Felix Haurowitz

Antigene, Antikörper und Immunität

SchlußbemerkungenDie vor 6 Jahren mitBreinl19 aufgestellte Anschauung über Bau und Bildung der Antikörper wird durch die Versuche der letzten Jahre gestützt. Die Versuche gestatten weitere Aussagen über die Reaktion zwischen Antigen und Antikörper, vor allem über das Wesen der zur Präcipitation führenden Vorgänge. Es ist anzunehmen, daß sich die an künstlichen, chemisch markierten Antigenen aufgedeckten Gesetzmäßigkeiten auch auf die natürlichen Antigene übertragen lassen; doch bedarf dies weiterer Prüfung an einem großen Material. Es ist der Zweck der vorliegenden Abhandlung, solche Versuche an klinischem Material anzuregen.

SOME STUDIES ON THE SPECIFICITY OF AMINO ACID ENTRY ROUTES IN PIGEON ERYTHROCYTES.

Abstract Inhibition of glycine entry into pigeon red cells by 33 amino acids, analogues, and derivatives was tested. Only N -methylglycine and N -ethylglycine inhibited glycine entry significantly. These were competitive inhibitors with ratios of inhibition constants to the Michaelis constant for glycine entry of 1.0 and 5.2, respectively. Derivatives with larger substituents on the nitrogen were essentially inactive as inhibitors, as were all the l -amino acids tested, two d -amino acids ( d -alanine and d -leucine), and several other glycine derivatives and analogues. The glycine route entry is evidently highly specific. An l -alanine entry route of high capacity was also observed. This was insensitive to inhibition by glycine and therefore distinct from the glycine entry route.

Mechanism of plastein formation

Abstract The formation by chymotypsin of plastein from concentration peptic hydrolyzates of egg albumin has been investigated in detail. Some properties of plastein are described. Free [14C]amino acids are not incorporated into plastein to an appreciable degree. However, 14C-labeled phenylalanine, tyrosine, threonine, aspartic acid, glutamic acid, leucine and isoleucine, and histidine are incorporated into plastein when their ethyl esters and chymotrypsin are added to the concentration digest. No incorporation of [14C]glycine, alanine, serine, cystine, proline and lysine was observed. Attempts by various methods to remove the incorporated radioactivity all failed; it is presumed that the labeled amino acids are bound in peptide linkage. Since the α-amino nitrogen remains unchanged during plastein formation, it is concluded that transpeptidation reactions are primarily responsible for plastein synthesis by chymotrypsin.

Biological Problems and Immunochemistry

MMUNOLOGY as a branch of biology and medicine started with the observation that human and animal organisms which had survived bacterial infections became immune to a later infection of the same kind. It was soon found that the serum of the immune organism is able to agglutinate the respective microorganisms, to lyse them or to render possible their phagocytosis. These actions of the immune serum are directed specifically against the microorganism whose invasion had been previously overcome. The unknown substance of the immune serum which reacts with the invading bacteria was called antibody, while the name antigen was chosen for the foreign agent which induced the formation of antibodies. This term antigen is not very suitable, however, because antigens, generally, are not directed against genes. The antigens used in older immunological work were bacteria and cells, chiefly red blood cells. Later, the important discovery was made that also uniform substances, such as crystalline proteins, are able to act as antigens, and that the antigenicity of cellular antigens is not due to the cell as a whole, but to the presence of several antigenic substances in the cell. Accordingly, each cell has to be considered as a mosaic of difYerent antigens giving rise to the formation of a variety of antibodies. Thus, the injection of pneumococci into rabbits leads to the formation of antibodies against the species-specific protein of pneumococci and of other antibodies against the type-specific capsular polysaccharides (Heidelberger and Avery, 1924). Both types of ant ibodies can be separated from each other A multiplicity of different antigens has also been found in red blood cells. Human erythrocytes contain the important blood group substances A and B. They are polysaccharides of the same type as those found in certain bacteria (Finland and Curnen, 1940; Kabat et al., 1948) and have also been found in saliva and in the gastric juice. The chemical nature of the M, N, and Rh antigens of human erythrocytes has not yet been elucidated. A still larger mosaic of different antigens has been found in the red blood cells of the dove (Irwin, 1947). The great importance of all these antigens for problems of anthropology, systematics, and genetics is discussed in the following papers, by Professors Boyd and Irwin, respectively, who have contributed signally to our knowledge about these things. If we want to get an insight into the mechanism of immunity, it is preferable to use uniform chemical substances as antigens. These are not only simpler than the complex cellular antigens, but can also be determined accurately by means of chemical analytical methods, and can be measured in the usual units of weight. The use of chemical substances as antigens has given rise to a new branch of immunology, to immuno-chemistry. It is obvious that this is the part of immunology from which essential contributions concerning the nature of immunological reactions can be expected. It is the task of this communication to report on advances in this field. The technique most frequently used in immilno-

Composition of antibodies against acidic and basic azoproteins.

