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HIGH-RESOLUTION ELECTROPHORESIS FOR THE ALLOTYPING OF HUMAN C-4 - PROPOSAL OF A RELATIONAL NOMENCLATURE

JOURNAL OF IMMUNOLOGICAL METHODS 1987年第1期96卷 69-76页

作者： KOLBLE, K RUKAVINA, V KALDEN, JR Institut und Poliklinik für Klinische Immunologie und Rheumatologie Universität Erlangen-Nürnberg Krankenhausstr. 12 D-8520 Erlangen F.R.G.

Among the major histocompatibility complex (MHC)-linked complement genes, the loci for C4A and C4B exhibit the most extensive structural polymorphisms. Therefore the differentiation of variant and complex C4 phenotypes often proves difficult in conventional immunofixation electrophoresis. To improve the available technique of C4 typing a closed horizontal electrophoresis system was combined with poly- and monoclonal alkaline phosphatase immunoprobing on contact diffusion blots. The high resolution and sensitivity of this method not only facilitated C4 allotyping but also revealed additional polymorphic variation. Relative electrophoretic mobilities specific for each C4 allotype were established by computerized remission densitometry and provided the basis for a quantitative denomination of C4 variants. Typing by high resolution electrophoresis and the proposed relational C4 nomenclature could be valuable for further immunogenetical studies of the C4 protein polymorphism.

关键词： C4 polymorphism High resolution electrophoresis Contact diffusion blotting Immunoprobing C4 nomenclature BCIP bromo-chloro-indolyl phosphate BF factor B of the alternative complement pathway C3 third component of complement C4 fourth component of complement EA cells sheep erythrocytes reacted with specific rabbit antibody EDTA ethylenediaminetetraacetic acid HLA human leukocyte antigen ISGN International System for Human Gene Nomenclature MHC major histocompatibility complex MoAb monoclonal antibody M r relative electrophoretic mobility NBT nitro blue tetrazolium SD standard deviation SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis SEM standard error of the mean

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CALCIUM-DEPENDENT AGGREGATION OF HUMAN-SERUM AMYLOID-P component

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BIOCHIMICA ET BIOPHYSICA ACTA 1982年第2期701卷 229-236页

作者： BALTZ, ML DEBEER, FC FEINSTEIN, A PEPYS, MB ROYAL POSTGRAD MED SCH DEPT MED IMMUNOL MED UNIT LONDON W12 0HS ENGLAND ARC INST ANIM PHYSIOL DEPT IMMUNOL CAMBRIDGE CB2 4AT ENGLAND

Normal human serum or isolated human serum amyloid P component was ultracentrifuged on density gradients containing either 10 mM EDTA or different concentrations of Ca2+ between 0.15-2.15 mM. In the presence of Ca2+ concentrations of 1 mM or more human P component sedimented more rapidly than it did in the presence of lower Ca2+ levels or of EDTA. This phenomenon was due to Ca2+-dependent aggregation of P component molecules and did not require the presence of any other serum constituents. It was completely inhibited by incorporating a physiological concentration (40 mg/ml) of serum albumin in the gradients, suggesting that free ionized Ca2+ is required to promote aggregation of the P component. P component from the mouse and the plaice (Pleuronectes platessa L.), a marine teleost, did not undergo the same Ca2+-dependent aggregation as human P component. These observations resolve a discrepancy existing in the literature concerning the sedimentation rate of human P component in density gradient ultracentrifugation and shed new light on its behavior with respect to Ca2+ which may be relevant to the deposition of P component in amyloidosis.

