ANA Staining Research Articles

OA17 An evolving case of Anti PM/Scl Anti body associated Aggressive Systemic Sclerosis

OA17 An evolving case of Anti PM/Scl Anti body associated Aggressive Systemic Sclerosis

Exosome circRNAs and ceRNA network profiles in different ANA sera.

Increasing evidences show that circRNAs are associated with some autoimmunity diseases either as a biomarker or therapeutic target. Exosomes containing nucleic acids and proteins are found in sera of series diseases and could serve as either diagnostic or therapeutic target. ANA serves as first common diagnostic test for autoimmunity disease, different ANA staining reflecting different types of autoimmunity disease. Till now, whether different ANA sera exosomes express different circRNAs and relevant ceRNA networks are still shortage of investigation. This study analyzed circRNAs, miRNAs, and their interaction networks in different ANA sera exosomes by high-throughput sequencing. It found no significant difference of total circRNAs and miRNAs amount across different ANA sera exosomes. However, significant differences were found of circRNAs, miRNA constituents, function analysis by KEGG and GO, and their ceRNA networks including miRNA-circRNA and miRNA-mRNA among different ANA sera exosomes, suggesting sera exosome circRNAs as either biomarker or mechanism of autoimmunity diseases.

Anti-Nuclear Antibodies Patterns in Patients With Systemic Lupus Erythematosus and Their Correlation With Other Diagnostic Immunological Parameters.

BackgroundAntinuclear antibodies (ANA) are major immunodiagnostic tools in systemic lupus erythematosus (SLE); however, their clinical and pathogenic roles are not yet elucidated and are a subject of controversy.ObjectivesThe aim of the study is to explore the pathogenic significance of ANA patterns among SLE patients, by analyzing their association with ANA titers, complement levels and other pathogenic immune markers, namely, anti-double-stranded DNA (anti-dsDNA), complements C3 and C4, rheumatoid factor (RF), anticardiolipin antibodies IgG (ACL IgG) and IgM (ACL IgM), Beta-2 Glycoprotein 1 Antibodies (β2-GP) IgG (β2-IgM) and IgM (β2-IgM), and lupus anticoagulant (LA).MethodA comparative cross-sectional study was conducted among 495 SLE patients, who were diagnosed and classified by consultant rheumatologists according to the new European League Against Rheumatism (EULAR)/American College of Rheumatology (ACR) 2019 criteria. SLE immunodiagnostic profiles were analyzed including the following parameters: ANA antibody titers and staining patterns, anti-dsDNA, C3 and C4 levels, aCL, and anti-β2-GP and LA.ResultThe most frequently observed ANA patterns were the speckled (52.1%) and homogeneous (35.2%) patterns, while other patterns were rare representing less than 7% of the patients each. ANA titers were highest in patients with mixed pattern followed by the speckled pattern. Of all the investigated patterns, the peripheral pattern showed the most pathogenic immune profile, namely, highest levels of anti-dsDNA, lowest levels of C4, and highest levels of aCL and β2-GP IgG and IgM.ConclusionThis retrospective study showed that speckled followed by homogeneous ANA patterns were predominant accounting for 52.1 and 35.2% of the patients. The ANA pattern showed several associations with other immune markers that are documented to have significant clinical implications in SLE. Peripheral, mixed, and speckled patterns were associated with higher profiles of immune markers indicative of a potential prognostic value of these patterns in SLE.

Are the Specific and Nonspecific ANA Staining Patterns of Behçet's Disease Patients Important?

