Gunnel Henriksson

Lymphoma and other malignancies in primary Sjögren’s syndrome: a cohort study on cancer incidence and lymphoma predictors

Objectives: To assess the risk of lymphoproliferative disease or other malignancy (standardised incidence ratios (SIRs)), in patients with primary Sjögren’s syndrome according to the American-European Consensus Criteria (AECC), compared with patients with sicca syndrome (non-AECC) and the background population. To identify predictors of malignancy and describe lymphoma types and survival probabilities. Methods: A linked register study using information from the Malmö Primary SS Register, Swedish Cancer Register, and Cause-of-Death Register for calculation of SIRs was carried out. Detected lymphomas were reclassified according to the WHO classification. Cox regression analysis was used to study the predictive value of clinical, laboratory, and histological findings at the time of diagnosis. Results: 507 patients with a median follow up of 8 years (range 1 month to 19 years) were included. SIRs (95% confidence interval (CI)) for malignancies in total and for non-Hodgkin’s lymphomas (NHL) were 1.42 (0.98 to 2.00) and 15.57 (7.77 to 27.85), respectively, in those fulfilling the AECC (n = 286). In non-AECC sicca patients (n = 221) SIR for malignancy of any kind was 0.77 (0.41 to 1.32); no lymphoproliferative neoplasms were detected. Significant predictors of lymphoproliferative disease were purpura/skin vasculitis (hazard ratio (HR) = 4.64, 95% CI 1.13 to 16.45), low complement factor C3 (HR = 6.18, 95% CI 1.57 to 24.22), low C4 (HR = 9.49, 95% CI 1.94 to 46.54), CD4+ T lymphocytopenia (HR = 8.14, 95% CI 2.10 to 31.53), and a low CD4+/CD8+ T cell ratio ⩽0.8 (HR = 10.92, 95% CI 2.80 to 41.83). 7/12 (58%) NHLs were diffuse large B cell lymphomas. Conclusion: A 16-fold increased risk for development of NHL was found. CD4+ T lymphocytopenia is an additional strong risk factor for developing lymphoma.

Autoantibodies Present Before Symptom Onset in Primary Sjögren Syndrome

Methods | All patients with primary Sjögren syndrome at Malmö UniversityHospital(Malmö,Sweden)havebeenincludedinaregistry since 1984.3 To obtain presymptomatic serum samples, the registry was linked with 3 biobanks containing specimens from 625 000 individuals submitted for microbiological analyses or population-based studies of healthy individuals.4 Date of symptom onset was determined retrospectively from the patient during the first office visit at which Sjögren syndrome was diagnosed. All cases provided written informed consent at inclusion in the registry. The study was approved by the ethics committee at Lund University. All patients with available samples who met consensus criteria for Sjögren syndrome1 were included. When multiple samples were available for a single patient, the earliest positive sample was used. Controls were randomly selected from the biobanks and matched by sex, age, and date of earliest sampling (within 60 days before or after) to each case. None of the controls were diagnosed with Sjögren syndrome. Those serum samples that were obtained from the microbiology biobank had been submitted due to symptoms unrelated to Sjögren syndrome (eg, pregnancy screening, suspected influenza). Samples were collected from 1976 through 2001 and Sjögren syndrome was diagnosed through 2011. Autoantibodies against Ro60/SSA, Ro52/SSA, La/SSB, Sm, RNP, Scl-70, Jo-1, ribosome P, and chromatin were detected using a multiplex immunobead assay (QUANTA Plex SLE Profile 8, INOVA Diagnostics) and analyzed on a Luminex 100 instrument (Luminex Corp). Antinuclear antibodies (ANAs) were analyzed by immunofluorescence with HEp-2 cells as the antigens. Immunoglobulin M-class rheumatoid factor (RF) was analyzed with an in-house enzyme-linked immunosorbent assay. Descriptive statistics and a 2-sided Friedman test were used for statistical analysis (Statistics version 20.0; IBM SPSS). P < .05 was considered statistically significant.

