Expression Of Membrane Receptors Research Articles

Histiocytosis-X cells express C3b, C3d, and C3bi receptor (CR1, CR2, and CR3) antigens.

Histiocytosis-X cells in the skin and other tissues share, with Langerhans cells (LC) of the epidermis, the membrane expression of the T6 and HLA-DR antigens [1, 2, 4] and the presence, in their cytoplasm, of an ultrastructural marker, the Birbeck granule [5]. In view of these similarities, hisfiocytosis-X cells are, at present, considered to represent an authentic proliferation of LCs or of cells derived from LCs. Histiocytosis-X cells and LCs express membrane receptors for the Fc fragment of IgG and for cleavage fragments of C3, as is demonstrated by the ability of these cells in suspension to form rosettes with antibodyand C3-bearing cellular intermediates [3, 10, 131. Using monoclonal antibodies (Mcab) specific for the human C3b receptor (CR1), C3d receptor (CR2), and the e-chain of the C3bi receptor (CR3), we demonstrated, by indirect immunofluorescence (IF) and immunoperoxidase (IPo) techniques, the presence of antigenic determinants of the three types of C3 receptors on the membrane of histiocytosis-X cells in the skin. Skin biopsies were taken from a 9-month-old infant, who had originally been referred to our department with Letterer-Siwe disease [i]. Initially, the child had diffuse lymphadenopathies and a typical papular eruption predominantly localized on the trunk and scalp. He presented crusty lesions coalescing to large plaques in the lumbar region; the papules were erosive and infiltrated in the inguinal region. After 8 weeks of follow-up, a granular interstitial infiltrate involving both lungs was seen on a chest X-ray; radiographic

Electrophoretic and cytochemical characterization of alpha-naphthyl acetate esterases in acute myeloid leukemia: relationships with membrane receptor and monocyte-specific antigen expression

Alpha-naphthyl acetate esterases (ANAE) were examined by cytochemical and isoelectric focusing (IEF) techniques in 48 cases of acute myeloid leukemia that were classified by conventional morphological criteria. Four main types of ANAE isoenzyme patterns were found by IEF, and comparisons with the expression of membrane receptors (Fc-IgG and C3b) and monocyte-specific antigens (UCHM1, UCHALF, and E11) suggest relationships between ANAE isoenzyme synthesis and distinct myeloid maturational stages. The results further indicate that the blast cells of acute myelomonocytic leukemia (AMML) may represent an immature variant of monocytic leukemia (AMoL) and that morphological examination alone is inadequate in the assessment of monocytic differentiation in acute myeloid leukemias. Inhibition studies of cytochemical ANAE activity with sodium fluoride (NaF) show that the presence of NaF- sensitive or NaF-resistant ANAE enzymes is often unrelated to the diagnostic category of acute leukemia. The results of this study are examined in relation to current concepts of myeloid differentiation, and the application of these findings to the subclassification of acute myeloid leukemias is discussed.

Electrophoretic and Cytochemical Characterization of Alpha-Naphthyl Acetate Esterases in Acute Myeloid Leukemia: Relationships With Membrane Receptor and Monocyte-Specific Antigen Expression

Alpha-naphthyl acetate esterases (ANAE) were examined by cytochemical and isoelectric focusing (IEF) techniques in 48 cases of acute myeloid leukemia that were classified by conventional morphological criteria. Four main types of ANAE isoenzyme patterns were found by IEF, and comparisons with the expression of membrane receptors (Fc-IgG and C3b) and monocyte-specific antigens (UCHM1, UCHALF, and E11) suggest relationships between ANAE isoenzyme synthesis and distinct myeloid maturational stages. The results further indicate that the blast cells of acute myelomonocytic leukemia (AMML) may represent an immature variant of monocytic leukemia (AMoL) and that morphological examination alone is inadequate in the assessment of monocytic differentiation in acute myeloid leukemias. Inhibition studies of cytochemical ANAE activity with sodium fluoride (NaF) show that the presence of NaF-sensitive or NaF-resistant ANAE enzymes is often unrelated to the diagnostic category of acute leukemia. The results of this study are examined in relation to current concepts of myeloid differentiation, and the application of these findings to the subclassification of acute myeloid leukemias is discussed.

