Macrophage heterogeneity in man. A subpopulation of HLA-DR-bearing macrophages required for antigen-induced T cell activation also contains stimulators for autologous-reactive T cells.

Utilizing somatic cell hybridization, we have developed a monoclonal antibody that interacts only with cells of the monocyte/macrophage (M phi) line and not with other myeloid or lymphoid cells. This antibody detects a 120,000-dalton determinant present on 37 +/- 2.8% of the peripheral blood M phi from several (HLA-DR)-disparate individuals and only depicts a subpopulation (approximately 30%) of HLA-DR-bearing M phi from any single subject. Cytolytic removal of this subpopulation of HLA-DR-bearing cells markedly diminished antigen-induced T cell reactivity, a deficiency that can be reconstituted with autologous M phi but not with either their soluble products containing lymphocyte-activating factor or with intact HLA-DR-disparate M phi. Whereas M phi bearing both the 120,000-dalton determinant and HLA-DR serve as effective stimulators for autologous mixed lymphocyte reactions. M phi bearing only HLA-DR determinants do not. However, this latter population of M phi can stimulate proliferation among alloreactive T cells. These studies indicate that the Mac-120 monoclonal antibody detects a subpopulation of HLA-DR-bearing M phi that is required for the genetically restricted presentation of conventional antigen to reactive T cells. Within the M phi population, these Mac-120+ cells constitute the most effective stimulators for autologous mixed lymphocyte reactions.

Decreased monocyte shedding of the migration inhibitor soluble CD18 in alcoholic hepatitis.

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.

T-lymphocyte sensitization in Graves' and Hashimoto's diseases confirmed by an indirect migration inhibition factor test.

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.

Directional migration of leukocytes: their pathological roles in inflammation and strategies for development of anti-inflammatory therapies.

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.

Human T cell subsets differ in their ability to migrate in vitro and to produce T cell migration inhibitory factor.

The spontaneous migration in vitro, production of T cell migration inhibitory factor (TIF) and the response to TIF of OKT4+ and OKT8+ human T cell subsets were studied. The OKT4+ lymphocytes migrated far better than the OKT8+ cells although the movement of both subsets was comparably inhibited by TIF. The OKT8+ subset was found to be a major source of TIF, while OKT4+ cells were responsible for macrophage migration inhibitory factor (MIF) production. The implications of lymphokine production by OKT8+ cells for the regulation of inflammatory responses are discussed.

Hypersensitivity of delayed type in hypertensive patients.

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.

Improvement of impaired leukocyte migration inhibition by thymosin in patients with head and neck squamous carcinoma.

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.

Evaluation of blocking mechanisms against immunological responses in patients with laryngeal carcinoma.

The specific tumor-induced leukocyte inhibition factor (LIF) production in laryngeal cancer patients has been investigated before and after the removal of adherent cells in order to evaluate the existence of a suppressor activity; 20 patients served as subject. The LIF production, after challenging the lymphocytes with 3MKC1 autologous tumor extracts, was significant in 12 patients and showed a further significant increase after the removal of adherent cells. In 3 patients with no previous significant LIF production, there was a conversion to significance when the adherent cells were removed. The other patients did not show any significant variation. These data seem to suggest the existence of a suppressor activity exerted by adherent cells in laryngeal cancer patients on LIF production.

Changes in the glomerular capillary wall induced by lymphocyte products and serum of nephrotic patients.

This study demonstrated that a soluble factor derived from patients with nephrotic syndrome caused changes consistent with the development of proteinuria when infused into the renal artery of a rat. The supernatants of cultures of stimulated lymphocytes of patients with the nephrotic syndrome containing autologous sera were infused into the renal arteries of rats and caused patchy spreading of the foot processes of epithelial cells and reduction of charge of the glomerular basement membrane of rat kidneys. The equivalent supernatants from normal subjects did not produce these changes.

In vitro induction of chronic myeloid leukemia associated immune reactivity in normal human lymphocytes by xenogeneic immune RNA.

