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H D Haubeck

Publications and source records attributed to H D Haubeck.

36 records · Page 2Linked to original sources
PubMed1994

Development of enzyme immunoassays specific for keratan sulphate- and core-protein-epitopes of the large aggregating proteoglycan from human articular cartilage.

In the course of chronic inflammatory and degenerative joint diseases proteoglycans are degraded by the action of proteases and oxygen radicals. Therefore, proteoglycan fragments, released from cartilage into the peripheral blood, might be useful markers of cartilage degradation. Sensitive enzyme immunoassays are useful for the detection of these proteoglycan fragments in serum. We therefore developed specific monoclonal antibodies against the large aggregating proteoglycan (aggrecan), which has been isolated and purified from human articular cartilage. Two monoclonal antibodies which recognize a novel cartilage-specific epitope on the keratan sulphate chain of aggrecan (mAb 4B3/D10) and an epitope of the core-protein of aggrecan (4G4/A10) were selected for the development of competitive enzyme-immunoassays. These assays allow the sensitive and specific detection of cartilage-derived proteoglycan fragments, not only in synovial fluid but also in serum. They can now be used for the study of inflammatory and degenerative joint diseases.

Adolescent↗
PubMed1993

Monoclonal, anti-domain and anti-peptide antibodies assign the molecular weight 160,000 granulocyte membrane antigen of the CD66 cluster to a mRNA species encoded by the biliary glycoprotein gene, a member of the carcinoembryonic antigen gene family.

mAb directed against the CD66 cluster of granulocyte differentiation Ag recognize Ag of the carcinoembryonic Ag family. A major Ag in extracts from granulocyte membranes bound by CD66 antibodies exhibits a relative molecular mass of 160,000. According to recent data, this Ag may be a product of the biliary glycoprotein (BGP) gene that belongs to the CEA gene family. As a result of alternative splicing, the BGP gene is transcribed into at least seven distinct mRNA species. To identify splice variants of BGP, antisera were raised against the A2 domain expressed in bacteria and to a peptide comprising the C-terminal 23 amino acids encoded by the 3' exon of the BGP gene. The antisera and an mAb specific for members of the BGP family were used to identify potential BGP splice variants in granulocyte membranes. For comparison, the binding of antibodies to Ag purified from human bile was investigated. In the membrane preparation from granulocytes, the only Ag identified by the mAb, the domain antiserum and the peptide antiserum, was the Ag of M(r) 160,000 recognized by a CD66 antibody. These results indicate that the M(r) 160,000 granulocyte membrane Ag of the CD66 cluster is the product of the BGP-specific mRNA containing all coding sequences of the BGP gene. Among two major biliary glycoproteins present in human bile, the M(r) 115,000 Ag contains the A2 domain, whereas the domain is lacking in the "classical" biliary glycoprotein of M(r) 85,000. None of the bile Ag bound the peptide antiserum.

Antibodies, Monoclonal↗
PubMed1991

Identification of membrane antigens in granulocytes and colonic carcinoma cells by a monoclonal antibody specific for biliary glycoprotein, a member of the carcinoembryonic antigen family.

The gene coding for 'biliary glycoprotein (BGP)' is a member of the carcinoembryonic antigen (CEA) gene family. A monoclonal antibody (MAb) was induced against a BGP-preparation isolated from human bile. The antibody did not crossreact with the carcinoembryonic antigen (CEA) and different non-specific crossreacting antigens. The anti-BGP MAb was used to identify BGP-related antigens in membrane extracts from granulocytes and the colonic carcinoma cell line HT-29. In granulocyte membranes, a single antigen of Mr 160,000 was bound. In membranes from HT-29 cells, a main antigen of Mr 85,000 was present. At high antigen concentration, an additional antigen of Mr 115,000 was identified. Since several transcripts of the BGP gene have been identified, the different BGP related antigens are probably products of alternatively spliced mRNAs.

Antibodies, Monoclonal↗
PubMed1990

Interleukin-1 beta induces synthesis and secretion of interleukin-6 in human chondrocytes.

Increased concentrations of interleukin-6 (IL-6) have been found in the synovial fluid of patients with osteoarthritis, rheumatoid arthritis and crystal-related joint diseases. It is therefore of great interest to identify the cells responsible for the production of IL-6, and to investigate whether IL-6 plays a role in the pathogenesis of degenerative or inflammatory joint diseases. Here we show that human interleukin-1 beta (IL-1 beta) induces IL-6 synthesis and secretion in differentiated human chondrocytes. In organ cultures resembling closely the in vivo system 10(6) chondrocytes incubated with 100 units of interleukin-1 beta per ml of medium led to the release of 6 X 10(3) units of IL-6 within 24 h. Chondrocytes cultured in agarose or as monolayers similarly incubated with IL-1 beta produced even higher amounts of IL-6: 70 X 10(3) units per 10(6) cells within 24 h. The induction of IL-6 synthesis by IL-1 beta was also shown at the mRNA level. IL-6 secreted by stimulated chondrocytes showed heterogeneity upon Western blot analysis.