Clinical significance of membrane receptors in ovarian carcinoma

The expression of membrane receptors was investigated at different human ovarian carcinomas. It could be demonstrated, that no specific binding sites of the gonadotropin hCG and the prostaglandins PGE2 and PGF2 alpha were found in the investigated specimens. However, 30% of ovarian carcinomas had receptors for the growth factor EGF. The determination of binding parameters resulted in an affinity constant (KD) between 1-6 X 10(-9)M, the number of binding sites (Bmax) varied between 5-250 fmol/mg). Further parameters such as histology, grade of differentiation, steroid hormone receptor status, stage and tumor rest after surgery were similar in the EGF receptor-positive and -receptor-negative groups. But, the response to chemotherapy was significant higher in the EGF receptor-positive group. The comparison of median survival time resulted in a better prognosis for patients with EGF receptor-positive carcinomas. Further investigations will clarify, if the EGF receptor status can be taken as a prognostic marker in the schedule of treatment in patients with ovarian carcinomas.

Growth-dependent expression of a cell surface glycoprotein

The expression of the hepatocellular membrane receptor for desialylated galactose-termining glycoproteins was studied during different proliferative stages of a human hepatoma cell line. Rapidly growing cells exhibited a reduced endocytotic rate of desialylated orsomucoid as compared to non-growing cells. This reduction was shown to be the consequence of a lower concentration of active cell-surface associated receptor protein in the dividing cells.

Fusion and biochemical expression of membrane receptors in foreign living cells

Successful transplantation of cell surface molecules from the membranes of one cell type to recipient cells of a different type is described. Plasma membranes purified from donor cells were fluoresceinated and fused to recipient cells using poly(ethylene glycol) and the fate of the transplanted membrane components was followed by fluorescence microscopy. In approximately 100 min the ‘foreign’ membrane components were seen to cluster and internalise. During this time, judged by the criteria of hormonal stimulation and immune cytotoxic killing, the cell surface of the recipient cell mimicked the cell surface phenotype of the donor cell.

Modifications of granulocyte subpopulations and of their adherence property during filtration leukapheresis in man

Summary. Polymorphonuclear leucocytes (PMN) isolated from the peripheral blood of 23 blood donors undergoing filtration leukapheresis were examined with regard to their number, expression of membrane receptors for rabbit IgG or human C3b and their ability to adhere to nylon fibres. All the donors showed an early granulocytopenia followed by a gradual return of PMN into the circulation, but differed in the number of PMN appearing in the rebound phase. Evaluation of rosette forming cells (RFC) and cells adhering to nylon fibres in this phase led to the recognition of two main groups of donors. In one of these groups the number of PMN, RFC and adhering cells remained unchanged compared to the pre‐apheresis values. The other group showed marked leucocytosis accompanied by a relative increase in the number of non‐RFC and non‐adhering cells. Possible mechanisms involved in the changes of PMN subpopulations are discussed.

Modifications of granulocyte subpopulations and of their adherence property during filtration leukapheresis in man.

Polymorphonuclear leucocytes (PMN) isolated from the peripheral blood of 23 blood donors undergoing filtration leukapheresis were examined with regard to their number, expression of membrane receptors for rabbit IgG or human C3b and their ability to adhere to nylon fibres. All the donors showed an early granulocytopenia followed by a gradual return of PMN into the circulation, but differed in the number of PMN appearing in the rebound phase. Evaluation of rosette forming cells (RFC) and cells adhering to nylon fibres in this phase led to the recognition of two main groups of donors. In one of these groups the number of PMN, RFC and adhering cells remained unchanged compared to the pre-apheresis values. The other group showed marked leucocytosis accompanied by a relative increase in the number of non-RFC and non-adhering cells. Possible mechanisms involved in the changes of PMN subpopulations are discussed.

Asynchronous expression of granulocyte membrane receptors in megaloblastic anaemia.

