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M Loos

Publications and source records attributed to M Loos.

At least 37 records · Page 2Linked to original sources
PubMed1994

The nature of X-ray-induced mutations in mature sperm and spermatogonial cells of Drosophila melanogaster.

Mutations at four X-linked visible loci (yellow, white, vermilion and forked) induced by X-irradiation of mature sperm and spermatogonial cells were analysed genetically and cytogenetically. In addition, a fraction of the intragenic vermilion mutations was analysed molecularly. Males of two wild-type strains (Amherst M56i and Berlin-K) were used. A total of 332,651 chromosomes of irradiated mature sperm and 311,567 of irradiated spermatogonial cells were scored. The ratio of F1 female sterile, F2 male lethal and F2 male viable mutations in mature sperm and spermatogonial cells is very similar. The cytogenetic analysis shows equal fractions of multilocus deletions and translocations among the mutations recovered from both stages of spermatogenesis. These data strongly suggest that the spectrum of X-ray mutations is similar in mature sperm and spermatogonial cells, including multilocus deletions and chromosome rearrangements. The molecular analysis of a number of intragenic vermilion mutations showed the presence of three small deletions (1-10 bp), one insertion of two nucleotides and seven single nucleotide changes.

Animals↗
PubMed1994

[Pathogenicity factors of enteric Salmonellae].

The number of cases of salmonellosis has steadily increased during the last decade. Until the end of the 70s, the serotype Salmonella typhimurium was most commonly isolated, but since 1986 the serotype S. enteritidis has become more common. Recent research has shown that the colon is more frequently infected than was earlier recognized, but most common is still an acute enterocolitis during an infection with salmonella. In severe cases, salmonella can spread from the gut, the site of infection, to cause a systemic disease. As in vitro studies have shown, gastroenteritic salmonellae seem to possess a number of so-called virulence factors that help them to evade the host's defenses against bacterial infection. Colonisation in the gut seems to be dependent on a 66 kD heat shock protein (HSP), which binds to a 15 kD glycoprotein present in gut mucus. For penetration into the mucosa, by invasion of the epithelial cells, bacterial invasions are newly synthesized. These proteins distort the epithelial membrane and induce a port of entrance. Macrophages will attack and engulf the bacteria, but salmonella can withstand these professional killers, and even grow intracellularly in them. In order to do so, not only LPS is required, but also specific outer membrane proteins (OMP), porins. Moreover, during their intracellular survival the bacteria need to adapt to the new microenvironment, and in reaction to stress, new HSPs are expressed. Finally, the O-specific sugar chains prevent the otherwise lytic action of complement in blood serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

C1q, the collagen-like subcomponent of the first component of complement C1, is a membrane protein of guinea pig macrophages.

C1q, a subcomponent of C1--the first component of complement, is synthesized by macrophages (M phi). Immunofluorescence and immunoperoxidase studies first indicated the presence of C1q on the surface of guinea pig (gp) and human peritoneal M phi (Loos, M., Storz, R., Müller, W. and Lemmel, E. M., Immunobiology 1981. 158: 213). In our study different methods for labeling of gp serum and gp M phi C1q were employed. The presence of C1q protein on the surface of gp peritoneal M phi is shown by cell surface labeling with the biotin derivative sulfosuccinimdyl-6-(biotinamido)-hexanoate and subsequent immunoprecipitation. The mechanism by which C1q is attached to the cell membrane was also investigated. Intact cells were treated with acid stripping-buffers or phosphatidylinositol-specific phospholipase C and separated membranes were extracted with a buffer containing 1 M KCl and 3 M urea. Regardless of which method was used, C1q remained attached to the membrane. When surface-labeled cells were cultured, they were found to release the C1q from their surface membrane into the culture medium. Lysates of biosynthetically labeled cells were used to show that, like secreted or serum C1q, cellular M phi C1q binds to immobilized homologous IgG. This implies that the globular regions of the cellular C1q are functionally active. The results reveal that (i) cellular M phi C1q is firmly located in the membrane throughout the biosynthetic pathway, such that it is comparable with an integral membrane protein, (ii) cellular M phi C1q is not reversibly bound to the cell surface via a receptor. We suggest that C1q, as a membrane protein of M phi, serves as an Fc binding factor that also is secreted into the environment.