The autoinflammatory character of Behçet's Disease has led researchers to investigate the role of autoantibodies. However, no significant positive result has been reported for autoantibody tests for the disease. To investigate the specific and nonspecific staining patterns of Behçet's Disease (BD) patients. 140 patients (87 females, 53 males) with an average of 41.9±3 years who were being followed up for Behçet's Disease, and a control group consisting of a total of 736 (464 females, 272 males) healthy volunteers made up of blood donors without any disease whose average age was 50.2±4 years were included in the study. Peripheral venous blood was collected from the patients and the sera were separated. Patient sera were studied by indirect immunofluorescence antibody test (IFA) at a dilution of 1/40 and 1/100. A total of 140 (87 females, 53 males) Behçet's Disease patients and 736 (464 females, 272 males) healthy controls were examined. The rate of ANA positivity was 11.6% in the control group and 10.7% in the Behçet's Disease group. In general, no difference was detected between the patients and the healthy controls in terms of autoantibody positivity (p>0.05). However, when examined in terms of patterns, the low detection of DFS70 and the observation of centriole staining type patterns in Behçet's Disease patients was noteworthy (p<0.05). Autoantibody tests, which hold an important place in classic autoimmune diseases, are not necessary for Behçet's patients, but they should be examined in terms of nonspecific patterns.

Anti-dense fine speckled (DFS) antibody: its staining pattern in indirect immunofluorescence and its clinical relevance.

Background: Dense fine speckled (DFS) pattern is defined by very intense, heterogeneous speckled staining of nucleoplasms of interphase HEp-2 cells and chromosomal areas of metaphase cells. The association of Anti-DFS70 and rheumatologic signs, symptoms, and diagnosis were evaluated.Methods: One-hundred-eight anti-DFS70 positives who got consecutively admitted to the Rheumatology clinic between January and June 2020 were analyzed. The clinical and laboratory findings of positives for anti-DFS70 antibody were compared with those with DFS pattern ANA IFA staining rates. Also, anti-DFS70 positivity rates and their correlation with the DFS staining pattern were analyzed retrospectively in 1016 CTD patients.Results: The most common complaint was joint pain seen in 77 (71.3%) and the most common laboratory abnormality was RF-positivity observed in 10/108 (9.3%) who had anti-DFS70 positivity. The most common ANA staining pattern was DFS (72/108; 66.7%); one-third had other than DFS. No statistical significance was found for the association of any of the rheumatological complaints and laboratory findings with the DFS staining pattern. ANA analysis was performed in a total of 964/1016 (94.88%) CTD patients and 44 (4.56%) of these positive for anti-DFS70. The correlation coefficient showed good correlations between the DFS pattern staining and anti-DFS70 antibody positivity (r=+0.773, p<0.001).Conclusions: Anti-DFS70-positives have a low rate of CTD. A low anti-DFS70 positivity rate was observed in patients with CTD. As such, it can be considered that anti-DFS70 does not predict CTD or even excludes it.

Are the specific and nonspecific ANA staining patterns of Behçet's Disease patients important?

BackgroundThe autoinflammatory character of Behçet's Disease has led researchers to investigate the role of autoantibodies. However, no significant positive result has been reported for autoantibody tests for the disease. AimsTo investigate the specific and nonspecific staining patterns of Behçet's Disease (BD) patients. Methods140 patients (87 females, 53 males) with an average of 41.9±3 years who were being followed up for Behçet's Disease, and a control group consisting of a total of 736 (464 females, 272 males) healthy volunteers made up of blood donors without any disease whose average age was 50.2±4 years were included in the study. Peripheral venous blood was collected from the patients and the sera were separated. Patient sera were studied by indirect immunofluorescence antibody test (IFA) at a dilution of 1/40 and 1/100. ResultsA total of 140 (87 females, 53 males) Behçet's Disease patients and 736 (464 females, 272 males) healthy controls were examined. The rate of ANA positivity was 11.6% in the control group and 10.7% in the Behçet's Disease group. In general, no difference was detected between the patients and the healthy controls in terms of autoantibody positivity (p>0.05). However, when examined in terms of patterns, the low detection of DFS70 and the observation of centriole staining type patterns in Behçet's Disease patients was noteworthy (p<0.05). ConclusionAutoantibody tests, which hold an important place in classic autoimmune diseases, are not necessary for Behçet's patients, but they should be examined in terms of nonspecific patterns.

Immune checkpoint inhibitor-induced inflammatory arthritis: a novel clinical entity with striking similarities to seronegative rheumatoid arthritis.

