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1.
Deletion of antigen-specific activity from leukocyte dialysates containing transfer factor by antigen-coated polystyrene 总被引:1,自引:0,他引:1
We have reported finding antigen-specific activity in human leukocyte dialysates (DLE) containing TF in the leukocyte migration inhibition (LMI) assay. To analyze this activity further, we have used polystyrene bound to antibody or to antigen as immunoadsorbent for DLE before pulsing nonimmune cells in the LMI assay. Candida-(CAN) immune or diphtheria toxoid-(TOX) immune DLE were depleted of all antigen-specific activity after absorption with specific antigen but not affected by absorption with specific antibody, respectively, and depletion of activity with antigen was abrogated by coating bound antigen with specific antibody before absorption of DLE. CAN-immune, TOX-immune DLE was selectively depleted for either CAN activity or TOX activity after absorption with CAN- or TOX-coated polystryrene, respectively, retaining its CAN-activity when absorbed with TOX and conversely retaining its TOX activity when absorbed with CAN; thus the antigen-specific activity binds to related but not unrelated antigen. The polystyrene-bound antigen-specific activity could be recovered by treatment with 8 M urea. We interpret these findings to suggest that such antigen-specific activity may be either a dialysable fragment of a T cell antigen receptor site, or a portion of the V-region, or a unique Ir gene product that assists in antigen presentation to other T cells. 相似文献
2.
We have detected new clues to the composition and function of “Transfer Factor” using the direct Leucocyte Migration Inhibition
(LMI) test as an in vitro assay of Dialysates of Leucocyte Extracts (DLE). This approach has revealed two opposing antigen-specific
activities to be present in the same >3500 <12,000 DA dialysis fraction — one activity is possessed of Inducer/Helper function
(Inducer Factor). The opposing activity is possessed of Suppressor function (Suppressor Factor).
When non-immune leucocyte populations are cultured with Inducer Factor they acquire the capacity to respond to specific antigen
and inhibition of migration occurs. This conversion to reactivity is antigen- specific and dose-dependent. When immune leucocyte
populations are cultured with Suppressor Factor their response to specific antigen is blocked and Inhibition of Migration
is prevented. 相似文献
3.
Unconjugated bilirubin inhibits VCAM-1-mediated transendothelial leukocyte migration 总被引:5,自引:0,他引:5
Keshavan P Deem TL Schwemberger SJ Babcock GF Cook-Mills JM Zucker SD 《Journal of immunology (Baltimore, Md. : 1950)》2005,174(6):3709-3718
During lymphocyte migration, engagement of VCAM-1 stimulates the generation of endothelial cell-derived reactive oxygen species (ROS) and activation of matrix metalloproteinases, facilitating endothelial retraction. Because bilirubin is a potent antioxidant, we examined the hypothesis that this bile pigment inhibits VCAM-1-dependent cellular events. The migration of isolated murine splenic lymphocytes across monolayers of murine endothelial cell lines (which constitutively express VCAM-1) is significantly inhibited by physiological concentrations of bilirubin, in the absence of an effect on lymphocyte adhesion. Bilirubin administration also suppresses VCAM-1-stimulated ROS generation and reduces endothelial cell matrix metalloproteinase activity. In a murine asthma model characterized by VCAM-1-dependent airway inflammation, treatment of C57BL6/J mice with i.p. bilirubin decreases the total leukocyte count in the lung parenchyma and lavage fluid, through specific inhibition of eosinophil and lymphocyte infiltration. Blood eosinophil counts were increased in bilirubin-treated animals, while VCAM-1 expression in the capillary endothelium and cytokine levels in both lung lavage and supernatants from cultured lymph node lymphocytes were unchanged, suggesting that bilirubin inhibits leukocyte migration. Conclusion: bilirubin blocks VCAM-1-dependent lymphocyte migration in vitro and ameliorates VCAM-1-mediated airway inflammation in vivo, apparently through the suppression of cellular ROS production. These findings support a potential role for bilirubin as an endogenous immunomodulatory agent. 相似文献
4.
