Production Of Migration Inhibitory Factor Research Articles

Macrophage Migration Inhibitory Factor (MIF) Production by Cord Blood Lymphocytes

Macrophage Migration Inhibitory Factor (MIF) Production by Cord Blood Lymphocytes

Inhibition of mitogen-induced lymphokine production by cyclosporin A.

The new antilymphocytic agent Cyclosporin A was found to inhibit the production and/or secretion of migration inhibitory factor (MIF) in human lymphocytes stimulated by Concanavalin A. Preincubation for one hour with the compound, followed by 8 hr restoration period of the cells in absence of the drug, resulted in moderate decrease in MIF synthesis and/or release. Cell viability was not affected. The agent was shown not to interfere with MIF action on the macrophage. We conclude that the molecular mechanism of action of Cyclosporin A is based, at least partially, on a blockade of synthesis and/or secretion of lymphokines from immunocompetent cells.

Lotus tetragonolobus fucolectin as a potential model for “MIF-like” modulation of macrophage function: Comparison of the interaction of Lotus fucose binding protein (FBP) and migration inhibitory factor (MIF) with macrophages in the migration inhibition assay

Fucose binding protein (FBP), the fucolectin from Lotus tetragonolobus, was compared with migration inhibitory factor (MIF) for its binding interactions with peritoneal macrophages in the migration inhibition assay and with concanavalin A (Con A) for its mitogenic properties for lymphocytes. FBP was nonmitogenic for guinea pig lymphocytes as demonstrated by its inability to stimulate incorporation of [ 3H]thymidine into TCA insoluble fractions or to induce production of MIF. FBP and MIF inhibited migration of responsive peritoneal macrophages in similar dose-response profiles, whereas refractory macrophages were significantly less responsive to both inhibitors. FBP and MIF were adsorbed correspondingly by responsive macrophages but not by refractory macrophage populations. FBP was bound to peritoneal macrophages during short-term pulse exposure to inhibit macrophage migration in a dose-response fashion similar to that of MIF. Enzymatic treatment of peritoneal macrophages with trypsin, chymotrypsin, or Pronase abrogated their response to both FBP and MIF. l-Fucose (0.1 M) significantly reduced FBP and MIF-mediated migration inhibition while d-fucose and l-rhamnose were ineffective. FBP was also found to augment MIF inhibition of macrophage migration under a variety of experimental conditions. Based on these findings, it is proposed that Lotus fucolectin may provide a useful model for studies on “MIF-like” modulation of macrophage function.

Effect of cimetidine on delayed hypersensitivity

Histamine influences the expression of delayed hypersensitivity skin tests through the histamine type 2 (H-2) receptor. Previous work has demonstrated this effect in vivo by local injection of H-2 receptor antagonists into the skin test site; and by incorporation of these agents into the culture media in in vitro studies. Cimetidine, a H-2 receptor antagonist, was given to guinea pigs before and during the period of sensitization to a hapten-protein conjugate. Delayed hypersensitivity was studied by skin testing and production of migration inhibitory factor (MIF). Increased MIF production was demonstrated in vitro; but no effect was noted in vivo. No good explanation exists for the failure to enhance skin test reactivity; but the presence of H-1 and H-2 receptors in vascular smooth muscle in the skin may explain skin test reactivity. Lymphocyte production of MIF is regulated by H-2 receptors on sensitized lymphocytes.

Regulation of granulomatous inflammation in murine schistosomiasis. In vitro characterization of T lymphocyte subsets involved in the production and suppression of migration inhibition factor.

Host granulomatous inflammation in murine schistosomiasis mansoni is a T cell-mediated immune response, which, at the chronic stage of the disease, undergoes T suppressor lymphocyte-dependent modulation. In the present study this phenomenon was further analyzed in vitro. Spleen cells of mice undergoing modulation (20 wk of infection) when mixed with spleen cells of animals exhibiting vigorous granulomatous responses (8 wk of infection) abrogated in vitro migration inhibition factor (MIF) production by the latter. Characterization of the delayed-type hypersensitivity T lymphocytes involved in lymphokine production showed that they belonged to the Lyt-1+ subset and did not express I region-encoded antigens. In contrast, T lymphocytes involved in the suppression of MIF activity belonged to the Lyt-2+ subpopulation of cells, which expressed I-J- and I-C-subregion determinants. These results suggest that the modulation of the granulomatous hypersensitivity response in mice is the result of T-T cell interaction with subsequent regulation of inflammatory lymphokine production.