To determine the clinical and serologic similarities and differences between inflammatory arthritis induced by immune checkpoint inhibitors (IA-irAE) and rheumatoid arthritis (RA). In this retrospective cross-sectional comparative study, 20 patients with IA-irAE were age and sex matched to 40 seropositive and 40 seronegative RA patients. Electronic medical records were reviewed from diagnosis of inflammatory arthritis through May 2019. Arthritis characteristics, treatment, and relevant laboratory and serologic studies were captured. Clinically, IA-irAE differed from seropositive and seronegative RA with respect to disease duration (4.18 versus 11.59 and 13.3months, respectively, p = 0.005 (IA-irAE vs seropositive RA), p = 0.002 (IA-irAE vs seronegative RA)), polyarticular joint involvement at presentation (75% versus 97.5% and 100%, p = 0.013, p = 0.003), absence of erosive changes (5.9% vs 43.6% and 53.8%, p = 0.005, p = 0.001), mean prednisone dose (24.7mg versus 16.53mg and 15.68mg, p = 0.008, p = 0.005), and use of methotrexate (5.0% versus 85.0% and 70.0%, p < 0.0001, p < 0.0001). Serologically, IA-irAE closely resembled seronegative RA. ANA positivity was seen in a minority of patients and did not differ significantly between all groups; however, the ANA staining pattern (speckled) was similar between IA-irAE and seronegative RA (100% versus 75%, respectively) and was not commonly observed in seropositive RA (18.2%). IA-irAE is a new subset of IA that resembles seronegative RA immunologically. Our findings suggest that further study of IA-irAE might provide a window into underlying pathogenic mechanisms of early-stage seronegative RA. Key Points • Comprehensive comparison of clinical features between inflammatory arthritis irAE (IA-irAE) and regular rheumatoid arthritis indicates IA-irAE as a new subset of inflammatory arthritis. • IA-irAE resembles seronegative RA immunologically, suggesting that study of IA-irAE may provide a window into underlying pathogenic mechanisms of early-stage seronegative RA.

Clinical characteristics and change in the antibody titres of patients with anti-MDA5 antibody-positive inflammatory myositis.

The aim of this study was to evaluate the clinical characteristics of patients with anti-melanoma differentiation-associated gene 5 (MDA5) antibody-positive inflammatory myositis, and the change in anti-MDA5 antibody titres before and after onset. For 105 PM/DM patients, newly diagnosed in our hospital within the period 2008-2016, serum anti-MDA5 antibody levels were measured at diagnosis and after treatment by ELISA using the MESACUP anti-MDA5 test. The relationships between anti-MDA5 antibody levels and clinical manifestations, laboratory data, and mortality were examined. Compared with patients who were anti-MDA5 antibody negative, those who were antibody positive demonstrated more frequent dermatitis, clinically amyopathic DM, interstitial lung disease and rapid-progressive interstitial lung disease, as well as significantly higher serum ferritin, significantly lower creatine kinase and aldolase, and significantly less frequent ANA (⩾1:160) and anti-cytoplasmic pattern of ANA staining positivity. Anti-MDA5 antibody titres were examined before disease onset in two patients; one showed antibody positivity with low titres 2 years earlier, while both exhibited increased titres at onset. Anti-MDA5 antibody titres declined significantly less in survivors than in non-survivors after treatment; however, there was no significant difference between the two groups when the rate was compared at 2 months after treatment. An initial decrease in anti-MDA5 antibody titre after commencement of treatment was observed in most of the patients, including in fatal cases, suggesting that this may not necessarily be a useful marker for treatment of patients with DM.

Relationship between certain T helper cytokines and ANA staining: who is the helper?

This study investigated some T helper cytokines' association with different ANA positive sera. Sera of 53 ANA-positive subjects (11 were stained of homogeneous pattern, 12 of speckled particle pattern, 9 of nucleolar pattern, 15 of centromere pattern, and 6 of the peripheral nuclear type) were collected and analyzed by ELISA for related cytokines and concentrations were compared with each other and 16 healthy donors. Investigated cytokines including IL-12(p70) and IFN-γ varied differently in the ANA sera. This study demonstrated that some T helper cytokines varied with different ANA-positive sera, suggesting that they may play different roles in related autoimmune diseases.

