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Objectives: During alcoholic hepatitis (AH) monocytes traverse the vascular boundaries and massively invade the liver. In principle, tissue extravasation can be limited through shedding of CD18 integrins from leukocytes, including monocytes. The soluble (s) product sCD18 conceals adhesion receptors on the endothelium, which reduces monocyte extravasation. In AH, monocytes are dysfunctional, but whether this involves their self-generated anti-migration is unknown. Our aim was, therefore, to investigate monocyte CD18 dynamics in AH., Methods: We studied 50 AH patients and 20 healthy controls. We measured monocyte expression and conformational activation of CD18, plasma (P)-sCD18, stimulated in vitro CD18 shedding and P-sCD18 in a short-term chronic-binge mouse model., Results: AH-derived monocytes had a 30-60% higher expression of active CD18 receptors (p < 0.01), but the sCD18 concentration per monocyte was reduced in vivo by 30% and in vitro by 120% (p < 0.01). Ethanol reduced the in vitro shedding of CD18 in the patients only. TNFα increased sCD18 concentration per monocyte, but less so in the patients (p < 0.04). P-sCD18 per monocyte was inversely related to disease severity. In early alcoholic liver disease, P-sCD18 was decreased in the mouse model., Conclusions: The monocyte CD18 integrins are highly activated in AH and the single monocyte shedding of CD18 was decreased favoring tissue extravasation. Alcohol in itself and altered monocyte responsiveness to TNFα may explain this lowered shedding., Translational Impact: The contribution of this mechanism to the excessive monocyte liver infiltration in AH should be further explored as it may serve as a potential therapeutic target to limit liver inflammation.

Directional migration of leukocytes is indispensable to innate immunity for host defense. However, recruitment of leukocytes to a site of tissue injury also constitutes a leading cause for inflammatory responses. Mechanistically, it involves a cascade of cellular events precisely regulated by temporal and spatial presentation of a repertoire of molecules in the migrating leukocytes and their surroundings (microenvironments). Here I will summarize the emerging evidence that has shed lights on the underlying molecular mechanism for directional migration of leukocytes, which has guided the therapeutical development for innovative anti-inflammatory medicines.

In the present study, camphor odor and intraperitoneal (i.p.) injection of cyclophosphamide (CY) were used as conditioned stimulus (CS) and unconditioned stimulus (US), respectively. In the unconditioned group, mice were exposed to camphor odor for 1 h followed by an i.p. injection of CY (75 mg/kg). On the next day, the above CS/US association trial session was repeated followed by smearing dinitrochlorobenzene (DNCB) on mouse abdominal skin for sensitizing the animal for delayed-type hypersensitivity (DTH) response. Five days after DNCB sensitization, mice were exposed to camphor odor (1 h), followed by an i.p. injection of CY, and then DNCB was smeared on the left ear of mice for the challenge of DTH response. Both the left/right ear weight ratio and the activity of leukocyte migration inhibitory factor (LMIF) were used as the index of DTH response, which was done 24 h after DNCB challenge. In the conditioned group, the treatment was the same as that in the unconditioned group, except that normal saline was injected on day 5 instead of CY. Furthermore, in order to analyze the mechanism of the conditioned response (CR), the mouse serum from the conditioned group (CR serum) was injected into normal mice 6 h prior to DNCB challenge. Results showed that in the conditioned group, left/right ear weight ratio and LMIF activity were statistically lower than that in the DTH group, and there was no difference between conditioned and unconditioned groups. Thus, an animal model of conditioned immunosuppressive response had been established. The results also showed that after CR serum was injected into normal mice, DTH response was also significantly suppressed. However, if CR serum was treated with dialysis (10,000 molecular weight cut-off), the suppressive effect of CR serum on DTH response disappeared. Taken together, the data suggested that a chemical compound(s) in serum, with a molecular weight less than 10,000, was important in mediating the conditioned immunosuppressive response. This may be a very important molecule(s) that could be very critical to our understanding of the interaction between the central nervous system and immune function., (Copyright 2000 S. Karger AG, Basel)

Leucocyte Migration Inhibition Factor (L-MIF) was measured in 41 children with marasmus, 19 with kwashiorkor, 5 with marasmic-kwashiorkor and 35 well-fed healthy children serving as controls. For L-MIF assay, two different antigens (live attenuated measles virus vaccine and diptheria pertussis tetanus (DPT) vaccine were used. Percentage migration indices obtained with the two antigens were significantly higher in the malnourished than in the well-fed healthy sex and age-matched controls (P < 0.01). The total serum protein and albumin concentrations were significantly reduced in the malnourished children compared with the controls (P < 0.01). Mean total leucocyte numbers were not significantly different in marasmic and marasmic-kwashiorkor children compared with the controls (P > 0.21).