Depressed leukocyte migration inhibition in response to streptococcal antigens in poststreptococcal glomerulonephritis

Leukocyte migration inhibition was examined in 14 patients with persistent disease following acute poststreptococcal glomerulonephritis (PSGN), in 9 individuals with previously documented beta hemolytic streptococcus infections which were not followed by glomerulonephritis, and in 13 normal controls, using as antigens streptococcal protoplast membrane (SPM) from type 12 (nephritogenic strain) and type 6 (nonnephritogenic strain), and streptokinase-streptodornase (SKSD). PSGN patients showed significantly less inhibition than the other groups in response to nephritogenic type 12 SPM antigens. No consistent differences were observed among the three groups in response to SPM antigen from type 6 or to SKSD. The data demonstrate that decreased production of leukocyte migration inhibition factor (LIF) in response to SPM antigen correlates with ongoing PSGN and is not merely a reflection of previous beta hemolytic streptococcus infection. The depressed cellular immune response to SPM in PSGN is not restricted to lymphocyte transformation as reported earlier and is probably specific for antigens derived from nephritogenic strains of streptococcus.

Lectin induction of lymphokines in cultured murine leukocytes

Antigens induce sensitized lymphocytes to undergo mitosis and to secrete soluble products, termed lymphokines, which modulate the immune response. Plant lectins are known to act as polyclonal lymphocyte mitogens and, in some cases, stimulate lymphocytes to produce lymphokines. In an effort to explore the relationship of specific cell surface glycoconjugates to the induction of mitosis and the production of lymphokine activities we have examined the ability of the mitogenic lectins, concanavalin A and Wistaria floribunda mitogen, and the nonmitogenic hemagglutinin from Wistaria floribunda seeds to stimulate the production of macrophage migration inhibition factor (MIF), macrophage chemotactic factor (CF), and lymphotoxin (LT). Concanavalin A causes lymphocytes to produce MIF and LT but no detectable CF activities. W. floribunda mitogen induces lymphocytes to produce soluble substances which exhibit all three lymphokine activities. The nonmitogenic W. floribunda agglutinin causes lymphocytes to produce MIF and CF but no observable LT activity. Within the sensitivity of the assays employed, the results indicate that mitogenesis is not a corequisite of the expression of either macrophage migration inhibition factor or lymphocyte-derived chemotactic factor but it may be associated with the induction of lymphotoxin. It is also apparent that the expression of each lymphokine activity is independent of the expression of the other lymphokines studied.

Immunological Assay for the Diagnosis of Coeliac Disease: Interaction between Purified Gluten Fractions

Increased leukocyte migration inhibition factor (LIF) production by coeliac patients' lymphocytes is a reliable immunologic test for gluten-sensitive coeliac disease providing the small peptide subfractions of gluten are used as mitogens. To investigate discrepancies in results obtained in various published studies, subfractions (B2 and B3) of crude gluten peptic tryptic digest [Frazer's fraction III (F3)] were used singly and in combination as mitogens in coeliac patients and compared in various doses with F3. There was significant increased production of LIF with B2 or B3 when tested separately as compared to a mixture of B2 +/- B3 or to F3. LIF production with B2 or B3 was 23.7 +/- 6.9%; with B2 + B3, it was 10.65 +/- 14.7% (t = 4.05; P less than 0.0005); and with F3, it was 9.2 +/- 10.2% (t = 5.8; P less than 0.0005). There was no difference in the response between B2 + B3 and F3 (t = 0.4; P = 0.32). Studies of LIF production following stimulation by various doses and combinations of gluten mitogens in the same patient gave the best results with B2 or B3 in 10 microgram doses, with a tendency to less LIF production with higher dosages or combinations of B2 + B3 or F3, even to abolition of LIF production in some coeliac cases. The LIF assay is a reliable immunologic test for gluten sensitivity, providing the B2 or B3 subfractions are used as mitogens.

