Detection of intracellular and cell surface immunoglobulin in non-Hodgkin's lymphoma.
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Biomedical subjects
Publications and source records attributed to A A Marucci.
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In lymph node tissue sections, six of 11 human cases of nodular lymphoma showed immunoglobulin within malignant nodules, and seven of nine cases of benign follicular hyperplasia showed immunoglobulin within follicles. In addition, distributions of lymphocyte cell membrane markers for T cells and B cells were determined in ten of 11 cases of nodular lymphoma. Lymphocyte suspensions in five cases contained monoclonal immunoglobulins and in three cases neoplastic cells showed a lack of surface membrane immunoglobulins. In two cases, the distribution of lymphocyte surface markers could not be distinguished from cells of benign lymph nodes. Combined data from intracytoplasmic immunoglobulin studies and lymphocyte surface marker assays indicated that eight of ten cases are of B cell lineage. Thus, the detection of intracellular immunoglobulin is not helpful in differentiating benign follicular hyperplasia from nodular lymphoma, but is complementary to lymphocyte surface marker assays in the determination of the origin of neoplastic cells in lymphoreticular malignancies.
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We describe a patient with multiple myeloma characterized by clinically evident extramedullary metastasis and marked anaplastic alterations of neoplastic cells. We describe the technique that detected the presence of intracytoplasmic immunoglobulin IgGk in mature plasma cells from paraffin-fixed tissue obtained from an iliac lesion and in anaplastic tumor cells infiltrating a lymph node from which a biopsy specimen was taken three years later. These data confirm that both tissues were composed of a cell with identical origin. In addition, the immunohistochemical technique is applicable for the identification of other antigens present in paraffin-embedded tissue.
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Rabbit antisera against highly purified L-asparaginase from Serratia marcescens and from Escherichia coli showed up to 60% inhibition of the catalytic amidohydrolysis of L-asparagine when combined with the homologous enzyme. This inhibition was diminished somewhat against the heterologous enzyme. Kinetic studies in the presence of these antisera showed an increased Kmapp for both homologous and heterologous enzymes using L-asparagine as substrate. In contrast, kinetic studies employing the poor substrate, L-glutamine, showed activation attributable to specific antibodies. This was seen in lower Kmapp values and up to twofold increases in the Vmax over the normal rabbit serum controls. The high degree of cross-inhibition (approximately 80%) and the low degree of cross-reactivity in the quantitative precipitin test (approximately 34%) suggest that these two enzymes possess structural similarities located mainly in the regions of the catalytic sites.
Two methods have been developed which permit use of the unlabeled antibody immunohistochemical technique for detection of human antibody, without the need for immunization of humans with peroxidase. Human antibody to herpes simplex virus (HSV) reacted with human cell cultures infected with HSV was the experimental system. In the first method an attempt was made to employ rabbit peroxidase-antiperoxidase (PAP) soluble complexes in connectin with human antibody. This was done by sequential addition to the HSV-infected cells of (a) human anti-HSV, (b) rabbit antihuman globulin, (c) guinea pig antirabbit globulin (the bridging reagent) and (d) rabbit PAP. Strong specific staining of HSV-infected cells was obtained; however, difficulties were encountered with nonspecific reactions on uninfected cells. In the second method PAP soluble complexes prepared with baboon antiperoxidase were bridged to the human anti-HSV antibody by rabbit antihuman globulin. Because of the phylogenetic relatedness of human and baboon globulins this resulted in firm binding which gave strong specific staining of HSV-infected cells without significant reaction in uninfected cells.
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A quantitative micro-complement fixation test capable of detecting nanogram quantities of antigen is described. The test is simple to perform and is highly reproducible. Typical results from three antigen-antibody systems are given.
Immunochemical properties of l-asparaginases from Serratia marcescens and Escherichia coli were compared by means of quantitative precipitin reactions and quantitative microcomplement-fixation reactions employing homologous and heterologous antisera. The enzyme preparations used in these tests produced one arc of precipitate upon immunoelectrophoresis. According to our results with the precipitin and micro-complement fixation reactions, both enzymes are related immunochemically. On the basis of these tests, distinct structural differences exist between the two enzymes. From results with the micro-complement fixation test, we estimated that the two enzymes could differ by 10 to 15 amino acid residues.
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