Time-related changes in specific cell-mediated immunity (CMI) and in humoral immunity were monitored in 20 bulls, aging 12 to 16 months, in four groups, each consisting of 5 animals. Group I was experimentally and group III-naturally infected with BHV 1 virus, groups II and IV serving as control animals. Tests for CMI included delayed type hypersensitivity (DTH), leukocyte migration inhibitory factor (LMIF) and granulocyte migration inhibitor factor (GMIF-LIF-buffy coat leukocyte migration inhibitory factor) in the presence of BHV I antigen while humoral immunity was tested by serum induced neutralization of the virus (SN) and by estimation of serum IgG, IgG1, IgG2, IgM and IgA levels. Immunologic, virologic and clinical studies were performed two days before infection and 17 timed within 91 days after the infection in bulls of group I and II and 7 times within 42 days after developing the disease in bulls of groups III and IV. The results showed that positive CMI reactions appeared 3 to 4 weeks earlier than positive antibody titers in SN test. The antibodies belonged most probably to IgG1 and IgG2 subclasses.

The aim of this study is to demonstrate the transference of PPD hypersensibility in an in vitro model, with dialysable colostral leukocyte extract (DCLE) of PPD+ and PPD-mothers, through measurements of leukocyte migration inhibition factor activity (LIF) from blood obtained of the umbilical cord of newborns from PPD+ mothers. The results show that DCLE PPD+ incubated with leukocytes of newborns from PPD- mothers had inhibition of leukocyte migration compared with migration of leukocytes incubated with DCLE PPD-. These results suggest that in this in vitro model, DCLE transfers hypersensibility to PPD.

The agarose migration technique was used for demonstration of delayed-type hypersensitivity to arterial vessel wall antigens in patients suffering from chronic essential hypertension. By means of this technique, it was demonstrated that the migration indices from the hypertensive patients differed significantly from the normotensive control persons, P less than 0.005. The significant difference was abolished when anti-LIF was added to the migration tests. This means that a hypersensitivity of the delayed type had developed in the hypertensive patients and the results indicated that the hypersensitivity was an autoimmunity to arterial vessel wall antigens.

Varied manifestations of herpes simplex keratitis are postulated to be related to alterations or paucity of protective immune response to the virus. In this study lymphocyte subpopulations and macrophage inhibition factor (MIF) assay are investigated in herpes simplex keratitis. Active T-lymphocytes are detected to be significantly low in active keratitis as compared to the healed stage (P less than 0.001) and normal control subjects (P less than 0.01). Active T-cells are also lower in bilateral keratitis than in unilateral keratitis (P less than 0.01), and in stromal keratitis than in epithelial keratitis (P less than 0.05). Total E-rosette-forming cells and leucocyte migration inhibition factor (MIF) assay demonstrates significantly lower values in bilateral keratitis than in unilateral keratitis, and in stromal keratitis than in epithelial keratitis. On healing, total E-RFC, active T-cells and MIF values improved and are comparable to those found in normal subjects. Enumeration of absolute lymphocyte counts and EAC rosette-forming cells (B-lymphocyte cells) did not yield any differences. Our observations demonstrate paucity of cell-mediated immune response in stromal keratitis. More marked deficiency is demonstrable in bilateral keratitis. Manifestations of herpes simplex keratitis and their healing is observed to be relatable to the level of cell mediated immune response.

The leukocyte migration inhibition test reveals in vitro the presence of lymphocyte sensitivity and, consequently, of cell-mediated immunity, to a given antigen. Applied in a variety of immune and allergic cases it proved to be useful for the positive diagnosis of the disease and/or for the detection of cell-mediated immune deficiency. The results obtained recommend the leukocyte migration inhibition test in the clinical practice.