Animals↗
PubMed1993

Complete functional C1q deficiency associated with systemic lupus erythematosus (SLE).

A complete functional deficiency of C1q is described in a patient suffering from SLE. From reduced plasma C1 activity of the parents a hereditary trait was assumed. The defective C1q molecule was haemolytically inactive, did not bind to immune complexes, and was not recognized by the monocyte C1q receptor. C1 activity in the patient's serum could be restored by the addition of purified C1q. Analysis by gel-filtration and ultracentrifugation experiments revealed an immunoreactive molecule of about 150 kD mol. wt, corresponding to one structural subunit of the C1q macromolecule, containing two A chain-B chain dimers and a C-C chain dimer. Applying Southern blot analysis with cDNA clones encoding for the three individual chains of the C1q molecule, no restriction fragment length polymorphism was detected, ruling out possible major alterations of the genetic information.

Adult↗
PubMed1993

The Fc-recognizing, collagen-like C1q molecule is a putative type II membrane protein of macrophages.

The Fc-recognizing, collagen-like C1q molecule, a subcomponent of the first component of complement, C1, is present on the cell surface of guinea pig peritoneal macrophages and human peritoneal and monocyte-derived macrophages. On closer examination, C1q appears to be a membrane protein (membrane C1q) of macrophages since it is i) anchored into the membrane throughout the biosynthetic pathway, ii) tightly and irreversibly bound to the cell surface and iii) only liberated if the intact membrane structure is disrupted by a detergent or repeated freeze/thawing. Additionally, the amino acid sequence of the A chain of human C1q displays properties that are characteristic for integral type II membrane proteins. The membrane C1q of guinea pig macrophages has a "lighter" B chain than serum C1q. Under physiological conditions in culture guinea pig macrophages release membrane C1q thereby converting it into the serum form. Moreover, membrane C1q appears to be involved in various cellular events such as binding of Fc, polyanions, lipid A and gram-negative bacteria to macrophages.

Animals↗
PubMed1993

Functional domains of the human C1q A-chain.

This brief review was inspired by discussions relating to the IIIrd. International C1 Workshop (this volume) and the realization that certain functional properties of the C1q molecule are limited exclusively to the A-chain. The collagen-like region of the A-chain contains a major binding site for non-immunoglobulin substances, which include C-reactive protein, serum amyloid P, LPS and DNA. This binding site is immediately adjacent to, and partially overlapping with, an arthritis-modulating epitope common to the C1q A-chain and various types of collagen, including cartilage type II collagen. At the N-terminal end of the C1q A-chain is a leader peptide sequence that anchors the intact C1q molecule firmly in the membrane of macrophages, the C1q molecule can thus be classified as a type II membrane protein, functioning as an additional receptor for molecules known to react with C1q in fluid phase such as the Fc region of IgG, LPS and polyanionic molecules (e.g. chondroitin sulphate, heparin, dextran sulphate etc.). The various domains within the A-chain, and their respective functions (or potential functions), are presented and discussed in the context of the intact C1 molecule and with regard to any wider functional relevance.

Amino Acid Sequence↗
PubMed1993

C1q in autoimmune diseases: rheumatoid arthritis.

Rheumatoid arthritis is an autoimmune disease involving stimulation of T cells and the production of autoantibodies. In this disease autoantibodies to collagen type II are believed to play a major role in inflammatory events ultimately resulting in joint destruction. However, collagen type II-containing cartilage has been discussed as one of the primary antigens (as evidenced by animal models), despite not being accessible. Evidence is presented here for the involvement of C1q, the collagen-like subunit of the first component of complement, in the pathogenesis of rheumatoid arthritis. The C1q A-chain contains an epitope exhibiting an identical sequence to part of an arthritis modulating epitope from collagen type II. Furthermore, preapplication of a synthetic peptide, representing the epitope on the C1q A-chain, has been shown to delay the onset and reduce the severity of collagen-induced arthritis in a DBA/1 mouse model. Since the complement system, in particular C1q (as part of the first component of the classical pathway), plays a major role in the inflammatory process, we propose that the collagen-like C1q molecule, altered during the inflammatory process, may result in the generation of autoantibodies which also recognize collagen type II, and may thus be considered as a link between the early inflammatory process in the joint and the only later occurring cartilage destruction.