The cAMP Response Element Binding Protein (CREB) Overexpression Induces Myeloid Transformation in Zebrafish

AbstractAbstract 4727 Introduction.Transcription factors has been widely confirmed to play a central role in acute leukemia, and among them the cyclic-adenosine monophosphate response element-binding protein (CREB) was identified to be involved in triggering acute myeloid leukemia. CREB protein overexpression has been found in the bone marrow of most pediatric patients with acute leukemia, and it has been shown to induce myeloid leukemia progression in vitro and in vivo by driving the upregulation of a series of target genes. CREB is evolutionarily conserved from invertebrates to human, and although the main focus of zebrafish research has traditionally been developmental biology, this model is currently used for cancer research. In fact, tumors induced in ZF have similar morphology and activated signaling pathways of human cancers. Aim.We aimed to develop a ZF harboring CREB overexpression in myeloid precursors, and by monitoring the induced myeloid malignancy, we will characterize CREB signaling and its involvement in the myeloid transformation process. Results.Using a Multisite Gateway System we constructed a vector containing human-CREB gene fused to EGFP, driven by an early myeloid promoter, zPu.1, to induce CREB overexpression specifically in the myeloid lineage of ZF. We injected the EGFP-CREB plasmid into one-cell stage zebrafish embryos, and monitored its expression during early development. Results showed that CREB was expressed in ZF zones typical of myelopoiesis, such as in the intermediate cell mass and anterior lateral mesoderm migrating through the yolk from 12 to 48 hours post-fertilization (hpf), and few cells circulating throughout the embryo from 24 hpf. CREB transcriptional activity on cAMP response elements (CREs) was measured and confirmed by luciferase assay. The co-localization of EGFP-CREB with the CRE-mCherry reporter was seen by fluorescence microscopy analysis. To test CREB target gene expression, embryos injected with CREB or Empty vector were subjected to RNA extraction and RQ-PCR. CREB over-expression was documented (up to 103 fold), and c-myb, stat3, rb, runx1, cyclins A, B, D1 and E2 were found upregulate at 24 and 48 hpf. By RNA whole mount in situ hybridization, we revealed pu.1, mpo, gata1 and fli.1 increased signals, suggesting a general disruption of the main hematopoietic factors.Ten CREB-injected zebrafish have been grown to adulthood and all of them displayed an abnormal/sick phenotype with abdominal enlargement and swelling from 10 to 12 months. Histochemical H&E staining performed on paraffin sections revealed an abdominal tumor and metastatic infiltration in kidney, skin, gills, muscles, adipose tissue. PAS staining identified the myeloid character of the tumor mass and kidney marrow. Wright-Giemsa and ANAE staining showed the predominance of clonal monocytes on appositions of tumor mass and kidney marrow of sick ZF. Tumor mass cell sorting displayed a clonal feature of the tumor with an enrichment of the myeloid-monocitic compartment. RNA extraction from the mass shows CREB overexpression as well as its target genes (c-myb, runx1, cyclin A1, cyclin B1, cyclin B2, cyclin E1), confirming CREB involvement in this tumor formation. Conclusion.We demonstrated that CREB overexpression recapitulates myeloproliferative disorder in ZF, supporting the ZF as a suitable model for studying CREB-induced leukemia. The discovery through which targets CREB would preferentially mediate the myeloid transformation will help to unravel leukemogenesis. ZF model might be useful to test CREB directed drugs. Disclosures:No relevant conflicts of interest to declare.

Antinuclear antibodies: A marker associated with steroid-dependence in patients with Ulcerative Colitis

