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Formation of antibody by isolated perfused lungs of immunized rabbits. The use of [ 14 C]amino acids to study the dynamics of antibody secretion

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... What then seemed surprising was that, after intravenous immunization, bone marrow and lung turned out to be potent antibody-forming organs (5). In fact, perfusion of lung yielded milligram quantities of antibody to PnPS (6). In these experiments we also found that the immunization induced not only specifi c antibodies but also high levels of antigen-nonspecifi c Ig. ...
... However, experimental studies ( Askonas and Humphrey, 1958;Humphrey, 1960) have shown that the rise in the y-globulins may be partly attributed to the synthesis of "non-specific" antibody. ...
... The concentration of the leucine will not change much during incubation, as essential amino acids are not formed anew, and, on the other hand, only a small proportion of the leucine was used. If further the crude assumption is made that the specific activity of the newly incorporated leucine is about half that of the free leucine in the medium (Loftfield & Harris, 1956;Steiner & Anker, 1956;Askonas & Humphrey, 1958), the mass of the leucine incorporated into serum albumin works out as 12-5x0-0264x2 = 0-66 jig. With a leucine content (by weight) of humanserum albumin of 11 %, deduced from data given by Hughes (1954) and Peters, Logan & Sanford (1958), the quantity of albumin newly formed in 48 hr. is computed as 6 ,ug./roller tube. ...
... What then seemed surprising was that, after intravenous immunization, bone marrow and lung turned out to be potent antibody-forming organs (5). In fact, perfusion of lung yielded milligram quantities of antibody to PnPS (6). In these experiments we also found that the immunization induced not only specifi c antibodies but also high levels of antigen-nonspecifi c Ig. ...
... At the height of the immune response to the capsular antigen, the animals were killed and the organs removed to determine their in vitro capacity to produce antibody. By using an assay based on the incorporation of 14C-labeled amino acids into 3,-globulin, intravenous immunization led to antibody production primarily in bone marrow and lungs and in some cases predominantly in the lung itself (18)(19)(20). ...
Article
In a search for possible genetic factors which may influence the immune response to the streptococcal carbohydrates, over 100 rabbits have been immunized with streptococcal vaccines, and representative examples of high and low response pairs mated. The concentration of precipitins to the group—specific carbohydrates has been measured in the antisera following primary intravenous immunization with heat-killed streptococcal vaccines, Group A, Group A-variant, and Group C. For the majority of rabbits, the concentration of precipitins varied between 1 and 10 mg/ml of antiserum; while in the minority, it was between 11 and 32 mg/ml. The offspring of rabbits with high antibody levels had a significantly higher concentration of antibody than was seen in the offspring of rabbits of low response parents. Such data suggest that the magnitude of the immune response to these carbohydrate antigens is under some form of genetic control. Not uncommonly in rabbits with hyper-γ-globulinemia following primary immunization, the group-specific precipitins are the predominant component of the γ-globulin. An unusual feature of such components is that they are electrophoretically monodisperse, and possess individual antigenic specificity. In this respect they resemble the myeloma proteins. When a response of this sort is not seen after primary immunization, it may occur after secondary immunization. Therefore, prior exposure to the same or closely related antigen may also have an influence on the occurrence of high concentrations of such uniform antibodies.
... (26,27) immunoglobulins are secreted from plasma cells without cell disruption . In contrast to the situation in many exocrine and endocrine cells, where secretory products are concentrated and temporarily stored in secretory granules, cells such as the plasmacyte appear to transport and discharge continuously without concentration and storage of their product (28,29,26) . At present, the mechanism of discharge is not understood, though two alternatives can be considered . ...
