Non-anaplastic peripheral T cell lymphoma in children and adolescents-an international review of 143 cases.

Peripheral T cell lymphomas (PTCL) are rare in children and adolescents, and data about outcome and treatment results are scarce. The present study is a joint, international, retrospective analysis of 143 reported cases of non-anaplastic PTCL in patients <19 years of age, with a focus on treatment and outcome features. One hundred forty-three patients, between 0.3 and 18.7 years old, diagnosed between 2000 and 2015 were included in the study. PTCL not otherwise specified was the largest subgroup, followed by extranodal NK/T cell lymphoma, hepatosplenic T cell lymphoma (HS TCL), and subcutaneous panniculitis-like T cell lymphoma (SP TCL). Probability of overall survival (pOS) at 5 years for the whole group was 0.56 ± 0.05, and probability of event-free survival was (pEFS) 0.45 ± 0.05. Patients with SP TCL had a good outcome with 5-year pOS of 0.78 ± 0.1 while patients with HS TCL were reported with 5-year pOS of only 0.13 ± 0.12. Twenty-five percent of the patients were reported to have a pre-existing condition, and this group had a dismal outcome with 5-year pOS of 0.29 ± 0.09. The distribution of non-anaplastic PTCL subtypes in pediatric and adolescent patients differs from what is reported in adult patients. Overall outcome depends on the subtype with some doing better than others. Pre-existing conditions are frequent and associated with poor outcomes. There is a clear need for subtype-based treatment recommendations for children and adolescents with PTCL.

The glucocorticoid receptor gene polymorphism N363S predisposes to more severe toxic side effects during pediatric acute lymphoblastic leukemia (ALL) therapy.

The survival rates in childhood acute lymphoid leukemia (ALL) have improved dramatically; however, patients still suffer from a variety of drug-related toxicities. Individualized therapy regimens promise the least toxic therapy regimen with the best hematologic outcome. Our aim was to investigate whether increased individual glucocorticoid sensitivity due to the N363S polymorphism of the glucocorticoid receptor increased susceptibility to steroid-related toxicities during ALL therapy. A total of 346 pediatric ALL patients were involved in the present study. N363S carrier status was investigated by allele-specific PCR. Clinical and laboratory signs of glucocorticoid-related toxicities, Day 8 prednisone response, and 5-year event-free survival were analyzed and compared retrospectively. Thirty-two of the 346 patients were heterozygous carriers (9.2 %). Hepatotoxicity (31.3 vs. 11.2 %, p = 0.004, carriers and non-carriers, respectively) and glucose metabolism abnormalities (18.8 vs. 3.8 %, p = 0.001, carriers and non-carriers, respectively) were significantly more frequent among carriers. There was no difference in the incidence of hypertension and encephalopathy/psychosis among carriers and non-carriers. Carriers were also more prone to have a combination of toxicities. All 363S carriers were good prednisone responders (100 %) and had significantly better 5-year event-free survival rates (93.1 vs. 71.86 %, p = 0.012), whereas among non-carriers there were more poor prednisone responders (8.28 %) and worse 5-year event-free survival rates. Patients with the N363S polymorphism in the glucocorticoid receptor are more prone to steroid-related toxicity during ALL therapy and should be monitored more closely. Patients with N363S polymorphism of the glucocorticoid receptor may be appropriate candidates for inclusion in the design of individualized therapies.

Glucocorticoid-induced apoptosis and treatment sensitivity in acute lymphoblastic leukemia of children.

Sensitivity of leukemic blasts to steroid therapy is one of the prognostic factors in acute lymphoblastic leukemia (ALL) in children. We examined the number of steroid receptors and the increase in the apoptotic index in peripheral blast cells after administration of prednisolone monotherapy in 21 children with ALL. A new diagnostic method was established based on determination of the apoptotic index in peripheral blood lymphoblasts to evaluate the steroid sensitivity of leukemic cells during the first day of therapy. The increase in apoptotic ratio, analyzed by morphologic and/or flow cytometric studies, was most expressed in the first 6 hours of treatment. The apoptotic ratio showed a good correlation with the clinical response. The number of steroid receptors (gcRs) on the blast cells was also examined, but it proved to be less informative than the in vivo steroid response itself.

