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[Detection of a factor suppressing leukocyte migration in the sera of allergy patients following antigen administration].

The effect on donor leukocyte migration of serum obtained from the patients with tuberculosis of the lungs, chronic pneumonia and healthy persons was studied after subcutaneous or intradermal injection of the microbial antigen (PPD, streptococcus and staphylococcus antigen). A factor inhibiting donor leukocyte migration appeared in the blood serum of sensitized individuals after the antigen injection. This factor proved to be localized in the serum fraction III obtained after the gel-filtration of sephadex G-200, and is sorbed by leukocytes.

Allergens↗

The role of cell-mediated immunity in the induction of inflammatory responses. Parke-Davis Award Lecture, 1977.

Reactions of cell-mediated immunity fall into two broad categories: those that involve direct participation of intact lymphocytes in the effector mechanism of the reaction and those that involve mediation by soluble lymphocyte-derived factors known as lymphokines. The first kind of reaction is essentially limited to lymphocyte-dependent cytotoxicity, although certain aspects of T cell-B cell cooperation may fall into this category as well. The second category appears to comprise the bulk of the so-called cell-mediated immune response and provides a link between this system and the inflammatory system. Various lymphokines have been shown to exert profound influence upon inflammatory cell metabolism, cell surface properties, patterns of cell migration, and the activation of cells for various biologic activities involved in host defense. Although substantial information is now available about various physicochemical as well as biologic properties of lymphokines, purification and characterization data are as yet too incomplete to allow us to ascribe all of these activities to discrete mediator molecules. Current work involving the development of antibody-based techniques for mediator assay may shed light on this issue. Information on the kinds of cells capable of lymphokine production is now available. Contrary to prior expectation, T cells are not unique in their capacity for lymphokine production. Under appropriate circumstances, B cells and even nonlymphoid cells can do so as well. The unique property of lymphocytes in this regard appears to relate to their ability to respond to certain specialized signals such as specific antigen or an appropriate mitogen. Mediator production per se may represent a general biologic phenomenon. Although lymphokines have been defined mainly in terms of in vitro assays, early speculations about their in vivo importance are proving correct. Evidence for the role of lymphokines comes from studies involving detection of lymphokines in tissues, studies involving injection of exogenous lymphokines, and studies involving suppression of in vivo reactions by various techniques. The use of antilymphokine antibodies has proven useful in the latter kinds of experiments. Work in many laboratories is beginning to relate these findings to clinically relevant situations. A major unsolved problem relates to the regulation and control of lymphokine production and activity. At present only a limited body of information is available on this point. This is a potentially fruitful area for future investigation since it may provide techniques for manipulating the immune system in ways that are clinically useful.

Animals↗

[Kinetic differences in induction of the killers and producers of the macrophage migration inhibitory factor (MIF) in primary and secondary mixed murine leukocyte culture test].

Conditions are selected for the killer induction during primary and secondary responses in the one-way MLC, stimulated by x-irradiated or killed allogenic lymphocytes. MIF is found in the MLC culture medium as a sharp peak on the 2nd or the 3rd day of the primary reaction, or as a plato from the 1st day of the secondary reaction. In contrast, the killers are shown to be induced much later reaching their maximum on the 5th and the 4th days, respectively. If heated, instead of irradiated, allogenic lymphocytes were used for stimulation of the primary MLC reaction, the MIF-producer kinetics was not changed, but the killer induction was found to be delayed.

Animals↗

The indirect assay for leukocyte migration inhibitory factor (LIF)--standardization and the effect of pH.

In the indirect assay for leukocyte migration inhibitory factor (LIF), lymphokine-rich supernatants were obtained by culture of stimulated lymphocytes and then tested for LIF activity in agarose plates using purified granulocytes as target cells. Studies on the standardization of the conditions under which LIF acts on the target cells are described, with emphasis on the use of "standard" supernatants of known LIF activity and the influence of pH on the action of LIF and the sensitivity of the assay. The observation that LIF activity is reduced when the ambient pH falls below 7.2 is suggested as an explanation firstly for the "escape" phenomenon seen particularly in capillary tube assays for LIF, and secondly for the reduced sensitivity of the capillarly tube assay in comparison with the corresponding agarose plate assay.

Granulocytes↗

MIF-CD74 axis facilitates MDSC infiltration in the tumor microenvironment of pancreatic ductal adenocarcinoma.

