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Biomedical subjects

R P Linke

Publications and source records attributed to R P Linke.

At least 127 records · Page 7Linked to original sources
PubMed1984

Monoclonal antibodies against amyloid fibril protein AA. Production, specificity, and use for immunohistochemical localization and classification of AA-type amyloidosis.

Monoclonal antibodies against amyloid fibril protein AA were produced by cell fusion of murine P3 X 63-Ag8.653 myeloma cells with spleen cells of immunized Balb/c mice. To increase immunogenicity, protein AA was coupled to horseradish peroxidase (HRP) or human high molecular weight kininogen (HMWK). Using micro-ELISA (enzyme-linked immunosorbent essay) seven hybridoma cell lines secreting antibodies that specifically bind to protein AA have been selected and cloned. When applied to formalin-fixed paraffin sections of a variety of different amyloid types using immunoperoxidase methods, five monoclonal antibodies bound specifically and strongly to amyloid only of the AA type. Since a series of different AA-amyloids could be stained, these reagents may be used to routinely diagnose AA-amyloidosis in tissue sections. A monoclonal antibody against HRP has also been produced that has been utilized to develop a monoclonal peroxidase-antiperoxidases (PAP) complex. When three immunoperoxidase methods were compared, the sensitivity of a conventional rat PAP was comparable to the monoclonal PAP complex, but the latter was easier to handle. Both methods were more sensitive than the indirect immunoperoxidase technique.

Amyloid↗
PubMed1984

Serological identification and separation of canine lymphocyte subpopulations.

In vitro treatment of bone marrow grafts with absorbed rabbit-antidog thymocyte globulin (ATG) prevented graft-versus-host disease in a substantial number of the dogs differing by one DLA haplotype. Absorbed ATG has now been used for serological identification of canine lymphocyte populations. Specific labeling of canine T-lymphocytes by absorbed ATG could be demonstrated by (a) a distribution of ATG-positive cells in suspensions of canine lymphoid organs similar to that of T cells observed in other species and by specific staining of paracortical thymus-dependent lymph node areas in immunohistochemistry, (b) complementary labeling of nylon-wool-separated cells by ATG and antiimmunoglobulin sera, and (c) correlation of ATG surface labeling with responder activities in mixed lymphocyte cultures.

Animals↗
PubMed1984

Immunohistochemical typing of amyloid on hydroxyethyl-methacrylate-embedded renal biopsies.

Hydroxyethyl-methacrylate (GMA)-embedded renal biopsies containing amyloid were tested by the indirect immunoperoxidase method with antibodies against the following purified amyloid fibril proteins: AA, three different A lambda preparations, Ak and AF. The anti-AA reagent was monoclonal (mc13), all the others were polyclonal. In four biopsies from cases with generalized amyloidosis, two were found to be of the AA and two of the A lambda type. Two control cases of membranous glomerulonephritis showed only a marginal reaction with anti-A lambda. These results demonstrate that GMA-embedded tissue sections are suitable for immunohistochemical classification of amyloid diseases, because proteinaceous antigenic determinants of amyloid fibril are preserved.

Aged↗
PubMed1983

[Familial Mediterranean fever with amyloidosis. Recent pathogenetic and therapeutic aspects].

Familial Mediterranean fever was diagnosed in a 34-year-old Turkish patient with severe nephrotic oedema. Immunohistochemical classification of a biopsy specimen showed amyloidosis of the AA-type. There was a definite increase of serum amyloid-A-protein (SAA). The typical recurrent fever, attacks of abdominal pain with symptoms of subileus and joint swelling could be treated successfully with colchicine, the oedema with diuretics. The progression of renal failure and proteinuria as indicator of the degree of amyloid-induced renal damage remained unaffected by this treatment. With dimethyl-sulfoxide (DMSO) a marked improvement in renal function and a lowering of the SAA level could be achieved. Thus this treatment inhibits the progression of amyloidosis of the AA-type in Mediterranean fever and may be considered for other forms of AA-type amyloidoses. It is possible that the lowering of the SAA-serum concentration and the improvement of renal function is due to an antiphlogistic effect of DMSO, the mechanism of action of which is so far unknown.

Adult↗
PubMed1983

Lymph node enlargement due to amyloid.

A case of a patient presenting with supraclavicular and cervical lymph node enlargement with demonstration of amyloid is presented. Histologic features are described. Amyloid deposition was not found in any other organ. Immunohistochemical typing with antisera raised against protein AA, A lambda, and A kappa showed a reaction only against A lambda. Thus, this case belongs to the immunoglobulin lambda light chain-derived types that occur in many other parts of the body, particularly the respiratory tract.

Amyloid↗
PubMed1983

Immuno-electron microscopic identification and classification of amyloid in tissue sections by the postembedding protein-A gold method.

