Variable expression of CD49d antigen in B cell chronic lymphocytic leukemia is related to disease stages.

We have investigated the expression of the dual specific adhesion molecule, VLA-4 (CD49d/CD29) on lymphocytes obtained from 62 patients with B-CLL and compared it with normal controls, patients with other hematological malignancies, and umbilical cord blood. The mean CD49d expression in patients with CLL was lower than the other group of leukemia and the CD19+, CD5+ cells of normal peripheral blood and umbilical cord blood (P < 0.001). The patients in RAI stage 0, I and II (early stage) had even lower CD49d expression, whereas patients in RAI stage III and IV (advanced stage) had relatively higher CD49d levels. In vitro adhesion of lymphocytes to fibronectin, being the extracellular matrix ligand of CD49d, was also investigated. Lymphocytes obtained from B-CLL were found to have lower adhesion to fibronectin than that from controls (P < 0.03). Furthermore, CD49d(low) B-CLL cells had lower adhesion to fibronectin, whereas CD49d(high) B-CLL cells showed normal adhesion ratios (P < 0.002). Further phenotypic analyses revealed the presence of myeloid markers (CD13 and CD33) in most of the advanced stage patients, although these were negative in early stage cases. Expressions of CD11a and sIgM were also low but CD11b was relatively higher in the early stages of the disease. On the basis of these results, we concluded that early stages of CLL are correlated with the expression of CD49d(low), CD11a(low), CD11b(high), CD13-, CD33-, sIgM(low) and also had lower fibronectin adhesion, whereas advanced stages of CLL are associated with CD49d(high), CD11a(high), CD11b(low), CD13+, CD33+, SIgM(high) and show normal fibronectin adhesion.

Glucose-6-phosphate dehydrogenase deficiency in Cukurova Province, Turkey.

The prevalence of red cell G6PD deficiency was studied by a fluorescence screening test in 5 different ethnic populations of Cukurova province. Analyses were performed on samples from the parents of each family unit. The frequency of occurrence in the population was calculated using family trees. 4 551 subjects representing 17 225 offspring from 18 villages were screened and a high incidence of G6PD deficiency was found in the Arabic speaking ethnic groups from Adana (10.4%) and Anthiochus regions (8.1%), whereas the deficiency was only 3.1% in Armenians, 0.8% in mixed villages and 0.5% in Turcomans. This study reveals that the high frequency of this deficiency found in some ethnic populations must be taken into account prior to the treatment of malaria with primaquine.

Mechanisms of smooth muscle antibody production: a clinical study in children with infections, haemolytic syndromes, and idiopathic thrombocytopenic purpura.

Sera from 530 children suffering from various diseases and from 64 controls were tested for smooth muscle autoantibodies (SMA) by indirect immunofluorescence. A high incidence of SMA (51-86%) was found in patients with viral and bacterial infections (viral hepatitis, infectious mononucleosis, measles, mumps, chickenpox, typhoid fever, and brucellosis), independently of liver invovlvement, and in patients with acute haemolytic anaemia due to G-6-PD deficiency (48%). By contrast, the incidence of SMA from patients with beta-thalassaemia major and idiopathic thrombocytopenic purpura was no higher than in the controls. The discrepancy in incidence in haemolytic anaemias due to different causes may reflect the effect of endogenous and extrinsic agents. In the viral infections, SMA were mainly of the IgM class and gave an 'SMA-V' staining pattern. In bacterial infections (typhoid fever and brucellosis), SMA were either IgG only or IgM and IgG, and the staining pattern was also mainly 'SMA-V'. In infections which affect or may affect the liver (viral hepatitis, infectious mononucleosis, typhoid fever, and brucellosis), SMA was present at high titres (1:80-1:320), whereas in infections not affecting the liver (measles, mumps, and chickenpox) the titres were lower (less than or equal to 1:80). In most patients SMA occurred transiently and without apparent pathogenetic significance. The antigen against which infection-induced SMA is directed is not actin; its nature has yet to be identified.

Mortality-lowering effect of systemic corticosteroid therapy in severe tetanus.

In a double-blind therapeutic trial, 32 patients suffering from severe tetanus were given prednisolone and 31 a placebo in addition to standard treatment. Mortality was lower in the prednisolone-treated group (31%) than in the placebo-treated group (55%). The women had more severe symptoms and a higher mortality than the men. The results suggest that the addition of a corticosteroid to standard treatment will lower mortality in tetanus, although meticulous nursing care is still the most important factor in treatment.

Double G0/G1 peak in the DNA histogram of aberrant marker positive acute leukemia patients is associated with a poor clinical outcome.