Abstract Rabbits were injected with azoproteins prepared by coupling bovine γ-globulin with diazotized arsanilic acid, anthranilic acid, or p -aminophenyltrimethylammonium chloride. The antibodies to these three antigens were isolated, purified, and analyzed. Their amino acid composition did not differ significantly. They contained approximately 3% carbohydrate. Their serological specificity may be due to differences in the amino acid sequence or tertiary structure, rather than to different amino acid content.

Zur Chemie des Hypophysenvorderlappen-Sexualhormons

ZusammenfassungDas Hypophysenvorderlappensexualhormon wird mit dem aus Schwangerenharn gewonnenen Stoff gleicher biologischer Wirkung verglichen. Die chemische Untersuchung ergibt für beide Stoffe Gleichheit im Verhalten bei der Adsorption und im Verhalten gegen Proteasen; beide werden durch Uranylacetat gefällt, durch Tonerde-Gel adsorbiert und aus den Fällungen durch wässerig-ammoniakalische Lösungsmittel eluiert; beide werden von proteolytischen Enzymen inaktiviert, und zwar von den peptonspaltenden Polypeptidasen. Sie dürften daher den Peptonen nahestehen. Durch ihr Verhalten gegen Enzyme ist ihre geringe Wirksamkeit bei Verabreichung per os erklärt.

Localization of antigens by autoradiography.

Summary Analyses of the distribution of isotopically labeled antigens in the animal were combined with autoradiography of the tissues. After intravenous injection of heavily substituted S35-sulfanilazo-bovine γ-globulin into mice, we found protein-bound S35 in the cytoplasm of the Kupffer cells of the liver. Similarly, activity of injected biosynthetically labeled S35-RbSA was concentrated in the cytoplasm of lymphoid cells of the spleen, particularly in the red pulp. After injection of lightly labeled S35-sulfanilazo-bo-vine γ-globulin into the foot pads of rabbits, the radioactive material was concentrated in the cytoplasm of the eosinophilic macrophages of the popliteal lymph nodes. In all these cell types, the protein-bound S35 was either present within the cytoplasm or adsorbed to the cellular membrane. Little intranuclear radioactivity was evident.

Peptide maps of antibodies against an antigen containing two different determinant groups

Abstract Bovine serum albumin doubly labelled by coupling with equivalent amounts of diazotized arsanilic acid (Ars) and p-aminophenyl-N-trimethyl-ammonium chloride (R 4 N) was injected into rabbits. Hapten-specific antibodies against the two determinants were separated from each other and from anti-BSA. Peptide maps of anti-Ars and anti-R 4 N differ from each other only in one or two spots. Large portions of the two types of antibody molecules seem to be identical. Since both haptens are bound to the same BSA molecules, we assume that they are initially incorporated into the same cells. It is not yet known whether anti-Ars and anti-R 4 N are formed in the same cells or whether the haptens, after fragmentation of the antigen, are transferred to different antibody forming cells.

Comparison of the peptide maps of antibodies against and acidic and a basic determinant group

Abstract Antibodies were isolated from the serum of rabbits injected with azophenyl-arsonate-BSA or azophenyltrimethylammonium-BSA. The structures of antibodies formed against the two haptens and also those formed against BSA were compared by finger-printing. No repoducible differences could be detected in the peptide patterns of the univalent fragments obtained by the papain digestion of the antibodies from pooled immune sera against each of the tow antigens. When the two hapten-specific antibody species were each prepated from single antisera, a difference involving a single peptide spot was observed. The same difference was found in the peptide maps of the BSA-specific antibody fragments. This suggests that the small difference between the fingerprints in the hapten-specific antibodies may arise from genetic differences in the globulin pattern of the two rabbits or from differences in the organ distribution of the two antigens rather than from the different immunological specificity of the two haptens. A small portion of the peptides obtained by tryptic digestion of the antibodies or their univalent fragments does not migrate upon electrophoresis or chromatography but remains at the origin. It was found that this immobolie material si rich in sulfur amino acids.

Serological properties of poly-L-tyrosine derivatives

Polytyrosin und Polytyrosylgelatine wurden mit diazotierter Arsanilsäure gekoppelt und in Lösung oder nach Fällung mit Alaun und Alkali Kaninchen injiziert. Die injizierten Substanzen wurden von keinem der Sera präzipitiert. Hingegen präzipitierte das Serum der mit gefällter Arsanilazo-polytyrosyl-gelatine injizierten Tiere Arsanilazo-Rinderserumglobulin. Wir schliessen daraus, dass Arsanilazopolytyrosin nicht als Antigen wirkt, dass es aber durch Bindung an Gelatine antigene Eigenschaften gewinnt.