bluuebunny/crossref_metadata_2025_split huggingface dataset:
Glycosaminoglycan profiles in cloned granulated lymphocytes with natural killer function and in cultured mast cells: their potential use as biochemical markers.
Authors: Bland C E , Rosenthal K L , Pluznik D H , Dennert G , Hengartner H , Bienenstock J , Metcalfe D D 
Published: December 2022

Abstract
Proteoglycans from three cloned, granulated lymphocyte cell lines with natural killer (NK) function (NKB61A2, HY-3, H-1) and one mast cell line (PT-18) were labeled with [35S]sulfate. [35S]proteoglycans were extracted in 1 M NaCl with protease inhibitors to preserve their native structure and were separated from unincorporated [35S]sulfate by Sephadex G-25 chromatography. [35S]proteoglycans from all four cell lines were chromatographed over Sepharose 4B and were found to have a similar range of m.w. The [35S]glycosaminoglycans from each cell line were then separated from parent proteoglycans by treatment with 0.5 M NaOH. The [35S]glycosaminoglycans from the three lymphocyte cell lines exhibited a similar m.w. as assessed by Sepharose 4B gel filtration, whereas the [35S]glycosaminoglycans from the mast cell line chromatographed as a smaller m.w. molecule. [35S )glycosaminoglycan charge characteristics were evaluated with DEAE C1-6B ion exchange chromatography. The consistency of the elution patterns was determined by using [35S]glycosaminoglycans obtained from radiolabelings of each cell line separated by 6 mo in culture. Each NK lymphocyte cell line reproducibly produced two distinct [35S]glycosaminoglycan chains that eluted in two regions well before the commercial heparin marker. The proportions of each chain were dependent upon the specific cell line. The mast cell line produced a single [35S]glycosaminoglycan chain, which eluted overlapping the internal commercial heparin marker, consistent with its higher charge characteristics. [35S]glycosaminoglycans from all cell lines were identified as chondroitin sulfates with the use of specific polysaccharidases. The NK lymphocyte glycosaminoglycans contained chondroitin 4-sulfate disaccharides. The mast cell glycosaminoglycans contained oversulfated disaccharides and chondroitin 4-sulfate disaccharides. Thus, each granulated NK lymphocyte cell line produced chondroitin sulfate glycosaminoglycans that were characteristic of that cell line and of different composition and less charge than those produced by cultured mast cells. These findings demonstrate that glycosaminoglycan profiles are useful biochemical markers in the characterization of diverse granulated cell lines including NK lymphocytes and mast cells.
The antibody specific to type 1 chain blood group A determinant.
Authors: Abe K , Levery S B , Hakomori S 
Published: December 2022

Abstract
Two monoclonal antibodies directed to the blood group A determinant have been established. One hybridoma antibody (AH16), obtained on immunization with human gastric cancer cell MKN45, was directed to the blood group A determinant carried by both type 1 and type 2 chains. The other antibody (AH21) was prepared by immunization with a purified type 1-chain A glycolipid, followed by selection of the hybridoma by positive reaction with the type 1, but negative reaction with the type 2-chain A glycolipid. Thus, the antibody specifically reacts to the A determinant carried by the type 1 chain, but not to the A determinant carried by the type 2 chain; an A determinant carried by type 1 chain differs significantly from that carried by type 2 chain in its conformational structure. The antibody AH16 agglutinates both A1 and A2 human erythrocytes, but AH21 agglutinates neither A1 nor A2 erythrocytes, which supports the results of methylation analysis that blood group A and H determinants in human erythrocytes are exclusively made of the type 2 chain and that A1 and A2 phenotypes are not related to the difference between type 1 and type 2 chains.
Arachidonic acid metabolism in purified human lung mast cells.
Authors: Peters S P , MacGlashan D W , Schulman E S , Schleimer R P , Hayes E C , Rokach J , Adkinson N F , Lichtenstein L M 
Published: December 2022

