Migration Inhibitory Effect Research Articles

Immunosuppressive properties of alpha 1-microglobulin.

alpha 1-Microglobulin (alpha 1m), a serum glycoprotein (26,000 d), was found to impede the proliferative response of human lymphocytes to purified protein derivative (PPD) and tetanus toxoid. The data suggest that alpha 1m operates through an unstable suppressor mechanism, which no longer can function after 24 h of preculturing. This effect of alpha 1m on antigen stimulation did not seem to be due to binding of alpha 1m to PPD or cells, to altered kinetics of the PPD response, or to non-specific cytotoxicity. In contrast, PPD-induced leucocyte migration was not reversed by alpha 1m. alpha 1m did not cause significant inhibition in experiments in which lymphocytes were stimulated by the mitogens phytohaemagglutinin or concanavalin A. Finally, alpha 1m had its own leucocyte migration inhibitory effect.

Species-restricted effects of human and mouse lymphokines on macrophages.

Lymphokines produced by antigenic or mitogenic stimulation of human, guinea pig and mouse lymphocytes were tested for their effects on monocytes or macrophages of the same and heterologous species, to determine whether there is any species restriction in their reactivity. Supernatants from lymphocyte cultures were tested for migration inhibitory activity in an indirect agarose microdroplet assay and for their ability to augument cytolytic activity of macrophages or monocytes in a [3H]thymidine-release assay. Supernatants of human peripheral blood mononuclear cells, stimulated with phytohemagglutinin or purified protein derivative of tuberculin (PPD) were able to strongly inhibit the migration of human monocytes and guinea pig peritoneal exudate cells (PEC), but had no detectable effect on mouse PEC. The human supernatants could also significantly augment the cytolytic activity of human monocytes, but had no effect on cytotoxicity by mouse peritoneal macrophages. Conversely, supernatants of PPD-stimulated spleen cells from mice immune to bacillus Calmette-Guérin (BCG) strongly inhibited the migration of, and significantly augmented cytolysis by, mouse PEC, but had no detectable effects on human monocytes. Moreover, supernatants of concanavalin A-stimulated lymph node cells from guinea pigs inhibited the migration of guinea pig PEC and human monocytes, but had no effects on mouse PEC. The migration inhibitory effects of the human and mouse supernatants did not appear to be mediated by interferon (IF), since partially purified type-1 IF had no detectable effect. In addition, supernatants of human lymphocytes stimulated by Corynebacterium parvum strain 5888, that induced little or no IF, were able to inhibit the migration of, and augmented cytolysis by, human monocytes. These C.parvum supernatants also showed migration inhibitory activity on mouse PEC but did not induce cytolytic activity in these cells.

Cell-mediated immunity to herpes virus type-1 in patients with recurrent corneal herpes simplex.

The proportion of T lymphocytes in the peripheral blood, their phytohaemagglutinin reactivity, and the leucocyte migration inhibitory effect of purified tuberculin protein were found to be equal in 25 recurrent corneal herpes simplex patients and 17 healthy individuals who had never suffered from clinically manifest herpes simplex virus (HSV) infection. Conversely, the HSV vaccine inhibited the leucocyte migration in the healthy group, but not in the recurrent herpes patients when the clinical symptoms were manifest. In 14 recurrent corneal herpes simplex patients, the examinations were repeated after a 3-month symptom-free period, and in these periods the patients corresponded equally to the healthy group in every respect.

Effects of autologous plasma from patients with malignant tumor on macrophage migration inhibition with tumor extract.

By the use of the indirect migration inhibition technique blocking or inhibitory effect of autologous plasma on the production of migration inhibitory factor (MIF) from lymphocytes contacted with autologous tumor extract was studied in 45 patients with malignant tumor. When autologous plasma was added to the lymphocyte culture incubated with autologous tumor extract in 44 of these patients, inhibition of migration was completely abrogated in 9 of these patients whose lymphocytes produced MIF when cultured in plasma-free medium. MIF activity was not abolished when autologous plasma was added to the supernatant of lymphocyte culture incubated with autologous tumor extract. Inhibition of migration was not affected by the addition of allogeneic plasma from a normal subject to the lymphocyte culture. In 15 patients, MIF was not produced when lymphocytes were cultured with autologous plasma alone. These results appeared to indicate that autologous plasma of patients with malignant tumor might block or inhibit the production of MIF from lymphocytes contacted with autologous tumor extract.

Assay of human lymphokines in vitro. Evidence for a migration stimulation factor (MStF) which interferes with the macrophage migration inhibition assay.

When cultured in vitro, human lymphocytes produced a range of materials which were able to reduce the migration inhibitory effect of human macrophage migration inhibition factor (MIF) and in some cases stimulated macrophage migration to greater than control levels. In addition, lymphocytes stimulated with antigen or mitogen produced a lymphokine-like migration stimulation "factor". Detectable production of this factor occurred 24--48 h after antigen stimulation. The migration stimulation factor (MStF) (MW 50-250,000; pI 6-5) was not chemotactic and could be separated from migration inhibition and chemotactic factors by chromatography and iso-electric focussing.