The effect of lactoferrin on the migration of human polymorphonuclear cells was investigated. High concentrations of lactoferrin (greater than 250 micrograms/ml) markedly inhibited the migration of granulocytes under agarose. This migration inhibition could not be neutralized by an antibody against leucocyte migration inhibitory factor (LIF), suggesting a direct effect of lactoferrin on the granulocytes. Low concentrations of lactoferrin were, however, indirectly capable of inhibiting neutrophil migration. The overnight culture of mononuclear leucocytes with low concentrations of lactoferrin (10 micrograms/ml) resulted in the release of granulocyte migration inhibiting factors in the cell free culture supernatant. Strong evidence indicating that the migration inhibiting factors were due to LIF, was obtained in experiments whereby the inhibitory activity could be completely neutralized by anti-LIF antibodies. The lactoferrin-mediated stimulation of LIF release by mononuclear leucocytes could be neutralized by an anti-lactoferrin serum, but not by an anti-albumin serum, whereas PPD-induced LIF release was not affected by either antiserum. These findings suggest that lactoferrin besides its well known anti-microbial properties, may also play a regulatory role in the migratory response of polymorphonuclear cells during inflammatory conditions.

Specific antitumor sensitization in patients with carcinoma of the stomach was revealed by means of the leukocyte migration inhibition test. After 24-hour preincubation at 4 degrees C in a serum-free medium the areactive leukocytes from patients with carcinoma of the stomach (stages III--IV) acquired the ability to react specifically to allogeneic antigens of the tumor of the same localization. Preincubation did not influence the inhibition of migration of leukocytes in non-tumor patients by stomach carcinoma antigens. The supernatant of preincubated leukocytes of tumor patients contained substances that inhibited migration of indicator leukocytes.

Jejunal biopsies from controls and coeliac patients were maintained in organ culture in the presence of gluten fraction III. The culture media were assayed for evidence of lymphokine activity in a migration inhibition test using normal peripheral blood leucocytes. Significant inhibition of migration was produced by media from untreated coeliac patients compared with controls (P less than 0.005) or treated coeliac patients (P less than 0.001), indicating the production of a leucocyte migration inhibition factor (LIF) by untreated coeliac mucosa in response to gluten fraction III. The degree of inhibition correlated with the preculture interepithelial lymphocyte count in the coeliac biopsies (P less than 0.02). In six coeliac patients studied when on a normal diet and on a gluten-free diet, LIF was produced while on a normal diet, but not while on a gluten-free diet. These results suggest that a local cell-mediated immune reaction to gluten is present in the mucosa of patients with untreated coeliac disease but that this is reversed by treatment with a gluten-free diet.

We examined neonatal lymphocyte production of the lymphokine, lymphocyte-derived chemotactic factor (LDCF). Supernatants from 1 neonatal lymphocyte cultures were paired with the supernatants from 10 adult lymphocyte cultures. Chemotactic activity was defined as the number of adult monocytes migrating toward phytohemagglutinin-stimulated supernatants minus the number of monocytes migrating toward nonstimulated control supernatants using a blind well chamber assay. Eight of the 10 neonatal lymphocyte cultures showed LDCF and five of the 10 adult lymphocyte cultures showed LDCF activity. The mean number of monocytes migrating toward neonatal supernatants was 13.0 +/- 1.5 and toward adult supernatants was 14.1 +/- 2.1. To determine if a quantitative difference in LDCF production did exist, six additional experiments were performed assaying multiple dilutions of supernatants. No evidence was found for a quantitative difference in neonatal LDCF production compared to adult production. Our studies show that neonatal mononuclear cells are functionally as competent as adult mononuclear cells to produce LDCF in response to a mitogen challenge.

Immunogenetic studies in rheumatoid arthritis have demonstrated an increased frequency of the HLA-DR4 alloantigen in individuals with both the sporadic and familial forms of the disease. To investigate the significance of this association, we ascertained whether the possession of DR4 is necessary for the expression of cellular and humoral immunity to native human type II collagen. Our results indicate that the presence of DR4 is not required for the development of autoimmunity to collagen in rheumatoid arthritis.