Abstract
Arachidonic acid metabolism has been explored in preparations of purified human lung mast cells prelabeled with arachidonic acid (AA). Cells were of 83 to greater than 96% purity, and each experiment was performed with four to six different preparations of mast cells. After overnight culture of the purified cells in the presence of 3H-AA, followed by extensive washing in buffer, mast cell uptake of labeled AA was 61.4 +/- 14.8 pmol/10(6) cells with 21 +/- 2.4% of the label in phospholipids, 73 +/- 2.1% in neutral lipids, and 3.6 +/- 0.8% as free AA. Analysis of the distribution of radioactivity in phospholipid classes revealed 51.4 +/- 5.5% of the label in phosphatidylcholine, 14.5 +/- 1.6% in phosphatidylinositol, 12.0 +/- 3.0% in phosphatidylethanolamine, and 9.1 +/- 2.4% in sphingomyelin, with the rest in other phospholipid classes. Challenge of these cells with an optimal concentration of anti-IgE led to the release of 20 +/- 4.0% of cellular histamine and to a reduction in labeled phosphatidylcholine and phosphatidylinositol to 75.5 +/- 8.8% and 84.2 +/- 4.5% of the control levels, respectively, (p less than 0.05); anti-IgE challenge produced no statistically significant change in the quantities of other labeled phospholipids. Activation of human lung mast cells with anti-IgE led to the release of 3.4 +/- 1.3% of the cellular 3H as AA and AA metabolites (1.5 +/- 0.6% as unmetabolized AA) in conjunction with 16 +/- 4.3% of the cellular histamine. Although activation of human lung mast cells with ionophore A23187 caused 70 +/- 1.1% histamine release, a similar quantity of AA and AA metabolites was released (a total of 4.0 +/- 0.8% with 2.3 +/- 1.5% as unmetabolized AA). Analysis of the released metabolites by liquid scintillation spectrometry after high performance liquid chromatography separation showed that approximately equal amounts of metabolites were produced after mast cell activation with anti-IgE and ionophore A23187. In this series of experiments approximately equal amounts of cyclooxygenase and lipoxygenase products were generated.(ABSTRACT TRUNCATED AT 400 WORDS)
Induction of chemiluminescence during interaction of tumoricidal effector cell populations and tumor cells is dependent on the presence of mycoplasma.
Authors: Köppel P , Peterhans E , Bertoni G , Keist R , Groscurth P , Wyler R , Keller R 
Published: December 2022

Abstract
A variety of host cells, such as activated macrophages, natural killer (NK) cells, and polymorphonuclear leukocytes (PMNL), are cytotoxic for an array of non-antibody-coated tumor cells. Because such effector cells appear to use oxygen-dependent mechanisms to effect tumor cell destruction in certain systems, the possibility of an involvement of toxic oxygen species has been considered. To investigate whether interaction of effector cells with neoplastic cells induces the generation of reactive oxygen species, resting and activated rat macrophages and rat spleen cells (as a source of NK activity) were exposed to viable tumor cells of varied origin, and chemiluminescence was monitored. This sensitive indicator of reactive oxygen generation was stimulated only when tumor cells or culture supernatants were contaminated with mycoplasma. Mycoplasma-free tumor cells and culture supernatants were in no case able to trigger chemiluminescence in any of these effector cell populations. On the other hand, tumor targets were equally susceptible to killing by effector cells irrespective of whether mycoplasma were present. The data suggest that generation of chemiluminescence during interaction of natural cytotoxic cells and neoplastic cells is an artifact and that reactive oxygen species do not function as an effector mechanism in antibody-independent natural killing effected by activated macrophages and NK cells.
Susceptibility of Giardia lamblia trophozoites to the lethal effect of human serum.
Authors: Hill D R , Burge J J , Pearson R D 
Published: December 2022

