Production and characterization of a monoclonal antibody (A1-3) that binds selectively to activated monocytes and inhibits monocyte procoagulant activity.

We describe the generation and characterization of a new monoclonal antibody, A1-3, which possesses two unique properties. First, A1-3 binds selectively to stimulated human monocytes. Secondly, A1-3 inhibits the procoagulant activity expressed by stimulated monocytes and by human brain tissue factor. Unstimulated human peripheral blood cells (granulocytes, lymphocytes, monocytes, red blood cells, and platelets), prepared in the absence of detectable endotoxin, express no procoagulant activity and fail to bind A1-3. Stimulation of peripheral blood monocytes. alveolar macrophages, or the monocyte-like cell line U937, however, results in the expression of procoagulant activity and the binding of A1-3. The surface antigen recognized by A1-3 was recovered from endotoxin-stimulated human monocyte vesicles by immune precipitation and demonstrated an apparent m.w. of approximately 52,000. It is proposed that the monoclonal antibody A1-3 detects a differentiation antigen on human monocytes that is expressed in response to stimuli for monocyte activation.

The effect of alphanapthylthiourea (ANTU)-induced acute injury on lung binding of antibody to angiotensin converting enzyme (ACE).

The effects of ANTU-induced acute pulmonary capillary injury on lung and serum ACE functional activity and the specific accumulation of radio-labelled anti-ACE in lung were explored. Rats were injected either with ANTU or the solvent and sacrificed at various intervals up to one week after injection. All ANTU-injected animals developed pulmonary edema and bilateral pleural effusions which resolved by the one week time point. At no time was there any significant change in serum ACE levels. The specific activity of total lung ACE however rose from 11.0 +/- .95 (mean +/- SEM) to 18.4 +/- 1.1 by two hours after ANTU; by 24 hours, however, solubilized lung ACE had fallen significantly to 6.9 +/- .79 (p less than .01). Total lung ACE had returned to control values by one week. In parallel groups of animals the accumulation of 125I-labelled anti-ACE (AA) or normal sheep immunoglobulin (NSG) was compared in control and ANTU-treated rats. The ratio of the radioactivity in the lungs of AA--injected animals to that in NSG--injected animals fell significantly after ANTU administration (5.0 +/- .88 to 1.2 +/- .28 at 2 hours) suggesting that immunoreactive ACE had fallen despite an increase in ACE functional activity. The decreased binding of AA at the early time points perhaps reflects internalization of endothelial cell ACE in response to injury and an inability of the antibody to interact with the enzyme. The reduction in binding at 24 hours (1.38 +/- .47) correlates with a reduction in total lung ACE. ANTI-ACE may be a useful reagent for quantitating endothelial cell damage following lung injury.

Myelomatous ascites.

Ascites is an unusual feature of multiple myeloma. We report a case of ascites occurring early in the course of a patient with myeloma in whom there was no evidence of intra-abdominal plasmacytoma, and the skeleton was relatively spared. The serum contained predominantly polymeric IgA, a feature not investigated in previous cases. We reviewed the relevant literature and will discuss the suggestion that human myelomas presenting with the triad of ascites, relative or absolute sparing of the skeleton, and an IgA paraprotein bear an analogy to mouse myelomas induced by intraperitoneal instillation of irritants. The relevance of polymeric IgA is discussed with respect to tissue origin of the paraprotein. Seventeen cases were identified consistent with a definition of "myelomatous ascites" (malignant myeloma in which plasma cells and/or monoclonal immunoglobulin can be demonstrated in ascitic fluid). IgA immunoglobulin class was present at three times the incidence seen in myelomas in general (five of seven cases were specified). In twelve patients there was no identifiable intra-abdominal plasmacytoma although liver infiltration was common. Amyloidosis was reported in only one case, and no cases of uncomplicated plasma cell leukemia were noted. Ascites was a presenting feature in five cases. In each of these five there was absolute or relative sparing of the skeleton, four had no evidence of plasmacytoma, and the paraprotein was IgA in three, IgG in one, and unreported in one. In no case was there a known history of chronic intra-abdominal irritation.