A cell migration inhibitory effect was evidentiated in normal guinea pig serum as compared with heat inactivated serum. Granulocytes when used as target showed a greater sensitivity to this effect than lymphomonocytes or macrophages. The migration inhibitory activity of GPS is abrogated or decreased by using complement destroying agents such as: heating at 56 degrees C for as little as 5 min, absorption on immune complexes in presence of calcium, on zymosan, on Sephadex G-50 or by adding EDTA or heparin to culture medium. The GPS dialysation fractions while exhibiting neither complement haemolytic effect nor migration inhibitory activity when tested alone, restored these functions by recombination. Absorption of GPS on homologous blood cells abrogated the migration inhibitory effect but retained the complement haemolytic function. When GPS absorbed on homologous blood cells mixed 1:5 with heat-inactivated serum (5 min at 56 degrees C), the migration inhibitory activity was regained, suggesting that the complement factors from the first sample were necessary for manifestation for the migration inhibitory activity from the heat-inactivated serum.

T-Lymphocyte sensitization in Graves' disease (GD) and Hashimoto's thyroiditis (HT) was studied by an indirect migration inhibition factor test using normal T-lymphocytes as second stage indicator cells. In the first stage, mononuclear cells or T-lymphocytes, fractionated by the standard Ficoll-Hypaque procedure from the blood of patients with untreated GD and HT, were cultured in Eagle's medium containing thyroid antigen, and their cell-free supernatants were saved. Normal T-lymphocytes as second stage indicator cells were packed in capillary tubes and placed in planchettes with the above supernatants to complete the indirect migration inhibition factor test. Inhibition of the migration of indicator T-lymphocytes was demonstrated when either GD or HT culture supernatants were employed. Moreover, there was a good correlation between the indirect using the culture supernatants and the direct migration inhibition factor test using mononuclear cells or T-lymphocytes. On the other hand, in both direct and indirect migration inhibition factor tests using mononuclear cells and mononuclear cell culture supernatants, respectively, in the presence of human liver antigen as a nonspecific antigen, there was no significant difference between controls and patients. From these results, we can conclude that GD and HT T-lymphocytes are sensitized to thyroid antigen and produce the lymphokine, migration inhibition factor, into the supernatant when exposed to this antigen.

The specific tumor-induced LIF production in 30 laryngeal cancer patients has been investigated before and after the removal of adherent cells to evaluate the existence of a suppressor activity. LIF production, after challenging lymphocytes with 3 M KCI autologous tumor extracts, was significant in 16 patients and showed a further significant increase after removal of adherent cells. A conversion to significance when the adherent cells were removed was shown in 6 patients, with no previous significant LIF production. These data suggest the existence of a suppressor activity exerted by adherent cells on LIF production in laryngeal cancer patients.

Mice of several inbred strains have been fed diets containing either large amounts of zinc (300 ppm Zn), small amounts of zinc (5 ppm Zn), or routine laboratory mouse chow. When the mice are fed on a high-zinc diet, murine strains, such as C3H/HeJ, AKR/J, and CBA/CaJ, which are normally susceptible to infection with Candida albicans and which normally release low titers of migration-inhibition factor (MIF) in vivo into the circulation, become more resistant to infection with C. albicans and release higher titers of MIF in vivo into the circulation. In addition, their capacity to elicit delayed type hypersensitivity responses may be enhanced. When the mice are maintained on a low-zinc diet, murine strains, such as C57Bl/10SNJ, which are normally resistant to infection with C. albicans and which normally release high titers of MIF in vivo into the circulation on appropriate antigenic challenge, become more susceptible to infection and release lower titers of MIF into the circulation. Under these conditions of low-zinc concentrations in the diet, their capacity to elicit delayed type hypersensitivity may be reduced. Thus, the concentration of zinc in the diet may have a pronounced effect on some in vivo parameters of cell-mediated immunity.

A prospective study involving 20 volunteers was designed to test relationships between the sensitivity to zymosan and the indicators of nonspecific immunity against streptococci (inhibitory activity of blood and production of mediators). The immunization with Mannozym produced a conversion of zymosan-induced MIF with a subsequent improvement of immunity indicators in 4 out of 7 primovaccinated and 2 revaccinated subjects. This effect was detectable in some vaccines within 2-4 days after immunization and persisted for up to 5 months. Initially detected changes, their kinetics in the vaccinated subjects as well as time-related variations in the pattern of indicators during a one-year period of observation seem to suggest that there was a relationship between the natural and induced cellular sensitivity to zymosan and the inhibitory activity of blood. The inhibitory activity of blood was effectuated through enhanced mediator-induced phagocytic activity and nonspecific opsonin mechanisms. The mediators fluctuated in their values with time and were demonstrable in immunized subjects. There were observed interlinked reactions of cellular immunity with zymosan, peptidoglycan and streptococci. The mechanisms found are believed to constitute a fundamental, relatively easy-to-stimulate means of human immunity which may explain the experimental and epidemiological inconsistencies in the strictly type-specific interpretation of the M - anti-M system. Under in vitro conditions the effect of nonspecific immunization was bacteriostatic, but the role of these nonspecific mechanisms under conditions in vivo remains unclear. It is recommended that effectiveness of nonspecific immunization be verified under conditions of natural infection in man.