Abstract
To define the potential role of complement and antibody in host defense against Giardia lamblia, the effect of human serum on axenically cultured G. lamblia trophozoites was studied. Sera from patients without a history of giardiasis and with no detectable antibody by an indirect immunofluorescence antibody (IFA) assay (IFA titers less than 1:2) killed from 8 to 76% of trophozoites (n = 23 sera). Whereas less than 10% of parasites incubated in phosphate-buffered saline alone were killed, 16 sera killed from 10 to 25% and six sera killed from 25 to 75%. One serum with an anti-G lamblia antibody titer of 1:128 killed greater than 98% of the parasites. The complement dependency of killing was demonstrated by abrogation of the lethal effect when serum was chelated with EDTA (7 mM) or was heat-inactivated (56 degrees C, 30 min), and, in an immunofluorescence assay, by detection of C3 on parasites killed by normal serum. When the classical complement pathway was selectively blocked by using serum chelated with Mg+2 (2 mM)-EGTA (8 mM), or serum congenitally deficient for the second component of complement, there was no killing; thus, killing was dependent on the presence of an intact classical pathway. In each of three sera from donors with negative IFA titers, an absorbable factor specific for G. lamblia, possibly antibody not detected by IFA, was required for classical pathway activation. To determine if alteration of the surface of G. lamblia would render it an activator of the alternative pathway of complement, trophozoites were studied after cell death or after treatment with neuraminidase or trypsin. In MgEGTA-chelated serum, dead trophozoites activated the alternative pathway as determined by consumption of Factor B and deposition of C3 on their surface. In contrast, untreated or enzyme-treated living parasites did not activate the alternative pathway.
Topologic mapping of protective and nonprotective epitopes on the variant surface glycoprotein of the WRATat 1 clone of Trypanosoma brucei rhodesiense.
Authors: Hall T , Esser K 
Published: December 2022

Abstract
Monoclonal antibodies were used in competitive antibody binding assays to define and map epitopes on the variant surface glycoprotein of the WRATat 1 clone of T. b. rhodesiense. By using a panel of 30 WRATat 1-specific monoclonal antibodies, 16 epitopes were defined that fall into four clusters, having 1, 1, 3, and 11 distinct epitopes respectively. All epitopes were easily classified as being 1) exposed uniformly on the surface of the trypanosome, 2) exposed only in the region of the flagellar pocket, or 3) "buried", based on the ability or inability of the monoclonal antibodies to bind living trypanosomes in a fluid phase immunofluorescence assay. Monoclonal antibodies that bind exposed surface epitopes are protective, whereas only three of seven that bind exclusively to flagellar pocket epitopes are protective. None of the nine monoclonal antibodies that recognize buried epitopes are protective. Also, antibody-mediated immunity to WRATat 1 trypanosomes is not associated with any particular subclass of antibody. The IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA subclasses each contain examples of protective monoclonal antibodies.
Adoptive immunotherapy of established syngeneic solid tumors: role of T lymphoid subpopulations.
Authors: Rosenstein M , Eberlein T J , Rosenberg S A 
Published: December 2022

Abstract
We have investigated the subpopulations of T cells necessary to mediate the cure of established tumors in two models of successful adoptive immunotherapy. In C57BL/6 mice bearing palpable and disseminated FBL-3 lymphoma, both Lyt-1+ and Lyt-2+ cells played a major role in mediating the regression and permanent cure of mice, whereas in BALB/c mice bearing the Meth A sarcoma the adoptive transfer of Lyt-1+2+ cells played a major role in mediating the regression of tumors and the curing of disease. Identical experiments performed in hybrid (BALB/c X C57BL/6) mice yielded similar results, further supporting our initial observation and indicating that in these two adoptive transfer model systems it is the tumor and not the variable expression of Lyt antigens by the host that determines which T cell subpopulation is required to cure mice of tumors.
Separation and isolation of rat natural killer (NK) cells from T cells with monoclonal antibodies.
Authors: Woda B A , McFadden M L , Welsh R M , Bain K M 
Published: December 2022

Abstract
It has been difficult to clearly differentiate rat NK cells from cytotoxic T cells. In this study we have shown that rat NK cells do not express the T cell protein defined by the OX 19 antibody. By using the FACS we have isolated the OX 19- OX 8+ lymphocyte subset that contains virtually all the NK activity. The simultaneous use of the OX 19 and OX 8 antibodies allows the separation of NK cells from T cells.
Interleukin 2 administered in vivo induces the growth of cultured T cells in vivo.
Authors: Cheever M A , Greenberg P D , Irle C , Thompson J A , Urdal D L , Mochizuki D Y , Henney C S , Gillis S 
Published: December 2022