Kinetics of amino acid transport by human-mouse myeloma hybrids--difference between human immunoglobulin producers and nonproducers.

Somatic cell hybridization techniques have allowed the preparation of interspecies hybrids that express the features of both parental cell lines. We have studied hybrids made with human myeloma cells fused to a continuous mouse myeloma cell line. In the present study we analyzed the kinetics of leucine influx and efflux in Ig producer and nonproducer hybrids. We found no statistical difference in amino acid influx; however, the rates of efflux were markedly increased in nonproducer hybrids as compared to the producers. The producer cells were tested further in puromycin known to inhibit protein synthesis. Under these conditions amino acid influx was not altered, but efflux was markedly increased resembling the findings in nonproducers. We conclude that hybrids that synthesize human immunoglobulins show decreased efflux of labeled leucine and this effect can be abolished by inhibition of protein synthesis. This difference in the efflux rate appears to be a consequence of immunoglobulin synthesis, rather than a component of a control mechanism of Ig synthesis.

Production of monoclonal antibodies to rat lung angiotensin-converting enzyme.

Several monoclonal antibodies to rat lung angiotensin-converting enzyme (ACE) have been produced. The antibodies are of the IgG class, do not inhibit ACE catalytic activity, and do not cross-react with the human or bovine enzyme. They bind in a curvilinear fashion to lung capillary endothelium by immunofluorescence microscopy, and radiolabeled antibody localizes in lung and other organs after intravenous injection into living rats. Each antibody tested appears to bind preferentially to lung rather than kidney ACE by ELISA, a finding supported by weak or absent immunofluorescence of kidney slices in vitro. These antibodies may be used to probe structural differences between ACE in various tissues and, by quantitating changes in accumulation of radiolabeled antibody in experimental models of lung injury, should complement functional measurements in determining the presence of subtle and progressive endothelial damage.

Combination chemotherapy for the treatment of metastatic basal cell carcinoma of the scrotum. A case report.

A case of metastatic basal cell carcinoma originating in the scrotum is presented. This tumour metastasized early in its course. It was treated with combination chemotherapy consisting of cis-platinum, bleomycin, and 5 fluorouracil, producing a partial remission with probable prolongation of survival. A review of the literature on chemotherapy of metastatic basal cell carcinoma is presented. This combination chemotherapy may be a good choice in the treatment of this rare complication of basal cell neoplasms.

Kinetics of human light chain synthesis by human-mouse myeloma hybrids.

Human myeloma cells from a patient suffering from a lambda Bence-Jones myeloma were hybridized with a continuous mouse myeloma cell line. Several hybrids were obtained, two of which produced the same lambda chain produced by the patient. They have proved to be stable over long periods of time in continuous cell culture. Kinetics of growth of this line proved to be linear over 72 h. This growth rate is independent of any possible nutrient depletion effect from the media. Rate of production of lambda chain was highest (45 micrograms/10(6) cells/24 h) when cell density was between 3-5 X 10(5)/ml and decreased as cell density increased. Efficiency of human light chain synthesis in these hybrids is similar to murine Ig synthesis by the producer parental mouse cells. Human-mouse myeloma hybrids that produce large quantities of human Igs can be established routinely, are stable over long periods of time, and can be used as a model system for the study of the control of synthesis of human Igs.

Metastatic basal cell carcinoma of the scrotum.

We report a rare case of basal cell carcinoma of the scrotum presenting with distant metastases. Rationale and treatment with bacillus Calmette-Guerin immunotherapy and subsequent combination chemotherapy are detailed.

Amino acid accumulation in mouse-human parental hybrid cells.

