of ANA was significantly higher in the oldest-old samples compared to blood donors. There was no association between health status and the presence of ANA in the oldest-old. The difference across age was most pronounced in men, with low levels at younger age, whereas the prevalence among the oldest-old men reached similar levels as in women. There were no associations between the presence of ANA and CD4/CD8 ratio or with CMV status in the oldest-old. Our findings confirm an increased prevalence of ANA in the oldest-old, and emphasize the importance of taking gender and age into consideration when evaluating ANA.     

Antinuclear antibodies in the oldest-old women and men

The aim of this study was to compare the prevalence of antinuclear antibodies (ANA) in very old individuals (≥86 years of age) with healthy younger (18–68 years) blood donors (n = 200) regarding gender, health status, ratio of circulating CD4/CD8 cells and cytomegalovirus (CMV) serology. Frozen plasma was used for ANA detection in two study groups, i.e. ‘OCTO’ (97 persons aged 86–92 years, 65% women) and ‘NONA’ (136 persons aged 86–95 years, 70% women). OCTO participants were recruited on the basis that they were healthy or moderately healthy according to a selection protocol. No exclusion criteria regarding health status were applied in the NONA sample. The prevalence of ANA was significantly higher in the oldest-old samples compared to blood donors. There was no association between health status and the presence of ANA in the oldest-old. The difference across age was most pronounced in men, with low levels at younger age, whereas the prevalence among the oldest-old men reached similar levels as in women. There were no associations between the presence of ANA and CD4/CD8 ratio or with CMV status in the oldest-old. Our findings confirm an increased prevalence of ANA in the oldest-old, and emphasize the importance of taking gender and age into consideration when evaluating ANA.     

Antinuclear antibodies specific for primary biliary cirrhosis

The serological hallmark of primary biliary cirrhosis (PBC) is the presence of antimitochondrial antibodies. However, antinuclear antibodies (ANA) are also detectable in approximately 50% of subjects with PBC. Most clinical laboratories use indirect immunofluorescence microscopy to detect ANA and two labeling patterns that predominate in PBC are 'punctate nuclear rim' and 'multiple nuclear dots.' Work over the past several years has shown that antibodies giving these patterns most often recognize nuclear pore membrane protein gp210 and nuclear body protein sp100, respectively. These ANA are highly specific for PBC and detected in approximately 25% of patients. Less frequently, ANA apparently unique to PBC recognize other proteins of the nuclear envelope and nuclear bodies. While antibodies against gp210, sp100 and some other nuclear proteins are very specific to PBC and may therefore be useful diagnostic markers, their connection to pathogenesis remains to be elucidated.     

Antinuclear antibodies detected by indirect immunofluorescence on HEp2 cells and by immunoblotting in patients with systemic sclerosis

The HEp2 cell cultures appeared highly sensitive in detecting the antinuclear antibodies (ANAb) in systemic sclerosis, principally anticentromere antibodies of the CREST syndrome. The immunoblotting used with either complex cellular extracts from HeLa and rabbit thymus or purified nuclear components (high mobility group (HMG) proteins and histones) is able to identify precisely the ANAb targets and to contribute to diagnosis. With nuclear extracts of HeLa cells, the sera from 75.8% of CREST syndrome subjects stained 18 and 22 kD proteins. Corresponding antibodies were also detected in 72.7% of these patients, on HEp2 centromers by indirect immunofluorescence. With the same extracts, 33.3% of sera from diffuse sclerosis/acrosclerosis patients contain antibodies staining 86, 73, 32 and 30kD. These sera also stain 77, 66 and 63kD from thymus extracts. Corresponding antibodies will be the anti-SCL-70 antibodies defined by double immunodiffusion. The anti-HMG antibodies were infrequent in systemic sclerosis, rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) and consequently without interest for diagnosis. The anti-whole histones antibodies which are less frequent in diffuse sclerosis/acrosclerosis (35.7%) than in SLE (41.3%) recognize especially H1 and H2A in the first diseases, H1 and H2B in SLE and H1 and H3 in RA.