不同底物检测抗核抗体的研究

以大鼠器官印片、人癌印片和人鳞状上皮癌细胞(HEp-2)作底物,采用间接免疫荧光法在59例各种结缔组织病患者中研究抗核抗体.人癌印片检测ANA的敏感性和特异性均高于动物器官底物,人癌印片所见的着丝点抗体经HEp-2细胞底物证实.     

Immunofluorescence studies in progressive systemic sclerosis (scleroderma) and mixed connective tissue disease

Immunofluorescence (IF) investigations of the skin were performed in thirty patients with progressive systemic sclerosis (scleroderma) and eight patients with mixed connective tissue disease (MCTD). The results show that speckled epidermal nuclear immunoglobulin deposition occurs not only in MCTD but also in true scleroderma. Granular IgM deposition at the dermo-epidermal junction of light-exposed skin was detected in both groups of patients, but six of eight MCTD patients also showed a granular IgM band in non-exposed skin. Antinuclear antibodies (ANA) were demonstrated in the sera of 96% and 100% of patients with scleroderma and MCTD respectively. The pattern of nuclear IF staining in scleroderma included dense fine speckles, large coarse speckles, threads, nucleolar and centromere staining. In MCTD, by contrast, the ANA staining pattern consisted of threads. The significance of ANA titres and immunological specificities for the in vivo reaction of serum ANA with epidermal nuclear antigens is discussed.     

Epithelium of the human palatal tonsil used for new test antigen in IIF examination (for pemphigus, pemphigoid antibodies and ANA).

The author presents the results of IIF examinations for pemphigus, pemphigoid antibodies and antinuclear antibodies (ANA) on the epithelium of the human palatal tonsil obtained by tonsillectomy in patients with chronic hypertrophic tonsillitis. Tonsil epithelium is more sensitive to intercellular antibodies (IC Ab), and antibodies of the basal membrane zone (BMZ Ab) than sample tissue of the monkey esophagus, while its reaction to ANA is the same or even weaker than monkey esophagus.     

Possible drug-induced pemphigus-like antibodies with the clinical manifestation of erythema multiforme

A patient with bullae and target lesions on the extremities and mucous membranes was seen with the clinical picture of erythema multiforme following an episode of pneumonia and a course of penicillin G potassium and tobramycin sulfate therapy. An unusually high titer of intercellular circulating (IC) antibodies was identified in the serum by indirect immunofluorescence (IF) microscopy, but direct lesional IF microscopy study results were negative. These IC antibodies were not true pemphigus antibodies and can best be termed pemphigus-like antibodies. These antibodies were characterized by their ability to fix complement, in contrast to pemphigus antibodies, which apparently fail to do so.     

Substrate specificity of antinuclear antibodies in scleroderma.

Studies of antinuclear antibodies (ANA) were carried out in 39 cases of systemic scleroderma and for comparison in 19 cases of systemic lupus erythematosus (SLE) and 4 of mixed connective tissue disease (MCTD) using indirect immunofluorescence (IF) methods under standard conditions. The results on three different substrates--monkey esophagus, guineapig lip and rat liver--are reported. In 48.7% of scleroderma cases ANA showed a substrate specificity. The highest percentage of positive results in scleroderma was obtained on monkey esophagus (97.4%) and the lowest on rat liver (61.5%). In SLE and MCTD, in contrast, only about 13% of the sera displayed such specificity. If only sera with substrate specificity are considered, the positive results on monkey esophagus and rat liver are 94.7% and 21.1%, respectively. Titers of sera reacting positively on 2 or 3 substrates were mostly in agreement, although some sera both in systemic scleroderma and SLE showed higher titers on monkey esophagus. The IF pattern was usually the same regardless of the substrate, Tests for ANA in scleroderma should be performed on at least 2 substrates simultaneously.     

Antinuclear antibodies in patients with polymorphic light eruption: a long-term follow-up study

