Release Of Migration Inhibition Factor Research Articles

Kinetics of migration inhibition factor release and macrophage response in experimental allergic contact dermatitis

Kinetics of migration inhibition factor release and macrophage response in experimental allergic contact dermatitis

Distinct alloantigens trigger proliferative or nonproliferative T lymphocyte activation in CBA/N, CBA/J, and C3H mice.

Lymphokine release and proliferation take place during one-way murine mixed leukocyte cultures (MLCs) when responder and stimulator cells differ in H-2 alloantigens, whereas a dissociation of these two functions can be observed where there are incompatibilities in non H-2 alloantigens, It is possible that this dissociation depends on different thresholds of activation of the two responses, or that some non H-2 alloantigens selectively activate lymphokine release or proliferation. To investigate this question we used H-2-matched CBA/N, CBA/J. C3H/HN, (CBA/N x CBA/J)F1, and (CBA/N x C3H/HN)F1 leukocytes, both as responder and stimulator cells. CBA/N mice and the male F1 hybrids obtained from CBA/N mothers carry an X-linked immune defect that results in an arrest of B lymphocyte maturation. In the majority of MLC combinations tested, migration inhibition factor (MIF) release and proliferation took place in parallel. However, CBA/N, but not F1 leukocytes, stimulated MIF release and not proliferation by CBA/J responders, whereas C3H/HN leukocytes stimulated proliferative responses but not MIF release by CBA/J responders. Since proliferation and MIF release have a similar threshold of activation, their dissociation indicates that different non-H-2 alloantigens can specifically activate distinct T cell functions. Moreover, previously unsuspected alloantigen differences between CBA/N and CBA/J are revealed by MIF release.

Demonstration of accelerated and increased migration inhibition factor release in vivo in a PPD retest reaction.

Sites at which positive skin-test reactions have been elicited exhibit an accelerated (so-called 'retest') reaction upon local reinjection of the antigen used for primary skin testing. When using a peritoneal cavity inflammation model in guinea pigs, local migration inhibition factor (MIF) release in vivo was studied in both primary and retest reactions to purified protein derivative. Increased retest reactivity could be demonstrated to coincide with an instantaneous and intense local MIF release. Estimation of both the absolute number of the antigen specificity of lymphocytes present in the peritoneal cavity at the time of retesting (170 h after primary testing) showed that the early burst of MIF release is primarily due to nonspecific local retention of increased numbers of lymphocytes.

Immunogenic fractions of Cryptococcus neoformans.

Cryptococcus neoformans cell and culture supernatant extracts were fractionated by ion exchange and gel filtration column chromatography. Various fractions were used to immunize mice, and to assess the release of migration inhibition factor, delayed type hypersensitivity, and protective immunity after challenge with C. neoformans. Results suggest that the C. neoformans fractions, which protect mice, contain a high molecular weight, predominantly carbohydrate antigen that can be distinguished from the capsular polysaccharide.

Suppressor T lymphocyte dysfunction in Graves' disease: role of the H-2 histamine receptor-bearing suppressor T lymphocytes.

The allo-suppressor effect of normal T lymphocytes on the production of migration inhibition factor by sensitized T lymphocytes of Graves' disease in response to human thyroid antigen has been studied further by a modified migration inhibition factor test employing purified T lymphocyte preparations. The production of migration inhibition factor was consistently abolished when normal T lymphocytes were mixed with the Graves' disease lymphocytes in various ratios (1:9, 2:8, and 5:5). However, pretreatment of the normal T lymphocytes with cimetidine (an H-2 histamine receptor antagonist) led to a demonstrable loss in their allo-suppressor properties, whereas pretreatment with chlorpheniramine (an H-1 histamine receptor antagonist) had no such effect. These studies indicate that a subset of normal T lymphocytes bearing H-2 histamine receptors suppresses the production or release of migration inhibition factor by sensitized T lymphocytes, and further suggest the possibility that there may be an abnormality in the H-2 receptors on Graves' disease suppressor T lymphocytes. It is conceivable that this defect is fundamental in the pathogenesis of Graves' disease.

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.

Granulocyte colony stimulating activity from lymphocytes: Separation from lymphokines by cytochalasin B

Lymphocytes from murine spleens released granulocyte colony stimulating activity (CSA), macrophage migration inhibition factor and lymphotoxin from 24–96 hr after stimulation with phytohemagglutinin (PHA). Cytochalasin B at 5 μg/ml completely inhibited the release of migration inhibition factor and lymphotoxin but significantly increased the release of CSA.