Increased subpopulations of CD16+ and CD56+ blood monocytes in patients with active Crohn's disease

Background Circulating monocytes may be subdivided according to the presence or absence of the Fc&ggr; receptor CD16 and the neural cell adhesion molecule CD56. Monocytes classified into these subpopulations are characterized by distinct phenotypic and functional features. We hypothesized that patients with active Crohns disease differ in their peripheral monocyte subpopulations. Methods Using flow cytometry we investigated the expression of CD16 and CD56 on circulating monocytes in 11 patients with active Crohns disease and 11 controls. These monocyte subpopulations were then analyzed for expression of the chemokine receptor fractalkine, CX3CR1, and the monocyte chemoattractant protein‐1, CCR2. Results We found a median 3.7‐fold increase in the number of CD16+ monocytes related to the population with high expression of the pattern recognition receptor CD14 compared to that in the controls (P < 0.001). By studying the percentage of monocytes expressing CX3CR1, and their relative fluorescence intensity (RFI), we found significant differences, with both the highest percentage and the highest RFI in the CD14lowCD16+ subpopulation, whereas the CD14highCD16+ subgroup represented an intermediate population. Inversely, CCR2 expression was highest in the populations with high expression of CD14, whereas the CD14lowCD16+ subpopulation showed the lowest percentage and the lowest RFI for CCR2. We found the percentage of CD14+CD56+ monocytes in patients with active Crohns disease to be increased 2.7 times compared to the controls (P = 0.011). Conclusions These results show that subsets of peripheral monocytes with a more mature phenotype are expanded in patients with active Crohns disease. (Inflamm Bowel Dis 2006)

Prediction of Sjögren's Syndrome Years Before Diagnosis and Identification of Patients With Early Onset and Severe Disease Course by Autoantibody Profiling.

Autoantibodies are highly characteristic of primary Sjögrens syndrome (SS) and represent important tools for studying its pathogenesis. Nonetheless, thus far, no systematic investigations have assessed the presence of autoantibodies before diagnosis. This study was undertaken to analyze how early and in what order autoantibodies appear, how predictive they are of primary SS, and whether they identify disease subsets.

Immunohistochemistry of the B-cell Component in Lower Lip Salivary Glands of Sjögren's Syndrome and Healthy Subjects

Serial sections of lower lip salivary gland (LSG) biopsies were examined by immunohistochemistry, using a battery of B‐ and partly T‐related antibodies (CD5, CD20, CD21, CD27, CD38, CD45RO, CD79a, Bcl‐2 and Bcl‐6) in different groups of subjects: healthy controls and clinically verified smoking or nonsmoking cases of primary Sjögrens syndrome (SS). The purpose was to characterize the B‐cell pattern of the lymphocytic foci and of the tiny perivascular infiltrates preceding the development of foci. Hyperplastic tonsil was used as stain control. In normal LSG, widely dispersed CD38+ and CD79a+ as well as some CD5+ cells are a normal constituent, with lack of staining with the other antibodies. In SS/LSG, the lymphocytic foci showed staining with all the antibodies, with variable degrees of overlapping or nonoverlapping. In SS/LSG of nonsmokers, CD20+ B cells make up a prominent part of the fully developed periductal lymphocytic foci, not overlapping with CD45RO. Also, CD20+ B cells did not overlap in the infiltrates with colocalized CD27+/CD38+ cells. CD20+ B cells and CD45RO+ T cells also occur as minute infiltrates perivascularly in areas of no foci in SS/LSG as well as in SS smokers lacking the typical foci. Smokers lack foci, but tiny infiltrates express CD20 as well CD45R0. Our findings suggest that CD20+ B cells and CD45RO+ T cells are early immigrants in the LSG of SS of smokers as well as nonsmokers and that another subgroup of CD27+/CD38+ B cells gradually mix with the first two to form the characteristic foci in SS/LSG. The simultaneous demonstration of CD20+ and CD27+ B cells in SS/LSG may constitute a significant diagnostic tool. Further, the findings suggest that the early immigrating lymphocytes may have been primed at a site remote from the glands before arriving via the blood to the gland tissue.

Sjögren's syndrome: Lymphoma predisposition coupled with a reduced frequency of t(14;18) translocations in blood lymphocytes

Sjögrens syndrome (SS) is a systemic autoimmune disorder with a strong tumor predisposition (a 44‐fold elevated incidence of non‐Hodgkins lymphoma has been reported). By polymerase chain reaction analysis of t(14;18), a key lymphomagenic event in peripheral blood lymphocytes, we found a lower frequency in a subset of 12 SS patients positive for SS‐A/SS‐B autoantibodies than in 21 healthy subjects and 20 SS patients lacking these SS marker autoantibodies (P < 0.001). All 14 mutants sequenced displayed signs typical of V(D)J recombinase activity. This perplexing result of a low rate of t(14;18) in a population strongly predisposed to t(14;18)‐associated tumor development may be explained by a constitutive deficiency in V(D)J recombinase leading to autoimmunity and increased lymphoproliferation. Mol. Carcinog. 24:226–231, 1999.