BACKGROUND: Previous studies have shown elevated titres of antinuclear antibodies (ANA) in 2.9-19% of patients with polymorphic light eruption (PLE). A diagnosis of lupus erythematosus (LE) was finally established in some of these ANA-positive patients. OBJECTIVES: To investigate whether the presence of ANA in patients with PLE merely represents an epiphenomenon or is associated with an increased risk of eventual progression to LE. METHODS: We identified 472 patients with PLE who had received prophylactic photo(chemo)therapy between 1986 and 2003 and were routinely tested for the presence of ANA. All ANA-positive (ANA titre of>or=1:80) patients were asked to attend for a follow-up examination comprising a medical history, complete skin inspection and a detailed laboratory analysis including ANA and antibodies against extractable nuclear antigens. RESULTS: Of all the patients, 55 (11.7%) were found to be ANA positive on one or several occasions, and three (0.6%) also had antibodies to SS-A/Ro. Thirty-nine (71%) of all ANA-positive patients including all Ro+ subjects were available for follow-up after a median follow-up period of 8 years (interquartile range 5-11.5). Twenty-five patients showed persistence of ANA positivity with a median titre of 1:160 (range 1:80-1:640), whereas in 14 patients ANA titres had returned to normal levels. None of the patients revealed additional clinical, histopathological or laboratory abnormalities suggestive of LE. CONCLUSIONS: After a median follow-up period of 8 years none of the ANA-positive patients developed LE. Our findings indicate that PLE is a benign disease without tendency to progress to LE.     

Positive direct immunofluorescence and autoantibody profiles in psoriasis patients

Psoriasis and systemic lupus erythematosus are common, but their coexistence is thought to be infrequent. Each of these diseases has a broad clinical spectrum, so that diagnosis may not be straightforward. This study aimed to investigate the following immunological parameters in psoriasis patients: (i) direct immunofluorescence (DIF); (ii) antinuclear antibody; (iii) anti-double-stranded DNA (dsDNA); (iv) anti-Ro; and (v) anti-nuclear ribonucleoprotein (nRNP). Of 300 cases, comprising 189 men (62.9%) and 111 women (37.1%), 17 (5.7%; 10 men, seven women) were positive for at least one immunological parameter. Nine of 300 (3%; seven male, two female) biopsy specimens from sun-exposed psoriatic lesions demonstrated bright continuous bands of granular IF along the dermoepidermal junction with immunoreactant immunoglobulin (Ig)G, IgM, C3 and fibrinogen. The intensity of IF at the dermoepidermal junction was graded 3+ and 2+. Three cases demonstrated IgM, two had IgG, two fibrinogen and six cases showed C3. Three cases demonstrated more than one immunoreactant. One case demonstrated C3 at the vessels. No specimen demonstrated IgA deposition. Three hundred sera were obtained from patients with psoriasis, of which five demonstrated elevated antinuclear antibody (ANA) titer; dilution titer varied from 1:80-640. Three had a homogeneous pattern and two had a speckled pattern. None had a peripheral pattern. Five (1.7%) of 300 demonstrated anti-Ro, two had negative ANA, and three were positive ANA, two of which were speckled and the other homogenous. Anti-dsDNA, anti-Sm and anti-nRNP were not detected. Ten patients had positive DIF but negative ANA, while five had positive ANA; all had negative DIF results. Thus, the incidence of psoriasis and lupus erythematosus coexistence is low and a baseline immunological screening test for psoriasis might not prove worthwhile.     

The prevalence of antinuclear antibodies in patients with apparent polymorphic light eruption

Polymorphic light eruption (PLE) is a very common photosensitive disorder, the most important differential diagnosis of which is lupus erythematosus (LE). One-hundred and forty-two patients with PLE were screened for circulating antinuclear (ANA), Ro and La antibodies over a 2-year period. Results were negative in 66 patients. Sixty-two patients had low-titre ANA of various patterns, ranging from trace to 1/80 without evidence of LE although one later developed subacute cutaneous LE. Fourteen had more significant findings, six with ANA ranging from 1/160 to 1/1280 but no anti-Ro antibodies, four with ANA ranging from 1/160 to 1/1280 and also with anti-Ro antibodies and four patients with anti-Ro antibodies but low-titre ANA, one of whom later developed discoid LE. Three of these 14 patients fulfilled the American Rheumatism Association criteria for the diagnosis of systemic LE, but it was not certain in any of the patients whether the PLE-like rash represented cutaneous LE or coincidental PLE. However the overall 10% incidence of definite or possible LE in patients with suspected PLE suggests that all PLE patients should be screened for LE.     

Clinical significance and correlation of antinuclear antibodies and anti-DNA antibodies.

The clinical significance and correlation of antinuclear antibodies (ANA) and anti-DNA antibodies was studied using 142 ANA positive sera from different patients having various diseases. High titers of ANA were found briefly in systemic lupus erythematosus and sometimes in scleroderma or mixed connective tissue disease. The peripheral pattern of ANA was seen exclusively in systemic lupus erythematosus and occasionally in mixed connective tissue disease. Anti-DNA antibodies could be found in systemic lupus erythematosus, discoid lupus erythematosus, scleroderma, chronic active hepatitis, but a high titer of anti-DNA (over 60 unit/ml) was present only in patients with systemic lupus erythematosus, especially those having lupus nephritis. There was little correlation between ANA and anti-DNA antibodies.     

