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SIMULTANEOUS DETECTION OF ALLOTYPES IN NATIVE AND ACTIVATED HUMAN C2 BY ISOELECTRIC-FOCUSING AND SILVER STAINING

JOURNAL OF IMMUNOLOGICAL METHODS 1989年第1期117卷 9-15页

作者： SEGURADO, OG ARNAIZVILLENA, A HOSP PRIMERO OCTUBRE E-28041 MADRID SPAIN

A simple and highly sensitive analytical isoelectric focusing (IEF) technique using immunofixation with anti-C2 serum followed by silver staining has been developed in order to study simultaneously the structural polymorphism of both native C2 and C2 activation fragments (C2a MW 74,000 and C2b MW 34,000). Structural C2 polymorphism of C2*B and C2*C allotypes (but not of C2*A1) was found to be associated with the C2a fragment, whereas C2b appears to display no polymorphism. Commercially available IEF gels and C2 antisera gave reproducible allotyping data and make this technique of general use for densitometric analysis of native and activated C2. In vivo C2 activation was studied through C2a/native C2 area ratios obtained by computerized densitometry. Significantly lower ratios were observed in healthy individuals than in patients with systemic lupus erythematosus (SLE), reflecting an abnormally high classical pathway activation of complement in SLE. This methodology may be of value for immunogenetic and functional studies of other complement components.

关键词： C2 polymorphism C2 activation Isoelectric focusing Silver staining Lupus erythematosus BF factor B of the alternative complement pathway C2 second component of complement EACA ϵ-aminocaproic acid EDTA ethylenediaminetetraacetic acid HLA human leukocyte antigen MHC major histocompatibility complex PBS phosphate-buffered saline SD standard deviation SLE systemic lupus erythematosus

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LACK OF BINDING OF C-3 TO IGG ANTIBODIES DURING THE ACTIVATION OF THE CLASSICAL complement PATHWAY ON THE RED-CELL

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MOLECULAR IMMUNOLOGY 1984年第3期21卷 191-195页

作者： CIRCOLO, A BORSOS, T Laboratory of Immunobiology National Cancer Institute National Institutes of Health FCRF Frederick MD 21701 U.S.A.Received 30 May 1983 Revised 30 September 1983 Available online 12 November 2002

The interaction between C3 [complement component 3] and natural human and rabbit anti-MTX [methotrexate] IgM and hyperimmune IgG antibody [Ab] bound to red cells to which MTX was covalently coupled. IgM Ab molecules bound to the cell surface were measured by their interaction with rabbit anti-human .mu.-chain IgG Ab;the bound anti-.mu.-chain or anti-MTX IgG was quantitated with 125I-labeled PA [protein A]. C3 uptake by .**GRAPHIC**. complexes from purified preparation of C3 was detected by the interaction of bound C3 with rabbit anti-C3 IgG Ab;the bound IgG was then measured with rabbiolabeled PA. In some experiments the uptake of C3 by the .**GRAPHIC**. complexes was measured by using 125I-labeled C3. To find out if C3 was bound to Ab molecules, anti-MTX Ab were eluted from the cells by excess fluid-phase MTX. After dialysis the eluted anti-MTX IgM or IgG Ab was than reattached to fresh EMTX. All cells were then analyzed for Ab and C3 content by a radioimmunoassay. When anti-MTX IgM or IgG was eluted from EMTX AC423 complexes no C3 was removed from the cells, and eluted IgM or IgG did not carry with them either C3 antigen or 125I-labeled C3. Thus, during activation of the classical C-pathway at the red cell surface, no C3 was bound to IgM (rabbit or human) or IgG (rabbit) antibody molecules.

关键词： C complement C1 first component of complement C4 fourth component of complement C2 second component of complement C3 third component of complement C1 … active form of C-components EDTA in VBS 0.01 M tri- N -ethylenediaminetetraacetate in veronal-buffered saline containing 0.1% gelatin, pH 7.4 IgM immunoglobulin of class M IgG immunoglobulin of class G E sheep red blood cell(s) MTX methotrexate E MTX sheep red blood cell(s) to which MTX was covalently coupled PA protein A of Staphylococcus aureus gp guinea-pig Ab antibody

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RECURRENT PYOGENIC INFECTIONS IN INDIVIDUALS WITH ABSENCE OF THE 2ND component OF complement

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JOURNAL OF CLINICAL IMMUNOLOGY 1982年第1期2卷 39-45页

作者： SAMPSON, HA WALCHNER, AM BAKER, PJ DUKE UNIV SCH MED DEPT PEDIAT DURHAM NC 27710 USA DUKE UNIV SCH MED DEPT MICROBIOL & IMMUNOL DURHAM NC 27710 USA

While deficiency of the terminal components of complement (C3-C8) has classically been associated with recurrent pyogenic infections, C2 deficiency evidently is also associated with recurrent infections in some individuals. The patient presented here had 2 major pyogenic infections prior to 1 yr of age and 2 lacked C2. Studies of alternative C pathway and humoral and cellular immunity were within normal limits. Family studies suggest an autosomal codominant pattern of inheritance for the C2 defect, which also corresponded to the inheritance of the HLA-A10, B18 haplotype. A review of the literature revealed 9 other cases of C2-deficient patients with well-documented recurrent infections. In these patients, Streptococcus pneumoniae is the organism most frequently implicated in infectious processes, whether or not their alternative C pathway is intact.

关键词： Immunodeficiency complement deficiency recurrent infections second component of complement

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DIFFERENT HLA-ANTIGEN ASSOCIATIONS FOR THE FUNCTIONALLY ACTIVE AND INACTIVE PRODUCTS OF THE complement C4F1 ALLELE

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HUMAN IMMUNOLOGY 1980年第1期1卷 23-30页

作者： ONEILL, GJ MINITER, P POLLACK, MS DUPONT, B Tissue Typing Laboratory Sloan Kettering Institute for Cancer Research 1250 First Avenue New York New York 10021 USA

The genetic polymorphism of the 4th human complement component, C4, was studied in 945 unrelated Caucasians. A 3rd allele of the C4F (Rodgers) locus, termed C4F1 was demonstrated. This allele is characterized using immunofixation electrophoresis, by the presence of an additional fast-moving anodal band of C4 which distinguishes it from the common C4F variant. The allelic frequencies fit the Hardy-Weinberg equilibrium assuming 3 alleles at the C4F locus: C4F, ***. and C4F1. The functional activity of the C4F variants was investigated using a specific hemolytic overlay technique for C4. In almost all individuals (75 of 78), the C4F1 allele codes for a functionally inactive C4 product only when it occurs on an HLA-B17 positive haplotype but the same allele codes for a functionally active fast variant of C4 when it occurs on an HLA-B37 positive haplotype (18 of 18). Very strong genetic linkage disequilibrium was observed for the C4F1 allele with HLA-B17 and B37. The active and inactive C4F1 variant also has marked nonrandom gametic association to different alleles of the Bf locus and to HLA-C locus determinants. No further variants of the C4S (Chido) locus were identified. Rodgers (Rg) typing by the plasma inhibition test of anti-Rg antiserum showed that plasma from individuals homozygous for the C4F1 allele is able to only partially inhibit anti-Rg whereas all C4F positive individuals totally inhibited the reaction.

关键词： Bf properdin factor B Ch Chido blood group antigen C2 second component of complement EDTA sodium (tetra) ethylenediamine tetraacetate C4 fourth component of complement GLO glyoxalase MgCl 2 magnesium chloride CaCl 2 calcium chloride Rg Rodgers blood group antigen

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