The activity of the leucocyte inhibitory factor (LIF) stimulated with Concanavalin A was examined in chronic uraemic, haemodialysed patients. It was found that the LIF activity decreased in chronic uraemic patients as compared with the normal controls. The absolute T lymphocyte count and active E rosette formation also decreased. In in vitro experiments the authors have also examined the inhibitory effect of the so-called "middle-sized molecule" (MSM) on uraemic materials separated from the serum and the haemofiltrate. They observed that the materials with a molecular weight of 1000-1500 isolated both from the serum and the haemofiltrate significantly inhibited the Con A stimulated LIF production of normal lymphocytes and decreased the ratio of the active E rosette-forming T lymphocytes. On the basis of their experiments the MSM uraemic materials are considered responsible for the decreased immune reactivity of uraemic patients. These uraemic toxins can be dialysed.

We have shown that hydrocortisone in physiological concentrations can inhibit the production of leukocyte migration inhibition factor (LMIF), but does not diminish the action of this lymphokine. Other agents tested failed to influence LMIF production. Inhibition of LMIF production by corticosteroids was influenced by the nature of the stimulus used for the production as an effect could be seen with PHA or Con A, but not Staphylococcal enterotoxin A (SEA). Production of LMIF was promptly restored after removal of the steroids. Furthermore, addition of a calcium ionophore to PHA restored the production of LMIF even in the presence of corticosteroids. In contrast, addition of exogenous IL-2 did not correct the defect in lymphokine secretion. We believe that inhibition of the production of LMIF by steroid may lead to defective granulocytic function and thus, predispose to infection.

In order to examine the suitability of leukocyte migration inhibition test (LMIT) in the capacity of in vitro assay system for dialyzable leukocyte extracts (DLE), the effect of DLE on hepatitis B and its antigen-specificity, the migration inhibitory activities to purified hepatitis B surface antigen (HBsAg) was measured using the leukocyte MIF test with DLEs obtained from HBsAb-positive or HBsAb-negative blood. The direct LMIT using agarose plate was modified according to the technique of Clausen et al. In spite of our assay system was dose-dependent for PPD, a significant response for purified HBsAg was not observed. However, some meaningful migration inhibition appeared when HBsAg and DLE were added simultaneously to the migration cells. From these results, it is concluded that DLE has antigen-specific and/or antigen non specific influences to the cell-mediated immunity for HBsAg Though some problems remain, we think our results are interesting, since the assay system for DLE has not been established and our study is closely related to the effect of DLE concerning hepatitis B.

In rheumatoid arthritis both lymphocyte-mediated and antibody-mediated immune reactions are important for the inflammatory lesions. In vivo activated B lymphocytes/plasma cells, T lymphocytes and monocytes/macrophages (Mo) are intimately involved in the disease process. Several clinical observations suggest an immunosuppressive action of gold salts. In humans, gold salts interfere with a number of Mo functions in vitro, including cellular interactions between Mo and T lymphocytes. Some workers have shown that the activation of human T lymphocytes is inhibited by gold salts, most probably secondary to an inhibition of Mo-T cell cooperation. Recent experiments indicate that gold salts also affect the in vitro differentiation of human B lymphocytes in response to polyclonal activators. Both the gold atom and the SH group seem to be important for the immunosuppressive actions of gold salts.

We have previously shown that the leukocytes of healthy EBV-seropositive (but not seronegative) donors respond with migration inhibition (LMI) when confronted with extracts of EBV-carrying (but not EBV negative) cells. In the present study, we have examined whether this EBV-specific LMI response is capable of detecting a membrane antigen on the surface of EBV-carrying virus nonproducer cells. Crude membranes from EBV-genome carrying and EBV-negative cell lines were used as antigen. Contamination with the EBV-determined nuclear antigen (EBNA) was ruled out. Membranes from EBV-genome carrying nonproducer cells inhibited the migration of leukocytes from healthy seropositive donors, whereas membranes from EBV-negative lines had no such effect. Seronegative donors did not show any LMI. The clear difference between the EBV-negative Ramos line and its EBV-converted sublines was particularly conclusive in showing that the membrane component is determined or induced by the viral genome.