关键词： Amyloid P component Ca 2+ dependence Protein aggregation (Human) C1 first component of complement C4bp C4-binding protein CRP C-reactive protein C3 third component of complement

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ISOLATION OF IA-LIKE ANTIGEN-BEARING CELLS FROM HUMAN PERIPHERAL LYMPHOCYTES THROUGH THE USE OF A MONOCLONAL-ANTIBODY TO FRAMEWORK DETERMINANTS OF IA-LIKE ANTIGENS

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JOURNAL OF IMMUNOLOGICAL METHODS 1980年第4期39卷 343-354页

作者： INDIVERI, F NG, AK RUSSO, C QUARANTA, V PELLEGRINO, MA FERRONE, S SCRIPPS CLIN & RES FDN RES INST DEPT MOLEC IMMUNOL LA JOLLA CA 92037 USA

A non-complement fixing monoclonal antibody (Q2/70) to framework determinants of human Ia-like antigens was used to develop a method for isolating Ia-like antigen bearing, i.e., Ia-like (+) cells and cells lacking these antigens, i.e., Ia-like (-), from human peripheral blood lymphocytes (PBL). The method was based on PBL sensitization with the antibody Q2/70, followed by rosetting with sheep (ShE) coated with purified rabbit anti-mouse Ig antibodies, differential centrifugation on a Ficoll-Hypaque gradient, and finally recovery of Ia-like (+) and Ia-like (-) cells from the bottom and the interface of the gradient, respectively. Marker analysis of the 2 cell subpopulations showed that the majority of the bottom cell fraction were Ia-like (+) and carried B cell markers such as membrane bound Ig (MbIg) and C3 receptors. The majority of the interface cell fraction were Ia-like (-) and carried T cell markers such as receptors for 2-aminoethylisothiouronium treated sheep erythrocytes (AETShE) and goat erythrocytes (GoE). Serological and functional studies showed that the Ia-like (+) cells: were useful targets in complement mediated cytotoxicity assays for HLA-DR typing;served as stimulator but not as responder in unidirectional mixed lymphocyte reactions (MLR);did not display lytic activity in natural killer (NK) cell cytotoxicity and in antibody dependent cellular cytotoxicity (ADCC);and proliferated in response to pokeweed mitogen (PWM) stimulation in the presence of helper T cells. The Ia-like (-) cells responded to but failed to stimulate allogeneic lymphocytes in the MLR;were highly active in NK and ADCC assays;and, provided helper activity in PWM stimulation of purified B cells.

关键词： ADCC antibody dependent cellular cytotoxicity AET 2-aminoethylisothiouronium bromide AET-E erythrocytes treated with AET C3 third component of complement C3d conversion product of C3 E erythrocytes EA antibody coated erythrocytes EAC1-3 mo EA coated with C1–C3 from mouse serum deficient in the fifth component of complement EAγ erythrocytes sensitized with IgG antibodies EAμ erythrocytes sensitized with IgM antibodies GoE goat erythrocytes MbIg membrane bound immunoglobulin MEM minimal essential medium MLR mixed lymphocyte reaction MoAb monoclonal antibody NK natural killer PBL peripheral blood lymphocytes PWM pokeweed mitogen ShE sheep erythrocytes Tγ T cells which express receptor for the Fc fragment of IgG Tμ T cells which express receptor for the Fc fragment of IgM

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APPEARANCE OF ACCEPTOR-BOUND C3B ON HLA-DR POSITIVE MACROPHAGES AND ON STIMULATED U937 CELLS - INHIBITION OF FC-GAMMA-RECEPTORS BY THE COVALENTLY FIXED C-3 FRAGMENTS

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MOLECULAR IMMUNOLOGY 1988年第3期25卷 295-303页

作者： ERDEI, A BAJTAY, Z FABRY, Z SIM, RB GERGELY, J UNIV L EOTVOS DEPT IMMUNOL JAVORKA S U 14 H-2131 GOD HUNGARY UNIV OXFORD DEPT BIOCHEM MRC IMMUNOCHEM UNIT OXFORD ENGLAND

The appearance and the functional role of acceptor-bound C3b during differentiation of human monocytes into macrophages were studied. Acceptor-bound C3b could be detected by the immune adherence (IA) test parallel to the expression of antigenic determinants specific to mature ***.i.e. on days 4-5 of culture. Consequently, the capacity of these phagocytes to fix C3b covalently via C3b-acceptors (C3Abs) can be considered as one of the signs of their activation/differentiation. All the mature macrophages positive in the IA test were also found to express HLA-DR antigens on their membrane. Using solubilized extracts of stimulated, 35S-cysteine-labelled cells of the human monocytic cell line, U937, we demonstrate that C3 synthesized by these cells can bind to C3bAs of the same cells. Covalently fixed C3 fragments were found to inhibit ***.-receptor-mediated ingestion of immune complexes and also antibody-dependent cellular cytotoxicity of monocyte-derived macrophages.