The autoimmune phenomena and the autoantibody profile have acquired a great importance in ulcerative colitis (UC). Few studies have explored antinuclear antibodies' (ANA) prevalence, but not its association with steroid dependence. We hypothesized that ANA could be a factor associated to steroid dependence and extraintestinal manifestations. We recruited a cohort of 110 consecutive patients with UC between October 2006 and July 2008. Among them, 2 patients with incomplete follow-up, 5 with proctocolectomy, and 6 with concomitant autoimmune disease were excluded. Finally, 97 patients were analyzed. ANA titers and staining patterns were determined by indirect immunofluorescence. Test results were interpreted as “positive” if they were above the ones reported for healthy Mexican individuals. In order to identify patients with non-diagnosed systemic lupus erythematosus we also performed anti-double stranded DNA (anti-dsDNA). Gender, age at diagnosis, C-reactive protein (CRP), disease extent, Mayo Score Activity Index, extraintestinal manifestations, and steroid dependence were analyzed in univariate and multivariate models. Numerical variables were analyzed with t-test, and the categorical with χ2 or Fisher's exact test. P value less or equal to 0.05 was considered as statistically significant. Forty-nine female and forty-eight male patients, 41.7 ± 22.2 years age, with 7.6 ± 6.8 years of evolution of the disease were studied. We found that 52 (53.5%) were positive for ANA, and 0 for anti-dsDNA. Prevalence of ANA was higher in steroid-dependent than in non-steroid-dependent patients (77.8% vs 48.1%, P = 0.020, OR = 3.8, CI 95% 1.1 - 12.5), and in those with uveitis (100% vs 51.1%, P = 0.040) or pyoderma gangrenosum (100% vs 51.6%, P = 0.078). No association was observed with gender, age, CRP, disease extent, and Mayo Score Activity Index. The multiple regression analysis model confirmed that only the presence of steroid dependence remained to be associated with ANA (P = 0.033, OR = 3.9, CI 95% 1.4 - 14.9). The prevalence of ANA in Mexican patients with UC is 53.6%. ANA are associated with steroid dependence in UC patients. Further studies are required to determine the role of ANA as serological marker in order to perform early therapeutic interventions with biological agents.

The presence of non-organ-specific autoantibodies is associated with a negative response to combination therapy with interferon and ribavirin for chronic hepatitis C.

BackgroundNon-organ-specific autoantibodies are found in a considerable number of anti-HCV positive patients. Previous studies investigated the clinical relevance of these antibodies in patients treated with interferon monotherapy, but not combination therapies.MethodsAnti-nuclear, anti-smooth muscle, anti-mitochondrial, anti-neutrophil-cytoplasmatic and anti-liver/kidney microsomal antibodies were determined in 78 consecutive anti-HCV positive patients by indirect immunofluorescence. The presence of these antibodies was related to demographic variables and to the outcome of antiviral combination therapy with interferon-α and ribavirin in 65 patients.ResultsIn our study, positivity for autoantibodies was associated with higher alanine aminotransferase levels and higher mean values for HCV-RNA (p < 0.01). Furthermore, negativity for non-organ-specific autoantibodies was associated with a favourable treatment outcome of combination therapy with at least one negative RT-PCR for HCV-RNA during treatment (OR 4.65, 95% CI 1.31 to 16.48, p = 0.02). ANA and SMA staining patterns and titers were not correlated to treatment response. With multiple logistic regression analysis, positivity for autoantibodies and HCV genotype were independently associated with outcome of antiviral combination therapy (p = 0.02).ConclusionsThe absence of non-organ-specific autoantibodies might indicate a significantly higher chance for viral clearance in response to combination therapy for chronic hepatitis C infection. Therefore, despite of an overall higher treatment response, the addition of the immunomodulatory drug ribavirin could accentuate immunological differences that affect treatment outcome and might have been less obvious in earlier studies analysing interferon monotherapy.

The composition of gingival inflammatory cell infiltrates in children studied by enzyme histochemistry

Abstract Gingival biopsies were obtained from 23 children, aged 5-11 years (8.6 ± 1.8 years). Specimens were taken from areas of the gingiva adjacent to the teeth which were to be extracted because of caries or its sequelae and which clinically had a gingival index score of at least 1. Staining for α-naphthyl acetate esterase with unspecific esterase at pH 5.8 (ANAE) permitted identification of T lymphocytes, monocytes/macrophages, plasma cells and non-reactive (ANAE-negative) cells. Cells which tentatively were identified as “natural killer” (NK) cells were also observed. Differential cell counting was performed for 10 specimens, selected on the basis of the presence of a well-defined inflammatory infiltrate, clear morphology throughout and good ANAE staining. Cell counts confirmed earlier studies showing that lymphocytes predominate in the inflammatory infiltrates in childrens' gingivitis. T lymphocytes dominated particularly in the periphery of the most densely infiltrated areas. Relatively few plasma cells were seen. It was concluded that T lymphocytes dominate in the inflammatory infiltrate in childrens' gingivitis.