The expression of Fc(IgG) and C3b membrane receptors by granulocytes and their precursors was examined in 23 cases of megaloblastic anaemia which were graded I-III according to morphological severity. Fractionated bone marrow and peripheral blood granulocyte receptors were assessed by rosette formation with optimally sensitized ox erythrocytes and the results compared with those found in 14 normal marrows. Promyelocyte Fc and C3b receptor activities in megaloblastic anaemia did not differ from normal whilst the number of Fc receptor positive myelocytes and later cells showed a significant increase (P less than 0.05 in Grade I, P less than 0.01 in Grades II and III) proportional to the severity of megaloblastosis. An increase in the number of C3b receptor positive granulocytes was seen in early megaloblastic anaemias and, in contrast to Fc receptor expression, showed the highest receptor activities in the Grade II cases. The most significant changes in receptor expression were seen at the metamyelocyte stage and appear to be related to the numbers of these cells found in the megaloblastic marrows. It is suggested that these alterations are primarily related to asynchronous granulocyte maturation and the application of these findings to the study of granulocytic disorders is discussed.

Immunologic evaluation of patients with polychlorinated biphenyl poisoning: Determination of phagocyte Fc and complement receptors

Monocyte and polymorphonuclear leukocyte (PMN) complement and immunoglobulin receptors were measured in peripheral blood samples obtained from 30 Yu-Cheng (PCB-poisoning) patients and 23 normal human subjects. Erythrocyte-antibody complexes (EA) and erythrocyte-antibody-complement complexes (EAC) were used to detect immunoglobulin and complement receptors respectively. Monocytes and PMNs obtained from patients had lower percentages of cells bearing immunoglobulin and complement receptors. The expression of membrane receptors is associated with cellular activities. Phagocytic cells are responsible for the elimination of the infectious microorganisms. The deficiency of phagocyte cell membrane receptors may be associated with defective resistance to infections found in Yu-Cheng patients.

Tumor-promoting phorbol esters induce macrophage differentiation from bone marrow precursors.

The tumor-promoting phorbol diester, 12-0-tetradecanoyl-phorbol-13-acetate (TPA), induces in liquid cultures of rat and mouse bone marrow cells a sequence of events strikingly similar to those initiated by colony-stimulating activity known to regulate growth. Changes include stimulation of DNA synthesis and induction of cell proliferation, enhancement of adherence to the substratum, increase in lysozyme secretion, the expression of plasma membrane receptors for the Fc portion of IgG and the capability of manifesting lymphokine-induced, immunologically nonspecific long-term cytotoxicity. These findings indicate that TPA selectively stimulates precursors of the mononuclear phagocyte lineage to proliferate and to differentiate into macrophages, thus mimicking the effects of macrophage colony-stimulating activity.

Multiple subsets of anti-tetanus toxoid antibody-producing cells in human peripheral blood differ by size, expression of membrane receptors, and mitogen reactivity.

Multiple subsets of anti-tetanus toxoid antibody-producing cells in human peripheral blood differ by size, expression of membrane receptors, and mitogen reactivity.

Defective Fc-mediated phagocytosis in C3H/HeJ macrophages. II. Correction by cAMP agonists.

Abstract Peritoneal macrophages from LPS hyporesponsive C3H/HeJ mice lose the capacity to bind and phagocytose opsonized sheep erythrocytes (EA) over a 48-hr culture period. This loss in Fc receptor capacity is markedly different from the progressive increase in phagocytic ability exhibited by cultured macrophages derived from LPS-responsive C3H/HeN mice. Since dibutyryl-cyclic adenosine monophosphate (DBcAMP) has previously been reported to modulate membrane receptor expression in lymphocytes and certain macrophage-like cell lines, we examined its effects on EA binding and phagocytosis by C3H/HeJ macrophages. DBcAMP not only reverses the binding defect in C3H/HeJ macrophages but also restores EA phagocytosis to the level of control C3H/HeN cultures. 8-Bromo-cAMP, as well as other agents known to elevate intracellular cAMP (i.e., isoproterenol plus isobutylmethylxanthine or prostaglandin E2) also corrected the phagocytic defect. Since the C3H/HeJ macrophage phagocytic defect can also be reversed by in vitro stimulation with a lymphokine-rich culture supernatant, we examined the effect of this treatment on intracellular cAMP levels. Lymphokine treatment produced a 60% increase in the levels of macrophage intracellular cAMP. These findings suggest that the C3H/HeJ differentiation defect may be secondary to some abnormality in a cAMP dependent pathway.