In vivo cutaneous antinuclear antibody positivity in palisaded neutrophilic and granulomatous dermatitis

Palisaded neutrophilic and granulomatous dermatitis (PNGD) is commonly associated with underlying systemic inflammatory and neoplastic diseases, infections, and drug reactions. In vivo cutaneous antinuclear antibodies (ANA) have been described in skin biopsies from patients with known autoimmune disorders, but not previously reported in the setting of PNGD. We present two patients with systemic lupus erythematosus (SLE) and histopathologically confirmed PNGD. Direct immunofluorescence (DIF) studies revealed in vivo cutaneous ANA positivity in both patients. DIF findings in the skin mirrored serum autoantibody results. ANA positivity in skin specimens is reported as highly predictive of systemic connective tissue diseases (SCTD), although specific testing is not currently recommended as part of the laboratory work-up or diagnostic criteria for these disorders. In this case report, positive ANA results in skin biopsies of PNGD reflect the serological findings and clinical evidence of SLE in both patients. In vivo cutaneous ANA positivity is an interesting and supportive finding in PNGD in the setting of SCTD.     

Terbinafine-induced subacute cutaneous lupus erythematosus

BACKGROUND: Recently, the induction of subacute cutaneous lupus erythematosus (SCLE) and exacerbation of systemic lupus erythematosus by terbinafine have been reported. OBJECTIVE: We describe 4 cases of SCLE, one associated with chilblain lupus, which occurred during therapy with oral terbinafine for onychomycosis. METHODS: Of 21 consecutive patients with SCLE attending the outpatient dermatology department at Muenster University clinic during a 1-year period, 4 patients with terbinafine-induced SCLE were seen. Patients were examined fully and photographed; histologic findings as well as serologic and follow-up data were evaluated. RESULTS: In addition to high titers of antinuclear antibodies (ANA) with a homogeneous pattern, anti-Ro(SS-A) antibodies were present; in 3 of 4 women, anti-La(SS-B) antibodies were also found. All patients had anti-histone antibodies as in drug-induced lupus and showed the characteristic genetic association of SCLE with the HLA-B8,DR3 haplotype; moreover, in 2 cases, HLA-DR2 was also present. After discontinuation of terbinafine, ANA titers decreased; anti-histone antibodies also became undetectable within 4(1/2) months in 3 patients concomitant with subsidence of the SCLE eruption in all patients. CONCLUSION: Terbinafine is a drug that appears to infrequently induce SCLE with high titers of ANAs and anti-histone antibodies in genetically susceptible persons.     

Serum soluble nucleosome and the broad family of antinucleosome antibodies are associated with organ and tissue damage in systemic lupus erythematosus in a Chinese population

Nucleosomes and the broad family of antinucleosome antibodies (ANAs; anti-double-stranded DNA, antihistone and antinucleosome antibodies) may contribute to the pathogenesis of systemic lupus erythematosus (SLE). We collected clinical information on 90 patients with SLE and 73 healthy volunteers and measured serum levels of the ANA family using a double-sandwich ELISA. The results showed that the levels of serum nucleosomes of patients with SLE was significantly lower and the levels of ANA were significantly higher than healthy controls. Negative correlations between serum nucleosomes and ANA, and positive correlations between individual ANAs were found. Patients with SLE with positive ANA had a significantly higher frequency of renal disorders than those with negative ANA. Determination of serum nucleosomes and ANAs contributes to SLE monitoring.     

Juvenile linear scleroderma associated with serologic abnormalities

We investigated 24 juvenile cases of linear scleroderma for the presence of systemic disease and serologic abnormalities. Thirteen of 24 patients had antinuclear antibodies (ANA) at titers of 1:40 or greater. Rheumatoid factor (titers greater than or equal to 1:20) was detected in seven of 17 patients tested, five of whom also had ANA. Two of five patients with ANA and rheumatoid factor had systemic diseases, such as nephritis and Raynaud's phenomenon. One patient with ANA developed typical dermatomyositis. Consequently, patients with linear scleroderma may be at some risk for developing systemic collagen-vascular diseases. On initial presentation, patients with linear scleroderma should give a complete history and receive a thorough physical examination as well as undergo laboratory evaluations for the presence of ANA and rheumatoid factor. Long-term observation with periodic reevaluation is appropriate for many members of this group.     