Progressive multifocal leukoencephalopathy (PML): in vitro cell-mediated immune responses to mitogens and JC virus.

Cell-mediated immunity was studied by in vitro tests in seven patients with progressive multifocal leukoencephalopathy (PML). Lymphocyte proliferation in response to mitogenic stimulation was reduced to a variable degree in all patients, indicating a general impairment of cell-mediated immune responsiveness, although mitogen-induced production of the lymphokine migration inhibitory factor (LIF) was normal in most cases. Cell-mediated immunity to JC virus (JCV) was assessed by LIF production in response to JCV antigen. In the six PML patients tested, LIF production with JCV antigen was absent despite the presence of antibody to JCV in serum. This contrasted with positive LIF production in seropositive normal individuals and patients and patients with other diseases. These results provide the first in vitro evidence of a depressed cell-mediated immune response to JCV in patients with PML, and support the hypothesis that PML is accompanied by a selective marked deficiency in cellular response to this virus in association with a general depression of cell-mediated immunity of variable severity.

Immune Competence of Newborn Lymphocytes

The immune competency of peripheral blood (cord) lymphocytes of newborn infants has been investigated by a battery of assays in vitro. Number of T cells was enumerated by E-rosette formation and a cytotoxicity assay by using an anti-human-thymocyte antiserum. Lymphokine production as an indicator of T cell function was evaluated by leukocyte migration inhibition factor production following stimulation with various mitogens, and "viral" and "immune" interferon production in response to stimulation with polyriboinosinic -cytidilic acid and phytohemagglutinin, respectively. Ability to respond to mitogens was also tested by means of 3H-thymidine uptake into DNA, and by measuring the early synthesis of protein by lymphocytes by means of 3H-leucine uptake. Lymphocyte cyclic adenosine monophosphate levels in resting cells and after trypsin treatment was also used as a test of cellular competency. Total B cells was evaluated by EAC-rosette formation, and class specific surface membrane immunoglobulin-bearing cells were assayed by the peroxidase immunoenzymatic method. Normal children and adults served as controls. The results indicated that no significant differences in immune competency could be shown between newborns and older people except for lymphocyte cyclic adenosine monophosphate levels which were lower in the newborns, both in resting and in stimulated cells. The relationship between this latter finding and the immune status is as yet not clear. It is concluded that neonatal lymphocytes are essentially immunocompetent, with the expression of their immune capabilities in vivo becoming apparent only after encounter with environmental antigenic stimuli.

Lack of suppression of lymphocyte MIF production by estradiol, progesterone and human chorionic gonadotropin

The hypothesis that hormones produced by the placenta contribute to immunosuppression during pregnancy is an attractive one. In order partially to evaluate this hypothesis, we utilized the Migration Inhibition Factor (MIF) Assay, which is an in vitro model of delayed hypersensitivity in that presensitized lymphocytes stimulated by antigen release a glycoprotein (MIF) which inhibits the normal migration of macrophages. The effect of placental hormones on this model of cellular immunity was tested using the direct and indirect MIF assays in pre-immunized guinea pigs. Estradiol (1–50 μg/ml), progesterone (1–50 μg/ml) and commercial grade or chromatography-purified hCG (1000–5000 IU/ml) did not affect normal macrophage migration and also failed to suppress production of MIF by guinea pig lymphocytes stimulated with antigen. None of the hormones either enhanced or suppressed macrophage response to preformed MIF. The results indicate that these hormones do not suppress lymphocyte reactivity as assessed by the production of lymphokines or the target cell response to the mediator. The reported effects of steroid hormones and crude hCG on other models of cellular immunity are discussed.

Cimetidine as an immunomodulator: chronic mucocutaneous candidiasis as a model.

