Börje Ridell

Recruitment of T cells into bone marrow of ITP patients possibly due to elevated expression of VLA-4 and CX3CR1

In idiopathic thrombocytopenic purpura (ITP), platelets are destroyed in the spleen, liver, and bone marrow (BM) by autoantibodies and cytotoxic T cells. In a DNA microarray screen of peripheral blood T cells, we found that VLA-4, CX3CR1, and CXCR4, involved in T-cell homing, had increased expression in ITP patients compared with controls. However, we only found increased protein expression of VLA-4 on T cells from peripheral blood by flow cytometry. To address a possible recruitment of T cells into the organs involved in platelet destruction, we analyzed T cells in BM. In BM, T-cell surface expression of VLA-4 and CX3CR1 was increased in ITP patients compared with controls. Furthermore, the number of CD3(+) T cells in BM, but not in blood, was increased in ITP patients compared with controls. This finding was confirmed by immunohistochemistry of BM biopsies. The number of regulatory T cells (CD4(+)/CD25(bright)) was decreased in the BM of ITP patients, whereas Fas expression was increased. In conclusion, ITP is associated with accumulation and activation of T cells in the BM. Recruitment of T cells into the target organ (eg, BM) is plausible and may be facilitated through increased VLA-4 and CX3CR1 expression. These molecules might serve as new treatment targets in ITP.

Trends in the incidence of chronic Philadelphia chromosome negative (Ph-) myeloproliferative disorders in the city of Göteborg, Sweden, during 1983-99.

Objective.  In the literature the incidence rates for the chronic Philadelphia chromosome negative (Ph‐) myeloproliferative disorders (MPD) are known to vary extensively; only a few studies have, however, been concerned with incidence trends over time. Therefore, the aim of the present work was to investigate possible trends as regards incidence rates over time for Ph‐MPD.

Expression of CD68+ tumor-associated macrophages in patients with diffuse large B-cell lymphoma and its relation to prognosis.

Tumor‐associated macrophages (TAM) have been ascribed both pro‐ and anti‐tumor properties, but the majority of clinical cancer studies have shown that the presence of a high number of TAM is related to poor prognosis, suggesting that TAM predominantly exert pro‐tumoral activity. The prognostic role of TAM in patients with diffuse large B‐cell lymphoma (DLBCL), however, is so far unknown. Therefore, TAM were immunohistochemically stained with a CD68 antibody in a retrospective, population‐based study including 176 DLBCL patients treated with curative intent. With the exception that patients >60 years of age had a larger number of CD68+ cells (1143 vs 1018 cells/mm2; P = 0.05), no significant differences were found between the number of CD68+ cells and other clinical factors. Similarly, germinal center B‐cell (GCB)/non‐GCB immunophenotype or low/high Ki‐67 percentage were not associated with CD68 expression. Finally, no significant correlation was found between the number of CD68+ cells and progression‐free survival (P = 0.34) or overall survival (P = 0.94). These data indicate that the pro‐tumor effect of TAM has limited clinical relevance in DLBCL patients, which could imply that therapeutic strategies aimed at enhancing their anti‐tumor activity are of continuous clinical interest.

Testicular lymphoma--a retrospective, population-based, clinical and immunohistochemical study.

From a population-based registry, 35 patients with histologically verified testicular lymphomas were identified: diffuse large B-cell lymphomas (DLBCL) in 33 and peripheral T-cell lymphomas in two cases. Twenty-two patients had localized disease (Pe stage I and II). Twenty-eight patients received systemic chemotherapy, 17 of whom also received intrathecal prophylaxis, and 12 out of these 17 also received radiotherapy to the contralateral testis. In the Pe stage I/II group, 7 out of 21 patients in complete remission (CR) relapsed. In 5 of them the CNS was involved (isolated CNS relapse in three). Remarkably late relapses occurred (up to 127 months). Intrathecal prophylaxis seemed to reduce the frequency of relapses involving the CNS, but the relatively short follow-up (median 45 months, range 34–88, for censored patients) prevents firm conclusions regarding efficacy. The outcome for the stage IV patients was poor, with only 1 out of 11 patients in continuous CR. Immunohistochemical analysis of the DLBCL tumours revealed that 31% had the germinal centre B-cell-like phenotype. CD44 was expressed in all the tumours of stage IV patients but in less than half of the Pe stage I/II patients. A high intratumoural microvessel density was correlated with a high degree of Ki-67 positive tumour cells and an inferior overall survival.

