Roald Matre

Human microglial cells have phenotypic and functional characteristics in common with both macrophages and dendritic antigen-presenting cells.

Resting microglia comprise up to 13% of the cells in human central nervous system (CNS) white matter. Their large number and dendritic morphology make them ideally suited to survey the CNS for noxious stimuli. Upon activation microglia gradually lose dendritic processes and transform into typical phagocytic macrophages. Microglia have been implicated as the main antigen presenting cell within the CNS, and appear to be of central importance as effectors and regulators of demyelination. To further characterize the capacity for immune reactivity within the human CNS, we have studied several characteristics of microglia, both in situ and in vitro. We find that human microglia have ultrastructural, phenotypic (CD11c, CD68, acid phosphatase), and functional (FcR and CR mediated phagocytosis) properties typical for cells of the monocyte lineage. Our data indicate that microglia also have properties in common with dendritic antigen‐presenting cells. Electron microscopy studies show extended dendritic cell processes on cultured microglia, and microglia are, like dendritic cells, negative for the monocyte markers nonspecific esterase, endogenous peroxidase, CD14, and RFD7. Microglia constitutively express HLA‐DR in situ, and express the dendritic cell marker RFD1 upon activation. Coculturing of microglia with CD4+ T cells results in clustering of T cells around microglia and initiation of a mixed lymphocyte reaction, both distinguishing features of dendritic cells. These functional properties of microglia may be of importance for the maintenance of an immunologic response in the CNS, an organ where dendritic cells, in contrast to other organs, have not previously been identified. J. Leukoc. Biol. 56: 732–740; 1994.

Reactive microglia in multiple sclerosis lesions have an increased expression of receptors for the Fc part of IgG

Receptors for the Fc part of IgG, FcRI (CD64), FcRII (CD32), and FcRIII (CD16) were studied by indirect immunoperoxidase staining of cryostat sections from normal and multiple sclerosis (MS) brains. Microglia in the parenchyma of normal white matter had a dendritic morphology, and were weakly stained by monoclonal antibodies (mAbs) to FcRI, FcRII, and FcRIII. In active MS lesions reactive microglia were strongly stained by the mAbs 32.2 (FcRI), IV-3 (FcRII), and 3G8 (FcRIII). Perivascular macrophages were stained by all anti-FcR mAbs in both normal white matter and in MS lesions, whereas endothelial cells were stained by the anti-FcRIII mAb only. The FcR on microglia and perivascular macrophages may be of functional importance in antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, and local immunoregulation. FcR on endothelium may be of importance in binding and transportation of immune complexes into the CNS. FcR mediated functions may consequently be highly relevant to the pathogenesis of MS.

Fc receptors for IgG on cultured human microglia mediate cytotoxicity and phagocytosis of antibody-coated targets.

We have utilized surgically resected human central nervous system (CNS) tissue to determine the expression and functions of Fc receptors (FcyR) on individual cell types found within the CNS. We observed all three classes of FcyR on microglial cells in situ and in vitro, but not on astrocytes or oligodendrocytes. Incubation of cultured microglia with immune complexes (antibody-coated red blood cells) induced phagocytosis, antibody-dependent cell-mediated cytotoxicity (ADCC), and oxidative bursts. We also found that microglia have the capability to produce T cell stimulatory soluble mediators after FcyR crosslinking. These functional responses were enhanced by pretreatment of the microglia with interferon-y (IFN-y). Our results implicate microglial effector responses triggered by interaction of FcyR with opsonized antigens as potential mediators of tissue injury within the CNS. Such injury may be particularly applicable to multiple sclerosis, an inflammatory demyelinating disease characterized by intrathecal production of immunoglobulins and cytokines.

Complement‐regulatory proteins in ovarian malignancies

Ovarian cancer has features that makes it well‐suited for MAb adjuvant immunotherapy. Several of the MAbs used in clinical trials mediate cancer cell destruction by activation of complement (C). In this study, therefore, we examined the ability of ovarian‐tumor cells to resist C attack. We found that the C regulators membrane cofactor protein (MCP, CD46) and protectin (CD59) were strongly expressed in the tumor cells in all 28 benign and malignant tumors examined. Decay‐accelerating factor (DAF; CD55) was more heterogeneously expressed, and only 75% of the tumors exhibited a moderate amount of DAF in the tumor cells. In adenoma cells, CD59 and DAF were preferentially located apically, while in adenocarcinoma cells they were expressed also at the basolateral cell surface. The ovarian‐carcinoma cell lines SK‐OV‐3, Caov‐3, SW626 and PA‐1 expressed both the 58‐and the 68‐kDa isoforms of MCP. DAF was present as a glycosyl‐phosphatidylinositol(GP1)‐anchored 70‐kDa glycoprotein. The surface‐expression level of DAF varied, and correlated with the vulnerability of the cells to C‐mediated lysis. CD59 was expressed as a GPl‐linked 19‐to 25‐kDa protein exhibiting multiple glycosylation variants. The surface expression of CD59 correlated with the amount of the main 1.9 + 2.1‐kb CD59 mRNA transcripts. Neutralization of CD59 with an anti‐CD59 MAb significantly enhanced C‐mediated killing of the cell lines. Low expression of C regulators on the PA‐1 teratocarcinoma cell line was associated with high sensitivity to C lysis. Thus, the expression of C regulators on malignant ovarian cells may constitute a tumor escape mechanism, and is a critical parameter to be examined when MAb therapy is being considered.