Activation of T cell cytotoxicity against autologous common acute lymphoblastic leukemia (cALL) blasts by CD3xCD19 bispecific antibody.

To develop an effective immunotherapy for B-lineage acute lymphoblastic leukemia (ALL), bispecific monoclonal antibodies (bsAb) were raised by cell fusion of two hybridoma cell lines secreting CD3 and CD19 antibodies. The resulting bispecific antibody contains two different specificities within a single antibody molecule. One binding site (CD3) activates the T cells via the T cell receptor complex, whereas the second binding site (CD19) targets the cytolytic T cells to malignant B cells. Leukemic blasts from children with B-lineage ALL showed stable and strong CD19 expression. CD3xCD19 bsAb were used to activate peripheral blood mononuclear cells (PBMC) from healthy donors or from patients with ALL during remission. Cytotoxic activity against autologous ALL cells by PBMC was induced upon addition of 100 ng/ml CD3xCD19 bsAb after 3 days of preincubation. Costimulation through CD28 increased T cell proliferation to some extent, but did not increase cytotoxic activity of PBMC against leukemic blasts. We present evidence for an effective and specific activation of resting human T lymphocytes by CD3xCD19 bsAb in vitro. Activation of cytotoxic effector T cells is feasible by preincubation with bsAb CD3xCD19 alone and does not rely on additional external costimulation. Thus, targeting of T cell cytotoxicity towards leukemic blasts via CD3xCD19 bsAb may represent a promising strategy for immunotherapy of B-lineage ALL.

Apoptosis as a possible way of destruction of lymphoblasts after glucocorticoid treatment of children with acute lymphoblastic leukemia.

Apoptosis (programmed cell death) is a physiologic phenomenon wherein the dying cell plays an active part in its own destruction. It has an important role in regulation of the balance of cell proliferation and cell death. The pharmacologic manipulation of apoptosis offers new possibilities for the prevention and treatment of cancer. One of the independent prognostic factors in the treatment of acute lymphoblastic leukemia is the sensitivity of the leukemic cells to corticosteroids. Apoptosis after glucocorticoid therapy is suggested as a prognostic factor in children with leukemia. Peripheral blood of children with acute leukemia was taken for morphologic and flow cytometric studies before and after the onset of prednisolone monotherapy. In most of the cases a positive correlation was observed between the decrease of blast numbers and the increase in apoptotic ratio in peripheral blood. In one case no response was observed either clinically or regarding apoptosis.

The diagnostic significance of autoantibodies against cartilage components or their counterparts in destructive articular diseases.

In this work four different types of antibodies to collagens, membrane protein of chondrocytes (MP), cartilage matrix proteins (CMP) and heat shock proteins (HSP-s) were tested in order to find an available clinical laboratory method for diagnostic and monitoring purposes in patients suffering from osteoarthritis. From the point of view of diagnostic efficiency, the estimation of antibody level to MP seemed to be the best and as a supplementary method the determination of rheuma factor was recommended. The anticollagen antibody estimation is less sensitive, anti CMP antibodies are not detectable. In spite of immunological crossreaction between HSP and cartilage matrix component the antibody level against HSP has no correlation with osteoarthritis. The appearance of humoral reaction, antibodies against different cartilage specific collagens and chondrocyte membrane proteins, is an epiphenomenon, however as the supposed immune complexes, trapped in a cartilage, play an important role in the damage of cartilage which may explain the self-perpetuating and chronic nature of cartilage degradation on osteoarthritis.

The clinical diagnostic significance of the investigation of receptor mediated pathway of LDL.

Some types of LDL-receptor investigations have been demonstrated which can also be carried out in routine clinical laboratory. Special attention has been layed on the testing of lymphocytes and monocytes which are available easier than tissue samples but, which are, however, according to our experiences appropriate for the LDL receptor analysis.