The effects of the macrocyclic lactone bryostatin-1 on leukemic cells in vitro.

The macrocyclic lactone bryostatin-1 was found to exert in vitro antineoplastic activity against several leukemic cell lines, including human K562 erythroleukemia, HL60 promyelocytic leukemia, REH and MOLT-4 lymphoblastic leukemias, CCRF-CEM lymphoma, KG-1 myeloid leukemia, and murine P388 lymphocytic leukemia. No statistically significant difference in sensitivity to bryostatin-1 was found between adriamycin-resistant P388 and K526 subclones and their sensitive counterparts. Freshly explanted clonogenic leukemic cells showed a variable sensitivity to bryostatin-1 in 10/12 tested samples. The IC50 of clonogenic leukemic cells was 4 x 10(-3) M bryostatin-1, and that of normal marrow CFU-GM was 10(-5) M. Leukemic cells exposed to bryostatin-1 showed a variable degree of monocytic differentiation as evaluated by ANAE staining and morphology. Bryostatin-1 is also able to inhibit the growth of CFU-GM from myelodysplastic marrow and to shorten the duration of dysplastic hematopoiesis in liquid culture. In conclusion, these data suggest that bryostatin-1 is a potent antileukemic agent in vitro that may be potentially useful for clinical studies.

The composition of gingival inflammatory cell infiltrates in children studied by enzyme histochemistry

Gingival biopsies were obtained from 23 children, aged 5-11 years (8.6 +/- 1.8 years). Specimens were taken from areas of the gingiva adjacent to the teeth which were to be extracted because of caries or its sequelae and which clinically had a gingival index score of at least 1. Staining for alpha-naphthyl acetate esterase with unspecific esterase at pH 5.8 (ANAE) permitted identification of T lymphocytes, monocytes/macrophages, plasma cells and non-reactive (ANAE-negative) cells. Cells which tentatively were identified as "natural killer" (NK) cells were also observed. Differential cell counting was performed for 10 specimens, selected on the basis of the presence of a well-defined inflammatory infiltrate, clear morphology throughout and good ANAE staining. Cell counts confirmed earlier studies showing that lymphocytes predominate in the inflammatory infiltrates in childrens' gingivitis. T lymphocytes dominated particularly in the periphery of the most densely infiltrated areas. Relatively few plasma cells were seen. It was concluded that T lymphocytes dominate in the inflammatory infiltrate in childrens' gingivitis.

Alpha-naphthyl-acetate-esterase activity in cerebrospinal fluid cells.

Mono-histiocytes and T-lymphocytes were assessed by the cytochemical alpha-naphthyl-acetate-esterase (ANAE) stain in 50 CSF samples of patients with various neurological diseases. The ANAE-activity of lymphocytes was decreased in multiple sclerosis and subacute sclerosing panencephalitis, while the activity of mono-histiocytes was increased in the group of infarctions and bacterial and viral infections of the central nervous system. In bacterial meningitis and viral meningo-encephalo-radiculitis the number of ANAE-positive lymphocytes increased after treatment and clinical improvement. ANAE staining appears to be a useful additional tool in CSF cytology in these conditions.

Immunohistological evaluation of alopecia areata treated with squaric acid dibutylester (SADBE)

A total of 19 patients with alopecia areata volunteered for serial biopsies of the scalp skin during SADBE treatment. Regrowth of terminal hair was seen in 12 of the 19 patients on the side of the scalp treated with SADBE for a minimum of four months, but not on a control side treated with sodium lauryl sulphate (SLS). Immunocompetent cells were characterized with ANAE staining and monoclonal antibodies in biopsy specimen showing marked peribulbar and perivascular inflammatory cell infiltrates. Inflammatory cell subclasses were repeatedly evaluated during SADBE treatment. No specific alterations in lymphocyte subclasses and macrophages were seen in relation to the hair growth response. In immunofluorescent studies it was found that in patients with regrowth of hair, immunoglobulins, fibrin and complement, appeared in the hair bulb and along the basement membrane during the treatment. Results were negative in patients without regrowth of terminal hair and in SLS-treated skin. As we could not demonstrate that any cell-mediated mechanism was involved, we suggest that SADBE may induce terminal hair growth through as yet uncharacterized mediators.

