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Biomedical subjects

T J Yang

Publications and source records attributed to T J Yang.

At least 73 records · Page 4Linked to original sources

Effects of horse and fetal calf serum on the expression of tumor-associated antigen and tumorigenicity of L5178Y leukemia/lymphoma cells.

A tumor antigen (TA) associated with murine leukemia-lymphoma L5178Y cells has been identified by the enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF) techniques. The antigen was present in both non-solubilized and 0.5% NP-40 solubilized membrane extracts. Rabbit anti-L5178Y lymphoma serum (RALS), extensively absorbed with normal mouse tissues, identified TA in extracts of L5178Y lymphoma and L5178Y leukemia cells grown in horse serum (L5178Y/HS), but not in extracts of L5178Y cells grown in fetal calf serum (L5178Y/FCS). Similarly, absorbed rabbit anti-L5178Y/HS serum specifically reacted with extracts of lymphoma and L5178Y/HS but not with L5178Y/FCS cells. Membrane IIF showed positive reactivity in 88% of lymphoma and 73% of L5178Y/HS cells, whereas splenic lymphocytes and L5178Y/FCS cells were negative. Goat anti-AKR virus serum reacted with soluble extracts of lymphoma, L5178Y/HS, and L5178Y/FCS as well as with normal DBA/2 tissues in the ELISA. However, goat anti-AKR virus serum did not block the reactivity of RALS to lymphoma in the blocking ELISA (BELISA). Expression of TA, but not murine leukemia viral antigen(s), was correlated with the in-vivo tumorigenicity of the L5178Y cells. The antigenic activity of lymphoma extract was reduced by incubation for 1 h at 56 and 65 degrees C, by trypsin digestion, and by exposure to pH 2.8 or 11.0 for 1 h. The antigen, sequentially purified by gel filtration and Lentil-lectin affinity chromatography, was a glycoprotein, with a molecular weight of approx. 64,000 daltons, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis.

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Production and characterization of a bovine T cell-specific monoclonal antibody identifying a mature differentiation antigen.

A monoclonal antibody (MAb), BLT-1, with specificity for bovine mature T cells was prepared by somatic cell hybridization of myeloma NS-1 and spleen cells from BALB/c mice hyperimmunized with bovine T lymphocytes. The MAb reacted with over 92% of nylon wool-nonadherent lymphocytes (T cells) but not with nylon wool-adherent EAC-positive lymphocytes (B cells) in the indirect immunofluorescence assay. It is an IgM, with kappa-light chains, which fixed complement well and killed over 95% of mature T cells in complement-mediated cytotoxicity assays. It reacted with the same proportions of peripheral lymphoid cells (peripheral blood, lymph nodes, and spleen) as the polyclonal goat anti-bovine thymocyte serum (GABTS), but only with 25% of GABTS-positive thymocytes. Immunoperoxidase staining of frozen tissue sections showed that the BLT-1-positive cells were located in the medulla of the thymus and in the T lymphocyte areas of lymph nodes. Western immunoblotting assays showed that the BLT-1-reactive membrane antigen is a 22,000 m.w. protein which was inducible in bovine thymocytes with bovine thymic hormones, thymosin fraction 5, thymosin alpha 1, and thymopentin ORF-18150, indicating that it is a mature T lymphocyte differentiation antigen. The thymosin alpha 1 and thymopentin were found to show additive effects on mature T cell antigen expression by bovine thymocytes.

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Regulation of leukocyte glass adherence and tube leukocyte adherence inhibition (LAI) reactivity by serum factors in dogs with progressing or spontaneously regressing canine transmissible venereal sarcoma (CTVS).

We examined the regulation of leukocyte glass adherence and tube leukocyte adherence inhibition (LAI) reactivity by serum factors in dogs with regressing or progressing canine transmissible venereal sarcomas (CTVS). Both regressor and progressor peripheral blood leukocytes (PBL), draining and non-draining lymph node cells (LNC), and splenic leukocytes were significantly responsive to CTVS antigen extract in tube LAI. In contrast, a significant decrease in basal glass adherence of progressor PBL, draining and non-draining LNC, and splenic leukocytes was observed. Normal glass adherence was restored to progressor leukocytes by extensive washing with warm serum-free media, while significant tube LAI responsiveness to CTVS antigen extract was maintained. Preincubation of regressor PBL and LNC with progressor sera in two-stage tube LAI decreased the basal glass adherence of treated leukocytes. This effect of progressor sera was heat labile, a characteristic of CTVS antigen. Collectively, these findings suggest that progressor leukocytes and progressor sera treated regressor leukocytes were activated by interaction with serum CTVS antigen and thus behaved in tube LAI as stimulated cells, even in the absence of CTVS antigen. Regressor but not progressor sera were shown to contain anti-CTVS IgG with specific arming activity for normal dog PBL, but not LNC in two-stage tube LAI. The nonadherent response of peripheral blood neutrophils in two-stage tube LAI was proportional to the concentration of arming IgG, whereas no change was observed in glass adherence of PBL. The results of this study define the role of progressor and regressor serum factors in the mechanism of tube LAI and demonstrate a relationship between leukocyte glass adherence and the clinical course of CTVS. These findings show that tube LAI is a simple and reproducible measure of active factors in the immune response to a tumor.