Four adult patients with chronic mucocutaneous candidiasis were studied to establish a possible role for cimetidine as an immunomodulator. These patients had negative baseline in-vivo and in-vitro cell-mediated immune response to candida antigen as measured by intradermal skin tests, lymphocyte transformation, and leukocyte migration inhibitory factor production to cimetidine, 300 mg by mouth, four times daily. Subsequently four of four patients developed strong (greater than 15 mm) intradermal skin test reactions, and two of four patients produced leukocyte migration inhibitory factor to candida antigen. Skin tests and leukocyte migration inhibitory factor production reverted to baseline negative values when repeated 4 weeks after discontinuation of therapy. After 4 additional weeks on cimetidine, four of four patients showed strong positive skin tests and leukocyte migration inhibitory factor production to candida antigen. Lymphocyte transformation was not affected by therapy.

Lymphokine Production in Primary Mixed Lymphocyte Reactions

The characteristics of the responder and stimulating cells involved in migration inhibition factor (MIF) production in primary 'one-way' mouse mixed lymphocyte reactions (MLR) were analyzed by using an indirect agarose droplet assay. T-lymphocytes are mainly responsible for MIF release, as shown by pretreatment with anti-Thy 1.2 serum plus complement or purification over a nylon wool column. On the other hand, macrophages and B-lymphocytes appear to be optimal stimulating cells. T-lymphocytes as stimulating cells induce MIF release, but to a much lesser degree than macrophages and B-lymphocytes. The kinetics of MIF production in MLR is related to the kind of stimulating cells employed. Lastly, the ability to release MIF is already present in the spleen of 1- to 2-week-old mice, lasts until 20 weeks of age and declines to undetectable levels at 50 weeks of age.

Modulation of lymphokine production by interferon.

Pretreatments with relatively low (25-250 U/ml) concentrations of interferon (IFN) enhanced while 5000-10 000 U/ml concentrations of IFN inhibited leukocyte inhibitory factor (LIF) and migration inhibitory factor (MIF) production of concanavalin A (Con A) stimulated human leukocytes. The lymphokine regulatory activity of IFN correlated with the antiviral activity of IFN preparations regardless of their specific activity and it showed species specificity. Modulation of lymphokine production depended not only upon the amount of IFN used but also on the length of IFN pretreatment. Blocking of lymphokine production by high concentration of IFN could not be explained by the effect of IFN on cell viability.

Impaired Cell-Mediated Immune Response in Patients with Congenital Rubella: Correlation with Gestational Age at Time of Infection

Lymphocyte transformation, interferon, and leukocyte migration inhibition factor synthesis were studied in purified lymphocyte cultures for 20 children with congenital rubella and 18 healthy children (seven susceptible and 11 immune to rubella). Lymphocyte transformation after phytophemagglutinin stimulation was significantly lower in children with congenital rubella as compared to healthy controls. Responses to purified rubella virus were absent in the susceptible controls and absent or at least two times lower in congenital rubella children than in immune controls. After purified rubella virus stimulation, leukocyte migration inhibition factor production was detected in all immune controls, but in none of the susceptible controls, or the congenital rubella-infected children. The results varied with gestational age of intrauterine infection: the impairment of cellular immune response, both after phytohemagglutinin or rubella virus stimulation, was more severe in the children infected in the first two months than in the latter stages of gestation.

Immunologic reaction of psychotic patients to fractions of gluten

Production of a leukocyte migration inhibition factor by peripheral blood lymphocytes in response to challenge with gluten fractions was studied in hospitalized patients with schizophrenia and other psychoses compared with normal individuals and with children and adolescents with celiac disease. The schizophrenic and other psychotic patients could be subdivided into two groups, one that responded in the leukocyte migration inhibition factor test as the celiac patients did and one that responded as the normal control subjects did. The psychotic and schizophrenic patients did not show any evidence of malabsorption. The authors speculate that gluten may be involved in biological processes in the brain in certain psychotic individuals.

