Cross-reactions between human and animal plasma proteins

Summary Cross-reactions between human plasma proteins and their homologues in primate blood were investigated systematically. From the three groups of proteins distinguished earlier [2] two have been especially examined; these findings are reported and discussed in the present communication. The Immunological Evolution Group (IEG) I, comprising IgA (-chain), IgD (-chain) and inter--trypsininhibitor, cross-reacts with pongid plasma only, IEG IIa, i.e. IgM (-chain), ₂-glycoproteins II and III and cholinesterase, does so with the pongid and cercopithecoid plasmas tested; IEG IIb, including acid _l-glycoprotein, _2HS-glycoprotein, _l-trypsininhibitor, haptoglobin and hemopexin, cross-reacts with pongid, cercopithecoid and cebus (platyrrhinian) plasma and finally IEG IIc, consisting of transferrin and Gc-globulin, does so with all primate plasmas tested, including prosimians. All the proteins named do not cross-react however with non-primate proteins as do those of IEG III. It is concluded, that the determinants reacting in the primate proteins increase in their evolutionary ages from IEG I over IEG IIa, IIb to IIc in the same way as the last common ancestors of man and the crossreacting species increase.

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Zusammenfassung Die Kreuzreaktionen zwischen menschlichen Plasmaeiweißen und ihren Homologen im Blut von subhumanen Primaten wurden systematisch untersucht. Von den drei früher voneinander abgetrennten Gruppen [2] wurden zwei für die vorliegende Versuchsreihe herausgegriffen; eine von ihnen konnte weiter unterteilt werden. Die erhobenen Befunde werden berichtet und diskutiert. Die Immunologische Evolutions-Gruppe (IEG) I, die IgA (-Kette), IgD (-Kette) und den Inter--Trypsininhibitor umfaßt, zeight Kreuzreaktionen nur mit den Plasmen von Pongiden. Die IEG IIa, zu der IgM (-Kette), die ₂ II und III und Cholinesterase gehören, kreuzreagiert mit den entsprechenden Plasmaproteinen der geprüften Pongiden und Cercopithecoidea, die IEG IIb—das sind saueres _l-Glycoprotein, _2HS-Glycoprotein, _l-Trypsininhibitor, Haptoglobin und Haemopexin — mit Pongiden, Cercopithecoidea und Cebus (Platyrrhini) und endlich die IEG IIc, die sich aus Transferrin und dem Gc-Globulin zusammensetzt, mit allen geprüften Primatenplasmen einschließlich denen von Prosimiern. Alle hier genannten Plasmaeiweiße zeigen jedoch keine Kreuzreaktionen mit Proteinen von Nichtprimaten, wie dies bei der IEG III der Fall ist.Aus den vorliegenden Befunden wird der Schluß gezogen, daß die Determinanten der Primatenproteine, die jeweils reagieren, in ihrem phylogenetischen Alter von IEG Iüber IEG IIa, IIb, IIc im gleichen Maße ansteigen, wie die letzten gemeinsamen Vorfahren von Mensch und den kreuzreagierenden Arten.

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(Chief: Prof. Dr. E. Krah)     

Cross-reactions between human and animal plasma proteins

Summary In continuation of earlier experiments the immunological cross-reactions between human plasma proteins and homologues from 9 non-eutherian mammals were investigated, using monospecific rabbit antisera to some 30 different human plasma proteins. The Ouchterlony method in our standard modification (Bauer, 1970a) was employed. 4 proto-and metatherian proteins were found to share antigenic structures with human homologues, namely albumin, -lipoproteins, fibrinogen and plasminogen.Preliminary experiments indicate that marsupial and monotrematal proteins have less antigenic determinants in common with man than any from the 24 non-human species, which belong to eutheria. The phylogenetical implications of our findings are discussed in some detail.

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Zusammenfassung In Fortführung früherer Versche wurden die Kreuzreaktionen zwischen menschlichen Plasmaeiweißen und ihren Homologen im Blut von 9 Proto- und Metatheria untersucht. Monospezifische Kaninchen-Antiseren gegen 30 verschiedene menschliche Plasmaproteine wurden getestet. Dabei fand die Ouchterlony-Methode in unserer Standard-Modifikation (Bauer, 1970a) Verwendung. 4 Proteine aus dem Blut der beiden genannten primitiven Säugergruppen, nämlich Albumin, -Lipoproteine Fibrinogen und Plasminogen, gaben Kreuzreaktionen mit ihren menschlichen Homologen.Vorläufige Ergebnisse zeigen darüber hinaus, daß die Proto- und Metatheria-Proteine weniger menschliche Determinanten tragen als irgendeines der 24 geprüften Eutheria. Die Bedeutung unserer Befunde bezüglich der molekularen Evolution der Plasmaproteine werden diskutiert.

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A preliminary report has been given on the cross-reactions of human plasma proteins with one kangaroo species (Bauer, 1971).     

Antipeptide antibodies discriminate between different SAA proteins in human plasma

Antipeptide antibodies raised against two distinct sequences of human serum amyloid A (SAA) discriminate between different plasma isoforms of this acute phase reactant. As different SAA gene products have meanwhile been identified in human plasma, the discrimination between these different isoforms may help to clarify further the time of appearance of these different forms in plasma and their potential amyloidogenicity.     

Precipitation of animal plasma proteins by puff-adder venom.