Two mechanisms of migration inhibition factor induction by tumour antigens

EVIDENCE has accumulated which stresses the importance of lymphokines in cell-mediated immunity both in vivo and in vitro1. But, the processes leading to release of lymphokines by antigen-activated lymphocytes, the mode of action of these lymphokines and their exact role in cell-mediated immunity are still not well defined. It has, however, been recognised that in vitro activation of immune lymphocytes by soluble antigens is macrophage dependent and that this activation is under the control of gene products of the major histocompatibility complex (H–2 in the mouse)2. Using a Moloney murine sarcoma virus (MSV)-induced tumour system, we have shown, in agreement with reports of some other systems3,4, that immune T lymphocytes required macrophages for migration inhibition factor (MIF) production and, in addition, only histocompatible macrophages could assist immune T lymphocytes for MIF release after stimulation with soluble tumour-associated antigens5. We report here that gene products of the H–2 complex regulated the macrophage-immune lymphocyte interaction for MIF release when soluble tumour extracts were used as the source of antigen. In contrast, when intact tumour cells were used as the source of antigen, a macrophage requirement was not detectable. Moreover, the direct lymphocyte-tumour cell interaction for MIF release was not under H–2 restriction, since allogenic as well as syngeneic tumour cells could activate MIF release from immune lymphocytes. These findings suggest the existence of at least two different pathways for MIF production, reflecting perhaps the activation of two distinct sub-populations of T lymphocytes or the activation of T lymphocytes at two different stages of differentiation.

Antigen Handling by Guinea Pig Macrophages: Further Evidence for the Sequestration of Antigen Relevant for Activation of Primed T Lymphocytes

Guinea pig macrophages can take up sufficient 2,4 dinitrophenyl guinea pig albumin during a brief in vitro exposure at 37 degrees C to trigger proliferation and lymphokine production with primed T lymphocytes on subsequent co-culture. Treatment of such antigen-bearing macrophages with trypsin, a procedure which removes surface antigen, does not alter the ability of such macrophage to initiate the release of migration inhibition factor from sensitized T lymphocytes. In addition, formation of antigen-specific rosettes between primed T cells and antigen-bearing macrophages is not blocked by high concentrations of antibody directed against the antigen mediating this interaction. Similarly, primed T lymphocyte DNA synthesis induced by antigen-bearing macrophages is not inhibited by specific antibody to that antigen. These data support the conclusion that the fraction of macrophage-associated antigen which is relevant to T lymphocyte activation does not reside on the macrophage surface but rather remains in a restricted compartment from which it is accessible to the T cell but unavailable to either blockade by specific antibody or removal by proteolytic enzymes.

The immunosuppressive mechanism of azathioprine. I. In vitro effect on lymphocyte function in the baboon.

The effect of azathioprine on in vitro baboon lymphocyte function tests was evaluated using the mitogen stimulation test, the mixed lymphocyte culture test, the migration inhibition factor test, the cell-mediated lymphocytotoxicity test and the antibody dependent cell-mediated cytotoxicity test. It was found that azathioprine inhibited phytohemagglutinin and Concanavalin A stimulation at lower concentrations than those required to inhibit pokeweed mitogen stimulation. It inhibited the MLC reaction with as little as 0.2 mug/culture in the microculture system. Azathioprine had no effect on (a) the release of migration inhibition factor, (b) the cell-mediated lymphocytotoxicity assay if presensitized cells were used, and (c) the antibody-dependent cell-mediated cytotoxicity assay. However, azathioprine inhibited CML if it was added during in vitro sensitization and induction of killer cells. These in vitro results suggest that azathioprine inhibits those reactions which require cellular division.

Production of migration inhibition factor (MIF) by human established B type cell lines derived from normal and malignant tissues: Studies of some factors affecting MIF release

Twenty-five established human lymphoid cell lines, initiated from peripheral blood leukocytes of normal donors or patients with various hemopoietic diseases, were found to release MIF into their culture fluid. These cells were also characterized by different rosette test procedures and by the presence of immunoglobulins. The percentage of EAC rosette-forming cells ranged from 30 to 90% and immunoglobulin bearing cells from 50 to 80%. No spontaneous rosettes were ever detected. Kinetic studies performed from the onset of the culture have shown that spontaneous MIF production is closely related to the establishment phenomenon. MIF activity detection in established cultures depends on various in vitro parameters including the cell growth pattern and the target cell used for the migration inhibition test. The significance of MIF production by permanent cell lines bearing B surface markers is discussed.

