Biomedical subjects

F A Anderer

Publications and source records attributed to F A Anderer.

At least 37 records · Page 2Linked to original sources

PubMed1989

Activation of natural killer cell cytotoxicity of human blood monocytes by a low molecular weight component from viscum album extract.

Viscum album extracts (Helixor) were investigated for their potency to influence natural killer cell (NK) cytotoxicity of human peripheral blood mononuclear cells (PBMC) in vitro. The samples investigated were unable to enhance NK cytotoxicity in PBMC/tumor cell co-cultures by direct short-term mediation but NK cytotoxicity of human PBMC was strongly stimulated when PBMC were pre-incubated for 72 h with a partly purified fraction (HM-BP) derived from extracts of V. album mali. Stimulation of NK cytotoxicity was not dependent from age and sex of PBMC donors and was directed against highly as well as moderately NK-sensitive human tumor cells. The responding effector cells were identified as monocytes/macrophages and stimulation of the NK cytotoxicity of these cells was not based on increased proliferation. The active component in the HM-BP fraction has a molecular weight of about 1000 Da or smaller and correlates with the structural criteria of an oligosaccharide. The activity was completely abrogated when the active fraction was treated with endoglycosidase F or alpha-glucosidase. Partial inactivation was observed after treatment with endoglycosidase D or hemicellulase. Moreover, the active fraction induced a reduction in tumor take incidence and tumor development in mice when applied before and after tumor challenge.

Animals↗

PubMed1988

Autoimmune expression of a cytoplasmic protein p60 in mice bearing metastasizing SV40-transformed tumors.

In STU mice bearing metastasizing SV40-transformed 51A-232B-M tumors, an immune response against a cellular 60kDa protein (p60) developed in about 50% of the tumor-bearing animals, in addition to the response against SV40 large T-antigen and cellular protein p53. The anti-p60 auto-immune response could be observed as early as 11 days after tumor challenge and was strictly linked with metastatic spread but was not a prerequisite for metastasis. Anti-p60 antibodies could not be detected in sera of animals bearing metastasizing Rous-sarcoma virus-transformed or methylcholanthrene-induced tumors, or in sera from human cancer patients with clinically confirmed metastatic spread. The anti-p60 auto-antibodies showed a broad cross-reactivity against components of similar size in a great number of cell lines of various species and in normal mouse tissue. The p60 auto-antigen is a cytoplasmic protein which is neither phosphorylated nor glycosylated in vivo. Immunoblotting performed with fresh cell lysates under non-reducing conditions using tumor-bearer sera revealed a diffuse p60 double band, but under reducing conditions only one sharp p60 band was observed. The reaction of p60 with anti-p60 auto-antibodies could be completely blocked by pre-treatment of fresh cell lysates with N-ethylmaleimide or p-chloromercuriphenyl sulfonate, or by oxidation in air prior to immunoblotting, indicating that the anti-p60 autoimmune response was directed against an epitope sensitive to SH-group-blocking reagents. Immunofluorescence studies with tumor-bearer sera showed only a very weak cytoplasmic fluorescence, possibly due to the nature of the p60 SH-groups in situ being masked. Immunoprecipitates with monoclonal antibodies against SV40 large T-antigen and p53 obtained from fresh cell lysates of SV40-transformed tumor cells contained no associated p60 auto-antigen. The p60 auto-antigen was purified from tumor cell homogenates with an enrichment factor of about 2,000; its iso-electric point is at pH 6.8. Determination of the biological half-life of p60 yielded a value of about 28 hr. The p60 auto-antibodies in pools of tumor-bearer sera taken at day 40 after tumor challenge all belonged to the IgG1 subclass.

Animals↗

PubMed1988

Casein kinase II accumulation in the nucleolus and its role in nucleolar phosphorylation.