In order to investigate the relationship between aberrant marker expression and DNA ploidy, 61 adult patients with acute leukemia (39 AML and 22 ALL) were studied. Aberrant marker expression was observed in 20 patients (16/39 of AML and 4/22 of ALL patients). In flow cytometric DNA analysis aneuploidy was observed in 18 patients (9/39 of AML and 9/22 of ALL patients). The incidence of aneuploidy in patients with aberrant marker expression was 35% whereas this was 26.8% in patients without aberrant marker expression. Furthermore, 7 patients with aberrant marker expression showed an aneuploid, double G0/G1 peaks appearance whereas the remaining 11 patients with aberrant marker expression had euploid DNA content. Double G0/G1 appearance was not observed in patients without aberrant marker expression. Further analyses revealed that this did not correlate with apoptosis. All 7 patients, who had both aberrant marker expression and double G0/G1 peak had a poor clinical outcome with a short survival and all died within three months whereas three-months survival was 67% for AML, 69% for ALL patients and 81% for patients with aberrant marker expression respectively (p<0.01). Our data indicate that the evaluation of the DNA ploidy in patients with aberrant marker expression may be of prognostic importance.

T and NK cell subset changes with microbial extracts and human HSP60-derived peptides in Behçet's disease.

OBJECTIVE: Microorganisms such as streptococcus and autoantigens such as 60 kD heat-shock protein (HSP60) are implicated in the etiopathogenesis of Behçet's disease (BD). METHODS: Peripheral blood mononuclear cells from patients with BD (n = 16) and healthy controls (HC) (n = 11) were cultured for 5 days with extracts of S. sanguis-KTH-1 (SS), E. coli (EC) and a mixed peptide combination from human HSP60 (aa 136-50, 179-97, 224-58 and 336-51) reported to be associated with BD. T and NK cell subset changes were determined by flow cytometry. RESULTS: In unstimulated 5-day cultures gammadelta+ (both CD4+gammadelta+ and CD8+gammadelta+), CD8+alphabeta+, CD4+CD56+ and CD8+CD11b+ cells were increased in BD compared to HC. In antigen-stimulated cultures of BD patients CD3+ and alphabeta+ T cells responded to HSP60 peptides whereas EC stimulated only CD16/ CD56+ NK cells. In the control group, similar to BD, alphabeta+ and CD4+ T cells responded to HSP60 peptides, however SS and EC mainly activated cytotoxic T cell subsets (CD8+CD11b and CD4+CD56+ T cells). CONCLUSION: Significant increases in unstimulated T cell subsets suggest the presence of an in vivo T cell activation in BD. In both patients and controls similar patterns of responses were observed against different microorganisms, however the role of human HSP60 peptides as immunodominant, crossreactive antigens could not be demonstrated.

IL-8 producing cells in patients with Behçet's disease.

OBJECTIVE: IL-8 is thought to be the principal chemokine responsible for neutrophil activation and tissue infiltration in patients with Behçet's disease (BD). In various studies serum levels of IL-8 were reported to be increased. METHODS: IL-8 mRNA was purified from both peripheral whole blood samples and separated lymphocytes, granulocytes and monocytes of patients with BD and compared to that from healthy (HC) and disease controls. IL-8 sequences were revealed by PCR amplification using appropriate sequence-specific primers. mRNA levels were determined semi-quantitatively using an image analyser in comparison with beta-actin. RESULTS: Although the differences did not reach statistical significance, BD patients tended to have higher IL-8 mRNA levels compared to HC in whole blood samples (2.0 +/- 1.4 vs 1.5 +/- 1.2) as well as in their lymphocytes (2.7 +/- 1.6 vs 1.5 +/- 0.9). No differences were observed between BD and HC in monocyte or granulocyte IL-8 mRNA levels. CONCLUSION: Our results suggest that the cellular source of IL-8 is diverse in BD with a possible major contribution by lymphocytes.

The effects of nitric oxide donors and inhibitors on neutrophil functions in Behçet's disease.

OBJECTIVE: The effects of nitric oxide donor SNAP and nitric oxide inhibitors L-NMMA and AG on the functions of neutrophils in patients with Behçet's Disease (BD) were investigated in vitro. METHODS: Oxidative burst and phagocytosis of neutrophils were evaluated by flow cytometry in patients with Behçet's disease (n = 32), inflammatory (n = 17) and healthy controls (n = 14), in the presence of L-NMMA, AG and SNAP. RESULTS: The stimulation index of oxidative burst was found to be significantly decreased following PMA stimulation in patients with active BD compared to inflammatory and healthy controls. Oxidative function of neutrophils were inhibited in all 3 groups in the presence of L-NMMA, AG and SNAP L-NMMA inhibited the oxidative burst of neutrophils obtained from healthy controls more than inflammatory controls and BD (80% vs 52% and 53% respectively, p = 0.001). No significant difference of phagocytosis inhibition was found with L-NMMA, AG and SNAP and there were also no differences between the groups (% 9-39). CONCLUSION: Nitric oxide donors and inhibitors may have a therapeutic role in Behçet's disease by suppresing neutrophil activity.