Receptors for cold-insoluble globulin (plasma fibronectin) on human monocytes.

This investigation focused on the role played by cold-insoluble globulin (CIg, plasma fibronectin) in monocyte function. Surface-bound CIg mediated a concentration-dependent of human blood monocytes to gelatin-coated surfaces. CIg also mediated the binding of gelatin-coated particles such as latex beads or tanned erythrocytes to surface-bound human monocytes. However, CIg did not mediate particle ingestion. Subfractionated CIg that was highly enriched in monomeric forms (zone II CIg, mol wt 190,000-235,000) was less effective than were fractions enriched in dimeric forms (zone I CIg, mol wt 450,000) in promoting monocyte attachment. Binding of CIg to a gelatin or plastic surface occurred in the absence of divalent cations, but monocyte attachment to CIg-coated surfaces required divalent cations, Mg++ being much more effective than Ca++. Cation-dependent cell attachment was reversible in that bound cells could be released by treatment with EDTA. Serum-mediated binding of monocytes to gelatin-coated plastic dishes was a result of its content of CIg because the binding activity was abolished by removal of CIg from serum, and could be restored by readdition of purified CIg. Treatment of monocytes with trypsin abolished subsequent cell attachment to CIg-gelatin surfaces or particles. Expression of certain other known monocyte membrane receptors (Fc and C3b) was markedly enhanced as a result of CIg-monocyte interaction. These several observations indicate that monocytes bear membrane receptors (termed receptor cold-insoluble globulin) for surface-bound CIg.

Surface Membrane Characteristics of Cells from Human Acute Monocytic Leukaemia

The clinical and laboratory features of 4 patients with acute monocytic leukaemia are described. Surface membrane receptor studies of these patients have shown increasing strength of receptor expression occurs with increasing morphological maturity of the monocyte. In addition, the study shows that expression of membrane receptors can vary from patient to patient and also with time and treatment in an individual patient. A further finding has been the loss of positivity with specific anti-monocyte serum when the disease becomes refractory to therapy.

Membrane receptors of mouse leukocytes. II. Sequential expression of membrane receptors and phagocytic capacity during leukocyte differentiation.

Analysis of four mature cell markers on mouse bone marrow leukocytes grown in vitro, demonstrated a distinct sequence of marker appearance during the terminal phases of granulocytic cell differentiation. A similar pattern of marker expression was also suggested by analysis of mature neutrophils and macrophages isolated from normal tissues. Among cultured neutrophils, receptors for the Fc portion of IgG (FcR) were first expressed on myelocytes and metamyelocytes, and then subsequently on more mature cells. Morphologically mature colony neutrophils (polymorphs) from agar cultures contained only FcR and complement receptor type two (CR(2)) (C3d receptor), and lacked both complement receptor type one (CR(1)) (C3b receptor) and the capacity to ingest latex, bacteria, or iron particles. Neutrophils from 2 and 3 wk liquid media cultures of marrow cells differed from agar grown neutrophils in that they had phagocytic capacity (particle ingestion) [Pi] in addition to FcR and CR(2). Furthermore, in the 4th and 5th wk of these continuous liquid cultures, CR(1) was also expressed, completing the surface marker profile of normal blood neutrophils. Based on these studies, the following order of appearance of these four markers on cells from the myelocytic series was proposed: FcR {arrow} FcR CR(2) {arrow} FcR CR(2) Pi {arrow} FcR CR(2) Pi CR(1). Differential studies of tissue leukocytes containing these same markers revealed that a heterogeneity existed among morphologically mature neutrophils. Even though 95 percent of blood polymorphs contained all four markers, the same was true of only half of spleen polymorphs and only 20 percent of bone marrow polymorphs. Cells of the monocyte-macrophage series were studies in parallel with neutrophils. Cultured marrow monocytes acquired the four mature cell markers so rapidly that the order of receptor appearance could not be determined. However, it was found that CR2 was lost during the terminal phase of monocyte maturation into activated macrophages.