Article
The subcellular sites of synthesis and route of intracellular transfer of immunoglobulin G (IgG) have been investigated by electron microscope radioautography with precursors used for the polypeptide chain (leucine-3H) and for the carbohydrate moieties (galactose-3H and glucosamine-3H). For this purpose, plasma cells from a mouse myeloma tumor were labeled with appropriate precursors and the distribution of radioautographic grains was determined at the end of the labeling period and after varying times of incubation in unlabeled medium. The results indicated that the polypeptide backbone is synthesized in a region of the cell occupied by the rough endoplasmic reticulum (RER) and is transported from there to the region of the Golgi complex. Galactose is incorporated in IgG primarily at the level of the Golgi complex, whereas the incorporation of glucosamine appears to take place both in the RER and in the Golgi complex. From the Golgi complex, the completed IgG molecules reach the plasma membrane and are discharged extracellularly. The latter route of transport and the mechanism of discharge are not understood but may be mediated via smooth-surfaced vesicles.
... At the height of the immune response to the capsular antigen, the animals were killed and the organs removed to determine their in vitro capacity to produce antibody. By using an assay based on the incorporation of 14C-labeled amino acids into 3,-globulin, intravenous immunization led to antibody production primarily in bone marrow and lungs and in some cases predominantly in the lung itself (18)(19)(20). ...
Article
In a search for possible genetic factors which may influence the immune response to the streptococcal carbohydrates, over 100 rabbits have been immunized with streptococcal vaccines, and representative examples of high and low response pairs mated. The concentration of precipitins to the group—specific carbohydrates has been measured in the antisera following primary intravenous immunization with heat-killed streptococcal vaccines, Group A, Group A-variant, and Group C. For the majority of rabbits, the concentration of precipitins varied between 1 and 10 mg/ml of antiserum; while in the minority, it was between 11 and 32 mg/ml. The offspring of rabbits with high antibody levels had a significantly higher concentration of antibody than was seen in the offspring of rabbits of low response parents. Such data suggest that the magnitude of the immune response to these carbohydrate antigens is under some form of genetic control. Not uncommonly in rabbits with hyper-γ-globulinemia following primary immunization, the group-specific precipitins are the predominant component of the γ-globulin. An unusual feature of such components is that they are electrophoretically monodisperse, and possess individual antigenic specificity. In this respect they resemble the myeloma proteins. When a response of this sort is not seen after primary immunization, it may occur after secondary immunization. Therefore, prior exposure to the same or closely related antigen may also have an influence on the occurrence of high concentrations of such uniform antibodies.
... However, there was evidence of considerable liver damage in the perfused liver preparations, so that these results may be artefactual. No significant catabolism of serum proteins has been found in the lungs or the pancreas (Askonas & Humphrey, 1958;Jarnum et al., 1966). The failure to identify a specific organ as the site of immunoglobulin catabolism suggests that catabolism may occur in a diffusely distributed organ system. ...
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Myotonic dystrophy (MyD), an autosomal dominant neuromuscular disease with multisystem abnormalities, is associated with hypercatabolism of IgG. The hypercatabolism is not related to structural abnormalities of the IgG molecule in MyD but appears to be due to a derangement of the serum IgG concentration-fractional catabolic rate relationship. Since the catabolic pattern of IgG is governed by the Fc portion of the molecule, the possibility of Fc receptor dysfunction in MyD has been explored. We have observed that although MyD patients have normal numbers of Fc receptor bearing leucocytes in their peripheral blood, MyD monocytes express significantly (P less than 0.02) greater numbers of Fc receptors (47.9 +/- 21.2 X 10(3) receptors/monocyte) than do monocytes of healthy subjects (29.1 +/- 9.6 X 10(3) receptors/monocyte). The mean affinity constants of the Fc receptors was lower in the MyD group (1.5 +/- 0.7 X 10(8)/M) than the normal control group (2.4 +/- 0.9 X 10(8)/M) but this difference was not statistically significant. MyD monocytes showed a propensity to shed Fc receptors in culture at 37 degrees C whereas no significant shedding was observed with control monocytes. Thus MyD monocytes may shed Fc receptors at physiological temperatures but at the same time express more receptors per cell than normal monocytes. This suggests that MyD monocytes may have an abnormally high turn-over of Fc receptors.