Amino Acid Sequence↗
PubMed1992

Isolation, sequence analysis and characterization of cDNA clones coding for the C chain of mouse C1q. Sequence similarity of complement subcomponent C1q, collagen type VIII and type X and precerebellin.

A mouse macrophage lambda gt11 cDNA library was screened using a genomic DNA clone coding for the C-chain gene of human C1q. Approximately 600,000 recombinant phage plaques were hybridized with peroxidase-labeled human C-chain probe and detected by enhanced chemiluminescence. Five positive clones were obtained. The size of the full-length cDNA is 1019 bp. The sequence identity of the nucleotide sequence with human C1q C chain is 79%, the identity of the deduced amino acid sequences is 73%. The mouse C1q C chain exhibits the same structural features as the human C chain, e.g. conservation of the cysteine residues. Like the mouse A chain, the mouse C chain has an RGD sequence that may be recognized by receptors of the integrin family. No RGD sequences have been found in any of the human C1q chains. The size of the C-chain mRNA (1.2 kb) and its tissue distribution (macrophages being the cell type with the highest mRNA concentration) are identical to the mRNA of the mouse A and B chains. Alignment of human and mouse C1q A, B and C chains exhibits two blocks of highly conserved residues within the C-terminal globular regions. Three other proteins, collagen type VIII and type X and precerebellin share this similarity with C1q, indicating the structural and probably functional importance of these regions within the non-collagenous domains of the molecules.

Amino Acid Sequence↗
PubMed1992

Purification and characterization of LPS-free porins isolated from Salmonella minnesota.

Porins of Salmonella minnesota, R595, were purified by anion exchange chromatography and subsequently isolated in their monomeric form by chromatofocusing. Two forms of porin could be isolated, both with an apparent molecular mass of 37,000, but of differing isoelectric points (pI 4.6 versus pI of 4.9). Porins with pI 4.9 did not contain any detectable LPS, but porins with pI 4.6 were found to contain trace amounts of LPS (1.3 x 10(-4) micrograms LPS/1 microgram porin) as measured using a highly sensitive limulus assay. Unlike the LPS-associated porins the monomeric porins were biologically inert with regard to pore formation, but they were still able to bind C1q, a subcomponent of the first component of complement.

Bacterial Outer Membrane Proteins↗
PubMed1992

Complement component deficiencies and infection: C5, C8 and C3 deficiencies in three families.

Three families are described with complement component deficiencies. In one family, five children had C5 deficiency; in a second family, two children had C8 deficiency and one child in a third family had C3 deficiency. The index cases were identified during screening of patients with recurrent pyogenic infections, recurrent meningitis and meningococcaemia. Two of the five C5 deficient patients had recurrent meningitis and meningococcaemia, two had recurrent respiratory tract infections and otitis and one was healthy. One of the C8 deficient patients had meningitis, meningococcaemia and pneumonia, whereas his sibling with the same deficiency was healthy. The patient with C3 deficiency had four episodes of meningitis and recurrent otitis.

Child↗
PubMed1992

Purification and characterization of a Salmonella typhimurium agglutinin from gut mucus secretions.

One of the earliest events in Salmonella typhimurium pathogenesis seems to be the interaction of the bacterium with the mucus of the gut. After exposing S. typhimurium to guinea-pig colon, we were able to demonstrate by electron microscopy that S. typhimurium bacteria were trapped on, or in, the mucus layer. Specific components isolated from crude mucus secretions were found to aggregate the bacteria. The degree of bacterial aggregation was dependent on the protein concentration of the crude mucus and on time. Aggregation of S. typhimurium could be abolished by sugars: L-fucose exhibited the strongest inhibition, followed by D-glucose, D-galactose and D-mannose. Lectins were also capable of inhibiting aggregation, the lectin of Ulex europaeus (UEA I), specific for L-fucose, was found to be a stronger inhibitor of bacterial aggregation than Con A. The agglutinin for S. typhimurium isolated from guinea-pig colonic crude mucus preparation was characterized as a 15 kDa glycoprotein. An affinity-purified anti-15 kD antibody inhibited, dose-dependently, the aggregation of S. typhimurium by crude mucus material.

