Biomedical subjects

M Blaszczyk

Publications and source records attributed to M Blaszczyk.

At least 55 records · Page 3Linked to original sources

PubMed1986

Monoclonal antibody-defined antigens of human prostate cancer cell line PC3.

Over 600 hybridomas were derived from the immunization of mice with live cells and aqueous extracts of the human prostatic carcinoma cell line PC3. A total of 26 hybridomas with restricted reactivities were selected, subcloned and antibodies tested on a variety of tumor and normal cells. Seven monoclonal antibodies showed reactivity for prostate cancer and other tumor cell lines, including breast carcinomas. Three of the antibodies obtained after immunization with live cells reacted with live cells only and three of the four antibodies obtained after immunization with cell extract reacted with cell extracts and spent culture media. The fourth antibody in the latter group was reactive only in the immunoperoxidase staining assay. Antibody PrS5 recognized a 90,000 molecular weight molecule from 125I-surface-labeled cells in immunoprecipitation analysis. Antibodies PrE3 and PrD8 detected a nonacid glycolipid pentasaccharide from PC3 cells and meconium, and a glycoprotein of 115,000 molecular weight from 125I-surface-labeled red blood cells. The similar patterns of reactivity in RIAs and antigen analysis suggest that antibodies PrE3 and PrD8 recognize the same molecule. The results emphasize the usefulness of immunohistochemistry in the testing of monoclonal antibodies and the impact of the form in which the antigen is presented on the resultant antibody specificity.

Animals↗

PubMed1986

Scl 70 antibody--a specific marker of systemic sclerosis.

Scl 70 antibodies were tested for in 107 patients with systemic sclerosis: 68 with acrosclerosis and 39 with diffuse scleroderma. Anticentromere antibodies (ACA) and other antinuclear antibodies (ANA) were tested for by indirect immunofluorescence on HEp-2 cells. Positive results for Scl 70 antibodies were obtained in 77% of cases of diffuse scleroderma and 44% of acrosclerosis. ACA and Scl 70 antibodies were found to be mutually exclusive. If acrosclerosis cases positive for anticentromere antibodies are excluded, the percentage of acrosclerosis cases positive for Scl 70 was 63%. ACA were found to be a marker of a benign, abortive subset of acrosclerosis with almost no cutaneous involvement (CREST), whereas Scl 70 did not discriminate between acrosclerosis and diffuse scleroderma. On HEp-2 cells Scl 70 positive sera gave a characteristic, fine speckled, almost homogeneous nuclear staining pattern.

Antibodies, Antinuclear↗

PubMed1985

[Plasmapheresis in the treatment of pemphigus].

The results of small-volume plasmapheresis were favorable in cases of severe pemphigus vulgaris and foliaceus if applied simultaneously together with corticosteroids and cyclophosphamide, both on lowered doses. The patients went into long-lasting remission. A special indication for this therapy are cases that do not allow the application of large doses of prednison because of severe side-effects of steroids. The effect of plasmapheresis does not appear to depend on the removal large quantity of antibodies. Small-volume plasmapheresis as a supplementary therapy of the combined treatment with steroids and immuno-suppressors showed better effects than large-volume plasmapheresis.

Adult↗

PubMed1985

Selection of monoclonal antibodies detecting serodiagnostic human tumor markers.

Monoclonal antibodies (MAbs) were selected for specific binding to spent media of cultured tumor cells. Out of more than 12,000 hybridomas screened, 19 were selected in preliminary inhibition assays for secretion of MAbs which detected antigens in cancer patients' sera. Antibody CO 29.11, which was studied in detail, bound to an antigen shed and expressed by adenocarcinoma cells of colon, stomach, pancreas and urinary bladder. CO 29.11 bound to purified sialylated Lewis a (Lea) antigen but to a different epitope and with a higher binding affinity than the MAb CA 19-9. CO 29.11 but not CA 19-9 bound weakly to unsialylated Lea antigen. In double-determinant radioimmunoassay with sera of patients with colorectal carcinoma, CO 29.11 was found to be a more sensitive marker than CA 19-9 for the detection in serum of sialylated Lea antigen.

Antibodies, Monoclonal↗

PubMed1985

Characterization of Lewis antigens in normal colon and gastrointestinal adenocarcinomas.

