MR imaging in children with dermatomyositis: musculoskeletal findings and correlation with clinical and laboratory findings.

OBJECTIVE: The purpose of this study was to describe the musculoskeletal MR findings in childhood dermatomyositis and to correlate MR findings with indicators of disease activity such as muscle strength and serum levels of muscle enzymes. SUBJECTS AND METHODS: This prospective study included 24 children: 19 children with dermatomyositis and five control subjects. The diagnosis of dermatomyositis was established by clinical findings and serum levels of muscle enzymes in all patients, electromyography in six patients, and biopsy in four patients. At the time of the initial MR evaluation, patients were classified on the basis of clinical findings as having active (n = 15) or inactive (n = 4) disease. A total of 44 MR evaluations of patients with dermatomyositis were included in the study: 19 initial MR examinations and 12 examinations repeated after 4-6 months of therapy in patients with active disease. An additional 13 examinations were performed on five patients. Conventional T1-weighted (SE 600/20) and T2-weighted (SE 2500/80) spin-echo and fat-suppressed MR images were obtained. The T2-weighted images (TE = 80) were used for comparison. In addition to the visual assessment, ratios between the signal intensity of muscles (gluteus, adductors, quadriceps, and hamstrings) and the signal intensity of subcutaneous fat in the same tomographic section were calculated. RESULTS: All patients with clinically active disease (n = 15) had abnormal findings on MR studies, whereas those with inactive disease (n = 4) had normal MR findings. Signal-intensity ratios of patients with active disease were greater than those in control subjects, whereas the ratios in patients with inactive disease were not different from those in control subjects. After 4-6 months of therapy, the average signal-intensity ratios of treated patients with repeated MR evaluations (n = 12) differed from ratios obtained before therapy in the same patients, but were not different from the ratios in control subjects. Other MR findings observed were perimuscular edema, enhancement of the chemical-shift artifact, and inflammatory changes of subcutaneous fat. Fat-suppressed imaging enhanced visualization of abnormalities. Markedly abnormal signal intensities of muscle were associated with marked elevations of serum levels of muscle enzymes; however, abnormal MR findings were visualized with normal serum levels of muscle enzymes. CONCLUSION: Findings of active childhood dermatomyositis on T2-weighted MR images include increased signal intensity in affected muscle, perimuscular edema, enhanced chemical-shift artifact, and increased signal intensity in subcutaneous fat. After therapy, signal intensity of muscle returns to normal. These MR findings are enhanced on fat-suppressed images.(ABSTRACT TRUNCATED AT 400 WORDS)

Data base analysis of protein expression patterns during T-cell ontogeny and activation.

We have developed a data base of lymphoid proteins detectable by two-dimensional polyacrylamide gel electrophoresis. The data base contains two-dimensional patterns and derived information pertaining to polypeptide constituents of unstimulated and stimulated mature T cells and immature thymocytes, single-cell-derived T- and B-cell clones, leukemia cells, and lymphoid cell lines. Using this data base, we have compared the protein constituents of mature T cells and immature thymocytes before and after mitotic stimulation. A subset of polypeptides that are induced in mature T cells following mitotic stimulation were found to be constitutively expressed in immature thymocytes. Other polypeptides exhibited differences in their expression between mature and immature thymocytes in a manner unrelated to proliferation. The identity of several constitutively expressed or mitotically induced proteins in lymphoid cells was established by microsequencing. These initial findings point to significant differences in the molecular pathways leading to proliferation between mature and immature T cells. The construction of this database should facilitate further studies of lymphoid differentiation and function.

PCNA levels in neuroblastoma are increased in tumors with an amplified N-myc gene and in metastatic stage tumors.

N-myc oncogene amplification in neuroblastoma has been found to be significantly associated with advanced stage disease and tumor progression. However, there is a lack of data on tumors, regarding the relationship between N-myc gene amplification and proliferation activity. Proliferating cell nuclear antigen (PCNA) is a proliferation-induced 36 kD nuclear protein that is the auxiliary component of DNA polymerase delta. PCNA levels in tissues have been found to correlate with proliferative activity. We have examined PCNA levels in neuroblastomas in relation to N-myc gene amplification and tumor stage. Statistically, significantly higher levels of PCNA were observed in tumors with an amplified N-myc gene relative to tumors with a single gene copy. The highest levels of PCNA were observed in advanced stage tumors with an amplified N-myc gene. Treatment of neuroblastoma cells in culture with retinoic acid, which induces differentiation, resulted in a substantial decrease in PCNA. Our results suggest that PCNA levels may reflect differences in proliferative activity between neuroblastomas, related to stage of the disease and to N-myc gene copy number.

Activation of resting peripheral blood lymphocytes through the T cell receptor induces rapid phosphorylation of Op18.

