Using the pulsed dye laser to influence scar formation after breast reduction surgery: a preliminary report.

Many patients undergoing bilateral breast reduction surgery develop problems. Foremost among these problems is scar hypertrophy and its attendant symptoms. Although there is as yet no firm prognostic indicator for hypertrophy, scars that become hypertrophic often have a particular blood vessel pattern, observable by transcutaneous microscopy, showing vessels that lay transversely across the incision line with minimal crosslinks between them. Hypertrophic scars that develop in incision lines become wide, with the final width of the scar dependent on the maximum thickness during the growth stages before maturation and resolution. Close monitoring of scars forming in the incision line using the transcutaneous microscope detected this aligned vessel pattern before overt hypertrophy was seen. Use of the Pulsed Dye Laser caused disruption in the vessel pattern, appeared to inhibit additional hypertrophic development, and promoted early maturation of scars.

Karyotypic abnormalities and immunoglobulin gene rearrangements in Hodgkin's disease.

Biopsy samples from seven patients with Hodgkin's disease (HD) were examined for cytogenetic abnormalities and rearrangement of the genes encoding the immunoglobulin chains and T-cell receptor chains. Three samples demonstrated clonal rearrangements of both IgH and IgL genes. No rearrangements of the TCR beta genes were detected in any of the samples. Karyotypic abnormalities were also found but only in the three cases where a clonal rearrangement of the immunoglobulin genes was shown. Two of these three cases had multiple karyotypic abnormalities, with the remaining patient being trisomic for chromosome 16 as the sole abnormality. These results are discussed and compared with previous reports in the literature concerning HD.

Establishment and characterization of a human CD4 positive cell bank for HIV related studies.

The human CD4 positive cell lines JM, CCRF, CEM, U937, HL60 and THP-1 have been cleared of mycoplasma contamination and defined by DNA fingerprinting and cell surface phenotype marker analysis. These cells have been banked and are now available as a source of standardized cell lines for HIV related research.

Analysis of T-cell receptor and immunoglobulin gene rearrangements in the diagnosis of Hodgkin's and non-Hodgkin's lymphoma.

The results of genotypic analysis of 29 cases of malignant lymphoma are reported and the application of this technique for differentiating between Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) is evaluated. Five cases with a differential diagnosis which included HD and NHL were analysed. These results are compared with those obtained for six B-cell NHLs, nine T-cell NHLs, and nine cases of HD. This report suggests that gene rearrangement analysis is useful in some cases in which the differential diagnoses includes HD and NHL as the absence of gene rearrangements is more consistent with a diagnosis of HD than of NHL. Two monoclonal antibodies reactive with the variable region of T-cell receptor beta-chain and molecular probes to the relevant variable region genes were used to assist in the diagnosis of T-cell lymphoma. This report confirms that genotypic analysis is useful diagnostically when the results are assessed in the context of the histopathological findings.

Distribution of a set of idiotopes detected by a monoclonal antibody panel in 42 cases of chronic lymphocytic leukaemia: definition of potential targets for immunotherapy.

Using a panel of 14 monoclonal V region reactive antibodies generated against a single IgG1 lambda paraprotein we have identified shared idiotopes in a group of 42 patients with chronic lymphocytic leukaemia (CLL). The specificity of cellular staining by indirect immunoperoxidase was confirmed in the majority of cases by an ELISA assay using secreted idiotypic immunoglobulin. In a few cases weak cellular binding by the panel antibodies could not be confirmed as specific for immunoglobulin. Four monoclonal antibodies specific for lambda chain determinants reacted with 5-29% of lambda expressing CLLs but the significance of this is uncertain as the antibodies may be recognizing one of the commoner V lambda subgroups. Two antibodies, which are only weakly reactive with normal serum immunoglobulin, detected heavy chain associated idiotopes which were expressed by 7% and 14% of CLL cases. One of these antibodies detects an idiotope which was expressed significantly more frequently by CLL associated immunoglobulin than by a panel of paraproteins. Such preferentially expressed idiotopes may be useful in investigating the biology of this disorder as well as providing attractive targets for immunotherapy.

Quantitation of expression of idiotopes recognized by a panel of monoclonal antibodies in normal serum immunoglobulin: use of a novel 4-stage ELISA assay.