The Lewis a antigen (Lea)- and Lewis b antigen (Leb)-active glycolipids and glycoproteins in normal colon mucosa, colon carcinoma tumors, and gastrointestinal tumor cell lines were studied by using monoclonal antibodies against Lewis determinants in solid-phase radioimmunoassay, chromatogram binding assay, and immunoprecipitation/PAGE. Lea-5 was a major component of the purified non-acid glycolipid fraction from normal colon mucosas from four patients examined and Lea-7 was a minor component in three of these four cases. Lea in colon carcinoma of the same patients was expressed as Lea-5 in all four cases and as both Lea-5 and Lea-7 in one of the four cases. All normal colon mucosa samples expressed Lea but not Leb. Leb-6 was expressed in all tumors obtained from the same patients as normal colon mucosa (Leb negative) samples and in the gastrointestinal tumor cell lines. These findings indicate that Leb is a colon adenocarcinoma-associated antigen and that Lea and Leb are coexpressed in tumor tissue. Evidence is presented for the coexistence of Lea and Leb carbohydrate determinants in both glycolipid and glycoprotein forms in the same cells.

Adenocarcinoma↗

PubMed1985

Isolation and characterization of anti-monosialoganglioside monoclonal antibody 19-9 class-switch variants.

Three different monoclonal antibody isotypes, IgG1, IgG2b, and IgG2a, were derived by selection of isotype-switch variants from the CO-19-9 hybridoma. All three antibodies retained their binding specificities and affinities and bound to the same epitope--as defined by the anti-idiotype analysis. Availability of gamma 1, gamma 2b, and gamma 2a Ig heavy chain variants directed against the same epitope on the monosialoganglioside antigen permitted detailed analysis of their Fc fragment receptor (FcR) binding affinities, their cytolytic activities (antibody-dependent, macrophage-mediated cytotoxicity) in vitro, and their tumoricidal activities in vivo. Analysis of the binding of three isotypes to the human FcR expressed by U937 cells induced by gamma interferon has shown that only IgG2a proteins bound to high-affinity FcR, but not IgG1 or IgG2b variants. Although all three isotypes were active in the antibody-dependent, macrophage-mediated cytotoxicity assay with murine thioglycolate-elicited macrophages, the IgG2a gave the highest percentage of lysis; similar results were obtained in the same assay with human monocytes as effector cells. In the nude mice experiment, only the IgG2a variant inhibited growth of human colorectal carcinoma, while IgG1 and IgG2b were ineffective. Thus, selection of isotype-switch variants resulted in the conversion of monoclonal antibody from noncytolytic to cytolytic with possible immunotherapeutic application.

Animals↗

PubMed1985

Anticentromere antibody: an immunological marker of a subset of systemic sclerosis.

Our clinical and immunological studies of 114 cases of systemic sclerosis, 54 of Raynaud's disease and 46 of other connective tissue diseases, centered on the diagnostic and prognostic significance of anticentromere antibodies (ACA). The ACA occurred in 21 of 84 patients with acrosclerosis, in four of 54 patients with Raynaud's disease but in none of 30 patients with diffuse scleroderma or transitional form, acrosclerosis-diffuse scleroderma, or 46 cases of other connective tissue diseases. The ACA-positive patients had no contracture or immobilization of the fingers, the indurations and/or indurative oedema were confined to fingers and usually no other types of ANA were detected. However, systemic involvement and the course of the disease were comparable in ACA-negative and ACA-positive acrosclerosis patients. The studies indicate that there is a subset of acrosclerosis with minimal indurations confined to the fingers, and ACA appears to be its serological marker. We propose to use the term CREST for this subset, which to date has not been exactly defined and is regarded by some authors as synonymous with acrosclerosis.

Adult↗

PubMed1984

Lewis blood group antigens defined by monoclonal anti-colon carcinoma antibodies.

Monoclonal antibodies directed against human cancer cells were prepared by the murine hybridoma technique. These antibodies detect Lewis blood group antigens as determined by indirect solid-phase radioimmunoassay, hapten inhibition studies, and chromatogram binding assay. One monoclonal antibody is specific for the Lea terminal carbohydrate of Gal beta 1----3Glc NAc(4----1 alpha Fuc) beta 1----3LacCer. Five monoclonal antibodies react with the Leb terminal carbohydrate sequence of Fuc alpha 1----2Gal beta 1----3GlcNAc(4----1 alpha Fuc) beta 1----3LacCer, and four of these antibodies are highly specific for this glycolipid and do not react with other similar di- and monofucosylated glycolipids. One of the anti-Leb antibodies cross-reacts with blood group H glycolipid and has binding properties similar to those of the previously described antibody NS-10-17 [M. Brockhaus, J. L. Magnani, M. Blaszczyk, Z. Steplewski, H. Koprowski, K.-A. Karlsson, G. Larson, and V. Ginsburg (1981) J. Biol. Chem. 256, 13223-13225]. Two antibodies react with both the Lea and Leb antigens, though both bind preferentially to Leb.

Antibodies, Monoclonal↗

PubMed1984

A fetal glycolipid expressed on adenocarcinomas of the colon.