A rabbit antiserum against highly purified casein kinase II from mouse tumor cells was used for immunolocalization of the enzyme in fixed, permeabilized mouse cells. Casein kinase II was highly accumulated in nucleoli compared to the extra-nucleolar space of the nucleus or to the cytoplasma. Casein kinase II samples highly purified from the cytoplasma, from the extra-nucleolar fraction of the nucleus or from nucleoli exhibited no differences with respect to structure and function. All samples originally had an alpha 2 beta 2 structure (alpha, 42 kDa; beta, 24 kDa) showing formation of the alpha'-chain (36 kDa) only in the late steps of purification. The isoelectric point of the alpha-chain of all three samples was pH 7.7 and that of the beta-chain was pH 6.4-6.6. Using ATP or GTP, all three casein kinase II samples gave the same results of maximum phosphorylation of purified nucleolar marker phosphoproteins pp105/C23, pp135 and B23, yielding pp135 as one of the most highly phosphorylated proteins with an incorporation of about 75 phosphate groups per molecule pp135. Studies on optimum conditions of phosphorylation of nucleolar phosphoproteins by casein kinase II revealed that each of the protein substrates individually responded to alterations of assay parameters such as pH, magnesium ion and sodium chloride concentrations indicating that predominantly individual structural criteria were responsible for optimum phosphorylation. The determination of the apparent Km of casein kinase II for purified nucleolar phosphoproteins yielded values of 0.15 microM (pp105/C23), 0.1 microM (pp135) and 1.0 microM (B23) identifying them as high-affinity substrates of casein kinase II.

Animals↗

PubMed1988

Monoclonal antibodies produced by in vitro immunization with sera of human-xenotransplant-bearing mice.

In vitro immunization procedures, using sera of athymic mice bearing human WOC ovarian tumors or CM III mammary tumors as immunizing antigen, induced a highly efficient formation of mABs (44% of antibody-producing clones) reacting with human ovarian and/or mammary tumor cells. More than half of these mABs showed cross-reactivity with mouse cell lines. Immunogenicity of normal mouse components in the sera from tumor bearers can be excluded since control immunization with sera of normal athymic mice yielded no mABs reacting with mouse or human cell lines. Furthermore, immunization with sera from tumor bearers did not induce mABs only reacting with mouse cells since 20% of the antibody-producing clones showed an exclusive specificity for the human tumor cells. On the basis of these results we concluded that the human-mouse cross-reacting mABs were induced by circulating human TAA with epitopes shared by mouse cellular components.

Animals↗

PubMed1987

Human SS-B/LA autoantigen contains a covalent protein-RNA linkage.

The chemical nature of association of RNA in immunoprecipitates of human SS-B/La ribonucleoprotein, an autoantigen expressed in various autoimmune disorders, was investigated. A fraction of RNA associated with SS-B/La immunoprecipitates was readily dissociated by SDS-polyacrylamide gel electrophoresis, yielding four main subfractions, R1-4, with chain lengths in the range of 90-130 nucleotides (R4), 140-175 nucleotides (R2 and R3) and above 200 nucleotides (R1). Moreover, the immunoreactive protein component, migrating with a molecular mass of 49 kDa, contained a very tightly bound RNA co-migrating with the protein unless the protein was proteolytically degraded. Most of the RNA molecules in this fraction, represented by about 20 components, had a free 3'-terminus but a blocked 5'-terminus and showed chain lengths between 10 and 125 nucleotides. After pretreatment with alkaline phosphatase and a mixture of ribonucleases T1 + T2 + A, adenosine 3',5'-biphosphate (pAp) was liberated by phosphodiesterase (Crotalus durissus) as the blocked 5'-end of the RNA. The chemical nature of the blockage was revealed after alternative treatment of the protein-pAp component with phosphodiesterase or nuclease S7 followed by acid hydrolysis and phosphoamino acid analysis which showed that a threonine residue must be directly involved in the RNA-protein linkage of 49 kDa SS/La antigen, indicating the presence of a covalent threonine-pAp bond.

Alkaline Phosphatase↗

PubMed1987

Multiple phosphorylation of human SS-B/LA autoantigen and its effect on poly(U) and autoantibody binding.

The metabolic turnover rates and the effect of in vitro phosphorylation on poly(U) and autoantibody binding of human SS-B/La ribonucleoprotein, an autoantigen expressed in various autoimmune disorders, were studied. The determination of the metabolic turnover rates of SS-B/La protein, SS-B/La protein phosphorylation and RNA binding yielded values of 12.1 h, 3.6 h and 3.7 h, respectively, indicating a possible functional correlation of RNA-binding and phosphorylation. This assumption was confirmed by studies of in vitro phosphorylation using purified SS-B/La protein and purified casein kinase type II as a model system. A high degree of phosphorylation of the SS-B/La protein (molecular mass 49 kDa) substantially diminished its binding capacity for poly[3H]U. However, binding of human autoantibodies against SS-B/La antigen increases 2-fold with increased SS-B/La phosphorylation. Complete phosphorylation in vitro led to partial molecular transformation, yielding an antigenically cross-reacting component with an apparent molecular mass of 51 kDa which could not be detected during in vivo phosphorylation.