Lymphokine-Like Factors Produced by Human Lymphoid Cell Lines with B or T Cell Surface Markers

In the present series of experiments we examined the ability of eight continuous cell lines derived from human lymphocytes to release lymphokines into their culture media. We have demonstrated that all could produce a macrophage migration inhibitory factor and a neutrophil chemotactic factor. This ability was independent of the B or T cell origin of the cell lines. As a further extension of previous reports, we report that the migration inhibitory effect does not seem to be due to cytotoxic effects as judged by trypan blue exclusion studies, and more important by the demonstration that the inhibition is reversible. The demonstration that neutrophil chemotactic activity is removable by treatment with a previously described and characterized anti-lymphokine antiserum suggests that this factor may be chemically similar to some of the lymphokines (migration inhibitory factor or chemotactic factor for macrophages) produced by antigen-activated guinea pig lymphocyte cultures. The failure to effect similar removal or inactivation of the macrophage migration inhibitory factor may simply reflect greater interspecies heterogeneity in this factor, or more interestingly, that this does not correspond to "classic" MIF. These results extend previous observations by defining further the capacity of lymphoid cell lines in continuous culture to produce mediators and by characterizing their relationship to conventional lymphokines produced by antigen or mitogen-activated lymphocytes.

Migration inhibitory effect of cell-free supernatants from PHA-activated cultures of human mononuclear leukocytes.

A two-step test for the demonstration of phytohaemagglutinin (PHA)-induced production of migration inhibitory factor (MIF) was investigated. In the first step, human purified mononuclear leukocytes were activated by incubation with PHA for 1 h. Then the cells were washed thoroughly and cultured for 1 to 4 days in TC medium without PHA. In the second step, the MIF-activity of the cell-free culture supernatants was tested on non-separated human leukocytes. The migration inhibition reaction was demonstrated by the agarose migration technique. The supernatants might have been contaminated by PHA, but it was shown that such contamination would be unable to cause any appreciable migration inhibition of the indicator leukocytes.

A study on the clinical application of a direct leukocyte migration test in chromium contact allergy.

A capillary tube leukocyte migration inhibition assay has been adopted as an in vitro method for the demonstration of chromium hypersensitivity in twenty-two subjects with clinically proven or suspected chromium allergy. Two complexes of trivalent chromium and human serum albumin, exerting different migration inhibitory effects, have been prepared and used as the antigen. In the presence of the chromium-albumin complex with strong inhibitory activity, sensitivity to chromium was demonstrated independent of the clinical condition of the skin in all the patch test positive subjects. An additional positive response to the second chromium-albumin complex was observed only in those patients who were clinically in a state of exacerbation of an allergic contact dermatitis in which chromium allergy was an active causative factor. The results were not influenced by skin allergic reactivity to compounds other than chromium and the method was found to be of practical clinical value for diagnosing chromium allergy.

Absence of leukocyte-migration inhibitory factor in serum from patients with mycosis fungoides.

Sera from 16 patients with mycosis fungoides at various stages of the disease were investigated for the presence of a leukocyte-migration inhibitory factor. The agarose gel techinque with human peripheral blood leukocytes as migrating indicator cells was used for the purpose. The migration inhibitory effect of patients sera, pooled AB-sera and sera from control persons was tested on peripheral blood leukocytes from patients and controls. Also upconcentrated sera were used. The presence of substances with migration inhibitory effect in serum from patients with mycosis fungoides could not be demonstrated. The significance of these findings is discussed.

Enhancement of migration inhibitory factor activity by plasma esterase inhibitors

The plasma esterase inhibitors alpha2-macroglobulin, alpha1-antitrypsin, C1-inhibitor, antithrombin-heparin cofactor, and, as previously described, soybean trypsin inhibitor (Kunitz) and diisopropylphosphorofluoridate (9) enhance the response of guinea pig macrophages to migration inhibitory factor. To obtain this effect, macrophages are incubated with inhibitors prior to assay. The data suggest that (a) the enhancement of migration inhibitory factor response is due to the inhibition of esterases associated with the macrophage through a distinct active site on the inhibitors, and (b) that the active sites of antithrombin-heparin cofactor and soybean trypsin inhibitor, which interact with the macrophage enzymes, are different from the active sites of these inhibitors which interact with thrombin and trypsin respectively. Chemical modification of the active site of antithrombin-heparin cofactor for thrombin and of soybean trypsin inhibitor for trypsin does not affect their capacity to enhance the migration inhibitory factor response. From studies with thrombin, it was known that antithrombin-heparin cofactor has a heparin binding site. Addition of heparin was found to prevent the migration inhibitory factor-enhancing effect of antithrombin-heparin cofactor. The present results suggest that plasma esterase inhibitors may play a regulatory role in the response of macrophages to mediators of cellular immunity.