Estimation of serum anticollagen and the antibodies against chondrocyte membrane fraction: their clinical diagnostic significance in osteoarthritis.

The serum anticollagen antibodies to collagen types II, IX, and XI, as well as the antibody level to chondrocyte membrane extract were investigated in patients suffering from severe osteoarthritis (n = 86) in comparison with patients free of primary arthritis (n = 33) and with control healthy patients (n = 44), respectively. Isolation and purification of cartilage antigens and their relevance to ELISA reaction have been outlined. Although the method for anticollagen antibodies to types IX and XI was more sensitive than that of type II, its sensitivity was very low (52%). The determination of the specific IgG fraction by affinity chromatography seemed to be more sensitive: in osteoarthritic patients the percentage of the "arthritogen" IgG rose to 10% of the total IgG. The determination of antibody level against chondrocyte membrane extract was adapted to human diagnostic purposes. In osteoarthritic patients the serum antibody level was significantly higher than in healthy controls. The specificity of this new test was proved by the facts that: (a) only the collagen-binding fraction of the membrane extract reacts with the patient's sera; (b) the ELISA reaction could be totally inhibited by the antigen; (c) patients suffering from noninflammatory joint diseases were characterized by low antibody levels.

The clinical diagnostic significance of serum antichondrocyte membrane antibodies in osteoarthritis.

In osteoarthritic patients significantly elevated level of antibodies against human chondrocyte membrane extract was found by double solid phase ELISA which seemed to be more specific for the disease than the antibody level of collagen types II, IX and XI. Altogether 86 patients with a mean age of 53.6 years and a ratio of 25% men and 75% women suffering from severe hip joint osteoarthritis were studied in comparison with 44 control persons with a mean age of 35.8 years and identical sex distribution. The specificity of reacting antibodies was proved partly by inhibition test and partly by the fact that the ELISA reaction could be produced only by proteins of the membrane extract bound to collagen (II) after affinity chromatography. The diagnostic importance of the methods is striking, in view of the lack of any other objective laboratory tests for the disease.

Effects of glycosaminoglycan polysulphate in the treatment of chondrocalcinosis.

The effect of intra-articular injections of glycosaminoglycan polysulphate (Arteparon) on pain, joint mobility, inflammatory reactions, cartilage calcification and the urinary excretion of inorganic pyrophosphate was studied in 12 patients with chondrocalcinosis. All cases were bilateral and the less affected homologous joint served as an untreated control. The patients were followed over a one-year period. The glycosaminoglycan polysulphate treatment brought about a significant reduction of pain (p less than 0.01) and improvement of joint mobility (p less than 0.001). This effect continued for the whole one-year follow-up period, but could not be seen in the control joints. After the treatment period of 2-7 weeks there was a marked decrease in cartilage calcification paralleled with an increase in the excretion of inorganic pyrophosphate.

[Effect of arteparon on the formation of calcium phosphate and calcium pyrophosphate crystals--comparative experimental study]. / Wirkung von Arteparon auf die Bildung von Calciumphosphat- und Calciumpyrophosphatkristallen--Vergleichende experimentelle Untersuchung.

The influence of Arteparon on crystal formation in vitro has been examined in calcium orthophosphate and in calcium pyrophosphate mixtures. Similar factors may be operative in the cartilage calcification processes, and thus in chondrocalcinosis. Authors have ascertained that Arteparon - like chondroitin sulphate - inhibits crystal separation. The optimal concentration for this inhibition lies in the range of the proteoglycan content of normal cartilage. Where there are physiological phosphate-pyrophosphate ratios and a low magnesium concentration, it is certain that Arteparon has an inhibiting effect. Apart from the spatial structure of the substance, the importance of negative charge density for the inhibition of separation of calcium phosphate and pyrophosphate crystals is thought to contribute to this effect of Arteparon.

The importance of minor collagenous chains in the articular cartilage.