Neutrophil activation in Behçet's disease.

OBJECTIVE: Neutrophils are implicated in the pathogenesis of Behçet's disease (BD). Various functions of neutrophils are studied to clarify this role. METHODS: The oxidative burst and phagocytic functions of neutrophils and surface molecules associated with neutrophil activation (CD10, CD14 and CD16) were investigated in BD patients by flow cytometric methods. Patients with inflammatory arthropathies, sepsis and healthy controls were also studied. RESULTS: In the oxidative burst experiments, after fMLP and PMA stimulation, stimulation index was found to be significantly decreased in patients both with BD and sepsis compared to healthy controls and inflammatory arthropathies (p < 0.001 and p < 0.01, respectively). The phagocytosis of labelled E. coli particles in patients with BD was not different from that of the healthy controls, while it was decreased in diseased controls (p < 0.001). The surface density of neutral endopeptidase (CD10) and the mean percentage of LPS receptor (CD14) was found to be significantly higher in both BD patients and diseased controls (p < 0.001). The mean percentage of CD16 expression was only low in patients with sepsis (p < 0.001), whereas CD16 intensity on cells was found to be lower in patients with BD as well as in sepsis (p < 0.01). CONCLUSION: These findings indicate the presence of in vivo pre-activated neutrophils in BD. A similar activation was also a feature of severe inflammatory disorders.

Remitting seronegative symmetrical synovitis with pitting oedema associated with chronic lymphocytic leukaemia.

We describe a case of remitting seronegative symmetrical synovitis with pitting oedema (RS3PE syndrome) in a 67-year-old man. Immunophenotyping studies and histology of biopsy specimens revealed chronic lymphocytic leukaemia (CLL). Polyarthritis and oedema were revealed by small doses of corticosteroids. The association of RS3PE syndrome with rheumatological and malign disorders are discussed.

Effect of interferon-alpha(2a) on neutrophil adhesion and phagocytosis in chronic myeloid leukemia and Behçet's disease.

Alpha-interferon (alpha-IFN) is implicated in a Behçet's disease (BD)-like syndrome observed in a small number of chronic myeloid leukemia (CML) patients. The effect of alpha-IFN on neutrophil adhesion and phagocytosis in CML patients, BD patients and healthy volunteers was investigated to clarify the reason for this observation. Ten subjects were studied for each group by incubating neutrophils with various doses of alpha-IFN. Basal neutrophil adhesions for CML patients, BD patients and healthy volunteers were similar. However, BD patients had greater basal phagocytosis than CML patients, and both groups had greater basal phagocytosis than healthy volunteers. Neutrophil adhesion and phagocytosis of CML patients increased following incubation with higher doses of alpha-IFN, and phagocytosis approached the high levels observed with BD neutrophils. This study provides evidence that alpha-IFN activates neutrophils in CML patients in a dose-dependent manner, and leads to a neutrophil function profile that resembles BD.

Anti-endothelial cell antibodies, endothelial proliferation and von Willebrand factor antigen in Behçet's disease.

Endothelial cell dysfunction has previously been demonstrated in Behçet's disease which has vasculitic features. In this study we investigated anti-endothelial cell antibodies (AECA) and von Willebrand factor antigen (vWF) levels in patients with Behçet's disease. In vitro effects of patient sera on endothelial cell proliferation were also evaluated. AECAs were present in 29% of 70 Behçet's disease patients (Binding Index: 25 +/- 29% vs 9 +/- 7% in normal controls, p < 0.005). 95% of AECA positive patients were clinically active compared to 74% of AECA negative patients (p = 0.04). Among specific organ manifestations only active arthritis correlated with AECA positivity (6 of 7 patients vs 14 of 63, p = 0.002). AECA positive patients had a significantly higher mean ESR (37 +/- 24 mm/h vs 21 +/- 17 mm/h, p:0.006). Mean vWF levels were also significantly higher in patients compared to controls (166 +/- 75% vs 84 +/- 34%, p < 0.0001). No correlations were observed between AECA titres and vWF levels. No significant differences were observed between patients and controls when endothelial cell proliferation was studied (Proliferation Index: 1.25 +/- 0.28 vs 1.12 +/- 0.25, p = 0.5). Our results suggest that AECA may be related to disease activity in Behçet's disease. The presence of vWF, even in patients in complete remission, might be related to factors other than endothelial damage for vWF release from endothelial cells.

Subcutaneous nodules caused by Pseudomonas aeruginosa: healing without incision and drainage.