This paper describes the application of a sensitive ELISA assay detecting as little as 10 ng/ml of a specific idiotope. Using this assay we were able to divide monoclonal anti-idiotypic antibodies generated using a single IgG1 lambda paraprotein as the immunogen into three distinct groups. Seven antibodies detected determinants which were expressed only by the immunogen. The remaining seven antibodies all reacted with normal serum immunoglobulin. Four antibodies reacted with "public" idiotopes which were strongly expressed by serum immunoglobulin, and three antibodies reacted with "restricted public" idiotopes which were only weakly expressed by serum immunoglobulin. This last group may be of significant interest as agents for the immunotherapy of B-cell malignancies. The conservation of expression of these idiotopes in many individual sera suggests that they have a physiological role in immune regulation and would therefore be excellent targets for immunotherapy of B-cell tumours while their quantitatively low expression by circulating immunoglobulin is unlikely to interfere with tumour cell specific binding in vivo.

Characterization of a new non-Hodgkin's lymphoma cell line (NCEB-1) with a chromosomal (11:14) translocation [t(11:14)(q13;q32)].

A new cell line, NCEB-1, was established by Epstein-Barr virus (EBV) transformation of peripheral blood mononuclear cells from a patient with centroblastic-centrocytic diffuse lymphoma expressing IgM lambda. The transformed cells were lymphoblastoid, with many cells showing a plasmacytoid morphology. The NCEB-1 cells had cytoplasmic Ig (CyIg), with loss of the surface Ig (SIg) expression. Cytogenetic analysis of the cell line demonstrated two clones with variations: a hypodiploid clone, with a complex karyotype including a t(11;14)(q13;q32) similar to the original tumor cells, and a near tetraploid clone with the same markers. Southern blot analysis of DNA from the patient's neoplastic cells and NCEB-1 demonstrated identical Ig heavy chain gene rearrangement, confirming the origin of the cell line. The cell line was not tumorigenic when tested in an in vitro assay using immunosuppressed mice. NCEB-1 has been in continuous culture for 9 months and will be valuable for the in vivo study of non-Hodgkin's lymphoma and EBV transformation.

T-cell lymphoma: morphology, immunophenotype and clinical features.

The histology, immunophenotype and clinical presentation of 43 cases of T-cell lymphoma are described. Cases were classified into nine types; T-lymphocytic lymphoma (three), mycosis fungoides (six), Sézary syndrome (two), T-zone lymphoma (13), angioimmunoblastic lymphadenopathy (AIL)-like T-cell lymphoma (five), pleomorphic medium cell (one), large cell immunoblastic (four), large cell polylobated (five) and lymphoblastic (four). The patients comprised 26 males and 17 females aged between 15 and 86 years. The majority showed disseminated disease at the time of diagnosis (18 stage IV, nine stage III, five stage II, eight stage I and three cases not staged). Thirty-one patients showed lymph node involvement. Cutaneous involvement was a common finding (18 cases, 10 cases excluding mycosis fungoides and Sézary syndrome). Details of therapy and clinical follow-up were obtained in 37 cases. With simple chemotherapy only one complete response (7%, 1/16) was obtained. With aggressive therapy 48% (13/27) of patients showed complete responses. Twenty patients died during the follow-up period. Life table analysis showed a 58% probability of surviving 1 year and 36% probability of surviving 3 years. There was a significant difference in survival probability between low/intermediate-grade (lymphocytic, Sézary syndrome, mycosis fungoides and T-zone lymphoma including AIL-type) lymphomas and high-grade (large cell immunoblastic and polylobated and lymphoblastic) lymphomas (P less than 0.025). However, when survival of T-zone and AIL-like T-cell lymphoma was compared with survival of large cell immunoblastic and polylobated lymphomas no significant difference was detected. Age (less than 50 years) and stage I or II disease were associated with significantly better survival (P less than 0.005 and P less than 0.05).

T-24.B-cell differentiation factor induces immunoglobulin secretion in human B cells without prior cell replication.