Agglutinins↗
PubMed1992

Modulation of type II collagen-induced arthritis in DBA/1 mice by intravenous application of a peptide from the C1q-A chain.

In this report we are able to show that intravenous (i.v.) application (day 0) of a nonapeptide (residues 26-34) from the human C1q A-chain (designated peptide A-C1q) prior to intradermal (i.d.) administration of chicken type II collagen (CII) in arthritis-susceptible DBA/1 mice (H2q), leads to abrogation of polymorphonuclear neutrophil (PMN) invasion into the joints. This nonapeptide exhibits epitope characteristics and high homology to residues 137-147 of CB11 (a cyanogen bromide fragment of chicken CII, known to contain both arthritis inducing and suppressing determinants). Arthritis index was lowest in animals pretreated i.v. with CII (as internal control), though animals pretreated i.v. with peptide K (residues 137-147 with an additional glycine residue from CB11) or peptide A-C1q exhibited comparative arthritic indices. Only in the arthritis-positive control group (day 0: PBS i.v.) did i.d. application of CII lead to invasion of PMN into the synovial layer and the joint space. Analysis of antibody (Ab) responses at day 48 after i.v. immunization (day 0) and CII challenge (day 7) revealed IgE-Abs to native CII and also to native C1q. IgG titers to CII were highest in animals pretreated with peptide A-C1q. Abs from this group, exhibiting activity to peptide A-C1q (immunizing antigen), were of mainly IgG1 and IgG3 isotypes. Evaluation of the immune response following i.v. application of peptide A-C1q or CII, prior to i.d. CII administration, in DBA/1 mice, revealed IgM responses to peptide A-C1q and peptide K, but not to CII. Intravenous application of peptide A-C1q led to generation of IgG3-Abs reacting only with peptide A-C1q and peptide K, but not with native CII. Additionally, i.v. application of peptide A-C1q elicited IgG responses to a pentapeptide, resembling amino acid residues 26-30 (K-G-E-Q-G) of the C1q A-chain. This five residue antigenic determinant is present in peptide K, in chicken and human CII as well as in human C1q. No specific IgE response to any of the antigens tested could be detected. Since a peptide from the C1q A-chain is both capable of eliciting immune responses and modulating CII-induced arthritis in mice, we postulate that the collagen-like complement component C1q is involved in the development of CII-induced inflammatory arthritic lesions, and may represent, in vivo, the early antigen responsible for inducing anticollagen antibodies prior to CII in hyaline cartilage becoming available as antigen.

Adjuvants, Immunologic↗
PubMed1992

A 66-kilodalton heat shock protein of Salmonella typhimurium is responsible for binding of the bacterium to intestinal mucus.

Salmonella typhimurium infections have increased during the last few years. However, the interplay of virulence factors in S. typhimurium pathogenesis is still poorly understood, particularly with regard to the mechanisms and components of the bacterium which are involved in its interaction with the intestinal mucus. We have observed that S. typhimurium is aggregated by incubation with colonic mucus (guinea pig model). To quantify this phenomenon, an aggregation assay was established. By using this assay, it was found that the aggregation profile of S. typhimurium strains freshly isolated from patients (age 9 and older) with salmonellosis correlated with the severity of the disease. An isolate with high aggregation behavior was chosen for characterization of the bacterial component involved in binding to colonic mucus material. The component of S. typhimurium responsible for aggregation was purified and characterized as a 66-kDa protein which was able to completely inhibit mucus-mediated bacterial aggregation. This protein was recognized by monoclonal antibodies against the 65-kDa heat shock protein (HSP) of Mycobacterium leprae. The 66-kDa protein of S. typhimurium was inducible by incubating the bacteria at 50 degrees C and was secreted into the supernatant, from which it could be isolated in both dimeric and polymeric forms. The monoclonal anti-HSP 65, as well as a polyclonal antibody against the 66-kDa protein of S. typhimurium, caused dose-dependent inhibition of the aggregation of S. typhimurium by crude mucus preparations. This is the first report showing that a bacterial HSP is involved in mucus-mediated interaction of pathogens with the host.