Op18 is a highly conserved major cytosolic phosphoprotein that has been implicated in signal transduction in a wide variety of cell types. Freshly isolated peripheral blood lymphocytes (PBL) constitutively express low levels of mostly unphosphorylated Op18. After mitogenic stimulation of PBL, Op18 synthesis is induced at a time when cells are entering S-phase. In this study, we have examined the phosphorylation of Op18 in freshly isolated PBL after activation of the T cell receptor by OKT3. Quantitative analysis of Op18 phosphorylation was undertaken by metabolic labeling with 32Pi and PhosphorImager analysis of two-dimensional gels. After 10 or 15 min of activation by OKT3, one of the three major phosphorylated forms of Op18, designated Op18c, increased approximately 10-fold, which represented a most pronounced change among a large number of phosphoproteins analyzed. In time course experiments, increased Op18 phosphorylation to yield Op18c was observed as early as 2 min. Continued OKT3-induced activation for 20 to 72 h resulted in a further increase in phosphorylated Op18 forms, which paralleled new Op18 synthesis and occurred at a time when cells were entering S-phase, as determined by [3H]-thymidine incorporation. Inhibitors of lymphoid proliferation, cyclosporin A and RPM, had no effect on early (less than 15 min) phosphorylation. Addition of calphostin C, a specific inhibitor of protein kinase C, 1 min prior to stimulation of resting T cells with OKT3 completely inhibited further phosphorylation of Op18. Incubation of PBL with calphostin C for 75 min decreased constitutive levels of phosphorylated Op18. In contrast, inhibition of cyclic nucleotide-dependent protein kinases with HA1004 had no effect on Op18 phosphorylation. Activation of cAMP-dependent protein kinase with Forskolin or 8Br-cAMP did not increase Op18 phosphorylation. Our results suggest that Op18 phosphorylation is mediated by protein kinase C activation as an early event in T cell activation through the T cell receptor.

Fat-suppressed MR imaging of myositis.

A hybrid fat-suppression sequence in magnetic resonance (MR) imaging was used to evaluate inflammatory muscle disorders in seven children: five patients with dermatomyositis, one patient with vasculitis, and one patient with viral myositis. Fat-suppressed multisection axial images obtained with the same repetition and echo times as those used to obtain standard spin-echo (SE) images enabled direct comparison of images, with little variation of T1 and T2 weighting. In six patients, the contrast on images obtained with T2 fat suppression was 15%-20% greater than contrast on conventional T2-weighted SE images. In all seven patients, the subjective judgment was that T2-weighted fat-suppression sequences improved visualization of muscle abnormalities. It is concluded that T2 fat suppression is useful in evaluation of inflammatory muscle disorders in children because it increases contrast and eliminates fat as a cause of muscle abnormality.

Serial magnetic resonance imaging in juvenile dermatomyositis.

Magnetic resonance imaging (MRI) was used to follow the course of juvenile dermatomyositis from the onset of disease through resolution of a primary relapse. The signal intensity of the T2-weighted image of involved muscles was elevated during periods of disease activity, and returned to approximately normal levels with effective suppression of disease activity. T1-weighted images of involved muscles remained approximately normal despite disease activity.

High levels of p19/nm23 protein in neuroblastoma are associated with advanced stage disease and with N-myc gene amplification.

The gene encoding a novel protein designated nm23-H1, which was recently identified as identical to the A subunit of nucleotide diphosphate kinase from human erythrocytes, has been proposed to play a role in tumor metastasis suppression. We report that untreated neuroblastoma tumors contain a cellular polypeptide (Mr = 19,000) designated p19, identified in two-dimensional electrophoretic gels, which occurs at significantly higher levels (P = 0.0001) in primary tumors containing amplified N-myc gene. The partial amino acid sequence obtained for p19 is identical to the sequence of the human nm23-H1 protein. An antibody to the A subunit of erythrocyte nucleotide diphosphate kinase reacted exclusively with p19. In this study, significantly higher levels of p19/nm23 occurred in primary neuroblastoma tumors from patients with advanced stages (III and IV) relative to tumors from patients with limited stages (I and II) of the disease. Even among patients with a single copy of the N-myc gene, tumors from patients with stages III and IV had statistically significantly higher levels of p19/nm23 than tumors from patients with stages I and II. Our findings indicate that, in contrast to a proposed role for nm23-H1 as a tumor metastasis suppressor, increased p19/nm23 protein in neuroblastoma is correlated with features of the disease that are associated with aggressive tumors. Therefore, nm23-H1 may have distinct if not opposite roles in different tumors.

Magnetic resonance imaging appearance of the muscles in childhood dermatomyositis.

Documentation of muscle involvement in a child thought to have dermatomyositis may require the performance of invasive procedures such as electromyography and/or muscle biopsy. We describe four patients with dermatomyositis in whom magnetic resonance imaging (MRI) demonstrated the muscle involvement. The involved muscles had increased signal intensity on the T2-weighted images (SE 2500/80) and normal appearance on the T1-weighted images (SE 600/20). The involvement of the muscles was not uniform. There was good correlation between the distribution of muscle involvement by MRI and functional testing. Follow-up MRI scans in patients with favorable outcome demonstrated that the affected muscles had returned to normal signal intensity. Although the MRI findings are not specific, in the proper clinical context they may be helpful in establishing the diagnosis of dermatomyositis. MRI may also be used in establishing an appropriate muscle biopsy site. In addition, MRI may be used for monitoring the progress of the disease.