Serologic abnormalities in patients with endemic pemphigus foliaceus (Fogo selvagem), their relatives, and normal donors from endemic and non-endemic areas of Brazil

Based on epidemiologic data, a current hypothesis states that Fogo selvagem (FS) may be triggered by environmental factors present in endemic areas of Brazil. Because the appearance of new cases is limited to those areas, we wanted to ascertain if the presence of the pemphigus autoantibodies was restricted to the patients. To further delineate the restriction of the autoantibody response in these patients we also investigated the presence of lupus-associated autoantibodies. Using indirect immunofluorescence (IF) we tested the sera of patients with FS (n = 196), their relatives (n = 138), their cohabitants (n = 13), and normal donors from endemic (n = 38) and non-endemic areas (n = 44) for pemphigus autoantibodies. Antinuclear antibodies (ANA) and anti-nDNA antibodies were determined by indirect IF against Hep-2 cells and Crithidia lucilliae, respectively. Autoantibodies against nRNP, Ro/SSA, La/SSB, and Sm were assayed by double immune diffusion in agarose gels. FS autoantibodies were present in the sera of all patients with active disease (n = 196, 100%, titers greater than 40 to 2560), but were not found in any sera from normal individuals in endemic or non-endemic areas. The titer of the FS autoantibody showed a rough correlation with the extent and activity of the disease. Furthermore, lupus-associated autoantibodies were not present in any of the tested samples. We conclude the FS antiepidermal autoantibodies are specific serologic markers of the disease and are not present in unaffected individual from the endemic areas. As such, they provide an important marker that should be useful in ongoing epidemiologic studies aimed at identifying putative etiologic agent(s).     

Antinuclear antibody studies in chronic cutaneous discoid lupus erythematosus.

The prevalence of antinuclar antibodies (ANA) in chronic cutaneous discoid lupus erythematosus (DLE) is influenced by both patient selection and test sensitivity. If one excludes serum samples from patients with DLE with a history suggesting extracutaneous disease and defines the significance of the ANA test by simultaneously testing serum samples from patients with well-characterized connective tissue diseases, then only a small percentage of the patients with DLE have ANA at significant titers. These patients with DLE do have a higher pervalence of positive ANA tests at low serum dilutions when compared with controls, but only a few have positive ANA tests at titers comparable to those seen in patients with active systemic connective tissue diseases.     

Clinicopathological evaluation of in vivo epidermal nuclear fluorescence

Background.  Nuclear fluorescence in keratinocytes is an occasional phenomenon, often present in autoimmune diseases, especially in connective-tissue disease (CTD); however, its clinical significance remains unclear.
Aim.  To investigate the profile of patients with positive nuclear staining on direct immunofluorescence (DIF) of skin samples.
Methods.  A retrospective analysis of 28 patient records from our immunodermatology laboratory was performed between May 2003 and June 2006. Inclusion criteria were the presence of autoantibodies (IgG, IgA or IgM) or complement (C3) binding keratinocyte nuclei on DIF.
Results.  The most prevalent diseases related to the nuclear keratinocyte DIF staining were systemic lupus erythematosus ( n  = 9), mixed CTD ( n  = 3), overlap syndrome ( n  = 3), Sjögren's syndrome ( n  = 1), and CREST (calcinosis, Raynaud's phenomenon, oesophageal dysmotility, sclerodactyly and telangiectasia) syndrome ( n  = 1). Serum antinuclear antibody (ANA) was positive in 20 of 28 patients, with titres varying from 1 : 160 to 1 : 1280. Of the 20 patients with positive anti-nuclear antibodies (ANA), 17 were positive for anti-extractable nuclear antigen antibodies, 12 had anti-SSA/Ro, 11 had anti-SSB/La and 8 had anti-ribonucleoprotein. Eight patients were negative for ANA. Positive predictive value of in vivo ANA for systemic CTDs was 75%.
Conclusion.  The present data suggest that in vivo ANA evaluation is an additional and feasible auxiliary tool for diagnosing CTDs.     

Antinuclear antibodies during Puva therapy.