Abstract
The capacity of exogenous IL 2 to induce the growth of antigen-activated T lymphocytes in vivo was evaluated. The in vivo growth of adoptively transferred T lymphocytes that had been previously cultured long-term with IL 2 was initially examined, because in vitro such T cells are exquisitely dependent upon exogenous IL 2 for proliferation and survival. Daily administration of IL 2 in vivo, beginning on the day of cell transfer, induced these IL 2-dependent long-term cultured T lymphocytes to proliferate in vivo, and the magnitude of in vivo growth was proportional to the dose of IL 2 administered. The capacity of IL 2 to induce the in vivo growth of antigen-activated T cells not previously exposed in vitro to exogenous IL 2 was similarly studied. T lymphocytes from the spleens of immune mice, activated by 5-day culture with tumor antigen before transfer, survived poorly in vivo when injected with antigen alone, but demonstrated marked proliferation in vivo in response to antigen and exogenous IL 2. By contrast, immune spleen cells transferred with antigen, but without prior culture, proliferated without supplementary exogenous IL 2. Moreover, the growth of noncultured donor T cells was not augmented by the administration of exogenous IL 2, implying that noncultured spleen cells immune to tumor antigens can produce sufficient amounts of endogenous IL 2 in vivo to sustain maximal T cell growth over the time period examined. Importantly, the ability of exogenous IL 2 to induce donor T cell growth in vivo correlated with its ability to function in vivo to augment the anti-tumor efficacy of specifically immune donor T cells in models for the adoptive therapy of disseminated antigenic murine leukemia. Thus, the current studies highlight the potential of exogenous IL 2 to induce T cell growth in vivo and suggest that the administration of IL 2 in vivo may be useful for augmenting T cell responses that are relatively deficient in the production of endogenous IL 2.
Dual parameter flow cytometric analysis of DNA content, activation antigen expression, and T cell subset proliferation in the human mixed lymphocyte reaction.
Authors: Williams J M , Loertscher R , Cotner T , Reddish M , Shapiro H M , Carpenter C B , Strominger J L , Strom T B 
Published: December 2022

Abstract
This study provides direct correlation via dual parameter flow cytometry (simultaneous assessment of immunofluorescence and DNA content) between mixed lymphocyte reaction (MLR) responder cell entry into the S/G2/M phases of the cell cycle with the kinetics of expression of two activation-associated cell surface proteins, Tac (IL 2 receptor) and 4F2 (unknown metabolic function). A small population of activated cells was identifiable by expression of both Tac and 4F2 antigens before peak DNA synthesis in the MLR. This population of activation antigen-positive cells expanded linearly in size from days 3 to 7 of culture. Treatment of immature MLR cultures with anti-4F2 Mab and complement (C) before DNA synthesis (treatment on day 3, peak DNA synthesis on days 5 to 6) resulted in blunted proliferation and activation antigen expression when the same culture was analyzed after maturation on day 6, indicating that the activated population had been previously detected and removed by anti-4F2 Mab + C. The 4F2 antigen was expressed on a greater percentage of cells in the MLR at all times (days 3 to 9) than was Tac, was present on virtually all S/G2/M phase responder cells, and a large fraction of cells remained intensely 4F2+ subsequent to peak DNA synthesis. In contrast, after initially preceding responder cell entry into the S phase of the cell cycle, the kinetics of Tac antigen expression closely paralleled the kinetics of responder cell proliferation. A subpopulation of cycling responder cells was noted in all MLR cultures studied that expressed Tac antigen weakly or not at all. Cells within both T4 and T8 cell subsets proliferate with similar kinetics in response to alloantigen. The possibility that activation antigens can be utilized to study effector cell generation in the MLR and that this flow cytometric technique may be utilized to analyze the response to various alloantigens is discussed.
A novel human leukocyte differentiation antigen: monoclonal antibody anti-D44 defines a 28 Kd molecule present on immature hematologic cells and a subpopulation of mature T cells.
Authors: Bernard A , Gay-Bellile V , Amiot M , Caillou B , Charbord P , Boumsell L 
Published: December 2022