The mechanism of action of soluble lymphocytic mediators: II. Modification of macrophage migration and migration inhibitory factor action by drugs, enzymes and cationic environment

The effect of a number of agents on the in vitro migration of guinea pig peritoneal macrophages and on the activity of macrophage migration inhibitory factor was studied. It was found that: 1. Dibutyryl cyclic adenosine monophosphate and theophylline cause pronounced inhibition of migration. Migration was not modified by cyclic guanosine monophosphate, noncyclic nucleotides or by prostaglandins E 1, E 2, and F 2α. Theophylline (10 −4 M), paradoxically, abolishes the migration inhibitory effect of migration inhibitory factor. 2. Migration was not influenced by subtoxic doses of actinomycin D, puromycin and cycloheximide, but macrophages escaped inhibition partially, when exposed to migration inhibitory factor in the presence of puromycin (1–2 μg/ml) and of cycloheximide (0.1 μg/ml). 3. The polyanions dextran sulfate and heparin promoted macrophage migration and dextran sulfate (40 μg/ml) partially abolished migration inhibitory factor action. 4. Treatment of macrophages with the proteolytic enzymes trypsin and pronase resulted in enhanced migration while exposure to neuraminidase and wheat germ lipase was without effect. 5. Macrophage motility was dependent upon the presence of serum and increased in proportion with raising serum concentrations. Migration inhibitory factor was inactive at high serum concentrations. 6. Macrophage migration was enhanced and migration inhibitory factor activity abolished in the absence of calcium and magnesium.

Migration-inhibitory effect of serum from patients with prostatic cancer.

Serum from patients with carcinoma of the prostate were observed to possess a factor(s) capable of inhibiting the migration of allogeneic leukocytes in the direct leukocyte migration test. Maximum inhibitory activity was obtained with those sera possessing an elevated level of an as yet unidentified factor(s) in the alpha2-globulin fraction of the serum of these patients. The presence of such a factor(s) may prevent interaction between leukocyte-inhibiting factor sensitive cells and tumour cells and emphasizes the role ofhumoral inhibitory or 'immunoregulatory' factors, such as alpha2-globulin, on immunologicresponses as blastogenesis and inhibition of leukocyte migration, as potential abrogators of mechanisms of host-resistance, e.g., immunologic surveillance.

Migration Inhibitory Effect of Cell-Free Supernatants from Tuberculin-Stimulated Cultures of Human Mononuclear Leukocytes Demonstrated by Two-Step Mif Agarose Assay

Abstract Agarose gel technique with human peripheral blood leukocytes as migrating indicator cells was used to demonstrate migration inhibition factor (MIF) in cell-free supernatants from cultures of tuberculin-stimulated human peripheral blood mononuclear leukocytes. In this two-step MIF agarose assay only a few hundred microliters of supernatant were used. MIF was produced if cells from Mantoux-positive persons were cultured with purified protein derivative of tuberculin (PPD), but also if the sensitive cells after preincubation with PPD were washed and cultured in medium without PPD. There was a good correlation between the migration inhibitory effect of supernatants from PPD-stimulated cultures and the Mantoux reaction. The two-step MIF agarose assay was more sensitive than the direct leukocyte migration agarose test and makes possible the testing of several pairs of supernatants and a standard of MIF activity on the same migrating indicator cell population.

Blocking of macrophage migration inhibitory factor action by microtubular disruptive drugs.

The microtubular disruptive drugs colchicine and vinblastine were found to have a strong enhancing effect on the <i>in vitro </i>migration of normal guinea pig peritoneal exudate macrophages. Both drugs were also found to protect macrophages from the migration inhibitory effect of supernatants of antigen-stimulated sensitized lymphocyte cultures. These findings suggest that colchicine and vinblastine may be of potential use as suppressors of the peripheral manifestations of delayed hypersensitivity.

Migration Inhibitory Effect of Cell-Free Supernatants from Mixed Human Lymphocyte Cultures

Abstract Cell-free supernatants from mixed human lymphocyte cultures inhibited the migration of non-separated human peripheral leucocytes and purified polymorphonuclear leucocytes. The maximum inhibitory effect was observed in 4-day cultures, but even after 2 days inhibitory effect was demonstrated. The migration inhibitory factor (MIF) from mixed lymphocyte cultures was non-dialyzable, 56°C heat-stable and non-sedimentable at 78,000 × G. MIF was produced also in mixed cultures of lymphocytes pretreated with mitomycin C although the inhibitory effect was diminished in most of these experiments. The migration inhibition was demonstrated with an agarose plate technique which permits examination of only a few hundred microliters of unconcentrated MIF-containing supernatant.

Cell-mediated tumor allograft immunity: in vitro transfer with RNA.

Specific inhibition of migration of spleen cells from C57B1/6J mice, which had rejected A/J sarcoma-1 tumors, occurred in the presence of A/J lymph mode antigens. The migration inhibitory effect was transferable to normal C57B1/6J spleen cells by RNA extracted from lymph nodes and spleens of immunized animals.