Cell membrane function during hybridization was assessed by examining the accumulation of four representative amino acids and a sugar in mouse-human hybrid cells, parental mouse cells and human cells. Qualitatively, accumulation in the hybrids was found to resemble the accumulation in parental mouse more than the human cells. In particular, the steady-state uptake of leucine and glycine in hybrids was similar to that in parental mouse cells and different than that observed in the human cells. The findings suggest that hybrids retain transport properties of mouse rather than human cells. These results are in keeping with our previous observation that the human mouse hybrids conserve most of the mouse chromosomes and the membrane proteins of the hybrids are very likely of mouse parental origin.

Non-random human chromosome distribution in human-mouse myeloma somatic cell hybrids.

The process of human chromosome segregation in human myeloma x mouse myeloma cell hybrids was investigated. The human myeloma cells were derived from two different patients. Fifty-eight independent hybrid lines were obtained. A representative number of cells from each line were karyotyped and isozyme markers for a number of the chromosomes retained were analyzed. Statistical analyses were performed on 113 cells from five different cell lines showing the largest complement of human chromosomes. A log-linear models approach to the statistical analysis was used to predict the expected chromosome segregation under a variety of assumptions concerning the dependency relationship between the pattern of loss and the specific chromosomes both within and between cell lines. Human chromosome segregation occurring after the hybrids were grown continuously in suspension for several months was also studied. Results indicated a statistically significant dependency among patterns of loss of specific chromosomes, with certain chromosomes preferentially retained and others lost more often than expected under the assumption of randomness of segregation.

Functional hybrids between human cytotoxic T and mouse myeloma cells.

The establishment of functional human cytotoxic T lymphocyte (CTL) hybrids was investigated. Human CTL, generated in a seven-day, one-way mixed lymphocyte-tumor cell interaction (MLTI) against an allogeneic melanoma cell line (DW) in the presence of a third-party helper cell line and crude interleukin 2 (IL2), were fused with a mouse myeloma cell line (P3-X63 Ag8). Following fusion in polyethylene glycol, the hybrids were examined for cytotoxic potential against the sensitizing target cells DW. Hybrids with detectable levels of cytotoxicity were cloned in soft agar. Two clones demonstrating stable activity were selected for analysis of lineage and specificity of cytotoxicity. Both clones expressed cytotoxicity in a reasonable stable manner without dependence on IL2 for growth or function. Interferon had no effect on the cytotoxicity of the hybrids against the natural killer (NK)-sensitive target cells K562 or the DW cells. The cytotoxic activities of the hybrids against the sensitizing target cells DW, however, could be markedly facilitated in the presence of IL2-containing supernatants in the assay medium and less so in the presence of lectin. The range of the cytotoxic activities of the two clones was identical and restricted to the DW cells and another melanoma cell line, suggesting the possibility of a shared target molecule(s) between these two target cells for these cytotoxic hybrids. These observations indicate that the hybrids might require a mediator present in IL2 supernatant for optimum expression of cytotoxicity and suggest that the hybrids express the cytotoxic specificity of the hybridized CTL. These hybrids offer unique opportunities for critical examination of the molecular mechanisms of cellular cytotoxicity and specificities exhibited by activated human CTL.

Choroid metastases from transitional cell carcinoma of the bladder: a case report and a review of the literature.

A case of choroidal metastases as the initial manifestation of disseminated bladder cancer is presented. Ocular metastases are an extremely rare manifestation of bladder cancer and when present, usually are a late manifestation. The pathophysiology of ocular metastases is discussed and literature is surveyed.

Membrane-associated ribosomes in producing and nonproducing mouse myeloma cells.

Mouse myeloma cell lines synthesize large amounts of immunoglobulin on their membrane-associated polyribosomes. Variants which no longer synthesize immunoglobulins have been studied and shown to have the same number of tightly bound membrane-associated polyribosomes as the parental cell lines. These polyribosomes are still active in the synthesis of nonimmunoglobulin proteins.