Stimulation of B lymphocytes from B-cell chronic lymphocytic leukaemia (B-CLL) with 12-0-tetradecanoylphorbol-13-acetate (TPA) has shown that these cells are capable of differentiation (Totterman, Nilsson & Sundstrom, 1980). Increases in the expression of different class II MHC antigens (Guy et al., 1983, 1986) and responsiveness to growth factors (Kabelitz et al., 1985; Suzuki, Butler & Cooper, 1985) have been studied. Supernatant from the human bladder carcinoma line T-24 contains a B-cell differentiation factor (BCDF) able to induce immunoglobulin secretion from CESS cells. We investigated the induction of proliferation and immunoglobulin secretion in human B cells by studying the effects of this factor on B-CLL cells, in both the presence and absence of TPA. We report here that this material (termed T-24.BCDF) causes immunoglobulin secretion to be initiated in these cells, and that this is not accompanied by detectable DNA synthesis. These observations were extended to normal human B cells and demonstrate that human B cells can secrete immunoglobulin in the absence of clonal expansion.

Prostatic infiltration in chronic lymphatic leukaemia.

Forty six men with chronic lymphatic leukaemia (CLL) were studied for up to seven years. Six patients required surgery for prostatic outlet obstruction. Histological examination of the prostatic chippings showed variable degrees of infiltration with small mature lymphocytes in all six patients, suggestive of a leukaemic origin for the cells. Patients with chronic lymphatic leukaemia who undergo prostatectomy may have a higher incidence of leukaemic infiltration than has been previously recognised.

The incidence, clonal origin and secretory nature of serum paraproteins in chronic lymphocytic leukaemia.

Immuno-isoelectric focusing (IIEF) showed a 61% incidence of serum paraproteinaemia in 56 patients with chronic lymphocytic leukaemia (CLL). A strong correlation between the serum paraprotein heavy chain isotypes and those of the cytoplasmic heavy chain immunoglobulins was observed with no discrepancy noted in light chain expression. Density gradient ultracentrifugation analysis of selected sera containing monoclonal IgM showed that the IgM paraproteins were mostly 19S, secretory IgM but one patient was found to have both 19S and 8S monoclonal IgM. When the cellular origin of the IgM and IgD paraproteins found in one patient was investigated, both paraproteins were found to share the same idiotype and originate from the neoplastic clone. These findings confirm the view that there is an incomplete maturation block in chronic lymphocytic leukaemia and that in vivo secretion of paraproteins by the neoplastic cells is a relatively common occurrence.

The role of immunoperoxidase techniques on paraffin embedded tissue in determining the histogenesis of undifferentiated thyroid neoplasms.

Twenty cases of undifferentiated thyroid tumours were reviewed histologically. In seven cases the histogenesis was difficult to determine using morphological criteria. Immunohistochemical staining with a panel of antibodies to lymphoid and epithelial cells, including monoclonal antibodies directed against the leucocyte common antigen, cytokeratin, and epithelial membrane antigen confirmed that four of these cases were lymphomas and that one was a medullary carcinoma. In the remaining two cases immunohistochemistry was unhelpful. In the thirteen histologically typical tumours, the immunohistochemical profile was in keeping with their histogenesis as determined by morphological criteria. Immunohistochemical staining with a panel of selected antibodies allows the reliable diagnosis of undifferentiated thyroid neoplasms, when this cannot be reached using routine histological techniques.

Expression of MHC class II antigens in human B-cell leukaemia and non-Hodgkin's lymphoma.

In this review we have summarized our experiences of serological analysis of MHC class II antigen expression in human B cell malignant disease. Cells from a large number of cases of B-cell chronic lymphocytic leukaemia (CLL) and non-Hodgkin's lymphoma (NHL) have been examined for expression of class II antigens. Using a number of monoclonal antibodies which in some cases are specific for class II subregion products (DP, DQ and DR), MHC class II antigens were detected by indirect immunofluorescence and fluorescent activated cell sorter analysis in CLL and by immunohistochemical staining in NHL. At the cell surface in many cases of B cell malignant disease, products of the different class II subregion genes are non-coordinately expressed. The most commonly occurring pattern of non-coordinate expression of class II molecules is of expression of DP and DR antigens in the absence of detectable DQ expression. These findings are in contrast to normal B lymphocytes where DP, DQ and DR antigens are expressed together at the cell surface. There is considerable heterogeneity among cases comprising individual histopathological categories of B cell malignancy, and in many instances heterogeneous class II phenotypes are also found on cells from the same tumour. In chronic lymphocytic leukaemia, class II antigen expression is inducible in vitro by treating the cells with the phorbol ester TPA. CLL cells treated with TPA have much increased levels of class II antigen expression at the cell surface and much increased steady state levels of class II specific mRNA transcripts detectable with complementary DNA probes. Aberrant class II antigen expression may be involved in the pathogenesis of B cell malignant disease.

