The binding and processing of monoclonal human IgG1 by cells of a human macrophage-like cell line (U937)

The goal of these experiments was to assess the relationship between the binding and processing of IgG by Fc-receptor-bearing cells. Cells of the U937 human macrophage-like cell line were incubated with 125I- labeled monomers, dimers, oligomers (composed of 2-4 IgG1 subunits), and HP (heavy polymers composed of 5 or more subunits per polymer) of monoclonal human IgG1 in vitro. Binding was assessed by spinning cells through a layer of phthalate oils. Internalization of IgG1 was assessed by quantitating residual binding to cells after surface-bound IgG was removed by a brief treatment with a solution containing 0.25 M acetic acid and 0.5 M sodium chloride. Catabolism was assessed by measuring the release of radioactive fragments of IgG1, which were not precipitated by 10% trichloroacetic acid. Unstimulated U937 bound about 10,000 molecules per cell of IgG1 monomer, with an equilibrium binding constant (Ka) of 5 X 10(8) M-1. After stimulation with a conditioned medium in vitro, binding per cell was increased 3-7--fold, and the Ka was decreased 2-4--fold. Both unstimulated and stimulated cells internalized and catabolized labeled IgG1 HP, but stimulated cells internalized and digested much more IgG1 HP per cell than unstimulated cells. Both monomers and dimers of IgG1 were internalized and degraded very slowly by stimulated cells, even though both preparations readily bound to cells. In contrast, oligomers and (to an even greater extent) IgG1 HP were internalized and degraded much more rapidly. Internalization of IgG1 HP was markedly inhibited by incubation at 4 degrees C, but not by incubation with a variety of metabolic inhibitors. Catabolism was inhibited by chloroquine and monensin (inhibitors of lysosomal acidification) and by cytochalasin (an inhibitor of microfilament polymerization). Binding to the surface of cells was not markedly inhibited by any agent tested. The capacity of cells to bind labeled IgG1 was markedly reduced by prior incubation in the presence of unlabeled IgG1. This reduction was in part due to the steric blockade of receptors caused by the avid, but reversible, binding of IgG1. In addition, IgG1 oligomers or HP (but not IgG1 monomers or dimers) also caused an irreversible reduction in the number of Fc receptors by a process analogous to receptor down-regulation, as observed in other receptor--ligand systems.     

Periosteal osteosarcoma--a European review of outcome

Periosteal osteosarcoma is a rare primary malignant bone tumour. Treatment is by surgical excision, but controversy remains about the value of chemotherapy. The members of the European Musculo Skeletal Oncology Society (EMSOS) collaborated to produce a dataset of 119 patients. The predominant site for the tumour was the femur, followed by the tibia. All but 2 patients underwent surgery, with 9 requiring amputation and the others having limb salvage. A total of 81 patients had chemotherapy, of whom 50 had neoadjuvant chemotherapy. There was no standard chemotherapy regime, but all patients receiving chemotherapy were given doxorubicin combined with at least one other agent. The overall survival was 89% at 5 years and 83% at 10 years. Eight patients developed local recurrence, of whom 5 died. Survival was related to appearance of local recurrence (P < 0.0001) but no other single factor. The use of chemotherapy was not shown to be a prognostic factor, but was used in two-thirds of the patients in this study.     

Possibility of selection against mtDNA mutations in tumors

Several studies of tumors have revealed substantial numbers of clonally expanded somatic mutations in mitochondrial DNA (mtDNA), not observed in adjacent intact tissues. These findings were interpreted as indicating the involvement of mtDNA mutations in tumorigenesis. Such comparisons, however, ignore an important confounding factor: the monoclonal origin of tumors as opposed to the highly polyclonal nature of normal tissues. Analysis of recently published data on the incidence of somatic mutations in nontumor monoclonal cells suggests that, contrary to the prevailing view, the process of tumorigenesis may be accompanied by active selection against detrimental mtDNA mutations.     

Endoprosthetic replacement of diaphyseal bone defects. Long-term results

We retrospectively studied 35 patients who underwent endoprosthetic reconstruction of diaphyseal bone defects after excision of primary sarcomas. The patients were treated between February 1979 and May 1999 and had more than 5 years follow-up. There were 22 males and 13 females and the median age at diagnosis was 29 (8–75) years. The bone defect measured a mean of 19 (10–27.6) cm. There were 29 femoral reconstructions, three tibial and three humeral. Cumulative overall survival for all patients was 65% at 10 years. Cumulative overall survival for prosthetic reconstruction, using revision surgery as an end point, was 63% at 10 years. Cumulative risk of failure of reconstruction, including infection, fracture, aseptic loosening, local recurrence and amputation, was 60% at 10 years. Tibial and humeral reconstructions fared less well than femoral. Endoprosthetic replacement is a useful method of reconstructing long intercalary defects, especially if situated in the femur.