Interferon in suction blister fluid from psoriatic lesions

Interferon (IFN) (at least 16 units/ml) was demonstrated in suction blister fluid obtained from lesional skin in nine of thirteen patients with psoriasis vulgaris and in two patients with allergic contact dermatitis, but not in blister fluid from unaffected skin. IFN was detected in only three of the sera from the patients with psoriasis. The difference in results between the blister fluid and the sera from these patients was statistically significant (P < 0·025). Evidence was obtained which indicated that the activity was probably IFN‐γ (immune IFN), but the additional presence of IFN‐α in some of the fluids could not be excluded. The data indicate that IFN is produced locally in the psoriatic lesions, most likely by activated T lymphocytes.

Associated disorders in myasthenia gravis: autoimmune diseases and their relation to thymectomy.

ABSTRACT— The prevalence of myasthenia gravis (MG) in the counties of Hordaland and Sogn & Fjordane on January 1, 1984 was 9.6 per 100,000 inhabitants. Other autoimmune diseases were found in 11 out of 48 MG patients. The occurrence of autoimmune thyroiditis (5 patients, 10.4%) and systemic lupus erythematosus (4 patients, 8.3%) in the MG patients was clearly higher than that reported in the general population. Rheumatoid arthritis was found in 2 patients (4.2%). The autoimmune diseases were mainly recorded among the nonthymectomized MG patients. In addition to those with definite diseases of autoimmune nature, 3 other MG patients had thyroid antibodies and 1 had antinuclear factor without clinical evidence of autoimmune disease. Seven MG patients (14.6%) had unspecific arthralgia during active periods of MG. Two MG patients had ankylosing spondylitis.

Ryanodine receptor antibodies related to severity of thymoma associated myasthenia gravis.

Ryanodine receptor (RyR) antibodies are detected in about 50% of patients with myasthenia gravis who have a thymoma. The RyR is a calcium release channel involved in the mechanism of excitation-contraction coupling in striated muscle. In this study the severity of myasthenia gravis assessed by a five point disability score was compared between 12 patients with myasthenia gravis, a thymoma, and RyR antibodies and 10 patients with myasthenia gravis and a thymoma but without such antibodies. Symptoms of myasthenia gravis were significantly more severe in patients with RyR antibodies. The mean (SD) disability scores were 3.7(0.5) in patients with antibodies and 2.7 (0.9) in those without at peak of illness, (p = 0.01) and 3.4(1.4) v 1.6(0.7) at the end of an average observation period of five years (p = 0.002). The number of deaths due to myasthenia gravis was five of 12 RyR antibody positive patients, and none of 10 RyR antibody negative patients (p = 0.04). RyR antibody levels correlated positively with severity of myasthenia gravis. The presence of circulating RyR antibodies seems to be associated with a severe form of thymoma associated myasthenia gravis.

In situ characterization of mononuclear cells in human dental periapical inflammatory lesions using monoclonal antibodies

Mononuclear cells in cryostat sections of human dental periapical inflammatory lesions were studied with the aid of murine monoclonal antibodies and with indirect immunofluorescence microscopy. T lymphocytes (OKT3-positive cells) made up a major part of the cells in the infiltrates. They were found mainly in clusters, although single cells were also seen. T helper cells (OKT4) were more numerous than suppressor/cytotoxic T cells (OKT8-positive cells), with a ratio of approximately 2:1. Langerhans cells (OKT6-positive cells) were not demonstrated: only a few scattered HNK 1-positive cells, probably natural killer cells, were detected. A large number of OKM1- and OKIa 1-positive cells were detected in the infiltrates. Their size and number varied considerably in the different areas of the sections. These cells are probably macrophages. Sheets of small OKIa 1-positive cells were also demonstrated, indicating the presence of B lymphocytes or activated T lymphocytes. The results indicate that immune reactions may be of importance in the pathogenesis of periapical inflammatory lesions.

Properative psychological variables predict immunological status in patients with operable breast cancer

In a prospective study of psychobiological responses in patients with operable breast cancer, psychological data were collected one day before surgery and immunological data one day before and seven days after surgery. Objectives: explore psychoimmunological correlates related to primary surgical treatment of women with operable breast cancer. Method: distress was assessed with Impact of Event Scale (IES), depression with Montgomery‐Aasberg Depression Rating Scale (MADRS), coping with Mental Adjustment to Cancer (MAC) scale, emotional suppression with Courtauld Emotional Control scale (CECS) and neuroticism with EPQ‐N. Number of lymphocytes and subsets of lymphocytes were analyzed using a flow‐cytometric method. Results: Intrusive anxiety and anxious preoccupation were statistically significant inversely correlated to number of lymphocytes, B, T total and T4 lymphocytes and depression to B and T4 lymphocytes using Bonferronis correction for multiple testing. Multivariate analyses including menopausal status, type of surgery and health behaviour variables demonstrated an independent, inverse effect of depression on total number of lymphocytes, T total and T4 lymphocytes 7 days after surgery. Pre‐post immune changes were influenced reversely by depression (decreased) and intrusion (increased). Conclusions: Psychological parameters have a statistically significant impact on the number of lymphocytes and subsets of lymphocytes.

Fc receptor for IgG (FcR) on rat microglia

Receptor for IgG (FcR) was demonstrated on rat microglia in vivo and in vitro by immunohistochemical staining with immune complexes of horseradish peroxidase (HRP) and rabbit IgG anti-HRP. Astrocytes, oligodendrocytes and neurons did not express FcR. Microglia in culture also showed FcR-mediated agglutination and phagocytosis of IgG-sensitized erythrocytes. A radiolabelled cDNA probe for rat FcRIII hybridized with a 1.4-kb RNA band in Northern blots prepared from total RNA from rat brain. FcRIII mRNA-positive cells in rat brain, presumably microglia, were demonstrated by in situ hybridization. FcR participates in the initiation of cytotoxic responses and of phagocytosis by microglia and is therefore likely to be important in mediating immune reactions in the brain.