Dr M. T. S. Peraçoli M. T. Rezkallah-Iwasso N. G. S. Mota M. R. Montenegro 《Mycopathologia》1993,121(3):149-156
The effect of dialysable leukocyte extracts (DLE) obtained from hamsters immunized withParacoccidioides brasiliensis (immune DLE) and from non-immunized hamsters (non-immune DLE) was studied in hamsters inoculated withP. brasiliensis by the intratesticular route. Treatment with immune or non-immune DLE was started during the third week of infection and was repeated at 7, 11, 15 and 19 weeks. A group of untreated infected animals was used as control. Animals were submitted to the delayed hypersensitivity skin test toP. brasiliensis antigen (PbAg) in vivo and assayed in vitro by the macrophage migration inhibition test in the presence of Phytohemagglutinin (PHA) and PbAg and by immunodiffusion for specific antibody. The animals were sacrificed at 4, 8, 12, 16 and 20 weeks. The morphology and extension of the lesions were studied at the inoculation site, and in lymph nodes, lungs, liver, spleen and kidneys. In contrast to the controls, animals treated with both DLEs maintained a positive cell-mediated immune response throughout the experiment and developed less extensive infection with a significantly lower number of fungi in the lesions. The results suggest that immune and non-immune DLE preparations modified the evolution of experimental paracoccidioidomycosis with equal efficiency. This similarity may be explained by the immunoregulatory activities of both extracts. 相似文献
5.
Heparin inhibition of human neutrophil and eosinophil-enriched leukocyte acid beta-glycerophosphatase 总被引:1,自引:0,他引:1
B C West 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1985,178(3):373-384
Heparin inhibited acid beta-glycerophosphatase (EC 3.1.3.2) from human blood leukocytes, eosinophil-enriched leukocytes, and neutrophils. The inhibition interfered in the hydrolysis of phosphorus from glycerophosphate, not in the formation or detection of colored complexes of phosphomolybdate in the second or color development step in two conventional assays. Heparin inhibited human hypereosinophilic syndrome leukocyte homogenate enzyme activity according to the equation: activity equals 0.946 - 0.087 ln heparin (units/assay) when heparin was varied from 1 to 100 units per assay. At 100 units of heparin per assay, 51% of the original activity remained. Enzyme activity was less in neutrophils than in eosinophils; moreover, the inhibition of neutrophil homogenate by heparin was considerably less than that seen in the eosinophil-enriched leukocyte preparations. In neutrophil homogenates containing 100 units of heparin per assay, 77.1% of activity without heparin was retained. When neutrophil lysates were utilized, less inhibition was observed: e.g., at 1 unit of heparin per assay, 91.7% enzyme activity was retained and at 1000 units, 76.2%; here, activity equals 0.289 - 0.007 ln heparin. The data allowed more precise consideration of the inhibition of acid beta-glycerophosphatase by heparin, and, while confirming quantitatively the greater content of acid beta-glycerophosphatase in eosinophil-enriched leukocyte preparations than in neutrophil preparations, provide experimental support for an acid beta-glycerophosphatase in human eosinophils, which is different from that in human neutrophils. It is more highly susceptible to heparin inhibition than acid beta-glycerophosphatase in human neutrophils from which it is apparently distinct. 相似文献
6.
Tani K Murphy WJ Chertov O Oppenheim JJ Wang JM 《Biochemical and biophysical research communications》2001,282(4):971-976
Cathepsin G is a neutrophil granule derived antimicrobial chymotrypsin-like enzyme. Our previous study showed that cathepsin G induces chemotactic migration of human phagocytic leukocytes and increases random migration of T lymphocytes. In this study, we investigated the capacity of cathepsin G to activate T lymphocytes and to modulate antigen-specific humoral responses in mice. We found that cathepsin G is mitogenic for and induces production of IFN-gamma by murine T cells in vitro. Injection of cathepsin G in BALB/c mice immunized with keyhole limpet hemocyanin (KLH) adsorbed to aluminum hydroxide resulted in a significantly increased production of KLH-specific IgG1 and IgG2a antibodies. There was a dose-dependent increase in KLH-specific proliferation of lymphocytes from draining lymph nodes from mice treated with KLH and cathepsin G when compared with those treated with KLH alone. Subsequent analysis of IFN-gamma and IL-4 release following in vitro re-stimulation of draining lymph node lymphocytes obtained from KLH-immunized mice suggested that cathepsin G augments KLH-specific Ig antibody production via activation of T cells, presumably involving both Th1 and Th2 pathways. Thus, neutrophil granule cathepsin G, in addition to its capacity to kill microbes and to enhance leukocyte motility, activates T lymphocytes and modulates humoral immunity. 相似文献
7.