The number of tumour‐infiltrating TIA‐1+ cytotoxic T cells but not FOXP3+ regulatory T cells predicts outcome in diffuse large B‐cell lymphoma

The prognostic significance of tumour‐infiltrating lymphocytes (TILs) in patients with diffuse large B‐cell lymphoma (DLBCL) remains controversial. Furthermore, the possible impact of regulatory T cells (Tregs) on survival in DLBCL is still unknown. We performed a retrospective study on the immunohistochemical expression of cytotoxic cells and Tregs, and their correlation with survival in 195 DLBCL patients. Patients with a small number of cytotoxic T‐cell intracytoplasmic antigen‐1 (TIA‐1)+ T cells (≤260 cells/mm2 tumour area; n = 52) had significantly better outcome than patients with a large number (>260 cells/mm2; n = 143); progression‐free survival (PFS) at 5 years was 67% vs. 50% (P = 0·03) and overall survival (OS) was 73% vs. 57% (P = 0·03). In multivariate analysis, the low TIA‐1+ group still had a better PFS (relative risk 0·75, 95% confidence interval 0·31–0·99; P = 0·05). The number of forkhead box protein 3 (FOXP3)+ Tregs had no influence on PFS (P = 0·89) or OS (P = 0·75). These results suggest that immunohistochemical analysis of cytotoxic T cells at time of diagnosis could provide additional prognostic information. The lack of correlation between the number of FOXP3+ cells and survival could possibly indicate that tumour‐infiltrating Tregs are of less clinical importance in DLBCL. However, these findings need to be explored in functional studies.

Rearrangement of chromosome No. 3 in a case of preleukemia with thrombocytosis

The clinical and cytogenetic findings of a patient with the preleukemic syndrome and a structural rearrangement involving both chromosomes No. 3 are described. The karyotypic abnormality consisted of an insertion of a part of the long arm of one chromosome No. 3 into the other, i.e., ins(3;3)(q27;q21q27). A prominent feature of the bone marrow was a marked megakaryocytic hyperplasia. The platelet count temporarily exceeded 1000 x 10(9)/liter. The findings of the present case, together with similar observations by others, suggest that the long arm of chromosome No. 3 may contain a region involved in the regulation of megakaryopoiesis.

Intratumoral microvascular density in malignant lymphomas of B-cell origin.

Sections of surgical lymph‐node biopsies of four types of malignant non‐Hodgkins lymphoma of B‐cell origin (B‐NHL) classified according to the R.E.A.L. terminology or lymphadenitis were immunostained in order to demonstrate endothelial CD34 (QBEnd 10) and to determine the microvascular density and vessel‐size distribution using an interactive image‐analysis technique. Only microvessels displaying a cross‐sectional area corresponding to a diameter of between 3.2 and 34.6 μm were included. The intratumoral microvascular density (iMVD) was found to be significantly higher in chronic lymphatic leukaemia (CLL, n=13) compared with the clinically more aggressive mantle cell lymphoma (MCL, n=9) and diffuse large B‐cell lymphoma (DLBCL, n=14). iMVD in CLL was also higher than in the follicular neoplastic parts (FL FOLL) of follicular lymphoma (FL, n=16). In FL FOLL the microvessel density was, moreover, significantly lower than in the surrounding non‐neoplastic FL tissue. In lymphadenitis (LA, n=10) the iMVD was higher than in DLBCL, FL FOLL and MCL. The data suggest that future studies focusing on the relationship between iMVD and the clinical outcome within each particular NHL group should be carried out in order to verify whether iMVD is a prognostic factor in NHL, as it is in carcinomas.

The impact of peripheral blood values and bone marrow findings on prognosis for patients with essential thrombocythemia and polycythemia vera