Abstract
Anti-D44 is a cytotoxic IgG2b monoclonal antibody (MAb), which reacts with two polypeptides of 28 and 30 Kd from human peripheral T cells after western blotting. The antigen is present in low density on bone marrow cells from the myeloid lineage, probably including most CFU-GM; the antigen is present on megakaryocytes but is not detectable on erythroid cells as well as pre-B and B cells. Growth of BFU-E appears to be partially inhibited after treatment with anti-D44 + complement, an effect that should be due to bone marrow T cells rather than to BFU-E destruction. With maturation, the antigen is barely detectable on mature polymorphonuclear neutrophils, and is not detectable on platelets and monocytes. In contrast, cells from the thymic cortex carry a high density of this molecule; medullary thymocytes carry a low density. Large thymocytes ("thymoblasts") distinct from epithelial or interdigitating cells also bear a high density of the antigen. In the periphery, a subpopulation of T cells displays a high density of D44 molecules. D44 antigenic density is not linked to cell growth or to T cell activation. Double labeling and cell sorting experiments have shown that 40% of E rosette-forming cells are D44(+)-CD4+, 10% are CD8(+)-D44+ and 10% are CD4(-)+CD8-, CD2+. In an accompanying report, correlation is shown between expression of the D44 defined molecule on T cell subpopulations and the function they exert. Immunoperoxidase studies of skin, brain, and kidney tissues, facilitated by the fact that the antigen is resistant to inclusion in Epon, which permits staining of semi-thin sections, have not revealed any positive nonhematologic cell.
Monocyte and NK cell cytotoxic activity in human adherent cell preparations: discriminating effects of interferon and lactoferrin.
Authors: Horwitz D A , Bakke A C , Abo W , Nishiya K 
Published: December 2022

Abstract
The comparative cytotoxic specificities of freshly isolated human adherent and nonadherent blood mononuclear cells were examined against seven established target cell lines in 4 and 18 hr chromium release assays. The relative sensitivity of each target cell line to the cytotoxic effects of both adherent and nonadherent effector cells in cultures was identical. Moreover, the relative enhancing effects of interferon on cytotoxicity by both effector cell types were also identical. These adherent cell preparations were contaminated with up to 6% NK cells, as demonstrated by OKM1 staining and flow microfluorometry. These NK cells were loosely adherent and could be removed by vigorous wash procedures. The remaining tightly adherent monocytes also had the capacity to kill K562 cells and Chang cells, but these cytotoxic effects could not be increased by interferon. Enhancement by lactoferrin, however, was consistently observed. Treatment of mononuclear cells with Leu-lla, a monoclonal antibody that reacts with all NK cells, also abolished the enhancing effects of interferon, but not lactoferrin. These studies suggest that caution must be exercised in attributing all cytotoxic activities in adherent cell preparations to monocytes, and that lactoferrin and interferon can be used as functional probes to detect two distinct blood mononuclear cell subsets with natural cytotoxicity.
Phenotypic and functional characterization of T cell clones derived from the cerebrospinal fluid of multiple sclerosis patients.
Authors: Santoli D , Defreitas E C , Sandberg-Wollheim M , Francis M K , Koprowski H 
Published: December 2022

Abstract
We describe here T cell cultures and clones established from the cerebrospinal fluid (CSF) of three patients with multiple sclerosis (MS) and one chronic meningitis patient with pleocytosis. Most of the cultures were activated with phytohemagglutinin (PHA) before growth in mitogen-free interleukin 2 (IL 2), and were never restimulated. Some of the clones obtained have been propagated for over 1 yr and are strictly IL 2-dependent. Immunofluorescence analysis performed with various monoclonal antibodies revealed that the CSF-derived lines had the characteristics of activated T cells with a stable expression of either suppressor/cytotoxic or helper/inducer surface antigens. Most of the clones established had a predominantly suppressor phenotype (OKT8+), except for the clones derived from one MS patient, which expressed only the helper phenotype (anti-Leu-3a+). Consistent with these data, the CSF-derived cultures displayed a variety of immunoregulatory functions, such as the ability to lyse nonspecific and PHA-stimulated target cells, to produce IL 2 upon mitogenic activation, and to modulate polyclonally induced Ig responses. The availability of long-term CSF T cell cultures derived from MS patients at various disease stages might provide a useful tool in investigating the factor(s) involved in the etio-pathogenesis of the disease.