[Back pain, bone metastases and medullary compression]. / Rygsmerter, knoglemetastaser og medullaer kompression.

A case of lung cancer and metastatic spinal cord compression is presented. Despite progressive backpain during two weeks and total loss of walking ability, the patient regained the ability to walk one week after radiotherapy for the malignant spinal cord compression. Guidelines for recommended diagnostic approaches are given.

Expression of polymorphic HLA-A,B epitopes in human urothelial cell lines.

Malignant human urothelial cell lines propagated in vitro have previously been demonstrated to express low amounts of monomorphic HLA-A,B,C as compared to premalignant urothelial cells. In this study the expression of polymorphic HLA-A,B epitopes in human urothelial cell lines have been investigated in greater detail. The expression of HLA-B locus coded epitopes in malignant TGrIII cells was demonstrated to be low or absent as compared to pre-malignant TGrII or slightly transformed TGrI cells, suggesting a mechanism by which malignant cells could escape from the host immune response. The extreme polymorphism of HLA-A,B,C antigens suggests that HLA typing could be used as a method to identify the origin of cell lines which is essential in the study of the process of malignant transformation in vitro, or when correlating in vitro data with clinical observations of the patient. Two urothelial cell lines classified as slightly transformed (TGrI) and two as pre-malignant (TGrII) could, according to their expression of polymorphic HLA-A,B epitopes, be identified as genuine independent cell lines. One TGrII cell line previously designated Hu1734 shared the same HLA-A,B phenotype as the genuine HCV29 (TGrII) cell line and is therefore suspected to be a subline of the latter. Out of 18 cell lines and sublines classified as TGrIII the fidelity of four (Hu1922 and three sublines of T24) was proved by their HLA-A,B phenotype. Mistaken identity either by contamination or false labelling of the cultures was suspected in samples of six TGrIII cell lines and sublines. In four of these the HLA-A,B type characteristic for the T24 cell line could be demonstrated, but in general HLA typing of TGrIII cell lines as a method to identify the origin of the individual cell lines failed, primarily due to the decreased expression of HLA-A,B,C antigens and the apparent selective loss of HLA-B locus coded antigens.

Reduced HLA-A,B,C expression in tumourigenic v-raf transfected human urothelial cells.

Tumourigenic (TGrIII) human urothelial cells grown in vitro have previously been demonstrated to have a markedly decreased expression of beta 2-microglobulin and HLA-A,B,C antigens as compared to non-tumourigenic (TGrII) human urothelial cell lines. Furthermore, during 'spontaneous' in vitro transformation of a non-tumourigenic (TGrII) human urothelial cell line Hu609 into a tumourigenic (TGrIII) subline Hu609T/LLH, changes in morphology and tumourigenicity have been demonstrated to be accompanied by a decreased HLA-A,B,C expression. After malignant transformation of the non-tumourigenic (TGrII) human urothelial cell line HCV29 by DNA transfection with the v-raf oncogene, four sublines could be isolated. In this study we have investigated these sublines for their expression of membrane bound HLA-A,B,C antigens and provide further evidence that an inverse relationship exists between tumourigenicity and monomorphic HLA-A,B,C expression. Treatment of the cells with recombinant human interferon alpha for 3 days increased the expression of HLA-A,B,C antigens by 50-150% indicating that at least some of the reduced HLA-A,B,C expression could be due to decreased synthesis of HLA-A,B,C antigens. All the transfected cell lines overexpress v-raf and c-myc.

Recombinant human interferon gamma exerts an anti-proliferative effect and modulates the expression of human leukocyte antigens A,B,C and DR in human urothelial cell lines.

In this study we have treated three malignant (TGrIII) and two pre-malignant (TGrII) urothelial cell lines with recombinant human interferon gamma (rHu-INF gamma). The malignant cells (HCV29-T112C1, Hu1703He and T24) were inhibited in growth by more than 50% after treatment with 100-1000 units of rHu-INF gamma/ml for 4 days as compared to untreated controls. The growth of the pre-malignant cell lines (HCV29 and Hu609) was not influenced to the same extent in the presence of rHu-INF gamma in the culture medium. Treatment with rHu-INF gamma increased the expression of monomorphic human leukocyte antigens (HLA) A,B,C as well as beta 2-microglobulin in all the cell lines tested, as demonstrated using a quantitative immunofluorescence assay. The tumourigenic cell lines increased their expression of HLA in a dose-dependent way, whereas treatment of the non-tumourigenic cells with higher concentrations of rHu-INF gamma than 10 units/ml, did not increase the HLA-A,B,C expression further. None of the cell lines expressed HLA-DR unless treated with rHu-INF gamma. No correlation between tumourigenicity and the dose of rHu-INF gamma required for "de novo" induction of HLA-DR could be demonstrated. After removal of rHu-INF gamma from the medium, the expression of HLA-DR gradually decreased in less than 14 days, indicating that the expression of HLA-DR is not constitutive but dependent upon the presence of rHu-INF gamma. We conclude that human urothelial cells grown in vitro are sensitive to the anti-proliferative and major-histocompatibility-complex-modulating effects of rHu-INF gamma, and that malignant urothelial cells are more sensitive than pre-malignant cells. Finally, our data indicate a possible role for rHu-INF gamma in the management of human bladder cancer.

Correlation between classification of human urothelial cell lines and HLA-A,B,C expression.

Quantitative changes in major histocompatibility class I antigen expression in tumour cells are believed to affect the host immune response against the tumour. In tumourigenic (TGrIII) human urothelial cell lines the apparent loss of polymorphic HLA-A,B epitopes has previously been demonstrated. In the present study, 3 non-tumourigenic (TGrII) and 6 tumourigenic (TGrIII) human urothelial cell lines have been investigated for their quantitative expression of monomorphic HLA-A,B,C and B2-microglobulin. Evidence is provided that an inverse correlation exists between tumourigenicity and HLA-A,B,C and B2-microglobulin expression. Furthermore, treatment of the cells with neuraminidase partly restored the expression of monomorphic HLA-A,B,C suggesting that at least some of the observed quantitative differences could be due to masking of the membrane bound HLA antigens by sialic acid-containing glycoconjugates.

Changes in HLA A, B, C expression during "spontaneous" transformation of human urothelial cells in vivo.

The immortalized but non-tumourigenic and non-invasive human urothelial cell line, Hu 609, known to express the appropriate HLA A,B antigens (A2,-; B5,-) has previously been demonstrated to undergo "spontaneous" in vitro transformation into an invasive and tumourigenic subline, Hu 609T/MV. This subline does not express the polymorphic HLA epitopes. In the present investigation we have followed two additional "spontaneous" transformations of the Hu 609 cell line into malignant sublines. Evidence is presented that the development of morphological changes and tumourigenicity were accompanied by a gradual loss of the expression of polymorphic HLA A,B epitopes and a reduction in the expression of monomorphic HLA A,B,C antigens. Antigens could be detected again after neuraminidase treatment. We conclude that the urothelial Hu 609 cell line after "spontaneous" transformation still possesses the HLA A,B epitopes. The observed quantitative differences in HLA expression between Hu 609 and its malignant sublines may be due to masking of the HLA antigens by sialic acid containing tumour-associated highly branched glycoproteins.