The Philadelphia chromosome‐negative (Ph‐) chronic myeloproliferative neoplasms include the three well‐known clinical entities polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Over time, patients with ET and PV may develop myelofibrosis (MF), and all three entities carry a risk of transformation into acute myeloid leukemia (AML). In a population‐based survey during 1983–1999, we studied a total of 358 patients who were diagnosed with ET and PV in the city of Gothenburg, Sweden. At the time of diagnosis, evaluable bone marrow biopsy material was available from 280 of these patients. The current work was aimed at investigating the impact of peripheral blood counts, spleen size, and bone marrow biopsy findings at diagnosis on long‐term survival and the risk of development of AML or MF in this well‐defined unselected population. The variables evaluated were venous blood hemoglobin concentration, packed cell volume, white blood cell count, platelet count, and splenic enlargement; as to bone marrow biopsies, interest was focused on reticulin content, focal or generalized collagen formation, bone marrow cellularity, and megakaryocyte profile number. Over the median observation time of 15 yr, the patients with ET did not demonstrate any significant difference as to survival compared to the normal Swedish population (hazard ratio, 1.23; 95% confidence interval, 0.97–1.51; p = 0.089). The patients with PV, on the other hand, had a significantly shorter survival compared to general population (hazard ratio, 1.66; 95% confidence interval, 1.38–1.99; p < 0.001). A lower hemoglobin concentration at diagnosis of ET predicted poorer survival (p = 0.0281), whereas patients with PV with splenic enlargement at diagnosis had a shorter survival (p = 0.037). In the patients with ET, the risk of transformation to either MF or AML was significantly associated with low hemoglobin concentration and high white cell count at diagnosis (p = 0.0037 and 0.0306, respectively). An increased reticulin content and hypercellularity in the bone marrow at diagnosis were also independent risk factors (p = 0.0359 and 0.0103, respectively). The risk of transformation in patients with PV was significantly associated with splenic enlargement and increase in bone marrow reticulin content (p = 0.0028 and 0.0164, respectively).

Tumour‐type‐specific capillary endothelial cell stainability in malignant B‐cell lymphomas using antibodies against CD31, CD34 and Factor VIII

The microvessel density (MVD) was assessed in lymph nodes infiltrated by diffuse large B‐cell lymphomas, mantle cell lymphomas, chronic lymphatic leukemia and follicular lymphomas, and in lymphadenitis. Serial sections of formalin‐fixed and paraffin‐embedded tissue were stained with antibodies against CD31, CD34 or Factor VIII. Using light microscopy and computerised image analysis, the number and size of individual immunostained vessel profiles within a preselected area size range corresponding to capillaries, postcapillary venules, small collecting venules and small arterioles were determined. A significantly larger number of vessels were registered following staining with anti‐CD34 than with anti‐CD31 or anti‐Factor VIII. Moreover, among the smallest capillary‐sized vessel profiles in all lesion types, there was a selective relative loss of stainability of anti‐CD31 and anti‐Factor VIII, resulting in a substantial total loss of visualised capillary‐sized vessels compared with anti‐CD34. In fact, the number of non‐detected capillaries following staining with anti‐CD31 and anti‐Factor VIII was significantly tumour type specific. These findings influence how we evaluate MVD data in B‐cell lymphomas and possibly also other tumour types, as well as data relating to capillary endothelium‐related functional variables of proliferation, apoptosis and maturation when different double‐labelling immunohistochemical techniques are used and different tumour types are analysed.

Incidence of chronic myeloproliferative disorders in the city of Göteborg, Sweden 1983-1992.

Abstract: An estimation of the incidence of polycythaemia vera (PV), essential thrombocythaemia (ET) and chronic idiopathic myelofibrosis (CIM) in the city of Göteborg, Sweden during the period 1983–1992 was made from a retrospective case analysis of patients registered as chronic myeloproliferative disorders (CMPD) at the Departments of Medicine and the Department of Pathology of the two major hospitals in the city. A total of 125 cases of PV, 56 males and 69 females were identified. The number of cases as well as the age‐specific incidence increased with age. The over all annual gender‐specific incidence was 2.69 cases per 105 male inhabitants and 3.12 cases per 105 female inhabitants. The incidence of PV in relation to the European Standard Population was 2.02 cases per 105 inhabitants and year. There were 72 cases, 20 males and 52 females, with ET. The age‐specific incidence was in all ages higher for females than for males and increased with age. The annual gender‐specific incidence was 0.96 per 105 male inhabitants and 2.35 per 105 female inhabitants. The incidence of ET in relation to the European Standard Population was 1.28 per 105 persons and year. There were 20 cases of CIM, 11 males and 9 females. The annual gender‐specific incidence of CIM was 0.53/105 male inhabitants and 0.41/105 female inhabitants. The incidence of CIM in relation to the European Standard Population was 0.31 per 105 persons and year. Seven persons, 2 males and 5 females, had a CMPD that could not be included in any of the above‐mentioned groups, but were registered as CMPD, unclassified.