Peripheral blood lymphocytes from normal human donors were treated with immune RNA (IRNA) prepared from lymphoid tissues of guinea pigs immunized with chronic myeloid leukemia (CML) cells, normal leukocytes (WBCs) and normal bone marrow (BM) cells, in order to study whether IRNA can confer specific immunoreactivity on normal lymphocytes. The IRNA treated lymphocytes were further exposed to solubilized membrane antigens from CML cells, normal WBCs and BM cells in a criss-cross fashion. The migration inhibition factor produced by these lymphocytes was tested in an in vitro leukocyte migration inhibition assay using normal leukocytes. The results indicated that IRNA prepared from guinea pigs immunized with all the three cell types could transfer the immune reactivity to normal cells in response to the respective antigens. The WBC IRNA incubated lymphocytes did not react with BM cell and CML cell antigens. A potent transfer of specific reactivity was shown by CML IRNA. IRNA produced against BM cells and CML cells demonstrated considerable cross reactivity perhaps due to shared immature cell antigens.

Specific LIF production in laryngeal cancer patients: evidence of suppressor activity exerted by adherent cells.

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.

[Current views on the mechanism of immunological disorders in Hodgkin's disease]. / Sovremennye predstavleniia o mekhanizme immunologicheskikh narushenii pri limfogranulematoze.

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.

Leukocyte migration-inhibition responses of nonvaccinated and vaccinated heifers to experimental infection with Brucella abortus.

The leukocyte migration agarose technique was used to show the leukocyte migration-inhibition responses of nonvaccinated and vaccinated heifers to experimental infection with Brucella abortus. All heifers had increased leukocyte migration inhibition after exposure to B abortus. Nonvaccinated heifers which aborted had the highest responses. The responses of the vaccinated heifers were significantly (P less than 0.01) lower than those of nonvaccinated heifers. None of the vaccinated heifers aborted.

The effect of uraemic "middle-sized molecules" on T lymphocyte functions.

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.

Effect of haemo- and peritoneal dialysis on the cell-mediated immune response in chronic uraemia.

Lymphocytopenia, decreased spontaneous rosette formation, and a decreased T lymphocyte count have been found in patients with non-uraemic glomerulonephritis (71 cases) and in different stages of uraemia (68 cases). In chronic glomerulonephritis and in the early stage of uraemia, cell-mediated hypersensitivity (lymphocyte migration inhibition) to glomerular basement membrane (GBM) characteristic of glomerulonephritis could be demonstrated. Hypersensitivity disappeared in the terminal stage of uraemia indicating endogenous immunosuppression.

Leucocyte migration inhibition factor (L-MIF) in malnourished Nigerian children.

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).

[Measles cellular immunity in patients with multiple sclerosis]. / Korevoi kletochnyi immunitet u bolnykh rasseiannym sklerozom.

The micromethod of leukocyte migration inhibition test was used to study cellular immunity in patients with multiple sclerosis (MS) to measles virus antigens and some other infectious (vaccinia virus, tuberculin) and noninfectious (brain white matter extract) antigens. In MS patients the reaction to measles antigen was weaker than in the control group, while the reactivity to the brain white matter extract was increased. As for the responses to vaccinia virus and tuberculin, these two grous did not differ statistically. The population of MS patients under study was not homogeneous in the intensity of measles cellular immunity (MCI). In the early stages of multiple sclerosis, MCI indices did not differ from those in the control group. MCI was the weakest in those subjects developing MS early in life who showed rapid worsening of the clinical status. Besides, MCI values were age-related: in older age groups they were low both in MS patients and in control subjects.

[Effect of 3-hydroxyanthranilic acid-antigen on the migration of peripheral blood leukocytes in patients with bladder tumors]. / Vliianie 3-OAK-antigena na migratsiiu leikotsitov perifericheskoi krovi bol'nykh s opukholiami mochevogo puzyria.

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.

[Studies of an assay system for dialyzable leukocyte extracts (DLE)--influence of DLE on leukocyte migration inhibition test by HBsAg].

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.