关键词： ADCC antibody-dependent cellular cytotoxicity BSA bovine serum albumin C3 third component of complement C3b ∗ nascent C3b C3b proteolytic fragments of C3 iC3b proteolytic fragments of C3 C3d proteolytic fragments of C3 C3bA, C3b acceptor structure capable of fixing nascent C3b covalently via the short-living metastable binding site of the complement fragment CR1 receptors for proteolytic fragments of C3 CR2 receptors for proteolytic fragments of C3 CR3 receptors for proteolytic fragments of C3 EA erythrocyte-antibody complex Fc γ R receptor reacting with the Fc-portion of IgG IA immune adherence binding of cells or particles possessing covalently bound C3b to CR1-bearing human red blood cells IAR IA-rosettes IC immune complex Mo monocyte MΦ macrophage PBS phosphate buffered saline PMA phorbol-12-myristate 13-acetate SDS sodium dodecyi sulphate RBC red blood cell Z zymosan

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Nutrition support in hospitalized patients with diabetes mellitus

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MAYO CLINIC PROCEEDINGS 1996年第6期71卷 587-594页

作者： McMahon, MM Rizza, RA MAYO CLIN & MAYO FDN DIV ENDOCRINOL METAB & INTERNAL MEDROCHESTERMN 55905

Many physicians will manage the care of hospitalized patients with diabetes mellitus who require parenteral nutrition or enteral tube feeding, The nutritional assessment, indications for nutrition support, estimate of nutritional needs, and biochemical monitoring guidelines for critically ill patients with diabetes are similar to those for nondiabetic patients, In general, a weight loss of up to 10% of body weight is well tolerated and, in the absence of severe stress, the provision of dextrose-containing crystalloid solutions and electrolytes is adequate for as long as 7 to 10 days, Studies that demonstrate a beneficial influence of nutrition support on clinical outcome administered nutrition for a minimum of 1 week, No data have established that support for a briefer duration is of clinical benefit, An important goal in the care of the hospitalized patient with diabetes is to avoid the extremes of hypoglycemia and hyperglycemia. Herein we provide our approach to achieving glucose control in stressed hospitalized patients with diabetes mellitus who are receiving parenteral and enteral nutrition, Although evidence is increasing that hyperglycemia impairs immune function, well-designed prospective randomized trials are needed to determine the risks, costs, and benefits of achieving glucose control and of providing nutrition support to hospitalized patients with diabetes mellitus.

关键词： C3 third component of complement PN parenteral nutrition

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IMMOBILIZATION OF C3B WITH RETENTION OF FUNCTIONAL-ACTIVITY

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JOURNAL OF IMMUNOLOGICAL METHODS 1990年第1期133卷 67-70页

作者： EDENS, RE LINHARDT, RJ WEILER, JM VET ADM MED CTR IOWA CITY IA 52240 USA UNIV IOWA COLL MED DEPT INTERNAL MED IOWA CITY IA 52242 USA UNIV IOWA COLL PHARM DIV MED & NAT PROD CHEM IOWA CITY IA 52242 USA

We compared eight commercially available, pre-activated affinity chromatography supports for ability to immobilize C3b that would retain functional activity. Pre-activated supports that we studied were: cyanogen bromide activated agarose, N-hydroxysuccinimide activated agarose, Reacti-Gel HW-65, Actigel A aldehyde activated agarose, thiopropyl activated agarose, 1,4-bis(2,3-epoxypropoxy) butane activated agarose, Reacti-Gel GF-2000 and tresyl activated agarose. The amount of C3b immobilized by each support varied from 81% for Actigel A aldehyde activated agarose to only 19% for Reacti-Gel GF-2000. We examined the functional capacity of the C3b immobilized on these various supports to participate in the alternative pathway. Immobilized C3b was mixed with factors D and B of the alternative pathway and examined over time for ability to consume factor B hemolytic activity. C3b immobilized on thiopropyl activated agarose consumed factor B at a rate comparable to unbound fluid phase C3b. C3b immobilized on other supports was less active in participating in factor B consumption. Thus, we have demonstrated the ability to immobilize C3b onto a solid matrix with the immobilized C3b retaining the ability to participate in the alternative pathway. This immobilized C3b can be used to fractionate substances with high C3b binding affinity.