Reactivity of neuraminidase-treated colorectal carcinoma cells with antibodies that detect the X-carbohydrate structure was greater than the reactivity of untreated cells. The same results were obtained with glycolipid extracts of meconium, a colorectal carcinoma cell line, three freshly excised human adenocarcinomas, and normal bronchial mucosa. The glycolipid was either not expressed or expressed in smaller quantities on the corresponding normal colon tissue. Further study showed that the major sialo-X glycolipid has six sugars including a single sialic acid which blocks X-antigenicity. These glycolipids were further analyzed by ion-exchange high-pressure liquid chromatography and thin-layer chromatography. These monosialo-X glycolipid antigens might serve as potential tumor markers.

Adenocarcinoma↗

PubMed1984

Extraction of circulating gastrointestinal cancer antigen using solid-phase immunoadsorption system of monoclonal antibody-coupled membrane.

An immunoadsorption system of monoclonal antibody immobilized on a polyolefin alloy fiber is described for extraction of serum gastrointestinal cancer antigen (GICA). Continuous circulation or single passage of plasma from gastrointestinal cancer patients through this antibody-fiber matrix resulted in 90% depletion of circulating GICA in 2 h using 0.6 mg immobilized antibody, and 90% depletion in 5 min using 8 mg antibody. Continual circulation resulted in total GICA removal in both cases. Desorption of antibody or of antibody-containing complexes was minimal. This methodology provides a selective and convenient means of removing any targeted substance by monoclonal antibody from the serum, and thus overcomes many of the shortcomings associated with conventional plasmapheresis.

Animals↗

PubMed1984

Increased levels of circulating HLA-DR antigen in sera of patients with acute lymphoblastoid leukemia.

Monoclonal antibodies that define HLA-DR antigen bind to a variety of human tumors, such as Burkitt lymphoma and melanoma cells grown in vitro and with the spent medium of these cultures. Two radioimmunoassays have been developed to detect HLA-DR antigen circulating in human sera. The inhibition assay is based on the inhibition of binding of monoclonal antibodies against HLA-DR to the target preparation; the double-determinant assay traces antigen bound by a solid-phase monoclonal antibody by the use of a second 125I-labeled antibody. Twenty-six of 39 sera from patients with acute lymphoblastoid leukemia, 2 of 29 sera from patients with acute myeloid leukemia, and 5 of 31 sera from patients with advanced metastatic melanoma showed increased levels of HLA-DR antigen, whereas none of 28 sera from patients with other malignancies had increased levels of HLA-DR antigen, and only 2 of 155 sera from healthy donors bound monoclonal antibodies to HLA-DR at detectable levels. The detection of circulating HLA-DR antigen in sera of cancer patients may be useful in monitoring patients with certain malignancies.

Antibodies, Monoclonal↗

PubMed1984

Identification and isolation of Lewis blood group antigens from human saliva using monoclonal antibodies.

Solid-phase radioimmunoassay, polyacrylamide gel electrophoresis and thin-layer chromatography were used to compare, identify, and characterize the Lewis antigens from human salivas, using monoclonal antibodies directed to the Lea and Leb determinants. Sialylated Lea glycolipid was detected in saliva from individuals with Le(a+ b+) and Le(a+ b-) phenotypes. Immunoaffinity chromatography of the saliva from individuals with different phenotypes revealed a glycoprotein of molecular weight greater than 200 kD bearing the Lewis antigenic determinants.

Antibodies, Monoclonal↗

PubMed1984

Circulating immune complexes in systemic scleroderma.

Circulating immune complexes were detected by the immunoelectrophoretic method in 18 of 29 (62 per cent) of patients with systemic scleroderma. The presence of immune complexes did not correlate with that of antinuclear antibodies to dsDNA, DNP, RNP, and Sm. The mean levels of immunoglobulins G, A, and M as well as of C3 were significantly higher in patients with systemic scleroderma than in blood donors.

Antibodies, Anti-Idiotypic↗

PubMed1984

Characterization of gastrointestinal tumor-associated carcinoembryonic antigen-related antigens defined by monoclonal antibodies.

Four major carcinoembryonic antigen-related glycopeptides (Mr 180,000, 160,000, 50,000, and 40,000) were detected in SW948 colon carcinoma cells and in colon adenocarcinoma tissue using a monoclonal antibody (C(4)20-32) generated by immunizing mice with SW1222 human colon carcinoma cells. Only the Mr 50,000 polypeptide was immunoprecipitated from normal colon mucosa by this antibody. Binding studies using other monoclonal antibodies and lectins indicated the different epitopes and carbohydrate attachment sites on each of the four polypeptides. Only monoclonal antibody C(4)20-32 recognized a common determinant on all four polypeptides which was revealed by its reactivity with each affinity-purified component.