Immune checkpoint inhibitors show insufficient efficacy against pancreatic ductal adenocarcinoma (PDAC). The tumor microenvironment (TME) has a remarkable influence on responsiveness to cancer immunotherapy. The aim of this study was to investigate immunosuppressive characteristics of TME in PDAC tissues. The flow cytometry (FCM) of PDAC surgical specimens revealed that the profile of tumor-infiltrating leukocytes was classified into myeloid cell- and T-cell-dominant subtypes; the myeloid subtype was associated with poorer patient outcomes. Myeloid-derived suppressor cells (MDSCs) showed the highest hazard ratio among various myeloid cell types. Single-cell RNA sequencing and FCM revealed that most MDSCs, but not lymphocytes, in PDAC tissues characteristically express CD74. Macrophage migration inhibitory factor (MIF), a CD74 ligand, was highly expressed in cancer-associated fibroblasts (CAFs) and cancer cells. Spatial transcriptomics demonstrated that the MIF-CD74+ myeloid cell interaction was recognized in CAF-dominant areas in PDAC tissue. CAFs expressing immune suppressor molecules such as MFAP5 and LRRC15 were consistent with MIF+ CAFs. Furthermore, MIF+ CAFs enhanced the migratory activity of MDSCs and promoted MDSC induction and activation. In the murine model, MDSCs were significantly increased in MIF-expressing PDAC tumors, as were CD74+ M-MDSCs per M-MDSC, confirming in vivo interaction between CD74 and MIF. MDSCs play a crucial role in creating an immunosuppressive TME in PDAC; the MIF-CD74 axis drives interactions between MDSCs and CAFs.

Humans↗

Abrogation of cell-mediated immunity by a serum blocking factor isolated from patients with infectious mononucleosis.

Lymphocytes from 80% of patients with infectious mononucleosis in this study failed to produce macrophage migration-inhibition factor in response to partially purified early antigen of Epstein-Barr virus or to tetanus toxoid, whereas lymphocytes from normal subjects did produce this lymphokine. Subsequent analysis of serum from the patients with infectious mononucleosis revealed a serum factor that completely abrogated antigen-specific inhibition of migration by human leukocytes as well as lymphocyte blastogenesis. The serum blocking factor was present in sera from 11 (73%) of 15 patients with infectious mononucleos but only in sera from two (13%) of 15 normal subjects. Samples of serum from five of the patients with infectious mononucleosis and five normal subjects were fractionated with use of Sephadex G-200 gel filtration, and the eluants were assayed for several substances known to inhibit cell-mediated immunity. Serum blocking factor activity could be demonstrated only in fractionated sera from patients with infectious mononucleosis. The serum blocking factor is postulated to be either a soluble immune complex or some as yet unidentified immunoregulatory globulin contained in the IgG fraction of human serum.

Antigen-Antibody Complex↗

Clinical study of a patient with lupus vulgaris before and after injection of dialyzable transfer factor.

This report describes the clinical improvement and acquisition of tuberculin skin-test sensitivity by a tuberculin-negative, drug-resistant patient with lupus vulgaris after a single injection of dialyzable transfer factor (TFd) from a tuberculin-positive healthy donor. The patient's lymphocytes showed a slight response to tuberculin in the leukocyte migration inhibition test and in the lymphocyte transformation test before TFd injection. The acquisition of cellular immunity to tuberculin was demonstrated in vitro by enhanced tuberculin-induced blast transformation. A good correlation between skin test and in vitro tuberculin sensitivity and clinical improvement was seen during the three years that the patient was observed.

Adult↗

Effects of lymphokines on fibrinolysis and kinin generation in human plasma.

Supernatants from Con-A-stimulated human lymphocytes containing leukocyte migration inhibitory factor, skin reactive factor and procoagulant activities were tested for their effect on fibrinolysis and kinin generation. The supernatants of stimulated lymphocytes activated the fibrinolysis and generated kinin in human plasma. These results are discussed from the aspect of cell-mediated (type IV) immune inflammation.

Animals↗

Cytokine production by blue tongue virus-infected fetal sheep cells.

The migration inhibition of guinea pig peritoneal macrophages by a factor(s) from media obtained from blue tongue virus-infected monolayer cultures was studied. Medium from blue tongue virus-infected sheep fetal cell cultures inhibited migration of guinea pig macrophages from agarose droplets. Medium from control cultures and stock virus did not inhibit macrophage migration. Medium containing migration inhibiting factor(s) in vitro induced an inflammatory reaction in the skin of a newborn sheep. The inflammatory reaction was observed 20 h after intradermal inoculation. The skin reaction consisted of infiltrates of mononuclear leukocytes in the superficial dermis. Control medium and stock virus caused no skin reaction.

Animals↗

Leukocyte migration inhibitory factor (LMIF) production in unidirectional mixed lymphocyte cultures.

Puromycin treatment of lymphocytes was used to develop a one-way test for leukocyte migration inhibitory factor (LMIF) production in the mixed lymphocyte culture (MLC) reaction. Lymphocytes incubated with this protein synthesis inhibitor induced a vigorous mediator production by nontreated allogeneic cells, being themselves unable to respond to stimulator cells. When puromycin-treated cells were stimulated with the mitogens PHA, ConA, or PWM, overall protein and DNA synthesis were significantly decreased with concomitant abolishment of LMIF production. Viability of stimulator lymphocytes was found to be essential for generation of the mediator in MLC reaction.

Cell Survival↗

[Effect of synovial fluid from rheumatoid arthritis patients on leukocyte migration].