关键词： Affinity chromatography complement C3b Agarose C3 third component of complement C3b cleavage fragment of complement CDI 1,1-carbonyldiimidazole CNBr cyanogen bromide DGVB 2+ dextrose gelatin veronal-buffered saline with calcium and magnesium EAC4b,3b, sheep erythrocytes antibody and complement components C4b and C3b EDTA ethylenediamine tetraacetate EDTA-GVB EDTA solution containing GVB GVB gelatin, veronal buffered saline HSA human serum albumin PBS phosphate-buffered saline Z number of lytic events per cell

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INFANTILE SYSTEMIC LUPUS-ERYTHEMATOSUS WITH ONSET SIMULATING CONGENITAL NEPHROTIC SYNDROME

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JOURNAL OF PEDIATRICS 1994年第1期124卷 27-31页

作者： MASSENGILL, SF RICHARD, GA DONNELLY, WH UNIV FLORIDA DEPT PATHOL GAINESVILLE FL 32611 USA

Two white female infants were seen with congenital nephrotic syndrome at age 6 weeks and 3 months, respectively. Both had hypocomplementemia, elevated antinuclear antibody and anti-double-stranded DNA titers, and diffuse proliferative glomerulonephritis with positive immunofluorescence in their initial renal biopsy samples. Although uncommon, infantile systemic lupus erythematosus should be considered in the evaluation of congenital nephrotic syndrome.

关键词： ANA Antinuclear antibody Anti-dsDNA anti-double-stranded DNA C3 third component of complement C4 Fourth component of complement CoNS Congenital nephrotic syndrome HLA Human leukocyte antigen IVIg Intravenously administered immuneglobulin SLE Systemic lupus erythematosus

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Leucocyte adhesion protein deficiency in Irish setter dogs

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Veterinary Immunology and Immunopathology 1992年第3-4期32卷 261-280页

作者： Trowald-Wigh, Gunilla Håkansson, Lena Johannisson, Anders Norrgren, Leif Hård af Segerstad, Carl Department of Medicine and Surgery Swedish University of Agricultural Sciences Uppsala Sweden Department of Clinical Chemistry University Hospital Uppsala Sweden Department of Pathology Swedish University of Agricultural Sciences Uppsala Sweden Department of Genetics Uppsala University Uppsala Sweden

Investigation of 12 Irish setter puppies from six litters with severe recurrent infections, neutrophilia and low body weight revealed a leucocyte adhesion protein deficiency with a total lack of CD11b and CD18. Their neutrophil function was severely impaired with a totally absent capacity to ingest C3b-opsonized particles, a significantly impaired capacity to ingest IgG-opsonized particles and significantly diminished adherence to nylon wool when compared with neutrophils from healthy control dogs. The chemiluminescence of patient neutrophils activated by C3b-opsonized particles was, consequently, significantly decreased compared with that of control neutrophils, while the respiratory burst assayed by phorbolmyristate acid (PMA) stimulated nitroblue tetrazolium (NBT)-reduction was normal in the patient group. Random migration and chemotactic responses of patient and control neutrophils, were similar. The etiology, pathogenesis and clinical manifestations of the Irish setter leucocyte adhesion deficiency were similar to that of the leucocyte adhesion deficiency in humans. © 1992.

关键词： C3 third component of complement CR3 complement receptor type III FCS fetal calf serum Hb hemoglobin LAD leucocyte adhesion deficiency syndrome NBT nitroblue tetrazolium PBS phosphate-buffered saline PMA phorbolmyristate acid RLU relative light units SEM scanning electron microscope WBC white blood cell

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