In a patient with multiple myeloma, numerous indurated, subcutaneous nodules and pyomyositis due to Pseudomonas aeruginosa were noted. These lesions resolved with ciprofloxacin plus ceftazidime therapy without surgical incision and drainage. Despite another course of cancer chemotherapy after total disappearance, there were no recurrences at the end of 3 months. Quinolones initially combined with other antipseudomonal beta-lactam agents may be the drugs of choice in the management of patients with subcutaneous nodules caused by P. aeruginosa.

The effect of 1,25 dihydroxyvitamin D3 on lymphocyte transformation in patients with chronic renal failure.

The changes in mitogen-induced lymphocyte response occurring in nine patients undergoing chronic hemodialysis and treated with 1,25 dihydroxyvitamin D3 (0.5 micrograms/day) were investigated. Prior to treatment the stimulation indices (SI) in patients with 15.4 +/- 4.2 for phytohemagglutinin (PHA) and 7.2 +/- 0.7 for conconavalin A (Con A). In the controls, stimulation indices were 44.4 +/- 13.5 and 20.2 +/- 5.3 for PHA and Con A, respectively. Following treatment, the stimulation indices increased to 36.9 +/- 6.2 for PHA (p < 0.05) and 18.6 +/- 3.9 for Con A (p > 0.05) indicating the beneficial effect of oral 1,25 dihydroxyvitamin D3 treatment on lymphocyte function in patients with chronic renal failure.

The muscle content and contractile capability of the common bile duct.

The supraduodenal portion of the human and canine common bile duct has been shown by indirect immunofluorescent staining with anti-actin sera to contain scattered bundles of smooth muscle arranged predominantly in a longitudinal direction with respect to the axis of the bile duct. Isolated strips of canine common bile duct in vitro can contact in response to stimulation with cholecystokinin. The force of contraction in a longitudinal direction is about three times that in a transverse direction but only 3% of that produced by isolated gallbladder strips. There appears to be a roughly proportional relationship between the amount of muscle demonstrated and the force of contraction in response to cholecystokinin.

Evaluation of chimerism with DNA polymorphisms in bone marrow transplantation.

Evaluation of chimeric status following allogenic BMT is an important tool for monitoring the replacement of host cells with donor cells and for determining the risk of relapse. Polymorphic DNA sequences can be used as powerful markers in identification of donor/recipient genotype differences, even between close relatives. Polymerase chain reaction (PCR) amplification of three variable number of tandem repeat (VNTR) loci and five single-locus polymorphisms (SLP) was used to identify chimerism in 40 recipient-donor pairs. Mixed chimerism was present in 11 patients, and complete chimerism in 29. This PCR method is a rapid and sensitive assay to detect engraftment and evaluate relapse potential, and thus is very useful in the clinical management of BMT patients.

Serum and urine levels of tumour necrosis factor in patients with genitourinary cancer and their relevance to disease status.

Serum and urine levels of tumour necrosis factor (TNF) which is a lymphokine was measured in patients with genitourinary cancer (either prostatic, bladder or testicular carcinoma) in order to find out whether there were elevated levels of TNF in those patients and if it was related to disease status. Although the preliminary results are not conclusive, further research on TNF may prove this immunoprotein to be a potential diagnostic and therapeutic agent.

Serum and synovial fluid adenosine deaminase activity in patients with rheumatoid arthritis, osteoarthritis, and reactive arthritis.

Adenosine deaminase activity was determined in paired samples of serum and synovial fluid taken from patients with rheumatoid arthritis (n = 12), reactive arthritis (n = 13), and osteoarthritis (n = 7), and the value of this investigation in the diagnosis of synovial swellings was assessed. Increased activity was found in the synovial fluid taken from patients with rheumatoid disease and reactive arthritis, though values were less raised in the latter. Synovial fluid taken from patients with osteoarthritis did not show significantly raised adenosine deaminase activity as compared with that of normal controls (n = 3).

Antibody to intermediate filaments of the cytoskeleton in patients with Behçet's disease.

Antibodies to 10-nm intermediate filaments (anti-IF) were determined in the sera of 30 patients with Behçet's disease (BD), in addition to C-reactive protein and C9, and an attempt has been made to determine whether the presence of anti-IF indicate disease activity. The vimentin type of anti-IF was found to be positive in 14 out of 30 patients with BD (47%), whereas it was positive in 35% of the patients with rheumatoid arthritis (20 cases), 16% of the patients with systemic lupus erythematosus (19 cases) and in only 9% of the normal controls. The anti-IF were predominantly IgG class and the titers in BD were significantly higher than those in normal controls. Out of the 14 patients with anti-IF, 10 showed significantly increased levels of serum C9 and 8 showed increased levels of CRP activity. Only one patient showed increased C9, but was negative for anti-IF and CRP. The presence of anti-IF in the patients' sera was found to be a more sensitive indicator, though not specific, for the clinical assessment of disease activity.