Chapter
A considerable amount of research has been done on antibody synthesis. This is first and foremost a result of its practical importance, which is reflected in the fact that a considerable portion of the population is vaccinated each year with the most varied antigens. It is also the reason for the comprehensive literature on the productive phase of antibody synthesis, i.e., the period commencing at the moment antibodies appear in the blood. However, at the present, many investigators are concentrating on the elucidation of processes which are initiated directly after administration of antigen to the organism and which ultimately lead to formation of antibody molecules. As in the biosynthesis of other proteins, nucleic acids can be expected to play an important role in the mechanism of antibody formation. Hence, it is not surprising that much attention is being devoted at the present time to synthesis of nucleic acids in antibody-forming cells and tissues.
Article
The subcellular sites of synthesis and route of intracellular transfer of immunoglobulin G (IgG) have been investigated by electron microscope radioautography with precursors used for the polypeptide chain (leucine-(3)H) and for the carbohydrate moieties (galactose-(3)H and glucosamine-(3)H). For this purpose, plasma cells from a mouse myeloma tumor were labeled with appropriate precursors and the distribution of radioautographic grains was determined at the end of the labeling period and after varying times of incubation in unlabeled medium. The results indicated that the polypeptide backbone is synthesized in a region of the cell occupied by the rough endoplasmic reticulum (RER) and is transported from there to the region of the Golgi complex. Galactose is incorporated in IgG primarily at the level of the Golgi complex, whereas the incorporation of glucosamine appears to take place both in the RER and in the Golgi complex. From the Golgi complex, the completed IgG molecules reach the plasma membrane and are discharged extracellularly. The latter route of transport and the mechanism of discharge are not understood but may be mediated via smooth-surfaced vesicles.
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Lung lesions characterized by extensive peribronchial and perivascular round cell infiltration and intrabronchial plugging with polymorphonuclear leukocytes were produced in 5 of 44 Ha/ICR gnotobiotic mice sacrificed 3 to 10 days after three intranasal inoculations of broth cultures of Mycoplasma pneumoniae. After 10 days, no significant lesions were seen, and the proportion of lungs positive for M. pneumoniae dropped off sharply. M. pneumoniae persisted for longer periods (up to 24 days) in the trachea, nasopharynx, and anterior nares. These findings would seem to represent a self-limited respiratory infection due to M. pneumoniae in gnotobiotic mice. Ring forms within granular alveolar pneumocytes were seen by electron microscopy in the lungs of triply inoculated gnotobiotic controls receiving sterile horse-serum broth as well as in the lungs of mice receiving broth cultures of M. pneumoniae. Ring forms must now be considered to be part of the nonspecific cellular reactions of pneumocytes to foreign substances in the lungs of mice rather than intracytoplasmic developmental forms of mycoplasma as previously proposed.
Article
Full-text available
In a search for possible genetic factors which may influence the immune response to the streptococcal carbohydrates, over 100 rabbits have been immunized with streptococcal vaccines, and representative examples of high and low response pairs mated. The concentration of precipitins to the group-specific carbohydrates has been measured in the antisera following primary intravenous immunization with heat-killed streptococcal vaccines, Group A, Group A-variant, and Group C. For the majority of rabbits, the concentration of precipitins varied between 1 and 10 mg/ml of antiserum; while in the minority, it was between 11 and 32 mg/ml. The offspring of rabbits with high antibody levels had a significantly higher concentration of antibody than was seen in the offspring of rabbits of low response parents. Such data suggest that the magnitude of the immune response to these carbohydrate antigens is under some form of genetic control. Not uncommonly in rabbits with hyper-gamma-globulinemia following primary immunization, the group-specific precipitins are the predominant component of the gamma-globulin. An unusual feature of such components is that they are electrophoretically monodisperse, and possess individual antigenic specificity. In this respect they resemble the myeloma proteins. When a response of this sort is not seen after primary immunization, it may occur after secondary immunization. Therefore, prior exposure to the same or closely related antigen may also have an influence on the occurrence of high concentrations of such uniform antibodies.