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Résumé Nous avons étudié rétrospectivement 35 malades qui ont subi une reconstruction endoprothétique après excision diaphysaire dun sarcome primaire. Les malades ont été traités entre février 1979 et mai 1999 et avaient plus de 5 ans de suivi. Il y avait 22 hommes et 13 femmes et lâge médian au diagnostic était de 29 (8–75) ans. Le défaut osseux mesurait en moyenne 19 (10–27.6) cm. Il y avait 29 reconstructions fémorales, trois tibiales et trois humérales. La survie totale cumulative pour tous les malades était 65% à 10 ans. La survie totale cumulative pour la reconstruction prothétique, en utilisant la chirurgie de révision comme élément final, était de 63% à 10 ans. Le risque cumulatif déchec en incluant: linfection, la fracture, le démontage aseptique, la récidive locale et lamputation étaient de 60% à 10 ans. Les reconstructions tibiales et humérales sont allées moins bien que les fémorales. Le remplacement endoprothétique est une méthode utile pour reconstruire de longues pertes de substance intercalaires, surtout si elles sont situées sur le fémur.

     

Possible metastasis of osteosarcoma to a remote biopsy site: a case report

In a patient who presented with an osteosarcoma of the proximal humerus, systemic staging revealed a lesion in the contralateral humerus, which was shown to be an enchondroma after open biopsy. The patient had a local recurrence develop, which necessitated forequarter amputation. Shortly thereafter, the patient had pain develop at the contralateral biopsy site, and examinations and repeat biopsy revealed metastatic osteosarcoma. We think it is likely that surgical trauma contributed to the development of this unusual metastasis. In this clinical situation, consideration should be given to doing the biopsy and definitive resection on separate occasions.     

MR imaging in the assessment of residual tumour following inadequate primary excision of soft tissue sarcomas

The purpose of this retrospective study was twofold: firstly, to assess the ability of MR imaging in confirming/excluding the presence of residual tumour following inadequate primary excision of soft tissue sarcomas; and secondly, to assess the accuracy of the original radiologists report as compared with a retrospective review of the scan hard copy in confirming/excluding. A total of 111 cases were identified that fulfilled the inclusion criteria of inadequate primary surgery followed by a MR scan and subsequent wide re-excision of the surgical field. The gold standard for the assessment of the MR imaging studies was histological examination of the re-excision specimens. Histological examination revealed residual tumour in 63 (56.7%) cases. In 48 cases the residual tumour was classified macroscopic (maximum diameter >10 mm) and 15 cases microscopic (maximum diameter 10 mm). The original radiologists reports failed to indicate the presence or absence of tumour in 7 (6.3%) cases. In the remaining 104 cases the diagnostic performance of MR imaging gave a sensitivity of 0.64, specificity of 0.93, positive predictive value of 0.93 and negative predictive value of 0.67. In 12 of the 21 false-negative scans the residual tumour was microscopic. Subjective assessment of the radiologists reports indicated that the proportion of equivocal reports was much higher in both the false-negative and false-positive groups as compared with the true groups. An unblinded retrospective review of the scan hard copies only differed from the original radiologists report in 8 (7.2%) cases. Contrast-enhanced sequences were not routinely obtained in this series. The results suggest that the poor negative predictive value can be attributed more to limitations of the MR scan and not to failures in observation or interpretation by the radiologists. Despite the low negative predictive value, MR imaging remains useful in planning the re-excision surgery by identifying the site and extent of the original operation and size of major residual tumour.     

A gene expression signature associated with metastatic outcome in human leiomyosarcomas

Metastasis is a major factor associated with poor prognosis in cancer, but little is known of its molecular mechanisms. Although the clinical behavior of soft tissue sarcomas is highly variable, few reliable determinants of outcome have been identified. New markers that predict clinical outcome, in particular the ability of primary tumors to develop metastatic tumors, are urgently needed. Here, we have chosen leiomyosarcoma as a model for examining the relationship between gene expression profile and the development of metastasis in soft tissue sarcomas. Using cDNA microarray, we have identified a gene expression signature associated with metastasis in sarcoma that allowed prediction of the future development of metastases of primary tumors (Kaplan-Meier analysis P = 0.001). Our finding may aid the tailoring of therapy for individual sarcoma patients, where the aggressiveness of treatment is affected by the predicted outcome of disease.