This is a review of the present knowledge on the newly-described minor cartilage collagens. The authors deal with the nomenclature, methodology, structure and function of the minor collagens based on both literary data and their own results. The authors were the first to describe the occurrence of minor collagens in adult human articular cartilage and the change of the minor collagen content with age and osteoarthrosis [25]. The methods used by different groups for fractionation of minor collagens are different. As the recommended pepsin digestion used for solubilization of these collagens gives different results for the size of one of the minor collagens called M-collagen, special attention should devoted to the method used. The differential salt fractionation of pepsin-solubilized collagen was found the best. In this case the molecular structure of the M-collagen remained intact. In the neutral salt extraction technique recommended recently by Burgeson et al. [5], yield of collagen from sample is high, M-collagen is degraded. The greatest solubilization of collagen was achieved by the cyanogen bromide technique. The mixture of peptides from different proteins, however, requires further fractionation. The authors propose to investigate minor collagens in tissue samples obtained from biopsies and in synovial fluid.

Minor collagens in arthrotic human cartilage. Change in content of 1 alpha, 2 alpha, 3 alpha and M-collagen with age and in osteoarthrosis.

The content and distribution of 1 alpha, 2 alpha, 3 alpha and M-collagens in human articular cartilage were studied. As controls, normal femoral heads and costal cartilage of autopsy material from newborn to 91-year-old persons were used. The osteoarthrotic cartilage was obtained from patients undergoing total hip replacement aged 45-80. The pepsin-digested cartilage collagen was fractionated by differential salt fractionation. The collagen content of the fractions was determined, and the fractions were separated by polyacrylamide slab gel electrophoresis. In the extracted collagen, the type II collagen varied from 82 to 97 per cent with increasing age. The 1 alpha, 2 alpha and 3 alpha chains decreased. M-collagen, especially of the high molecular weight components, disappeared with age. In osteoarthrosis three types of change - degeneration, new fibrocartilage formation on the surface of osteophyte and reparative cartilage - were separately studied. In all types of osteoarthrosis, an increase of minor collagens was found. In newly formed fibrocartilage, the reappearance of M-collagen was conspicuous. It is proposed that the three types of osteoarthrotic cartilage may be characterized on the basis of content and distribution of minor collagens.

The effect of proteoglycans of cartilage and over-sulphated polysaccharides on the development of calcium-hydroxy-apatite (CHA) crystal formation in vitro.

A coulometric model system is described which facilitates the quantitative study of the kinetics of transformation of amorphous calcium phosphate (ACP) into calcium-hydroxy-apatite (CHA) crystals. Proteoglycans of high molecular weight and over-sulphated polysaccharides (Arteparon, dextran sulphate) delayed CHA crystal formation. The results have enabled us to characterize the structure activity relationship of inhibitors of CHA formation, and to postulate a general structural requirement for molecules with inhibitory effect. As working mechanism, binding of calcium ions by sulphate groups of polyanions was supposed, which might reversibly impair "the critical nuclei formation", and/or further deposition of calcium ions in the CHA crystals. The clinical, therapeutical significance of the determination of the threshold concentration of different compounds is discussed.

The effect of inorganic pyrophosphate (PPi) anions on the in vitro collagen fibril formation.

This paper presents a study on the effect of sodium salt of pyrophosphate (Na-PPi) in different concentrations (1-10 mMol/l) on the in vitro collagen fibril formation. Collagen was mixture of type I and III collagen dissolved in neutral buffer containing sodium chloride of 1.0 mol/l. Na-PPi added to collagen delayed the fibrillary precipitation of collagen from the solution. Electronmicroscopically the fibrils are of native collagen type, but they are thicker and show a pattern of fine substriation. Calcium and other divalent cations showed differences in their influence on the effect of Na-PPi. This experimental model was developed in order to interpret the supposed role of PPi in the inhibition of apatite crystal formations as well as the fibrogenic effect of its calcium salt in PPi-arthropathy. The effect of PPi on the collagen fibril formation should also be taken into consideration.