Adenocarcinoma↗

PubMed1984

Monoclonal antibody localization of Lewis antigens in fixed tissue.

Monoclonal antibodies that bind specifically with Lewisa (Lea) and Lewisb (Leb) antigens were used in an immunoperoxidase assay to characterize Lewis (Le) antigenic profile of a variety of fixed tissues. Representative sections of normal and malignant tissue from the stomach, colon, pancreas, and kidney were examined and compared. Lea antigen was expressed more often in gastric adenocarcinomas (80%) than normal gastric mucosa (40%). In gastric tumors with concomitant expression of Le antigens, larger areas within an individual tumor expressed Lea antigen rather than Leb antigen. Expression of Leb antigen in normal colonic tissue was seen only in the proximal colon and not distal colon; colon carcinomas, on the other hand, expressed Leb antigen (64%) regardless of where the primary arose. Leb antigen was expressed in large pancreatic ducts (three of three) more often than Lea antigen (one of three); exclusive expression of Lea antigen was demonstrated in the proximal convoluted tubules of normal kidney (three of three) and in renal cell carcinomas (six of eight).

Animals↗

PubMed1984

Identification of melanoma-associated antigens using fixed tissue screening of antibodies.

Early culture supernatants from hybridomas that were obtained through fusions of mouse myeloma cells with lymphocytes of melanoma-immunized mice were screened for their reactivity with a paraffin-embedded cell block of a melanoma cell line, using a biotin:avidin immunoperoxidase procedure. Eleven monoclonal antibodies were derived that define several new melanoma-associated antigens. The antigens include a neutral glycolipid, gangliosides, membrane-associated proteins, cytosolic proteins, and strongly secreted proteins. These antibodies, which detect antigens that withstand tissue fixation and embedding procedures, were tested for reactivity in fixed cell lines, as well as in melanoma biopsies. These antibodies may provide powerful tools in diagnostic studies of human malignant melanoma biopsy material.

Animals↗

PubMed1983

A simple procedure for determining Lewis phenotypes in human saliva.

We have selected 4 murine monoclonal antibodies that specifically bind to Lewis type blood group haptens to establish an assay in which antigens present in human saliva are bound to polystyrene beads and then detected by monoclonal antibodies in radioimmunoassay or in peroxidase-antiperoxidase immunoassay. Two of these antibodies detect both the Lea and Leb determinants (Leab), 1 antibody detects only Lea, and 1 only Leb. All Lewis phenotypes (Lea-b-, Lea+b-, Lea-b+ and Lea+b+) are easily detectable in this assay. Of the 60 individuals tested, 1 was Lea+b+, 4 Lea-b-, 12 Lea+b- and 43 Lea-b+.

Animals↗

PubMed1983

Mouse monoclonal antibodies against human cancer cell lines with specificities for blood group and related antigens. Characterization by antibody binding to glycosphingolipids in a chromatogram binding assay.

Solid phase radioimmunoassay and a chromatogram binding assay were used to characterize the binding specificities of five monoclonal antibodies generated from mice immunized with human tumor cell lines when tested against various glycolipids. Four antibodies derived from mice immunized with pancreatic carcinoma cells detected specifically the human blood group B determinant, Gal alpha 1 leads to 3Gal (2 comes from 1 alpha Fuc). These antibodies preferred type 2 (Gal beta 1 leads to 4GlcNAc) glycolipids. No reactivity was detected with a rat B determinant based on GalNAc. An antibody derived following immunization with a rectal carcinoma cell line was shown to have binding properties identical with those of an antibody that reacts specifically with the stage-specific embryonic mouse antigen (SSEA-1), bearing the determinant Gal beta 1 leads to 4GlcNAc (3 comes from 1 alpha Fuc) (Gooi, H. C., Feizi, T., Kapadia, A., Knowles, B. B., Solter, D., and Evans, M. J. (1981) Nature 292, 156-158). Thin layer chromatography was used to detect the binding of a monoclonal anti-tumor antibody recently shown to react with a sialylated Lea glycolipid (Magnani, J. L., Nilsson, B., Brockhaus, M., Zopf, D., Steplewski, Z., Koprowski, H., and Ginsburg, V. (1982) Fed. Proc 41, 898) and an anti-Leb antibody (Brockhaus, M., Magnani, J. L., Blaszczyk, M., Steplewski, Z., Koprowski, H., Karlsson, K.-A., Larson, G., and Ginsburg, V. (1981) J. Biol. Chem. 256, 13223-13225) to mixtures of glycolipids from normal and tumor tissues.

Adenocarcinoma↗