Adolescent↗
PubMed1991

Gene expression of the A- and B-chain of mouse C1q in different tissues and the characterization of the recombinant A-chain.

Immunoscreening of a mouse macrophage cDNA library with an anti-mouse C1q-antibody resulted in the isolation of cDNA clones. The deduced amino acid sequence was homologous to the A-chain of human C1q. Homology on the DNA level was found to be 76% and on the protein level 72% thus it appeared the clones coded for the mouse C1q A-chain. An immunoblot of murine serum C1q separated by SDS-PAGE was detected with an A-chain specific antibody that had been affinity purified on recombinant mouse C1q A-chain expressed in Escherichia coli. The antibody preparation reacted exclusively with the mouse C-chain (as defined by SDS-PAGE). Northern blot analysis with strand-specific cDNA probes coding for the A- and B-chain of murine C1q showed that mouse peritoneal macrophages produced the highest concentration of C1q gene transcripts. RNA from mouse spleen, thymus, heart, and brain gave substantial hybridization signals, whereas RNA preparations from liver, kidney, lung, and small intestine appeared to contain only trace amounts of C1q mRNA. In a Northern blot analysis of different guinea pig cells and tissues, only RNA preparations from peritoneal macrophages hybridized with the mouse C1q probes. These results indicate that macrophages are a major site of C1q biosynthesis.

Amino Acid Sequence↗
PubMed1991

C1q-bearing immune complexes detected by a monoclonal antibody to human C1q in rheumatoid arthritis sera and synovial fluids.

Using a monoclonal antibody directed against the C-chain of human C1q, we detected C1q-bearing immune complexes (IC) in sera and synovial fluids of rheumatoid arthritis (RA) patients. In a sandwich-ELISA, C1q-bearing IC were captured by the solid-phase monoclonal antibody and then detected with peroxidase-labeled F(ab')2-antibodies to either human IgG or IgM. The results of this assay were compared to an ELISA-modification of the C1q-solid-phase binding assay (C1q-SPBA). C1q-bearing IC were detected in 81.1% of RA-sera and the 65.2% of RA-synovial fluids. IgG as well as IgM was present in 72.6% of the sera and 70% of the synovial fluids which were positive in both assays. Most RA sera that were only positive for C1q-bearing IC, contained IgG alone (81.5%). The corresponding synovial fluids showed IgG alone (53%) or both IgG and IgM (41.1%). IgM alone (25%) could be detected in sera, e.g. in juvenile forms of RA. The levels of IC were higher in synovial fluid than in paired serum. In comparison to normal human serum (NHS) and patients with osteoarthritis, complement activity (CH50 titers) and C1q-values in patients with RA were frequently elevated. Since the formation of C1q-bearing IC is an indicator for the classical complement pathway activation, an assay with monoclonal anti-C1q antibody may be a useful tool in the diagnosis of rheumatoid diseases.

Antibodies, Monoclonal↗
PubMed1990

Studies on the interaction of C1q, a subcomponent of the first component of complement, with porins from Salmonella minnesota incorporated into artificial membranes.

Purified outer membrane proteins (OMP) of Salmonella minnesota, Re-form, were incorporated into liposomes. These induced in macrophages a chemiluminescence signal identical to that of the intact Re-form. This signal was abolished by preincubation of porin-containing liposomes with purified C1q. Incorporation of isolated OMP into black lipid membranes (BLM) resulted in channel-formation which could not be inhibited by isolated C1q. Additionally, incubation of OMP-containing liposomes with BLM resulted in pore-formation within the BLM. This was amplified when lipid A was present within the liposomes. Preincubation of OMP-containing liposomes with purified C1q abolished pore-formation within the BLM.

Animals↗