T and B lymphocytes, macrophages and Langerhans´ cells during the course of contact allergic and irritant skin reactions in man

The in situ distribution of lymphocyte subclasses, macrophages and Langerhans´ cells was analysed with histochemical (ANAE staining), immunohistochemical (immunoperoxidase) and electron microscopic methods during the course of epicutaneous allergic and irritant skin reactions. Serial biopsies were taken during the first 5-72 hours following the allergen (nickel and chromium) or irritant (non-anoic acid and dithranol) application. Empty and petrolatum-filled Finn chambers were used as controls. No significant differences in the number of mononuclear inflammatory cells or their subclasses were found in the early recognition and elicitation phases of the allergic or irritant-induced reactions. Nor was there any difference in the frequency of lymphocyte-Langerhans´ cell contacts, although exocytic cells were most frequent in dithranol reactions.

Characterisation of blood and synovial fluid lymphocytes from patients with rheumatoid arthritis and other joint diseases by monoclonal antibodies (OKT series) and acid alpha-naphthyl esterase staining.

Mononuclear cell preparations from peripheral blood (PBL) and synovial fluid (SFL) of 27 Patients with rheumatoid diseases (15 patients with definite rheumatoid arthritis (RA), 10 with other inflammatory joint diseases (OJD), 1 with sarcoid arthritis (SA) and 1 with traumatic arthritis (TA) were examined for lymphocyte subpopulations determined by monoclonal antibodies of the OKT series and by the dot-like, acid alpha-naphthyl esterase staining (ANAE) activity. In patients with classic, active RA, blood T cells carrying the OKT8+ (suppressor/killer) phenotype were significantly reduced leading to an elevated OKT4/OKT8 ratio of 4.1 +/- 0.4 compared with 2.1 +/- 0.1 in healthy controls. In 10 patients with OJD this diminution of OKT8+ cells in peripheral blood was less pronounced or absent. As regards SFL subpopulations, patients with RA and OJD exhibited a similar distribution pattern with an elevation of OKT8+, Ia+ and ANAE negative cells and a similar OKT4/OKT8 ratio of 1.5 +/- 0.3 and 1.6 +/- 0.4, respectively. Similar results were also obtained in the only patient with TA, whereas the patient with SA and one RA patient with relapse after surgical synovectomy exhibited high OKT4/OKT8 ratios, both in synovial fluid and peripheral blood. Neither the OKT markers nor the dot-like ANAE staining pattern were significantly correlated to parameters of systemic or local disease activity as estimated by erythrocyte sedimentation rate and a local disease activity index.

Acid alpha-naphthyl acetate esterase staining of T lymphocytes in human skin

T lymphocytes were stained in order to disclose alpha-naphthyl acetate esterase activity (ANAE staining) by adopting the method described by Mueller et al. In ANAE-staining of frozen sections from human lymph nodes, more than 90% of the lymphocytes in paracortical areas (T cell areas) were ANAE-positive, but in cortical follicles (B cell areas) less than 5% of the cells were positive. Lymphocytic infiltrations in various dermatoses (lichen ruber planus, psoriasis, SLE, atropic dermatitis, erythema multiforme, poikiloderma atrophicans vasculare, and Sézary syndrome) were investigated. A high percentage of ANAE-positive lymphoid cells (greater than 80%) was found in most cases. One patient with chron. lymphatic leukaemia, however, had a smaller proportion of ANAE-positive lymphocytic cells in an erythema multiforme skin infiltrate. ANAE staining seems to be an easy method for the identification of T lymphocytes in skin sections. The results of this investigation support the hypothesis that T lymphocytes have an affinity to skin.