During PUVA therapy 7 patients out of 34 with severe psoriasis developed circulating antinuclear antibodies (ANA) (21%). Before treatment only 3 patients of 50 (6%) considered for PUVA had detectable ANA. The ANA titres were usually low. Antibodies against native DNA as studied with the Crithidia luciliae test, were not found, and blood and urinary screening for collagenosis was negative. All 7 patients responded well to the PUVA treatment. The significance of these findings remains to be determined.     

Is minocycline therapy in acne associated with antineutrophil cytoplasmic antibody positivity? A cross‐sectional study

BACKGROUND: Minocycline (MN), one of the commonly prescribed therapies for acne, is known to be associated with autoimmune disorders including drug-induced lupus. However, data are sparse regarding the prevalence of autoimmune disease in acne or in patients with acne treated with MN. OBJECTIVES: To establish the prevalence of antinuclear antibodies (ANA), antineutrophil cytoplasmic antibodies (ANCA) and new autoimmune syndromes in an MN-exposed and unexposed population with acne. METHODS: In a cross-sectional study, 252 patients with acne vulgaris were assessed. Sixty-nine per cent had been exposed to MN at some point or were taking the drug at the time of the interview. Data recorded included duration of disease (acne) and drug history as well as possible side-effects of drugs, in particular joint symptoms (pain and swelling). In addition, blood was taken for ANA, ANCA, liver function tests and HLA analysis. RESULTS: There was no statistical difference in the prevalence of ANA positivity between patients exposed (13%) or not exposed (11%) to MN. However, higher titres of ANA (1/160 or higher) were found in the MN-exposed group (45% compared with 12% in the unexposed group). ANCA positivity was found in 7% of the MN-exposed group but no positivity was found in the unexposed cohort (P = 0.022). In 58% of cases, the ANCA detected were of the perinuclear pattern (p-ANCA) with myeloperoxidase specificity, and this finding was associated with clinical symptoms in the majority of cases. Two p-ANCA-positive patients were thought in retrospect to have developed a drug-induced lupus syndrome. CONCLUSIONS: ANA positivity is seen in patients with acne irrespective of exposure to MN; however, p-ANCA appear to be a serological marker for developing autoimmune disease in patients receiving MN.     

Specificity of antinuclear antibodies in scleroderma-like chronic graft-versus-host disease: clinical correlation and histocompatibility locus antigen association

Summary Chronic graft-versus-host disease after bone marrow transplantation presents, in a few cases, as mild to severe scleroderma-like changes. Patients with chronic graft-versus-host disease with and without sclerodermatous skin changes were analysed for antinuclear autoantibodies (ANA) and antinucleolar autoantibodies (ANoA) and the results correlated with disease symptoms and histocompatibility locus antigen (HLA) pattern. Nineteen patients with chronic graft-versus-host disease and scleroderma-like skin changes. 18 with chronic graft-versus-host diseae without scleroderma. and 17 controls on immunosuppressive treatment were screened for ANA and ANoA using enzyme-linked immunosorbent assay, immunodiffusion and immunoblot techniques. Four patients with severe scleroderma had antibodies to topoisomerase I. two had antibodies against PM-Scl. both characteristic serological findings in idiopathic systemic scleroderma. One patient had La/SSB antibodies and, in three cases, antibodies to the nucleolar antigen C23 (nucleolin) could be identified. A possible correlation between antinucleolin antibodies and disease activity was observed. HLA-Al. -B1. and -B2 were found significantly more often in patients with scleroderma-like symptoms in comparison to patients without scleroderma-like symptoms. Chronic graft-versus-host disease with seleroderma-like manifestations can be associated with the occurrence of ANA specific for idiopathic scleroderma. The development of seleroderma after bone marrow transplantation might have a HLA-linked genetic background.     

Nuclear antibodies as serologic markers in progressive systemic scleroderma

In all, 36 patients with progressive systemic sclerosis (29 women, 7 men) were studied clinically and immunologically; 15 patients had acrosclerosis (type I) and 21, sclerosis extending beyond the wrist (type II). The sera of all patients were evaluated for ANA (HEp-2-cells), Scl-70, centromere and other ENA antibodies. The centromere antigen was characterized by immunoblotting. All patients had high-titer ANA antibodies (100%); 36% of patients had the Scl-70 antibody (a marker antibody for PSS); and in 22% of our patients a centromere antibody was detected. In all cases the anti-centromere sera reacted with a 19.5-kd polypeptide and in 2 cases they reacted with 23- and 25.5-kd proteins in addition. In patients with centromere antibodies there was increased organ involvement (heart, lung, kidney) compared with patients who had anti-Scl-70 or other nuclear antibodies.