Expression of MHC class II antigens in human B-cell leukaemia, and increased levels of class II antigens and DR-specific mRNA after stimulation with 12-O-tetradecanoyl phorbol-13-acetate.

Cells from the peripheral blood of B-cell chronic lymphocytic leukaemia (CLL) patients were examined serologically for the expression of cell surface MHC class II antigens with monoclonal antibodies (mAbs) specific for the products of HLA-DP, -DQ and -DR genes, and mRNAs from the cells of three patients were analysed with a cDNA probe specific for DR beta chain genes. In 12 cases of CLL studied by indirect immunofluorescence and FACS analysis, a variable proportion of cells failed to express detectable levels of HLA-DP and HLA-DQ antigens at the cell surface, although greater than 90% of the cells had detectable expression of HLA-DR antigens. In all cases, greater than 90% of the cells expressed MHC class I antigens and the majority of cells reacted with the Leu-1 (CD5) mAb. Cells from different patients expressed variable levels of MHC class II antigens, and this was reflected in the finding of variable levels of mRNA detectable with the cDNA probe. Culture of cells with the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA) induced much increased levels of expression of MHC class II antigens. HLA-DP and -DQ antigens were expressed on greater than 90% of the cells in all cases studied after culture of the cells with TPA, and MHC class II specific mRNA transcripts were correspondingly increased. In a single case of plasma cell leukaemia studied, MHC class II antigens were not detectable at the cell surface and their expression was not induced after culture of the cells with TPA.

Deficient expression of MHC class II antigens in some cases of human B cell leukaemia.

Cells from the peripheral blood of patients with B cell chronic lymphocytic leukaemia (B-CLL) and acute lymphoblastic leukaemia (ALL) were examined for the expression of MHC class II antigens, using a number of monoclonal antibodies (MoAb) including L243 (anti-DR) and TU22 (anti-DQ). There was wide variation in expression of MHC class II antigens in CLL, both from patient to patient and among cells from the same individual. In a number of subjects a significant proportion of the cells had detectable levels of expression of DR antigens but not of DQ antigens. In some cases of ALL although almost all cells were MHC class II positive, DQ expression was undetectable. Differentiation of CLL cells, induced by culturing the cells with 12-0-tetradecanoyl phorbol-13-acetate (TPA), was accompanied by increases in MHC class II expression at the cell surface of up to more than 20-fold, and resulted in detectable expression of DQ antigens on greater than 90% of the cells in all the subjects studied.

Immunophenotype analysis of malignant histiocytosis of the intestine.

Five cases of malignant histiocytosis of the intestine and one case of true histiocytic lymphoma were studied using immunohistological techniques. In paraffin sections tumour cells in all cases were shown to contain alpha-1-antitrypsin and to express the leucocyte common antigen. Four of the five cases of malignant histiocytosis of the intestine and the case of histiocytic lymphoma expressed the epithelial membrane antigen. Cryostat sections in four cases of malignant histiocytosis of the intestine showed that most tumour cells reacted with anti-T cell monoclonal antibodies. Only a minority expressed a typical monocyte macrophage phenotype.

Surface glycoproteins as markers of the cellular status of B chronic lymphocytic leukaemia lymphocytes.

The phenotype of B CLL cells is investigated with respect to their surface glycoproteins. These glycoproteins are identified by vectorial tritiation followed by 1 and 2 dimensional gel electrophoresis, and by lectin and MoAb binding using immunoprecipitation and flow cytometry. The profiles of the CLL cells are compared with those of normal B cells, prepared from tonsils, and T cells from peripheral blood. The CLL cells show many similarities with T cells, particularly the expression of glycoproteins which bind the MoAbs gpL 115, F10-44-2 and EZB 52, and a complex set of binding sites for Helix pomatia lectin. The significance of these observations in terms of the cellular origins of the leukaemic lymphocytes is discussed.