Autoantibodies↗

PubMed1987

Mediation of human NK-activity by components in extracts of Viscum album.

Viscum album extracts (Iscador) were investigated for their potency to influence NK cytotoxicity in vitro. In vitro short term cytotoxicity assays (4 h) with human peripheral mononuclear cells (PMNC) and human K 562 tumor cells showed a drastic enhancement of NK cytotoxicity in the presence of V. album extracts. The presence of the V. album components during tumor cell lysis was essential since preincubation of PMNC with V. album extract followed by thorough washing did not lead to enhancement of NK cytotoxicity. One responding effector cell was identified as a member of the large granular lymphocyte (LGL) family carrying both Leu 7 and Leu 11 surface markers. Furthermore, monocytes depleted of LGL, but not differentiated macrophages, showed a weak enhancement of their cytolytic activity in the presence of V. album extract. Fractionation of V. album extracts revealed two active fractions one (C1) with about 3-4000 D and the other (C2) less than 1000 D. Both components enhanced NK cytotoxicity of LGL (Leu 7+, Leu 11+) as well as of monocytes showing enhancing effects also against moderately NK-sensitive tumor cell lines.

Adjuvants, Immunologic↗

PubMed1987

Quantitation and potential function of nucleolar phosphoprotein pp 105 in mouse tumor cells, embryonic cells and normal tissues.

1. Nucleolar phosphoprotein pp 105 was determined in various mouse cell and tissue extracts using a highly sensitive ELISA. The results indicate that the highest relative amounts of pp 105 correlate with cells and tissues of high growth rate such as tumor cell lines, solid tumors and embryonic tissues. 2. The specific phosphorylation of pp 105 was compared in a 1 min endogenous phosphorylation assay with native cell and tissue extracts. 3. The mitogenic activity of highly purified pp 105 was demonstrated in cultures of resting mouse embryonic cells and mouse thymocytes.

Animals↗

PubMed1986

[The doubling time of circulating CEA as an individual prognostic criterion of recurrence in patients with gastrointestinal cancers].

In a prospective study of 928 patients with gastrointestinal cancer registered for primary resection, the postoperative carcinoembryonic antigen (CEA) time courses were analysed in connection with disease recurrence. Only patients with established diagnosis of disease recurrence and complete follow-up to death entered the evaluation of prognostic criteria of the CEA time course. In this group the CEA time courses of 103/201 patients with recurrent disease exhibited an exponential increase of the serum CEA concentration, i.e. a linear relationship of log CEA and time, which allowed the calculation of the CEA doubling time. All 103 patients developed metastatic spread and generally exhibited CEA doubling times ranging between 10 and 158 days, in patients developing peritoneal carcinosis up to 343 days. The individual CEA doubling times of patients with recurrent disease who received no treatment (n = 71) correlated well with the times of individual survival after the initial CEA increase of the log CEA phase (rs = 0.812; P less than 0.001) thus confirming the results of a previous retrospective study. When the survival time is expressed in multiples of the individual CEA doubling time (IDT), no patient survived longer than 10.8 IDT. The median value of survival was 5.4 IDT. Patients with metastatic spread who underwent various treatments of recurrent disease (n = 32) survived distinctly longer showing survival times up to 32.6 IDT. This could be confirmed by comparing the observed survival after the initial CEA increase of treated and untreated patients (life table method) exhibiting highly significant differences (P less than 0.001).

Carcinoembryonic Antigen↗

PubMed1986

Prognostic value of preoperative serum CEA level compared to clinical staging. IV. Histological grading and tumor type in colorectal and gastric cancer.

In a clinical investigation of observed postoperative survival, 410 patients with colorectal cancer and 269 patients with stomach cancer have been registered for primary surgical treatment connected with a long-term follow up. Histologic grading and tumor typing were examined as potential prognostic factors and compared with the prognostic information covered by operability, tumor extension, and the preoperative CEA level. Statistical treatment of the data revealed no prognostic significance of the tumor types adenocarcinoma, mucinous, and anaplastic tumours in gastric cancer. Histologic grading specified ranges associated with significant differences in survival of gastric and colorectal cancer patients. However, histologic grading did not provide prognostic information in addition to operability and tumor extension. However, histologic grading gave additional prognostic information to preoperative CEA levels in the range of 0-5 micrograms CEA/1 serum but not in the range greater than 5 micrograms CEA/1. The results indicate that the prognostic information of preoperative serum CEA level is not directly linked to the histologic grade of a tumor.