Expression of DNA-dependent protein kinase in human granulocytes

ABSTRACTHuman polymorphonuclear leukocytes (PMN) have been reported to completely lack of DNA-dependent protein kinase (DNA-PK) which is composed of Ku protein and the catalytic subunit DNA-PKcs, needed for nonhomologous end-joining (NHEJ) of DNA double-strand breaks. Promyelocytic HL-60 cells express a variant form of Ku resulting in enhanced radiation sensitivity. This raises the question if low efficiency of NHEJ, instrumental for the cellular repair of oxidative damage, is a normal characteristic of myeloid differentiation. Here we confirmed the complete lack of DNA-PK in PMN protein extracts, and the expression of the truncated Ku86 variant form in HL-60. However, this degradation of DNA-PK was shown to be due to a DNA-PK-degrading protease in PMN and HL-60. In addition, by using a protease-resistant whole cell assay, both Ku86 and DNA-PKcs could be demonstrated in PMN, suggesting the previously reported absence in PMN of DNA-PK to be an artefact. The levels of Ku86 and DNA-PKcs were much reduced in PMN, as compared with that of the lymphocytes, whereas HL-60 displayed a markedly elevated DNA-PK concentration. In conclusion, our findings provide evidence of reduced, not depleted expression of DNA-PK during the mature stages of myeloid differentiation.

Ku Protein and DNA Strand Breaks in Lip Glands of Normal and Primary Sjogren's Syndrome Subjects: Lack of Correlation with Apoptosis

The aim was to examine tissue expression of Ku protein in lower lip salivary gland (LSG) biopsies from cases of primary Sjögrens syndrome (SS) and from normal subjects. Methods: immunohistochemistry was used with antibodies to Ku70/86 and also Ki67, PCNA and p53. In addition, the Klenow method was applied in order to detect evidence of apoptosis. Sections of hyperplastic tonsil served as additional controls. Results: in normal controls, LSG acinar cells stained negatively whereas LSG excretory duct cell nuclei stained positively with Ku and Klenow and occasionally with PCNA but negatively with Ki67 and p53. In LSG focal sialadenitis of SS cases, some lymphocytic cells showed staining with Ku, Ki67, PCNA, Klenow and p53. In addition to duct cell Ku and Klenow as well as PCNA staining which was not much different from normals, a few ductal epithelial and also mononuclear cells stained with p53. In focal sialadenitis, some acinar cells showed staining with PCNA as well as with Klenow. Conclusions: our findings in LSG biopsies of SS cases added little to an increased understanding about the pathogenetic mechanisms in the development of focal sialadenitis in SS. However, in normal LSG, ductal epithelial but not acinar cells seem to express a constitutively specific Ku protein and Klenow profile, suggestive of DNA strand breaks but not clearly associated with ongoing apoptotic events. It may reflect an enhanced stress response, which may be pathogenetically important in the early events of focal sialadenitis development in primary Sjögrens syndrome.

A reduced level of multiple mutation in a shuttle vector passaged in Sjögen's syndrome cells

Sjögrens syndrome is a systemic disorder with unknown etiology, displaying many signs of autoimmunity. Although the basic mechanism of this disease is unknown, a defect in somatic mutagenesis of antibody genes has been suggested. Using a shuttle vector plasmid, we here show that the number of vectors with multiple base changes in a marker gene was reduced in B cell lines from two patients with Sjögrens syndrome (8% in both), as compared with values reported for cell lines from normal human donors (16-27%). This finding suggests that a reduction of the rate of somatic mutagenesis may influence the development of symptoms in Sjögren patients.

Autoantibody profiling can predict primary Sjögren's syndrome years before diagnosis and identify those with early onset and severe disease course.

Autoantibodies are highly characteristic of primary Sjögrens syndrome (SS) and represent important tools for studying its pathogenesis. Nonetheless, thus far, no systematic investigations have assessed the presence of autoantibodies before diagnosis. This study was undertaken to analyze how early and in what order autoantibodies appear, how predictive they are of primary SS, and whether they identify disease subsets.