By means of the migration inhibition test, the influence of the synovial fluid of rheumatoid arthritis patients on normal blood lymphocytes was investigated. There was the hypothesis that the migration inhibitory factor is already formed in the synovial membrane of rheumatoid arthritis patients. In 35 cases a migration inhibition could be demonstrated, in 3 cases the migration of normal lymphocytes remained uninfluenced, in 6 cases a migration enhancement occurred. The demonstration of the rheumatic factor in the synovial fluids used was partly positive, partly negative. Especially the demonstration of a migration enhancement--this phenomenon could be reproduced repeatedly--cannot yet be interpreted unequivocally and requires further investigations.

Arthritis, Rheumatoid↗

Antibodies to guinea pig lymphokines. IV. Suppression of the mixed leukocyte culture reaction by anti-lymphokine globulin.

An antiserum, (anti-lymphokine globulin, ALyG) directed against highly purified products of activated lymphocytes, inhibits the proliferation of responding cells in guinea pig mixed leukocyte cultures ((MLC). This serum recognizes three newly synthesized lymphocyte products (one of them being migration inhibition factor (MIF) which are involved in the mediation of delayed hypersensitivity reactions in vivo. Since ALyG does not appear to contain cytotoxic antibodies against guinea pig lymphocyte antigens and its inhibitory activity cannot be removed by absorption with lymphoid cells it seems likely that the inhibition of MLC reactivity is not mediated by the lysis of stimulator or responder cells and its target is not the lymphocyte per se but possibly some factor elaborated during MLC response. By contrast, antisera with specificity for histocompatibility (H) antigens can inhibit the MLC when the appropriate H antigens are present on the responding and/or stimulating cell population. However, this inhibitory of the antisera can be effectively absorbed with lymphoid cells bearing the appropriate H antigens. The addition of ALyG even 48 hr after the initiation of culture results in a marked inhibition of MLC reactivity. This finding is consistent with the elaboration of a mitogenic factor or signal during the first 48 hr of culture and the delivery of this signal to the responding cell population. Thus, ALyG does not appear to interfere with the synthesis of this factor but must be present after its release in order to block responder cell proliferation. Furthermore, the stimulatory effect of this MLC mitogenic factor, but not of PPD-induced mitogenic factor on "third party" cells can be completely inhibited when the cells are cultured in the presence of ALyG. These findings suggest that the MLC-mitogenic factor in this system is identical to or cross-reactive with one or both of the newly synthesized molecules recognized by the ALyG.

Animals↗

[In vitro liberation of lymphocytic factors inhibiting migration of human leukocytes induced by gold, cadmium and mercury chlorides].

The presence of 300 microgram gold and cadmium and 10 microgram mercury chloride per millitre of survival medium, during the process of T.I.M.L., results in the release by human lymphocytes of soluble factors which inhibit migration of leucocytes (L.I.F. and M.I.F.). This stimulation of cellular immunity suggests that the anti-infectious therapeutic effect of gold and mercury inorganic salts results from activation of the histiocytary system, through the release of these lymphokins.

Cadmium↗

[The inhibitor factor of leukocyte migration in acne vulgaris].

The inhibition factor of the leukocytic migration was studied in 26 patients with Acne Vulgaris, stages III and IV, being positive in all the cases. The LIF positive shows the mobilisation of lymphocytic population, answering an active and constant immunologic conflict. As the PA is antigenic, because it produces a blastification (TTL positive), we can say that it acts as antigen provoking immunologic mechanisms with help to explain the inflammatory process at the pilosebaceous follicle in the AV.

Acne Vulgaris↗

Human leukocyte migration inhibitory factor (LIF). I. Effect of synthetic and naturally occurring esterase and protease inhibitors.

The activity of leukocyte migration inhibitory factor (LIF) obtained from Sephadex-G-100-chromatographed supernatants of concanavalin-A-stimulated human lymphocytes was suppressed by two synthetic serine esterase and serine protease inhibitors (di-isopropylfluorophosphate (DEP) and phenylmethylsulfonyl fluoride (PMSF)). LIF activity was also reduced by the naturally occurring protease inhibitors soybean trypsin inhibitor and aprotinin. The observed effect of DFP and PMSF was irreversible, since elimination of the inhibitors by dialysis did not restore LIF activity. The effect of PMSF was dose-, time-, and temperature dependent, and hydrolytic products of PMSF as well as sodium fluoride were inactive in blocking LIF. These results suggest that LIF may act as a serine esterse or a serine protease, or both of these, and that this putative enzyme is present in an activated form in supernatants from mitogen-stimulated mononuclear cells.

Aprotinin↗

Studies of assay conditions for macrophage migration from an agarose droplet.

The agarose microdroplet method is a relatively simple and economic technique to determine migration inhibition of leukocytes or macrophages in vitro. In the present study, further cultural and technical requirements of this method for the determination of macrophage migration inhibition have been defined: influence of macrophage handling before the assay, kinetics of migration and dependence on the pH of the medium. Considering defined conditions, the agarose microdoplet assay gives highly reliable and reproducible results. In comparative experiments, it proved to be as sensitive and valid as the capillary tube technique.

Animals↗