Adult↗
PubMed1990

Glucocorticoid-induced appearance of the macrophage subtype RM 3/1 in peripheral blood of man.

The effect of intravenous administration of the glucocorticoid prednylidene on the macrophage subtype RM 3/1 in the peripheral blood of man was studied. Injection of 60 mg steroid resulted in an increase in the proportion of RM 3/1 positive monocytes 12 h after application from the basic level to about 80%. After 24 h the number of RM 3/1 positive cells decreased but remained elevated over the basis rate for a period of at least 15 days. Similar results were obtained after administration of 30 or 6 mg prednylidene even if the peak value occurred with a delay of 12 h after 6 mg. A dose- and time-dependent induction of the RM 3/1 positive subtype could also be demonstrated in vitro by the addition of prednylidene or dexamethasone to cultured purified human monocytes/macrophages. Deoxycortone or indomethacin had no effects. These results suggest that glucocorticoids exert their influence on cells of the mononuclear phagocytic lineage by inducing a distinct monocyte/macrophage subpopulation which seems to be associated with anti-inflammatory functions.

Adult↗
PubMed1988

Mitogenic activity of high molecular polysaccharide fractions isolated from the Cupressaceae Thuja occidentale L. I. Macrophage-dependent induction of CD-4-positive T-helper (Th+) lymphocytes.

The arborvitae or Thuja occidentale L., one of the Cupressaceae, has rarely been investigated until now. Several authors have demonstrated that allopathic extracts of this plant could be used as strong antiviral agents directed against plant and animal viruses. Polysaccharide fractions with molecular weights ranging between 20,000 and 1,000,000 and higher have been isolated from the aqueous alkaline extract of the herbal parts of T. occidentale L. by ethanol precipitation and fractionation by ultrafiltration. High molecular subfractions of Thujapolysaccharides (TPS) proved to be highly mitogenic on peripheral blood leukocytes. It was demonstrated by the alcalic immune phosphatase-antiphosphatase and Pappenheim staining methods that the mitogenic and cluster-forming activity of TPS cause T cell induction, in particular, of CD 4-positive T-helper/inducer cells as opposed to B cells. The CD-4+ T-helper/inducer cell induction is connected to an increased production of IL-2. The cluster-forming ability and mitogenity of TPS correlates well with the 3H-thymidine uptake and seems to be IL-1 and IFN-gamma dependent as could be shown by blocking the mitogenic effect using anti-IL-1- and anti-IFN antibodies. Whether it is possible to use these polysaccharide fractions as an adjuvant in the therapy of immune deficiency syndromes and cancer must now be further investigated.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1988

Modulation of cellular anti-tumour responses by tumour-specific T suppressor lymphocytes.

Tumour-specific Ts cells, induced by a protocol which simulates early stages of tumourigenesis in BALB/c mice by the syngeneic plasmacytoma ADJ-PC-5, suppress the generation of specific cytotoxic T lymphocytes (CTL) in a primary in vitro mixed lymphocyte tumour culture (MLTC) of BALB/c spleen cells against ADJ-PC-5. The influence of these Ts cells on the generation of non-specific killer cell activity has now been analysed. The data show that non-specific killer cells generated during in vitro MLTC lyse both ADJ-PC-5 and YAC-1 cells. They are different from tumour-specific CTL as shown by cold target inhibition experiments and on the basis of their different phenotype. The generation of non-specific killer activity during MLTC can be completely suppressed by ADJ-PC-5 specific Ts cells. Activation of non-specific killer cells by recombinant interleukin-2 (r IL2) alone is not influenced by these Ts cells. The in vitro data are best explained by assuming that the target for suppression are ADJ-PC-5 specific T helper cells. Their inactivation will result in depletion of IL2 in the culture, which is required to activate non-specific killer cells. Prevention of activation of specific and non-specific killer cells in vitro by activated tumour-specific Ts cells in the presence of the tumour, suggests that similar mechanisms might operate in vivo and that inactivation of tumour-specific T helper cells might be of central relevance.

Animals↗
PubMed1988

Selective in-vitro activation of BALB/c T suppressor lymphocytes specific for syngeneic ADJ-PC-5 plasmacytoma cells.