VARIABLE PHENOTYPIC EXPRESSION IN A FAMILY WITH “CANDIDA ENDOCRINOPATHY SYNDROME”

The Candida endocrinopathy syndrome (CES), an autosomal recessive disorder, has been associated with endocrinopathies and a variety of cell mediated immune defects (CMI). The cell mediated immune responses of a family of 9 members, 3 of which are affected with CES have been investigated by 1) in vivo skin reactivity to Candida extract (CE), phytohemagglutinin (PHA) and dinitrochlorobenzene (DNCB); 2) T and B cell resetting; 3) in vitro lymphocyte response to PHA and CE; 4) release of migration inhibition factor (MIF) in response to PHA and CE; 5) the effect of patients' sera on the in vitro response of normal lymphocytes to CE. Results are summarized as followsNo abnormal CMI responses were detected in other non-affected members. All the affected individuals had defects in CMI functions. Although it has been reported that family members with CES demonstrate the same defects in CMI, we have demonstrated a variety of defects in this sibship, indicating variable expressivity of the gene involved.

Bronchial lymphoid tissue.

The role of the respiratory tract in the defense against potential pathogens has been a subject of interest to microbiologists, virologists and immunologists for many years. The description of the secretory IgA system, common to mucosal surfaces throughout the body, has extended the interests of immunologists in the mechanisms of mucosal resistance (1). Like the gastrointestinal tract, the respiratory tract is characterized by the predominance of IgA containing cells in the lamina propria, by the presence of secretory IgA in bronchial and nasal secretions, and the demonstration of secretory component in the glandular epithelia. Further, the demonstration of local antibody, mainly of the IgA class, following immunization by nose drops or aerosol with a variety of antigens has shown that this system must play a major role in immunological responses to local antigen (2,3). Resistance to infection by certain organisms has been correlated with the presence of some local antibody and a lack of correlation exists between resistance and serum antibody. That immunity to infection in the respiratory tract is not only mediated by humoral antibody responses has been clearly shown in experiments of local immunization in animals and man in which specific release of migration inhibition factor has been found in lymphocytes from bronchial washings (4,5).

Autoimmune thyroiditis, adrenalitis and oophoritis

Described here is a young woman suffering from autoimmune thyroiditis, adrenalitis and oophoritis. This patient was carefully investigated by endocrine studies, with humoral antibodies to thyroid and adrenal, and the release of migration inhibition factor by her lymphocytes when cultured with thyroid, adrenal and ovarian antigens. Cell-mediated immunity appears to be the most important factor in the pathogenesis of these closely related disorders. A discussion of the interrelationship of these organ-specific autoimmune endocrine gland disorders is presented.

Mechanisms of Immunosuppression: Effects of Cyclophosphamide on Cellular Immunity

Abstract A course of cyclophosphamide administered to guinea pigs after immunization with mycobacteria led to a transient inhibition of cutaneous hypersensitivity to tuberculin protein. This treatment resulted in generalized depletion of lymphoid cells, including lymphopenia, and a substantial reduction in the number of macrophages found in induced peritoneal exudates. Results of in vivo cell transfer studies indicated that cyclophosphamide-treated recipients were rendered transiently unresponsive to purified tuberculin protein (PPD). However, sensitized lymph node cells from drug-treated donors were capable of transferring immunity to normal recipients. In vitro tests of cellular immunity with lymph node cells from drug-treated animals yielded discordant results. The proliferative response to both PHA and PPD was significantly impaired. However, these lymphoid cells functioned normally in assays for migration inhibition factor (MIF). Thus, these data suggest that cyclophosphamide can limit proliferation without impairing intermitotic functions of sensitized lymphocytes, such as the release of MIF. Results of this study suggest that cyclophosphamide does not impair development of a population of specifically sensitized T-type lymphocytes. However, several components of the expression phase are affected; the resultant anergy is probably due to a summation of effects on lymphocytes and macrophages.

Staphylococcal enterotoxin B induced release of macrophage migration inhibition factor from normal lymphocytes

Staphylococcal enterotoxin B (SEB), a potent lymphocyte mitogen, inhibits migration of peritoneal exudate cells from most guinea pigs but does not inhibit migration of purified macrophages. Experiments were designed to test the ability of highly purified SEB to induce normal lymphocytes to release migration inhibition factor (MIF). Supernatants of lymph node lymphocytes cultured with SEB inhibited the migration of purified macrophages, indicating the release of a migration inhibition factor. Mitomycin-C blocked the SEB-induced release of MIF. SEB-induced MIF localized in the albumin fraction on Sephadex G-200 chromatography. Antibody to SEB specifically blocked the inhibitory effect of SEB on migration of normal guinea pig peritoneal exudate cells.