1. Plasma and serum samples obtained from various animals never previously exposed to snakes or snake venom were diffused against different concentrations of puff-adder, Bitis arietans, venom using the double immunodiffusion technique. 2. Depending upon venom concentration, two precipitin arcs could be produced in the case of all plasma samples used. No serum samples showed any arcs except pigeon serum, where one precipitin line was observed. 3. By altering the concentration of snake venom between 1% and 10% when immunodiffusing against plasma a change in position of the precipitin lines was observed and also the disappearance of one or both of the two bands at higher concentrations. This indicates that the arcs observed are in all probability due to precipitation of plasma protein fractions. 4. Previous results indicated that one of the two bands observed when diffusing venom against plasma was due to the precipitation of fibrinogen. By diffusing snake venom against heparin we have now shown that the second band involves this molecule and is not due to another coagulation factor as was suggested previously.     

Cross-reactions between Staphylococcus epidermidis and 23 other bacterial species

By quantitative immunoelectrophoretic methods, 43 antigens were found in a mixture of sonicated preparations of four Staphylococcus epidermidis strains, using corresponding rabbit antiserum. Two of the antigens were identified as cell wall teichoic acid and a peptidoglycan antigen, respectively. Using this antigen/antibody reference system, cross-reactions between S. epidermidis antigens and antigens from other bacterial species were investigated. Fourteen of the S. epidermidis antigens cross-reacted with antigens from all S. aureus strains investigated. Only few cross-reactions were found between S. epidermidis and bacteria not belonging to the Micrococcaceae. The antigenic relatedness, expressed as a matching coefficient, seems promising for taxonomic work.     

Genetic studies of low-abundance human plasma proteins

Summary Genetic variation in the C1R subcomponent of the first complement component C1 was investigated in U.S. whites by isoelectric focusing and immunoblotting. In addition to the previously described two alleles, the products of a new and rare third allele designated C1R ^*3 were detected. The expression of the new allele is consistent with autosomal codominant inheritance, which is confirmed by family data. The frequencies of the C1R ^*1, C1R ^*2 and C1R ^*3 alleles in 201 randomly selected U.S. whites are: 0.908, 0.090, and 0.002, respectively.     

Binding of lithium and boron to human plasma proteins

The binding of lithium and boron, at normal physiological levels, to plasma proteins has been investigated by the techniques of precipitation with ethyl alcohol and gel chromatography. Assays of lithium and boron were made by thermal neutron activation and mass spectrometric assay of³He and⁴He. Results of alcohol precipitation experiments for plasma from two apparently healthy donors showed that 13 ± 4% and 16 ± 3% of the lithium in plasma is protein bound, but essentially no boron is bound under the conditions used. We believe that because of denaturation of proteins which occurs during alcohol precipitation, these percentages represent lithium and boron tightly bound to protein molecules. The results of the gelchromatography experiment, on the other hand, showed that lithium and boron are bound to a wide range of plasma proteins, from low (∼ 60,000 amu) to high (∼ 1,000,000 amu) molecular weights, and to very low- (∼ 6000 amu) molecular-weight ligands. Although a clear identification of the specific proteins which bind lithium and boron cannot be made at present, some possibilities can be suggested. Certain commercial equipment, instruments, or materials are identified in this article to specify adequately the experimental procedure. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or equipment identified are necessarily the best available for the purpose.     

Cross-reactions between the antigens of healthy pulmonary tissue and Moraxella catarrhalis

The study of cross-reactions between healthy pulmonary tissue antigens and Moraxella catarrhalis with the use of SDS-electrophoresis and immunoblotting revealed that in the component of healthy pulmonary tissue with a mol. wt. of 40 kD epitopes existed to which antibodies were produced, capable of cross reaction with the components of M. catarrhalis with a mol. wt. of 35 kD and 70 kD. In addition, the presence of cross-reactions between cytokeratin-8, protein contained in healthy pulmonary tissue, and M. catarrhalis antigens was established.     

Cross-reactions between engineered xylose and galactose pathways in recombinant Saccharomyces cerevisiae

Background

Bioethanol can be produced from sugar-rich, starch-rich (first generation; 1G) or lignocellulosic (second generation; 2G) raw materials. Integration of 2G ethanol with 1G could facilitate the introduction of the 2G technology. The capital cost per ton of fuel produced would be diminished and better utilization of the biomass can be achieved. It would, furthermore, decrease the energy demand of 2G ethanol production and also provide both 1G and 2G plants with heat and electricity. In the current study, steam-pretreated wheat straw (SPWS) was mixed with presaccharified wheat meal (PWM) and converted to ethanol in simultaneous saccharification and fermentation (SSF).

Results

Both the ethanol concentration and the ethanol yield increased with increasing amounts of PWM in mixtures with SPWS. The maximum ethanol yield (99% of the theoretical yield, based on the available C6 sugars) was obtained with a mixture of SPWS containing 2.5% water-insoluble solids (WIS) and PWM containing 2.5% WIS, resulting in an ethanol concentration of 56.5 g/L. This yield was higher than those obtained with SSF of either SPWS (68%) or PWM alone (91%).

Conclusions

Mixing wheat straw with wheat meal would be beneficial for both 1G and 2G ethanol production. However, increasing the proportion of WIS as wheat straw and the possibility of consuming the xylose fraction with a pentose-fermenting yeast should be further investigated.