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Detection of immune complexes in sera of dogs with canine transmissible venereal sarcoma (CTVS) by a conglutinin-binding assay.

The canine transmissible venereal sarcoma (CTVS) is capable of extended growth in an allogeneic host. Since immune complexes can enhance allograft survival in other animal models, we used an enzyme-linked conglutinin-binding assay to determine the presence and amount of circulating immune complexes in dogs with CTVS. With the conglutinin-binding assay, 23 of 64 dogs (36 per cent) bearing CTVS had concentration of immune complexes 3 standard deviations greater than those detected in normal canine serum. When dogs with circulating immune complexes were separated into 3 groups based on whether the size of CTVS had increased (progressor), decreased (regressor) or remained the same (steady-state) during the week before collection of serum, no significant difference was found in the amounts of immune complexes in sera from progressor dogs. In 3 regressor dogs that received a second transplant of CTVS, the mean concentration of circulating immune complex was significantly greater than the mean for progressor dogs. In progressor dogs, amounts of immune complexes decreased with increasing tumour volume. Following sucrose density gradient ultracentrifugation analysis of sera from a normal and progressor dog, 27S complexes containing both IgG and IgM were detected in serum from a progressor dog. Thus, it appears that the conglutinin-binding assay is a useful and sensitive method for detecting immune complexes in canine serum.

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Role of cyclic AMP in antiserum-induced growth inhibition of murine leukemia L5178Y cells.

Heat-inactivated rabbit antiserum, in the absence of complement, induced a 1.5-2-fold increase in cyclic AMP levels in target cells L5178Y leukemia lymphoblasts within 10-20 min after the experiment. This change preceded the previously reported delayed inhibitory effects of antiserum on cell growth such as inhibition of RNA, DNA, and protein synthesis and cell proliferation, suggesting that cyclic AMP may be one of the mediators of the antigen-antibody reactions which occur at the cell surface. Furthermore, the addition of cyclic GMP or excess calcium to either antiserum or cyclic AMP-treated cultures alleviated the growth inhibitory effects of either antiserum or cyclic AMP, substantiating further the hypothesis proposed.

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The pineal body of the dog.

A technique is described for finding the pineal body of the dog. The posterior half of the skull is cut a little behind the parietofrontal suture, through the occipital condyles. The cerebral hemispheres and cerebellum are carefully sliced, disclosing the pineal at the frontal edge of the colliculi. Two types of cells are present, those with completely round nuclei, and others with vesicular and variably shaped nuclei. In the histological pattern, ependymal cells were observed on the edges, pinealocytes and glial cells within the gland.

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Canine transmissible venereal sarcoma: quantitation of T-lymphocyte subpopulations during progressive growth and spontaneous tumor regression.

The levels of T-lymphocyte subpopulations expressing IgMFc and IgGFc receptors (i.e., T mu lymphocytes and T gamma lymphocytes, respectively) were quantitated for T-lymphocytes obtained from peripheral blood, draining and nondraining lymph nodes, and the tumor mass during progressive growth and spontaneous regression of the canine transmissible venereal sarcoma. Analysis of the T-lymphocyte subpopulations in these lymphoid compartments demonstrated that distinct profiles of T mu and T gamma lymphocytes correlated respectively with the growth and regression statuses of the tumor. The percent of T gamma lymphocytes in the peripheral blood of dogs with progressing, steady-state, and late-regressing tumors was significantly increased over that of control dogs (P less than .05, P less than .001, and P less than .001, respectively) and over that of dogs with early-regressing tumors (P less than .05, P less than .01, and P less than .02, respectively). The percent of T gamma lymphocytes in the lymph nodes draining the tumor was observed to have a significant increase in dogs with progressing (P less than .01), steady-state (P less than .01), and late-regressing (P less than .001) tumors compared with that in control dogs. The percentage of T gamma lymphocytes was observed to have a significant increase in the nondraining lymph nodes of dogs with steady-state and late-regressing tumors compared with that of control dogs (P less than .01 and P less than .002, respectively) and that of dogs with progressing tumors (P less than .001 and P less than .0005, respectively). The percent of tumor-infiltrating T gamma lymphocytes was lowest in tumors that were growing progressively. A significant increase in T gamma lymphocytes was observed in steady-state (P less than .05), early-regressing (P less than .001), and late-regressing (P less than .05) tumors. Early-regressing tumors contained significantly (P less than .005) greater levels of T gamma lymphocytes than did late-regressing tumors.