Article
The catabolism of IgG2a was followed in rats by the use of 125I, 14C and antibody labels. The organ and subcellular distribution of 125I and 14C was studied following the intravenous injection of [125I]IgG2a and [14C]IgG2a. The distribution of [125I]IgG2a and [14C]IgG2a after their incubation in vitro with cell suspensions of rat spleen and lymph nodes was also studied. The results show a close relationship between the liver, spleen and lymph nodes, and IgG2a catabolism. Intermediate products and metabolites of IgG2a were found intracellularly in the spleen and lymph nodes.
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Immunoglobulin synthesis and secretion have been studied in the rabbit lower respiratory tract, both in the normal state and after infection with Diplococcus pneumoniae or Listeria monocytogenes. In vitro synthesis of immunoglobulin and specific antibody was assessed by incorporation of 14C-labeled amino acids into protein. Lower respiratory tract secretions and serum were analyzed for immunoglobulin and antibody against the infecting organism. Normal respiratory tract produced small quantities of immunoglobulin, most of which was IgG. After bacterial infection of the lower respiratory tract, there was a marked increase in local synthesis of immunoglobulin, especially IgG. Specific antibody of IgG class was produced in all lungs infected with listeria by the 11th day, and in lungs infected with pneumococcus by the 8th day. Secretions from all normal and infected lower respiratory tracts contained IgA and IgG. The IgA to IgG ratios in secretions of normal animals, and animals infected with listeria or pneumococcus, were 2.3, 2.5, and 2.6, respectively. Sera of animals infected with L. monocytogenes contained specific antibody of IgG class but lacked IgA antibody, whereas secretions had both IgA and IgG class antibody against listeria. Similarly, sera of animals infected with D. pneumoniae had IgG class antibody but no IgA antibody, whereas only IgA antibody was found in secretions. The evidence that locally synthesized immunoglobulin (especially IgA), including specific antibody, is secreted into the lower respiratory tract lumen is discussed. Further definition of the role of "local" antibacterial antibody in the respiratory tract is of considerable importance.
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The chapter presents a discussion on effects of injury on plasma proteins. The chapter states that injury in different forms is regularly accompanied by acute changes in plasma proteins. The changes mostly studied comprise a fall in plasma albumin and transferrin, a rise in plasma fibrinogen, haptoglobin, and ceruloplasmin, and the appearance of C-reactive protein in the plasma. An increase in total plasma mucoprotein and in protein-bound carbohydrate suggests that other individual proteins are also affected. The chapter mentions that the changes are independent of the type of injury. They occur in man and in animals after surgery, after burns, after accidental trauma, after γ-irradiation, after myocardial infarction, and after injury because of bacterial and other infections, and start to develop within the first 24 hours of injury. The chapter explains that the acute effects of injury on plasma protein level are best explained, mainly in terms of an increase in metabolic turnover of the protein concerned. All the proteins acutely affected are formed in the liver, which is stimulated to increase protein synthesis through the action of some factor generated by injury. The net effect differs from protein to protein. The chapter highlights that the net transfer of protein from the intravascular to the extravascular compartment contributes to the fall in plasma albumin and transferrin, but in most injuries this is likely to be a minor factor only.
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Gamma globulin formation in vitro by various tissues was studied using the incorporation of C14-L-lysine into a protein precipitable by a specific anti rabbit gamma globulin serum prepared in a sheep. It was demonstrated that the rate of gamma globulin formation in similar numbers of spleen cells is much higher if taken from an immunized rabbit at the height of antibody formation than that in normal spleen cells. Besides spleen, other tissues shown to form gamma globulin in normal adult rabbits were: peripheral lymph nodes, bone marrow, lung, mesenteric lymph nodes, appendix, and thymus. In tissues from newborn rabbits gamma globulin formation could not be demonstrated. Rabbits 1 week old already showed a beginning of significant gamma globulin formation in appendix tissue, followed approximately 3 weeks later by gamma globulin synthesis in spleen and thymus. Histological observations on these tissues were described and correlated with findings on gamma globulin formation. In the discussion an attempt has been made to relate these observations on newborn and immature rabbits to those available in the literature on antibody formation in newborn animals.