Interleukin 1 activity in normal human urine 总被引:17,自引:0,他引:17
E S Kimball S F Pickeral J J Oppenheim J L Rossio 《Journal of immunology (Baltimore, Md. : 1950)》1984,133(1):256-260
Human leukocyte dialysates contain components capable of amplifying cutaneous delayed-type hypersensitivity (DTH) reactions. In the present study, two such amplifiers, both less than 3500 m.w., were partially purified from human leukocyte dialysates by gel filtration on Sephadex G-10 followed by high pressure reverse-phase liquid chromatography. These amplifiers of DTH were examined for their effects on production of the migration inhibitory lymphokines leukocyte migration inhibition factor (LIF) and macrophage migration inhibition factor (MIF). The amplifiers were found to increase LIF and MIF production by antigen- or alloantigen-stimulated human peripheral blood lymphocytes in a dose-dependent fashion. Further analysis demonstrated that although antigen-stimulated T4 and T8 cell subpopulations could produce LIF activity under the assay conditions employed, amplification of lymphokine production by modulator was only observed with the T4 subset. 相似文献
8.
We have investigated the transfer of specific cell-mediated immunity (CMI) to osteosarcoma-associated antigens (OSAA) to hamsters with dialyzable leukocyte extracts (DLE) from OSAA-immunized rabbits. The transfer of specific CMI was determined by leukocyte adherence inhibition (LAI) assay and skin testing. DLE was prepared from rabbits immunized with OSAA, purified protein derivative (PPD), or fibrosarcoma cell plasma membrane preparation (FSM). Control DLE was prepared from rabbits injected with 0.85% NaCl. Significant leukocyte adherence inhibition was observed with leukocytes from hamsters that had received OSAA-specific, PPD-specific, and FSM-specific rabbit DLE, when OSAA, PPD, and FSM were used as antigens, respectively. Similarly, significant ear swelling after injection of OSAA, PPD, or FSM was observed only in hamsters that had received DLE from rabbits immunized with OSAA, PPD, or FSM, respectively. These results suggest that CMI specific for OSAA, PPD, or FSM can be transferred to normal hamsters by DLE from immunized rabbits. 相似文献
9.
J.Timothy Harrington 《Cellular immunology》1977,30(2):261-271
Macrophage migration inhibition by peritoneal leukocytes was studied in BALB/c mice bearing intraperitoneal allogeneic EL-4 lymphomas to explore the role of this immune effector function in allograft rejection. The nonadherent peritoneal leukocyte population harvested between 8 and 10 days after allograft inoculation inhibited migration of nonimmune murine macrophages as demonstrated by both direct and indirect migration assays using the agarose droplet method. This host response also contained large numbers of adherent macrophages which others have shown to be cytotoxic to EL-4 target cells. These findings provide direct evidence for lymphokine activity in allograft rejection and suggest that lymphocyte mediators may attract and activate the cytotoxic macrophages observed in this response. 相似文献
10.
S Albrecht L T Thuy G Pasternak B Schlott J Reinh?fer E Lunow G Loewe H J Gütz 《Acta biologica et medica Germanica》1978,37(11-12):1747-1757
The sensitization of lymphocytes from patients with different tumors was tested against a 3 M KCl-extract of fetal tissue (3.--6. month) by the leukocyte adherence inhibition assay (LAI) and by the leukocyte migration inhibition assay (LMI). Sensitization was compared with the reactivity of controls without any detectable tumor. In the LAI assay the leukocytes of 13/15 patients and 2/12 controls showed an inhibition of the adherence. In the LMI-assay 11/17 tumor-bearing patients and 6/18 controls reacted positive in the presence of the antigen preparation. The two methods demonstrated that patients bearing tumors of different histology are sensitized to fetal antigens. 相似文献
11.
Leukocyte inhibitory factor (LIF), which was derived from human peripheral blood lymphocytes by stimulation with concanavalin A ad partially purified by Sephadex G-100 gel filtration, inhibited the in vitro spontaneous migration and chemotaxis of human PMN leukocytes as assessed in a Boyden chamber micropore filter assay. The inhibitory activity was attributed to LIF, a principle defined in terms of its inhibition of PMN leukocyte migration from glass capillary tubes since it was preferentially directed to PMN leukocytes as compared to mononuclear leukocytes, exhibited a size comparable to LIF by gel filtration, and was inactivated by diisopropyl fluorophosphate in parallel with LIF. Incubation of PMN leukocytes with LIF released additional inhibitory activity, distinct from LIF, which resembled the neutrophil-immobilizing factor (NIF) by virtue of its approximate m.w. of 4000 by filtration on Sephadex G-25, inactivation by trypsin digestion, and preferential noncytotoxic inhibition of spontaneous migration and chemotaxis of PMN leukocytes as compared to mononuclear leukocytes. Thus LIF inhibits PMN leukocyte migration both by a direct action on the cells and by an amplification pathway that is mediated by low m.w. chemotactic inhibitors similar to NIF. 相似文献
12.