T suppressor (Ts) lymphocyte activation is the first reaction of the immune system under conditions which simulate early stages of tumourigenesis. There is evidence that the Ts cell inducing antigen on ADJ-PC-5 plasmacytoma cells of BALB/c origin is a self antigen, which is also expressed on normal BALB/c spleen cells. In order to study this Ts cell inducing determinant in more detail we have developed an in vitro system which allows the selective activation of ADJ-PC-5 specific Ts cells in the absence of activation of cytotoxic T (Tc) cells. The system now allows us to study the modulation of Ts cell induction and function.

Animals↗
PubMed1986

Evidence for a T suppressor cell-inducing antigenic determinant shared by ADJ-PC-5 plasmacytoma and syngeneic BALB/c spleen cells.

The BALB/c plasmacytoma ADJ-PC-5 induces in early stages of tumorigenesis specific T suppressor (Ts) cells apparently as the first reaction of the immune system towards the growing tumor. In attempts to apply the concept of early induction of Ts cells in neoplasia for the induction of tolerance to allografts we have tried to induce Ts cells against alloantigens by injecting allogeneic spleen cells in doses increasing exponentially from 10 to 10(5) cells/mouse. Experiments using various strain combinations failed to demonstrate induction of specific Ts cells by this procedure. To explain the apparent discrepancy between these and the tumor data the capacity of the ADJ-PC-5 tumor cells to induce Ts cells in allogeneic combinations was analyzed. ADJ-PC-5 cells are capable of inducing specific Ts cells in CBA/J mice. These Ts cells specifically suppress the induction of a primary cytotoxic T cell response of CBA/J spleen cells against BALB/c, BALB.B and (BALB/c X C57BL/6)F1 cells but not one against B10.D2 or C57BL/10 target cells. The ADJ-PC-5-induced CBA/J Ts cells have been further analyzed using a panel of different target or responder cells. From these experiments the following conclusions can be drawn: the CBA/J Ts cells are specific for an antigen which is shared by ADJ-PC-5 and normal BALB/c spleen cells. The antigen in question is not coded for by the H-2d major histocompatibility complex. The data suggest that the Ts cell-inducing antigen on ADJ-PC-5 tumor cells is not a tumor associated neoantigen but a self antigen also present on normal BALB/c lymphoid cells.

Animals↗
PubMed1986

Regulation of immune responses against the syngeneic ADJ-PC-5 plasmacytoma in BALB/c mice. IV. Tumor-specific T suppressor cells, induced at early stages of tumorigenesis, act on the induction phase of the tumor-specific cytotoxic T cell response.

The BALB/c plasmacytoma ADJ-PC-5 induces specific non-cytolytic T suppressor (Ts) cells under experimental conditions which closely resemble early stages of tumorigenesis. The phenotype of these Ts cells is Thy-1.2+, Lyt-1.2-, Lyt-2.2+, I-Ad-. They inhibit the induction of the cytotoxic T cells in a primary mixed lymphocyte tumor culture (MLTC) against the ADJ-PC-5 plasmacytoma. Specificity of suppression can be demonstrated by testing them in MLTC against a panel of syngeneic control tumor. Only the induction phase of a primary MLTC can be suppressed, and within 24 h of MLTC, cytotoxic T (Tc) cells become resistant towards suppression. ADJ-PC-5-specific Tc cell clones are also resistant to suppression. The fact that Ts cell-mediated specific suppression can be overcome by addition of semipurified IL2 to the MLTC suggests that Th cells, and not Tc cell precursors, are the target. The findings are discussed with respect to the potential use of Tc effector cells in cancer therapy.

Animals↗
PubMed1985

Isolation and characterization of in vitro and in vivo functions of a tumor-specific T suppressor cell clone from a BALB/c mouse bearing the syngeneic ADJ-PC-5 plasmacytoma.

A spontaneously transformed T suppressor (Ts) clone, A12-D11/t, is described which arose from the antigen-specific Ts clone A12-D11 isolated from cells of the inguinal lymph node of a BALB/c mouse bearing the syngeneic plasmacytoma ADJ-PC-5. A12-D11/t Ts cells suppress in vitro specifically a primary syngeneic cytotoxic antitumor response with the consequence that no ADJ-PC-5-specific cytotoxic T cells can be generated. The in vivo effects of A12-D11/t Ts cells were studied by injecting them into BALB/c mice. Spleen cells from those mice subsequently failed to respond against ADJ-PC-5 plasmacytoma cells, whereas their response against other syngeneic BALB/c tumors remained unaffected. By using a model system which allows us to study the host's immune reactions to the antigenic load corresponding to initial stages of tumorigenesis, it has been shown previously that ADJ-PC-5-specific Ts cells are activated before the antigen threshold for the activation of cytotoxic T cells is reached. Regarding its phenotype and specificity, the A12-D11/t Ts clone seems to be the exact counterpart of such a Ts cell, and is therefore of special interest in the study of the role of Ts cells preventing immunity against a growing tumor.