Staphylococcal protein A is a bacterial cell wall product that binds human immunoglobulin G and thereby interferes with opsonization and phagocytosis of Staphylococcus aureus by neutrophils. Phagocytic cells are also responsive to various non-immunoglobulin lymphocyte mediators. We utilized the detection of a newly recognized mediator, a neutrophil migration inhibition factor from T-lymphocytes (NIF-T), to show that aggregates of staphylococcal protein A and immunoglobulins G could inhibit the responsiveness of neutrophils to NIF-T. That such aggregates may alter the responsiveness of neutrophils to lymphocyte mediators that amplify or modulate phagocytic functions may have important pathogenetic implications in staphylococcal infection.

In a preliminary study of the effects of SK-SD and thymosin on leukocyte migration inhibition in patients with squamous carcinoma of the head and neck, the cancer patients had significantly lower leukocyte migration inhibition of SK-SD than normal subjects. Thymosin increased the inhibition to SK-SD in the cancer patients to levels similar to those in normal subjects, and decreased the inhibition in normal subjects. These results confirm and extend the results of previous studies of the effects of thymosin in vitro, which show restoration of immune reactivity in patients with impaired cellular immunity and either no effect or a decrease in immune reactivity in subjects with normal cellular immunity. These combined observations provide a rationale for determining the clinical effects of thymosin in immunoincompetent patients with head and neck cancer and suggest that immunorestorative agents such as thymosin be used with caution in patients with normal cellular immunity.

Experimental allergic orchitis (EAO) was induced in rats following the injection of homologous testicular homogenate (THr) emulsified in Freund's complete adjuvant. Immune RNA (iRNA) was extracted from the spleen and lymph nodes. Normal guinea-pig peritoneal exudate cells (GP-PEC), when incubated in vitro with iRNA, were able to specifically recognise and respond to the immunising antigens (THr and PPD) as assessed by the direct migration inhibition reaction (MIR). Positive MIR's were also observed when incubated peritoneal exudate cells were tested against testicular homogenates from mice and guinea-pigs. This would confirm the presence of a common testicular antigen(s) in the three species studied. The reaction would appear to be organ-specific, kidney homogenate being unable to cause migration inhibition. GP-PEC incubated with iRNA which had been pretreated with RNAse did not show antigen-specific migration inhibition. Similarly, GP-PEC treated with RNA extracted from non-immunised donor rats did not respond.

Tests for the production of migration inhibitory factor by peripheral blood leukocytes in response to ubiquitous bacterial and fungal antigens were carried out in patients with untreated Graves' disease and in healthy control subjects. Dose-response studies, tests for the production of this factor after 72 hours' stimulation with phytohemagglutinin as a test for reserve, and tests before and after 24 hours' preculture to deplete suppressor cells were also performed in some patients. The antigens used were Candida, Trichophyton-Oidiomyces-Epidermophyton, mumps live attenuated virus and purified protein derivative of tuberculin. The production of migration inhibitory factor was measured by the agarose microdroplet method.The production of migration inhibitory factor in response to all the antigens except mumps virus was slightly greater in the patients than in the control subjects, although the differences were not significant. The dose-response characteristics and the production of migration inhibitory factor after stimulation with phytohemagglutinin were similar in the two groups. The production of migration inhibitory factor in response to suboptimal concentrations of Candida, Trichophyton-Oidiomyces-Epidermophyton and mumps virus was not enhanced in either group after 24 hours' preculture apart from a slight increase in response to mumps virus in the patients.These results fail to support the suggestion that patients with Graves' disease have a deficiency of suppressor cells.

Certain clinical and immunological symptoms detected in patients with lymphogranulomatoses are studied for their interrelation with immunological processes which occur in the lymphoid tissue affected by the lymphogranulomatosis. A hypothetic scheme of such an interrelation is presented.