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Immunoelectron microscopic localization of a tumor-associated antigen (TAA) in the canine transmissible venereal sarcoma by anti-TAA-antiferritin hybrid antibodies.

Immunoferritin and immunoperoxidase reagents were used to localize a tumor-associated antigen (TAA) of the canine transmissible venereal sarcoma (CTVS). Round tumor cells from the CTVS at different stages of growth, i.e., progressive, steady state, and regressing, had TAA diffusely distributed throughout the cytoplasm. In general, TAA was not found on the plasma membrane, within the nucleus, between inner and outer membranes, in cytoplasmic vacuoles, or specifically with any part of the cytocavitary system. Transitional tumor cells, which are intermediate cell types between round cells and spindle-shaped cells and which appear in the tumors at steady state and regressing stages, contained less TAA in their cytoplasm than did the round cell type. The microvilli of tumor cells also contained TAA, suggesting that, in addition to whole cell lysis, shedding of all or parts of these processes may be a mechanism of TAA release as evidenced by the presence of antigenic activity in the extracellular material.

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Amplification of lymph node cell tube leukocyte adherence inhibition (LAI) reactivity by leukocyte adherence inhibition factor (LAIF).

This investigation examines the immunologic basis for specific antigen-induced tube leukocyte adherence inhibition (LAI) reactivity of draining lymph node cells (LNC) from dogs with canine transmissible venereal sarcoma (CTVS). CTVS regressor LNC, macrophage-depleted LNC, and enriched T lymphocyte fractions, but not enriched B lymphocyte fractions, were specifically reactive to CTVS antigen extract in direct tube LAI. In addition, regressor LNC amplified tube LAI responses by generating supernatants with leukocyte adherence inhibition factor (LAIF) activity for normal dog indicator LNC and enriched peripheral blood mononuclear cells (PBMC) in an indirect tube LAI assay. However, macrophage-depleted LNC and enriched T lymphocyte fractions failed to generate supernatants with LAIF activity, suggesting that macrophage accessory cells play a central role in the amplification of tube LAI. Interestingly, CTVS regressor peripheral blood leukocytes (PBL) and PBMC, which were specifically reactive in direct tube LAI, also failed to generate supernatants with LAIF activity. These findings demonstrate a distinction between LAIF-mediated amplification and direct tube LAI reactivity, and suggest that leukocyte populations with differing cellular proportions and from different immunologic compartments may participate in tube LAI via different mechanisms.

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Increase in the haemolytic complement activity of dogs affected with cyclic haematopoiesis.

Cyclic haematopoiesis (CH) is an inherited disorder which occurs in both humans and Grey Collie dogs. The disease is characterized by fluctuations in the numbers of peripheral blood leucocytes, reticulocytes and platelets at regular intervals and is associated with a variety of clinical signs. The most prominent cycle observed is that of neutrophils. The 12-day neutropenic cycle includes a period of relatively normal neutrophil counts, a period of neutropenia and generally a period in which neutrophil counts greatly exceed the normal range. In this study the daily serum haemolytic complement activity (classical pathway CH50) of CH and normal Collie dogs was assayed. The serum CH50 of normal Collie dogs was relatively stable throughout the test period. In contrast, the serum CH50 of CH dogs fluctuated extensively and the mean serum CH50 of CH dogs during the neutropenic cycle greatly exceeded that of normal Collie dogs over the same test period. A close temporal relationship between the stage of the CH neutropenic cycle and the serum CH50 was observed. The mean serum CH50 during neutropenia was not significantly different from that observed when neutrophil counts were within normal range, both values being significantly higher than that of normal dogs. However, the mean serum CH50 during the period of neutrophil rebound was significantly (P less than 0.01) higher than that during the period of neutropenia or normal neutrophil counts. These data suggest that alterations in the production of complement components or regulatory proteins occur at regular intervals in CH dogs.