Guinea Pig Lung Lavage Cells After Intranasal BCG Sensitization

Recent studies have suggested that intranasal administration of antigen can induce local cell-mediated immunity in lung lavage cells. The present study was designed to examine the changes in composition of lung lavage cells and their capacity to produce the lymphokine migration inhibitory factor after intranasal immunization with BCG in guinea pigs. Results indicate that guinea pigs responded to respiratory tract BCG infection with an increase in immunocompetent cells in the bronchoalveolar tract and with production of migration inhibitory factor. After local pulmonary BCG administration, the total number of cells increased as compared with that of the uninfected animals, the increase being statistically significant within 2 weeks. This marked increase in the total cell population is due to a more than doubling of the number of macrophages in the lavage fluid. Animals also developed at this time positive delayed hypersensitivity to intradermally administered purified protein derivative. A significant increase in the total lymphoid cells and macrophage population was observed again at 6 weeks after sensitization, suggesting that the response is biphasic in nature. At 6 weeks, however, there was also a significant rise in total lymphocytes and T cell population in addition to macrophage numbers. This increase in T cells correlated with an increase in production of migration inhibitory factor in the presence of purified protein derivative. These data suggest that the immune response of the respiratory tract after BCG challenge involves increased recruitment of immunocompetent cells locally at the site of infection and that these cells are capable of producing effector molecules in terms of the elaboration of migration inhibitory factor.

Role of the thymus in regulation of production of macrophage migration inhibitory factor in mice of different genotypes

The effect of the thymus on production of macrophage migration inhibitory factor (MMIF) was studied in C57BL and CBA mice thymectomized at the age of 4–6 weeks. MMIF production in response to stimulation by tuberculin was estimated at the peak of the immune response on the 1st–21st days after the operation. This factor was found to be produced as early as the 1st day after thymectomy. MMIF production is absent in athymic nude mice. Thymectomy also stops the immune response in the early stages of its development. Spontaneous migration of macrophages also is modified in immune and nonimmune animals. These changes are more marked in C57BL mice.

Macrophage migration stimulation factor, migration inhibition factor and the role of suppressor cells in their production

Guinea pig lymph node lymphocytes and human peripheral blood lymphocytes when stimulated by specific antigen or mitogen will release factors that affect in vitro macrophage migration. Migration inhibition factor production appears to be under the control of suppressor cells which are T lymphocytes. When suppressor cells are generated by stimulation with Con A for 4 days, migration stimulation factor (M.St.F.) activity is found. In other situations where M.St.F. is found this is thought to be due to increased suppressor cell activity. For example, young adults produce this lymphokine when stimulated with Con A, whereas aged individuals produce MIF. Concanavalin A appears to be the mitogen of choice for M.St.F. production, and phytohemagglutinin for MIF production. The release of this putative factor M.St.F. from suppressor T cells helps to explain some of the difficulties that have existed in studies of macrophage migration inhibition.

Immunosuppression by mycobacterial arabinomannan: Ellner, J. J. and Daniel, T. M. (1979). Clinical Experimental Immunology, 35, 250

The means by which mycobacteria elude host defence mechanisms to cause human disease is unknown. This report provides the initial demonstration of a soluble product of Mycobacterium tuberculosis with immunosuppressive activity which may contribute to the pathogenicity of this organism. Mycobacterial D-arabino-D-mannan (AM), purified from culture filtrates of M. tuberculosis by Concanavalin A-Sepharose affinity chromatography did not itself induce 3H-thymidine incorporation or migration inhibitory factor production by peripheral blood mononuclear cells from tuberculin skin-test-positive healthy humans although it produced weak delayed skin-test reactions in these subjects. However, this mycobacterial cell wall polysaccharide suppressed the response of human lymphocytes to mycobacterial and other antigens as assessed with these in vitro correlates of delayed hypersensitivity. Immunosuppression by arabinomannan was not contingent upon prior exposure of the individual to M. tuberculosis, since it occurred with lymphocytes from tuberculin skin-test non-reactive as well as reactive subjects. Nor was it due to cytotoxicity, an antimitotic effect of the polysaccharide preparation or generation of suppressor cells. Rather, AM seemed to suppress macrophage-dependent, antigen-induced activation of human lymphocytes. This immunosuppressive substance elaborated by mycobacteria has potential relevance to the pathogenesis of human tuberculosis and the nature and limitation of the resulting host response.