Inhibition of Fc receptors on a murine lymphoid cell line by cholera exotoxin

AFTER binding to its receptor, Gml ganglioside1,2, cholera exotoxin has been demonstrated to be a potent activator of the adenyl cyclase system in several cell systems. The response to exotoxin is related to an increase in intracellular cyclic 3′, 5′ adenosine monophosphate (cyclic AMP). Agents that increase intracellular cyclic AMP have been shown to suppress certain immunological responses, including histamine release3, lysosome degranulation4, mitogenic stimulation5, T lymphocyte rosette formation6 and antibody-dependent cellular cytotoxicity7. We now report that cholera exotoxin can inhibit the expression of plasma membrane receptors for the Fc portion of IgG in a murine lymphoid cell system.

Receptors for IgG molecules on human lymphocytes forming spontaneous rosettes with sheep red cells.

A mixed rosette technique with sheep red blood cells (SRBC) and ox erythrocytes heavily coated with rabbit antibody (EA) was employed to simultaneously identify human peripheral blood T lymphocytes and IgG receptor-bearing cells. The findings of a noticeable proportion of "mixed rosettes" in peripheral blood lymphocytes freshly drawn from normal individuals and of an even higher number of these mixed rosettes in cell suspension kept in culture media supplemented with fetal calf serum provide evidence for the capacity of human T cells to express membrane receptors for antigen-antibody complexes.

Expression of adenylate cyclase, catecholamine receptor, and cyclic adenosine monophosphate-dependent protein kinase in synchronized culture of Chang's liver cells (S phase-membrane receptors-NaF stimulation).

In Chang's liver cells synchronized by exposure to thymidine, both cyclic adenosine monophosphate-dependent and cyclic adenosine monophosphate-dependent and cyclic adenosine monophosphate-independent protein kinase activities increased throughout the cell cycle in a manner that corresponded approximately to the overall increase in cellular protein. Thus, the catalytic and the cyclic adenosine monophosphate-binding regulatory subunits of protein kinase appear to be temporally coexpressed. Adenylate cyclase activity stimulated by catecholamine, as well as basal activity, decreased markedly after release of cells from thymidine blockade during the S phase of the cell cycle; this was followed by recovery and overall net increase to fully expressed activity by the peak of mitosis. After initial decrease early in the S phase, cyclase activity assayed in the presence of fluoride ion began to rise before the rise in basal and catecholamine-stimulated activities, indicating that regulation of membrane receptors for catecholamines and for basal activity may be at least in part separate from that for catalytic activity of adenylate cyclase. The differential expression of membrane receptors for adenylate cyclase during the Chang's liver cell cycle affords a possible mechanism for control of response to hormones during the cell cycle.

The preparation and properties of macrophage-L cell hybrids.

The plasma membrane of the mouse peritoneal macrophage has specific receptors which enable the cell to bind IgG or complement-coated sheep red cells and is also rich in a divalent cation-dependent adenosine triphosphatase (ATPase) activity. L cells lack these macrophage membrane markers. The question of macrophage membrane receptor expression was investigated in DBA/2 mouse macrophage x mouse LMTK(-) cell hybrids produced with the aid of Sendai virus. Three independent clones and one mass culture were isolated by their ability to grow in hypoxanthine, aminopterin, and thymidine (HAT) selection medium. These hybrids retained 85-100% of the sum of two parent cells' chromosomes and expressed several genes derived from both parents, including glucose phosphate isomerase isozymes and H-2 antigens. The hybrids displayed ATPase activity which was intermediate between that of the macrophage and L cell. The macrophage specific receptors for antibody or complement-coated red cells could not be demonstrated on hybrid cells. The selective absence of these receptors is probably because of a failure in gene expression rather than to loss of genes.