Carcinoembryonic Antigen↗

PubMed1986

Phosphoprotein pp135 is an essential component of the nucleolus organizer region (NOR).

The association of phosphoproteins pp135 and pp105 with distinct substructures of the nucleolus was studied by cytochemical and immunological methods at the light microscopic and electron microscopic level. Both phosphoproteins exhibited a very high affinity for silver and Giemsa staining compared to other nucleolar proteins. Immunolocalization of pp135 and pp105 during mitosis by light microscopy revealed a tight association of pp135 with the silver staining nucleolus organizer region (NOR), whereas pp105 (cross-reacting with C23) appeared to be only partially associated with the NOR, exclusively at telophase. At the immunoelectron microscopic level the distribution of pp135 and pp105 was investigated in interphase nucleoli. Phosphoprotein pp135 was located in the fibrillar shell and pp105 in the fibrillar shell and the granular zone. The fibrillar centers were essentially free of both phosphoproteins..

Anaphase↗

PubMed1986

Activation of human monocyte and natural killer cell-mediated tumour cell killing by two dialysable thymic factors.

Components of calf thymus extract dialysable under acid conditions contained two natural killer (NK) cytotoxicity-stimulating factors, CSFa and CSFb, which could be separated by ion exchange chromatography. NK cytotoxicity of human peripheral blood mononuclear cells (PBMC) against human K562 tumour cells was strongly enhanced after 72 h pre-incubation with the factors. The CSFa/b-specific stimulation of PBMC required the presence of monocytes. The cytotoxic effector cells activated during pre-incubation of PBMC with the CSF were identified as monocytes and as NK cells present in the fraction of large granular lymphocytes (LGL). Selective cell depletion studies with LGL-containing subpopulations (free of monocytes) allowed factor-specific discrimination of the activated LGL. Pre-incubation of PBMC with CSFa stimulated NK cytotoxicity of LGL (Leu 7+11-; T8-), whereas pre-incubation with CSFb resulted in stimulation of LGL (Leu 7+11-; T8+). The biological effects of CSFa and CSFb could be further distinguished by analysis of surface marker expression during incubation of PBMC. CSFb scarcely influenced T4 expression, but strongly enhanced the expression of T8 and that of transferrin receptor, whereas CSFa had no significant influence on the expression of these three surface markers. Both factors induced a drastic reduction of tumour take incidence or tumour development in mice when applied before and after tumour challenge.

Animals↗

PubMed1986

Characterisation of nucleolar proteins as autoantigens using human autoimmune sera.

Sera from 271 patients with systemic rheumatic diseases were screened for antinucleolar antibodies using immunofluorescence. Antinucleolar antibodies were found in the sera of 73% of patients with progressive systemic sclerosis, 4% of patients positive for autoantibodies with rheumatoid arthritis, and 8% of patients with rheumatoid arthritis associated with sicca complex, but not in patients with systemic lupus erythematosus, Sjögren's syndrome, undifferentiated connective tissue disease, or in healthy donors. The antinucleolar sera (n = 20) were analysed by immunoblotting techniques. In four sera antibodies against nucleolar proteins with molecular weights of 35 kD, 37 kD, 69 kD, 92-93 kD, and 93 kD could be immunodetected. The nucleolar autoantigens were extractable from the nucleoli together with the preribosomal particle fraction and could be solubilised by ribonuclease (RNAse) treatment. Their presence in the nucleolus was sensitive to actinomycin D treatment of cells. The 37 kD autoantigen could be identified as nucleolar phosphoprotein B23.

Animals↗

PubMed1985

[Carcinoembryonic antigen: diagnosis and tumor progression in gastrointestinal tumors].

Tumour progression in 340 patients with resected gastrointestinal primary tumours was monitored using the gradual increase in carcinoembryonic antigen (CEA) in serum. The commencement of the rise in CEA generally preceded clinical detection of the cancer by several months. The degree to which the rise in CEA correlated with the recurrence of cancer was investigated. There was a marked difference in the distribution of the rises in CEA between local tumour growth and distant metastases. CEA increases of more than 1 microgram CEA/l serum in 10 days occurred exclusively in patients with distant metastases. There was a further marked difference in the distribution of the CEA increase between the group with liver metastases and the groups with peritoneal carcinomatosis or other metastases. The site of the primary tumour had no influence on the CEA increase during formation of metastases.