Amyloid deposits from 3 patients with amyloid of the AA, A lambda, or A kappa type were investigated on ultrathin sections using the protein-A gold method and affinity purified amyloid-type-specific immunoglobulins. Anti-AA, -A lambda, and -A kappa reacted specifically with the corresponding amyloid fibril visualized by the agglomeration of the gold colloid particles. Therefore, this method can identify and classify amyloid fibrils and can distinguish them from other fibrillar proteins at the electron microscopic level.

Aged↗
PubMed1983

Identification of amyloid A protein in a sporadic Muckle-Wells syndrome. N-terminal amino acid sequence after isolation from formalin-fixed tissue.

The amyloid of a patient (WAL) with a sporadic Muckle-Wells syndrome was analyzed in tissue sections and after it had been isolated from formalin-fixed tissue. The predominant amyloid fibril protein was the amyloid A (AA) type determined by the following criteria. (a) Only antiserum against protein AA gave a strong specific reaction, whereas the other antisera with specificity against amyloid of immunoglobulin origin, i.e., A-lambda and A-kappa, did not stain using the indirect immunoperoxidase technique on formalin-fixed, paraffin-embedded tissue sections. (b) In immunodiffusion, the predominant amyloid fibril protein from WAL isolated in pure form from formalin-fixed tissues precipitated in a line of identity with anti-AA and a purified and chemically identified protein AA from another patient. (c) Amyloid fibril protein from WAL had a molecular weight of 8000 to 9000 in sodium dodecyl sulfate polyacrylamide gel electrophoresis and, thus, is in the same range as the commonly found protein AA. (d) The N-terminal amino acid sequence analysis was that of protein AA. In addition to the pure amyloid fibril protein AA (WAL), proteins of higher molecular weights with AA-antigenic determinants were also isolated. These proteins may represent protein AA in a complex form or protein AA linked to other proteins. Since an inflammation is the most likely cause of amyloidosis complicating the Muckle-Wells syndrome an antiinflammatory therapy (as in other AA-type amyloidoses) is recommended.

Adult↗
PubMed1982

Primary structure of the variable part of an amyloidogenic Bence-Jones Protein (Mev.). An unusual insertion in the third hypervariable region of a human kappa-immunoglobulin light chain.

The complete amino acid sequence of the variable region of Bence-Jones protein Mev. from a patient suffering from multiple myeloma and generalized amyloidosis is presented. The amino acid sequence of the Bence-Jones protein Mev. is related to other human kappa-immunoglobulin L-chains of subgroup I. With valine established in Position 191 of the constant region, it is of the Inv (3) allotype. Two types of the Bence-Jones protein Mev. were found, one beginning with the typical N-terminal aspartic acid, and another lacking the N-terminal tripeptide and commencing with methionine in Position 4. A unique insertion of glutamic acid after Position 95 was found in the Bence-Jones protein. This is the position where the V- and J-gene segments join. The J-region of Bence-Jones protein Mev. exhibits some marked differences to the five J-regions recently established by nucleic acid sequencing. This suggests, that there must be considerable polymorphism in human kappa-J-genes. The amyloid fibril protein from the same patient (A Mev.) has also been sequenced up to Position 27. It was found to be identical to the sequence of Bence-Jones protein Mev. commencing with aspartic acid. The molecular mass of the amyloid fibril protein was found to be between 11 000 and 12 000 Da as estimated by gelfiltration and dodecyl sulfate-polyacrylamide electrophoresis.

Amino Acid Sequence↗
PubMed1982

Immunohistochemical identification and cross reactions of amyloid fibril proteins in senile heart and amyloid in familial polyneuropathy. Lack of reactivity with cerebral amyloid in Alzheimer's disease.

Amyloid fibril proteins in senile cardiac amyloid (ASc1), in familial amyloid polyneuropathy (AF), in congophilic angiopathy, and in Alzheimer's disease were investigated in tissue sections by the indirect immunoperoxidase method using antisera against ASc1, AF, and serum prealbumin (PA). Control antisera were tested against amyloid fibril proteins of the AA- (serum amyloid A-derived), A lambda-, and A kappa-(immunoglobulin light chain-derived)amyloid types. Senile cardiac amyloid was specifically identified in tissue sections with antisera against ASc1, AF, and PA, but not with anti-AA, -A lambda, and -A kappa, thus showing cross-reactivity of ASc1 and AF with each other and with PA, but antigenic differences to the other amyloid types. Likewise, amyloid in AF syndromes, including the Japanese, the German, and the Portuguese types, were stained with anti-AF, anti-ASc1, and anti-PA, but not with the antisera against AA, A lambda, A kappa, again showing common antigenic determinants of AF and ASc1 and PA. This is in agreement with chemical and immunochemical data obtained by others. Cerebral amyloids in microangiopathy and in Alzheimer's disease have also been investigated with these antisera and no reaction was found. The cerebral amyloids, therefore, would appear to be antigenically different from ASc1, AF, AA, A lambda, and A kappa types of amyloid.