The role of the macrophage in the guinea pig mixed leukocyte culture was investigated. Macrophages obtained from oil-induced peritoneal exudates, peritoneal wash-out cells, spleen, and alveolar washings were found to be effective stimulators of allogeneic lymph node and splenic lymphocyte DNA synthesis. The stimulatory properties of macrophages proved radioresistant but viability dependent. Unfractionated lymph node cells or adherence column purified lymph node lymphocytes and thymocytes were only minimally active as stimulators, even in the presence of macrophages syngeneic to the responder lymphocytes. Allogeneic fibroblasts, polymorphonuclear leukocytes, L2C leukemia cells, and xenogeneic (murine) macrophages failed to simulate. These data provide evidence that the macrophage is the predominant stimulator of the mixed leukocyte culture in the guinea pig. 相似文献
13.
Fernández-Ortega C Dubed M Ramos Y Navea L Alvarez G Lobaina L López L Casillas D Rodríguez L 《Biochemical and biophysical research communications》2004,325(3):1075-1081
According to UNAIDS, the global HIV/AIDS epidemic increased to 40 million the number of people living with the virus around the world. Dialyzable leukocyte extract obtained by our group is a low molecular weight dialyzable material from peripheral human leukocytes previously in vitro induced with Sendai virus (DLE-ind), and more recently, from non-induced leukocytes (DLE n/i). Previous results have shown the ability of DLE-ind to inhibit HIV in vitro replication in MT4 cell; to reduce TNFalpha secretion, and to delay in vivo progression to AIDS in early stage of HIV infection. In this work we present evidences that DLE n/i also inhibits HIV in vitro replication and reduces TNFalpha secretion in human whole blood like DLE obtained from induced leukocytes. Taking together these results show that both properties of DLE, HIV in vitro inhibition and TNF production modulation, are not dependent on in vitro Sendai virus induction of leukocytes. 相似文献
14.
Roselynn A. Dynesius-Trentham Donna Rowland Elizabeth Laroche David E. Trentham 《Cellular immunology》1981,63(2):362-373
Rat lymph node cells (LNC) produce a mediator which exhibits functional and physicochemical similarities to human leukocyte inhibitory factor (LIF). Measurement of LIF can be used to quantify cellular sensitivity in rats to foreign protein antigens or to a heterologous antigen in vitro. Concanavalin A-induced rat LIF has a molecular weight of 100,000-60,000 daltons and retains activity after heating to 56 °C for 30 min. Rat LIF is not synthesized in the presence of puromycin and appears to be a protein, since it is inactivated by treatment with chymotrypsin. Moreover, the activity of rat LIF is susceptible to the serine esterase inhibitor, diisopropylphosphofluoridate, but it is resistant to neuraminidase treatment. Finally, rat LIF preferentially inhibits the migration of rat and human polymorphonuclear leukocytes but not that of rat or guinea pig peritoneal exudate cells enriched for macrophages. Except for the more dispersed size of rat LIF, these properties are analogous to those described for human LIF. LIF activity is generated, in an antigen-specific manner, by LNC sensitized to ovalbumin or purified protein derivative of tuberculin when cultured with the antigen used for sensitization. LNC from rats rendered arthritic by prior intradermal (id) injection of native chick type II collagen in incomplete Freund's adjuvant produce LIF in response to this heterologous antigen. These studies delineate a new assay for cellular sensitivity in rats and provide additional evidence that cellular reactivity to type II collagen is present in this animal model of arthritis. 相似文献
15.
D K Novikov A P Shmakov I D Borisenko 《Biulleten' eksperimental'no? biologii i meditsiny》1978,86(11):568-572
Blood serum of patients suffering from cancer of the stomach and urinary bladder inhibited in vitro migration of autologous leukocytes, leukocytes of donors and control patients, and also guinea pig macrophages in over half of cases. In chromatography of these sera on Sephadex G-100 the activity inhibiting the leukocyte migration was revealed in fraction I (Mol. wt. over 100000) and in fractions IV and V (Mol. wt under 30 000). The blood serum and its fractions from cancer patients failed to eliminate the leukocyte migration inhibition caused by the tumour antigens in comparison with the leukocyte migration in the medium with control serum without any antigens. As suggested, the activity of fraction I inhibiting the leukocyte migration was due to the antigen-antibody complex, and of fraction IV and V--to a factor similar by its properties to the factor produced in vitro by lymphocytes stimulated by the antigens or mitogens. 相似文献
16.