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Establishment of asparaginase- and guinea pig complement-resistant subline of murine lymphoblastic leukemia L5178y cells.

Asparaginase-resistant subline L5178Y/Asp of murine lymphoblastic leukemia L5178Y cells was established from resistant clones isolated from soft agar cultures treated with Escherichia coli asparaginase. The L5178Y/Asp cells were found to be over 1000-fold more resistant to E. coli asparaginase, co-resistant to inhibitory effect of guinea pig serum (complement), more tumorigenic for syngeneic DBA/2 mice, and to grow much better (higher plating efficiency) in soft agar medium. With the use of the resistant subline in immunocytotoxicity-cytostasis test in the presence of guinea pig complement, we have been able to demonstrate the presence of tumor-associated antigen(s) on the L5178Y cells which is/are also demonstrable by enzyme-linked immunosorbent assay.

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Canine transmissible venereal sarcoma: tumor cell and infiltrating leukocyte ultrastructure at different growth stages.

Electron microscopic study of the canine transmissible venereal sarcoma (N = 26) at different stages of growth showed that tumors in a progressive phase of growth (N = 12) were comprised mostly of large, round cells with prominent nuclei and nucleoli, a few spindle-shaped cells and collagen fibers, focal areas of necrosis, and lymphocyte infiltrates. Regressing tumors (N = 9) had the fewest viable round cells, greater numbers of infiltrating leukocytes, and were comprised almost entirely of connective tissue (collagen bundles) in the latest stages of regression. The infiltrating leukocytes consisted primarily of lymphocytes, some eosinophils, and neutrophils, but very few macrophages. Round-cell nuclei contained numerous perichromatin and interchromatin granules. Their cytoplasm often had annulate lamellae complexes and large deposits of glycogen. Loosely packed tumor cells had extensive proliferation and elongation of cytoplasmic extensions (filopodia). Evidence for cell-mediated tumor cell lysis was demonstrated by degenerative changes in tumor cells which were closely apposed to lymphocytes. Spindle-shaped cells in regressing tumors had intravacuolar collagen fragments, suggesting a possible degradative capacity by these cells.

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Identification and quantitation of bovine T cell antigen with enzyme-linked immunosorbent assay (ELISA).

The enzyme-linked immunosorbent assay (ELISA) was used to identify and quantitate T cell antigen(s) in bovine thymocytes and peripheral blood T cells. The ELISA, employing non-ionic detergent-solubilized lymphocyte membranes as antigen, and goat anti-bovine thymocyte serum (GABTS) and horseradish peroxidase-conjugated rabbit anti-goat immunoglobulin (PORAG) as direct and indirect reactants, respectively, showed that thymocytes and peripheral blood T cells separated by erythrocyte-antibody-complement (EAC) rosetting technique had high concentrations of T cell antigen. In contrast, bone marrow cells and EAC-positive peripheral blood B cells had minimal T cell antigen. 92.0 +/- 3.5% of EAC-positive bovine lymphocytes were found to be B cells and 89.5 +/- 5.8% of EAC-negative lymphocytes T cells, as determined by SmIg and anti-T cell immunofluorescence (IF) techniques. In both IF and ELISA, pre-absorption of rabbit anti-goat IgG serum was needed to obviate cross-reactions with bovine B cells. A competitive ELISA developed for quantitating T cell antigens, showed that peripheral blood T cells contained only 46% of the antigen present in thymocytes.

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Morphologic changes following intraarticular inoculation of Mycoplasma bovis in calves.

Intraarticular inoculation of Mycoplasma bovis into the joints of six-week-old calves caused severe arthritis in five inoculates and mild arthritis in a sixth. Intraarticular inoculation of killed M. bovis did not cause arthritis. Arthritic calves had fever, joint swelling, lameness, neutrophilia, and intercurrent pneumonia from which M. bovis could not be recovered. Gross lesions were massive fibrinosuppurative synovitis and tenosynovitis, erosion of cartilage, and its replacement by polypoid granulation tissue. Histologic lesions were extensive ulceration of synovial membranes, leukocytic infiltration of the subsynovium, congestion, hyperemia, and thrombosis of the subsynovial vessels. Cartilage erosion was accompanied by chronic osteomyelitis and formation of pannus tissue. The presence of thrombi and platelet aggregates suggests that the inflammatory process in the synovium may arise from the interaction of M. bovis with the vasculature and the coagulation process of the host.

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