Adolescent↗
PubMed1981

Amphiphilic properties of the low molecular weight component of serum amyloid-A protein shown by charge-shift electrophoresis.

To test whether the low molecular weight component of serum amyloid-A protein (SAAL) is amphiphilic in character, serum amyloid-A protein (SAA)-containing sera and isolated SAAL were subjected to charge-shift electrophoresis by the technique of Helenius and Simons (Helenius, a. and Simons, K. (1977) Proc. Natl. Acad. Aci. U.S.A. 74, 529-532). When compared to six hydrophilic and two amphiphilic serum proteins, SAAL was shown to behave like the latter in this test. Therefore, SAAL displays properties of apolipoproteins and integral membrane proteins. In the same test, amyloid-A protein (AA) also was found to be amphiphilic, confirming a previous report. The fact that protein AA displays a larger charge shift than that of protein SAAL suggests that the main hydrophobic site of SAAL resides in its N-terminal AA-portion rather than in its C-terminal (SL) part.

Amyloid↗
PubMed1980

[Classification of amyloidoses at biopsy by the immunoperoxidase method (author's transl)].

Three chemically defined amyloid types (AA, A lambda, A kappa) were clearly identified by means of antisera against purified amyloid fibril proteins in tissue sections, using the indirect and the unconjugated immunoperoxidase methods. The two methods gave equally good results both in the cryostat and in the paraffin sections. In contrast to micro-extraction, they have the advantage that fixed material can also be investigated and even the smallest amount of amyloid visible in the tissue section is accessible to typing. It is consequently possible to classify amyloid deposits and amyloid degenerations into uniform, chemically defined disease pictures with the possibility of amyloid type-specific diagnosis and therapy.

Amyloidosis↗
PubMed1980

Idiopathic amyloidosis in the stone marten (Martes foina): identification of amyloid fibril proteins in tissue sections using the immunoperoxidase technique.

Amyloid fibril proteins isolated from a spleen of a wild stone marten (Martes foina, Exleben) with idiopathic amyloidosis show resemblance to protein AA by amino acid analysis. An antiserum directed against these proteins can be used to identify the marten's amyloid in formalin-fixed tissue paraffin-embedded sections using the immunoperoxidase method.

Amyloid↗
PubMed1979

Hairy cell leukaemia: surface markers and functional capacities of the leukaemic cells analysed in eight patients.

In eight patients with hairy cell leukaemia (HCL) peripheral blood cells and in two patients also spleen cells were analysed for surface markers and functional capacities. Only cells containing the tartrate resistant isoenzyme 5 of the acid phosphatase were considered. Hairy cells (HC) of all patients were found to adhere spontaneously to glass and plastic surfaces and to spread after adherence like monocytes. They ingested latex particles of more than 1 micron diameter, but, in contrast to monocytes, did not phagocytose erythrocytes sensitized either by IgM or by IgG antibodies. HC of all patients bore Fc-receptors with a high binding affinity for aggregated IgG. Using 125 I-labelled F(ab')2-fragments of monospecific antibodies in autoradiography, only one light chain type was detected on HC of individual patients. In four patients mu- and delta-chains were simultaneously expressed on HC, whereas in two patients only gamma-chains and in one case only mu-chains were observed on HC. One patient showed a combination of gamma- and delta-chains on his HC. A great variation in density of surface immunoglobulings of HC was observed within individual patients. After removal by capping, surface immunoglobulin reappeared on HC during cell culture, but more slowly than on normal B-lymphocytes. As shown in two patients by internal labelling, HC secreted immunoglobulin light chains, but no heavy chains. On the basis of these findings the classification of HC as belonging to the B-cell lineage, rather than to the monocytic lineage, seems to be justified.

Adult↗
PubMed1979

Temperature-induced dissociation of serum amyloid protein SAA.

Serum amyloid A protein (SAA) has been shown to be unstable at elevated temperatures. While in agreement with earlier reports, molecular weights of approximately 180,000 daltons were determined by gel filtration at 37 degrees C and below, above 58 degrees C additional AA-antigenic activity was found in the void volume and at the position of approximately 12,000 daltons. The latter polypeptide resembled in size and immunoreactivity SAAL in that it had AA-antigenic determinants exposed at 4 degrees C (unlike SAA). Because the release of SAAL or a similar molecule could not be prevented by the enzyme inhibitor PMSF, a temperature-dependent dissociation of SAA is proposed. In view of the known occurrence of amyloid following febrile conditions, the change in size and immunoreactivity of AA-antigenic proteins in vitro may indicate that a similar mechanism in vivo is important in the genesis of AA-type amyloidosis.

Amyloid↗