Animals↗
PubMed1985

Natural killer cell-mediated cytotoxicity does not depend on recognition of mannose 6-phosphate residues.

Interaction of mannose 6-phosphate-specific receptors with their ligands has been suggested to be essential for natural killer cell (NK)-mediated cytotoxicity. Indeed, mannose 6-phosphate-specific receptors and ligands bearing mannose 6-phosphate residues are demonstrable on human peripheral blood leukocytes with NK activity as well as on K-562 NK target cells, allowing at least in principle such an interaction. It can also be shown that NK activity of human peripheral blood leukocytes is inhibited by mannose 6-phosphate. The following observations, however, exclude an essential role of the mannose 6-phosphate receptor-ligand system in NK cell-mediated cytotoxicity. 1) NK cytotoxicity is sensitive to a broad range of structurally unrelated sugar phosphates. 2) NK activity is normal in patients with I cell disease (mucolipidosis II), which due to a genetic defect are unable to synthesize the ligands for the mannose 6-phosphate-specific receptor. 3) NK cytotoxicity is not inhibited by an antiserum against the mannose 6-phosphate receptor, which blocks the receptor function.

Cell Line↗
PubMed1982

Regulation of immune responses aginst the syngeneic ADJ-PC-5 plasmacytoma in BALB-c mice. III. Induction of specific T suppressor cells to the BALB/c plasmacytoma ADJ-PC-5 during early stages of tumorigenesis.

Initial stages of tumour growth are not easily accessible to investigation. Therefore an experimental procedure was developed to mimic tumorigenesis as closely as possible. BALB/c mice received intraperitoneally exponentially increasing numbers of irradiated syngeneic ADJ-PC-5 plasmacytoma cells. The initial injection began with two cells per mouse and according to the generation time of this tumour, subsequent doses were doubled until mice had received up to 10(5) tumour cells. At various stages of treatment, peritoneal exudate cells (PEC) and spleen cells (SC) were tested for either cytotoxicity or specific suppression of induction of a primary in vitro T-cell cytotoxic response (CTL) of BALB/c spleen cells against ADJ-PC-5 plasmacytoma cells. No cytotoxic PEC were found. Instead, PEC from mice in which the final tumour cells number had reached or exceeded 10(3) irradiated ADJ-PC-5 cells, induced complete suppression of this primary in vitro CTL. Specificity was found both in the induction and effector phase of suppression. Specific suppression was mediated by Thy-1.2+ cells and amplified by non-specific suppression through adherent cells. The data arae discussed in context with previous findings on the in vivo immunogenicity and tolerogenicity of the ADJ-PC-5 plasmacytoma. They suggest that induction of T suppressor (Ts) cells might be an early event in tumorigenesis.

Animals↗
PubMedDate not supplied

A large keratan sulfate proteoglycan present in human cervical mucous appears to be involved in the reorganization of the cervical extracellular matrix at term.

OBJECTIVE: To elucidate the function of keratan sulfate proteoglycan (KS-PG) in the human uterine cervix, we analyzed its distribution with respect to physiologic conditions. METHODS: Immunohistochemistry was used to localize KS bearing proteoglycans (mAb 5D4) and decorin (mAb 6B6) in the lower uterine segment. Proteins present in cervical mucous were labeled with biotin, glycosaminoglycan chains were digested enzymatically, and the samples were analyzed by Western blot. RESULTS: Decorin was detected throughout the extracellular matrix, in tissues from menstruating nonpregnant women, in early pregnancy, from women who had cesarean at term, at postpartum hysterectomy, and from postmenopausal women. In menstruating nonpregnant women, in early pregnancy (first trimester), and in postmenopausal women, KS-PG was detectable only in epithelial, mucous-producing cells. Interestingly, in samples obtained either at the time of cesarean at term (lower uterine segment) or after postpartal hysterectomy, KS-PG was detectable throughout the extracellular matrix, indicating that the expression of KS-PG is associated with reorganization of the tissue. Biochemical analysis of the KS present in mucous revealed a core protein in the range of 220 kDa, suggesting an identity with the large KS-PG described previously. CONCLUSION: At parturition, a large KS-PG, which is virtually exclusively present in the cervical mucous of either early or nonpregnant women, was detected in the extracellular matrix. This finding indicates that cervical ripening is accompanied not only by quantitative but also by qualitative changes in the composition of the extracellular matrix.

Adult↗