By means of the leukocyte migration inhibition test a significant depression of migration index (indicating increased immunity) was found in 10 untreated coeliac patients compared with 24 control subjects with the dietary antigens bovine serum albumin (BSA) and egg white but not with milk. The degree of immunity was similar to that obtained with gluten fraction III as antigen. Treatment with a gluten-free diet led to a decrease in immunity to egg white and BSA, but immunity to gluten fraction III was increased in the early months of treatment.

Supernatants from cultured mouse and human tumour cells, but not mouse or guinea-pig fibroblasts, inhibited the production of a lymphokine, macrophage chemotactic factor, by PHA-stimulated mouse spleen cells. The supernatants affected spleen cells from old, but not young, mice. They were most active if added at the start of the spleen cell culture and did not act by binding phytohaemagglutinin (PHA). The active material had an approximate molecular weight, on membrane filtration, of 1000-10,000 and could be bound to and eluted from Con A-Sepharose. Tumour supernatant factor(s) of similar molecular weight inhibited the production of interleukin 1 (lymphocyte activating factor) in response to lipopolysaccharide by stimulated thioglycollate-induced peritoneal exudate macrophages, but not by Corynebacterium parvum-activated macrophages. Similar tumour-produced material has been found to inhibit the early phase of delayed-type hypersensitivity reactions in older mice. It is suggested that this effect is due, at least in part, to inhibition of interleukin 1 production leading to inhibition of lymphokine production.

Neutrophil migration inhibition factor from T lymphocytes (NIF-T) purified by antibody affinity chromatography and gel filtration chromatography was radioiodinated and identified as a 26,000-MW protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). NIF-T was identified by elution of biological activity from gel fractions and selective adsorption of a radioiodinated mediator by HL60 cells differentiated in the presence of dimethyl sulfoxide (DMSO) to develop receptors for NIF-T. A goat neutralizing antibody for NIF-T neutralized and immunoprecipitated migration inhibition activity in the conditioned medium from Mo T-lymphoblast cells and human peripheral blood lymphocytes (PBL) cultured with concanavalin A (Con A), but not from RPMI 1788 B-lymphoblast cells with leukocyte migration inhibitory factor (LIF) activity. These studies distinguish NIF-T both chemically and immunologically from LIF.

Natural, humoral and cellular immune mechanisms have all been implicated in the pathogenesis of chronic inflammatory periodontal disease. However, confusion still exists as to the role played by each of these immunological mechanisms. Recently, characterization of the cell types within the progressive lesion has been established, in which four recognizable zones were described. Immediately subjacent to the epithelium lining the periodontal pocket both polymorphonuclear leukocytes (PMN's) and macrophages were seen, while cells deeper in the tissues had the morphological appearance of lymphocytes. The majority of these lymphocytes had a B-cell phenotype although a few T-cells and macrophages were found. On the advancing front of the lesion the cells had the morphological appearance of plasma cells, the majority of which contained IgG. Other cells found in this region had the morphology of plasma cells yet contained no cytoplasmic immunoglobulin, but they did contain substantial amounts of lysosomal enzymes. Similar cells have previously been described in periodontal disease; their frequent association with fibroblasts may suggest that they are important in the pathogenesis. Deposits of IgG and fibrin were found in the fibrous tissue band surrounding the lesion. These results are reviewed and, although the zones described were not anatomically distinct, by describing the lesion in this way it was possible to establish a convenient model to explain the immunopathogenesis of progressive chronic inflammatory periodontal disease. In this respect, progressive chronic inflammatory periodontal disease in man should be considered as a B-cell lesion.

Immuno-reactivity to tumor extracts in patients with colorectal cancer was evaluated by the direct leukocyte migration inhibition assay. Hypertonic potassium chloride extracts were prepared from a group of synchronous colorectal cancers and from single colorectal cancers. Both groups of extracts were tested in patients with single colorectal cancers. All patients were tested preoperatively, none were previously treated, and none had distant metastases. Synchronous extracts showed no significant migration inhibition in allogeneic assays. Single colorectal cancers showed highly significant inhibition to autologous extracts only and not to allogeneic extracts. These data support the contention colorectal cancers are antigenically dissimilar.

The effect of 3-hydroxyanthranilic acid-antigen on peripheral blood leukocyte migration inhibition was studied in 75 patients with bladder malignancies. The antigen appeared to modify the said migration. A correlation was established between stage of tumor progression and the effect of 3-hydroxyanthranilic acid-antigen on leukocyte migration.