The indirect agarose technique of leukocyte migration inhibition has been used to measure the response of human peripheral blood lymphocytes to several viruses. Using commercially available viral antigens, the indirect assay was found to be more sensitive than the direct agarose technique. Supernatants from cultures of sensitive lymphocytes with virus contained a nondialysable factor which inhibited the migration of polymorphonuclear leukocytes (PMN). Under strict conditions of assay, whereby all culture supernatants were tested together on the same PMN preparation, the degree of migration inhibition obtained in response to mumps virus correlated well with the size of the skin test reaction to mumps. A similar relationship was shown for PPD. A good correlation existed also between the degree of migration inhibition and the lymphocyte transformation response for each of these two antigens. 相似文献
17.
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. 相似文献
18.
Supernatant fluid obtained after centrifugation of the suspension of viable lymph node cells of immunized animals proved to induce in vivo in the lymph node cells of intact mice sensitivity to lysis with a specific antigen in vitro. This property was possessed after chromatography of the supernatant fluid on Sephadex G-200 by the 3rd fraction (MW about 30000 dalton). DNA-ase, trypsin or deproteinization failed to influence whereas RNA-ase inactivated this fraction in respect to the inducing properties. 相似文献
19.
Chondroitin sulfate (Structum) interacts with human leukocyte elastase, a potent mediator of articular cartilage degradation, producing a partial inhibition of the enzyme activity (60% at saturation). Kinetically, the inhibition mechanism can be classified as simple intersecting, hyperbolic noncompetitive and is almost identical to that found earlier for similar compounds. The best inhibitory activity of chondroitin sulfate was found in fractions having at the same time a high proportion of chondroitin-6-sulfate relative to the corresponding 4-isomer and a high molecular mass. Thus, a fraction with high Mr and containing 92% of isomer 6 inhibited leukocyte elastase with Ki = 1.8 micrograms/ml, whereas a fraction with low Mr and almost equal composition of the 4- and 6-isomer had Ki = 140 micrograms/ml. Ki for unfractionated chondroitin sulfate was 3.4 micrograms/ml. It is suggested, that the modulation of the extracellular activity of cartilage-degrading enzymes by cartilage-derived factors may explain, at least in part, the beneficial effects of some therapeutically used chondroprotective agents. 相似文献
20.
A functional role for circulating mouse L-selectin in regulating leukocyte/endothelial cell interactions in vivo 总被引:7,自引:0,他引:7
Tu L Poe JC Kadono T Venturi GM Bullard DC Tedder TF Steeber DA 《Journal of immunology (Baltimore, Md. : 1950)》2002,169(4):2034-2043
L-selectin mediates the initial capture and subsequent rolling of leukocytes along inflamed vascular endothelium and mediates lymphocyte migration to peripheral lymphoid tissues. Leukocyte activation induces rapid endoproteolytic cleavage of L-selectin from the cell surface, generating soluble L-selectin (sL-selectin). Because human sL-selectin retains ligand-binding activity in vitro, mouse sL-selectin and its in vivo relevance were characterized. Comparable with humans, sL-selectin was present in adult C57BL/6 mouse sera at approximately 1.7 micro g/ml. Similar levels of sL-selectin were present in sera from multiple mouse strains, despite their pronounced differences in cell surface L-selectin expression levels. Adhesion molecule-deficient mice prone to spontaneous chronic inflammation and mice suffering from leukemia/lymphoma had 2.5- and 20-fold increased serum sL-selectin levels, respectively. By contrast, serum sL-selectin levels were reduced by 70% in Rag-deficient mice lacking mature lymphocytes. The majority of serum sL-selectin had a molecular mass of 65-75 kDa, consistent with its lymphocyte origin. Slow turnover may explain the relatively high levels of sL-selectin in vivo. The t(1/2) of sL-selectin, assessed by transferring sera from wild-type mice into L-selectin-deficient mice and monitoring serum sL-selectin levels by ELISA, was >20 h, and it remained detectable for longer than 1 wk. Short-term in vivo lymphocyte migration assays demonstrated that near physiologic levels ( approximately 0.9 micro g/ml) of sL-selectin decreased lymphocyte migration to peripheral lymph nodes by >30%, with dose-dependent inhibition occurring with increasing sL-selectin concentrations. These results suggest that sL-selectin influences lymphocyte migration in vivo and that the increased sL-selectin levels present in certain pathologic conditions may adversely affect leukocyte migration. 相似文献