Carcinoembryonic Antigen↗

PubMed1985

Monoclonal antibody-defined circulating human tumor-associated antigen with epitope shared by cytokeratins.

Sera of human colonic carcinoma xenografted rnu/nu rats were used to immunize rnu/+rats in order to obtain an immune response against circulating human tumor-associated components. After fusion of rat spleen cells with mouse myeloma cells monoclonal antibody MAB 108 could be established which reacted with two 40 and 45 kD cytokeratins as well as with vimentin, with a soluble 37 kD protein apparently derived from the 45 kD protein and with a 37 kD protein released by tumor cells. The MAB 108-specific epitope was also detected in tissue polypeptide antigen (TPA), a human tumor-associated antigen originally described by Björklund et al. (22).

Animals↗

PubMed1985

The clinical validity of circulating tumor-associated antigens CEA and CA 19-9 in primary diagnosis and follow-up of patients with gastrointestinal malignancies.

The clinical validity of monitoring the tumor markers carcinoembryonic antigen (CEA) and CA 19-9 were investigated in 602 patients with colorectal, gastric, and pancreatic carcinomas. Sensitivity and specificity of the tests were evaluated preoperatively as well as in the postoperative follow-up for early detection of disease progression and recurrence. At a 95% level of specificity as calculated from a group of 150 patients with benign diseases, the CEA test with monoclonal antibody had a preoperative sensitivity of 39% in colorectal cancer and 21% in gastric cancer. On the other hand, CA 19-9 had a sensitivity of 19% in colorectal cancer, 21% in gastric cancer, and 89% in pancreatic cancer. In the postoperative follow-up it was found that a combination of both tumor marker tests was most profitable in gastric carcinomas, yielding an increase of sensitivity from 59%-94%, showing a high degree of complementarity. The gain in sensitivity provided by the CA 19-9 test over the CEA-test in colorectal cancer was very low. The gain in sensitivity, however, provided by the CEA test over the CA 19-9 test in pancreatic carcinoma was also very low. On the basis of these results it has to be recommended that cases with pancreatic carcinoma are to be monitored most efficiently with the CA 19-9 test, whereas in cases with colorectal cancer the CEA test should be used primarily. However, in gastric cancer the combined use of CEA and CA 19-9 represents a highly valuable basis for monitoring the course of disease.

Antibodies, Monoclonal↗

PubMed1985

Prognostic value of preoperative serum CEA level compared to clinical staging: III. An approach to scoring of prognostic factors in colorectal cancer.

In a clinical study of observed postoperative survival of colorectal cancer patients, we investigated the application of a risk score based on tumor-related prognostic parameters. Six hundred seventy-four patients have been registered for primary surgery of colorectal cancer since 1974 who did not receive further postoperative treatments. The prognostic parameters included operability, tumor extension, and preoperative serum carcinoembryonic antigen (CEA) level. The scoring system was based on the average death-rate ratios of subgroups of patients and their age and sex-matched reference groups derived from the general life table of the population of the Federal Republic of Germany. The individual score sums of the patients exhibited score sum ranges which characterized groups of patients with entirely different observed survival. The prediction of individual survival after primary operation was only partly possible. In the plot of individual survivals vs individual score sums, a marginal risk zone was obtained which evidently represents the zone of maximum expected survival of patients who do not receive further postoperative treatment.

Aged↗

PubMed1985

Immunodetection of nitrocellulose-adhesive proteins at the nanogram level after trinitrophenyl modification.

A method for protein detection on nitrocellulose membranes based on modification with 2,4,6-trinitrobenzenesulfonic acid and reaction with anti-trinitrophenyl (TNP) serum as first antibody followed by peroxidase-conjugated second antibody is described. Protein quantities between 1 and 3 ng can be detected in the dot test. This method was used in a double immunodetection procedure after electrophoretic transfer of proteins localizing first a distinct antigen with its specific antiserum followed by visualization of the complete protein pattern on the same blot by the TNP/anti-TNP method as described above. As only water-soluble reagents are employed no shrinkage of the membrane occurs. Furthermore, the method can be used in a simultaneous immunodetection procedure visualizing the specific antigen together with TNP marker proteins using a mixture of the specific antiserum and the anti-TNP serum as first antibody.

Animals↗