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The production of individual serum proteins by individual kinds of tissue is difficult to prove or to disprove in experiments with intact animals or with surviving organs, since these contain a multi-tude of different kinds of tissue, often of over-lapping activities and capable of mutual inter-action. It is therefore preferable to investigate uniform tissue in isolation, i.e. in culture. Such tissue proliferates, in principle, indefinitely. Its biochemical potentialities presumably do not exceed those of the same kind of tissue in vivo, though they might be more restricted; thus the positive proof that a particular protein is synthe-sized by tissue in culture indicates that such syn-thesis may also proceed in vivo. Intact animals or surviving organs are impreg-nated with fluids containing serum proteins, e.g. blood. It is therefore necessary when using them in experiments on protein synthesis to distinguish between pre-existing protein and newly synthe-sized protein. This can be done by the use of labelled amino acids, when only the newly formed protein will contain the label. Even in experiments with tissue cultivated in a medium originally free of serum proteins, however, the distinction between preformed and newly formed proteins is still needed in view of the protein contents of the cells themselves. For this reason, the use of isotopes is almost indispensable in this case as well. Moreover, the use of radioactive isotopes gives a method where the sensitivity of measurement is very high. Three kinds of tissue were used for our experi-ments, namely mesenchyma tissue from chick embryos, epithelial-tumour HeLa tissue and a transformed human-liver-cell strain HLM. The tissue was incubated with nutrient medium con-taining [14C]amino acid, and subsequently the soluble proteins of the tissue and medium were separated and fractionated by selective precipita-tion with ethanol. Electrophoresis in starch gel, followed by staining, was carried out as a test for the purity of the fractions. Finally, the radio-* On leave from the Clinical Pathology Department, 14 activity of the fractions separated with ethanol was measured; in one series (Table 3) the radioactivity of the eluates from the electrophoresis gels of such fractions was measured. The purity of the prepara-tion of serum albumin has also been tested radio-immunochemically. Various control experiments were needed to make sure that the results were not falsified by adsorption or other processes simulating genuine incorporation of amino acid. The most striking result of our work is the proof that both chick-mesenchyma tissue and HeLa-tumour tissue make serum albumin. A preliminary report of some of our results has been given (Abdel-Samie, Broda, Kellner & Zischka, 1959).
Article
THE problem of sites of synthesis of gamma-globulins and of their relative contribution to the total synthesis of these proteins in the normal animal nas received little attention compared with that devoted to similar studies on the synthesis of antibody globulins. This problem becomes specially interesting if one considers the controversial views which have been put forward on the origin and nature of gamma-globulins1.
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This chapter focuses on the immunology of schistosomiasis. Consideration of innate immunity to schistosomes begins with the host specificity shown by the parasitic species. It is well recognized that of the three common schistosomes of man, S. japonicum is the least host-specific. This species meets little innate resistance in a variety of vertebrate animals, especially dogs, goats, sheep, pigs, and cattle, which as domestic animals gain importance as reservoir hosts. Although a variety of mammals have been implicated as hosts of the schistosomes of man, the degree of infection tolerated by each host species varies considerably. The degree of susceptibility of a host can be determined by the percentage of parasites developing, the growth and structural development of the worms, the ability of the worms to produce eggs, the infectivity of miracidia for suitable snails, and the sex ratio and location of the worms in the host. However, only those animals, which become infected with large numbers of worms and emit numerous viable eggs have epidemiological significance as potential reservoirs of infection. The mechanisms of host susceptibility are discussed in the chapter and the nature of acquired immunity is reviewed.
Article
Monoclonal anti-streptococcal group antibodies can readily be induced by intravenous hyperimmunization. A vaccine of heat-inactivated bacteria is used for immunization. Such antibodies show all the criteria of uniformity: electrophoretically, they migrate as sharp and narrow bands; their polypeptide light chains are restricted to one or two bands in polyacrylamide disc electrophoresis; they are homogeneous as to class, type and allotype; their variable regions exhibit individual antigenic specifity, and amino-terminal Edman degradation of the light chains yields a single amino acid residue per degradation step. These antibodies are therefore valuable counterparts to Bence Jones proteins and myeloma proteins. They allow for most detailed studies of the structure-function relationship of antibodies. The induction of high homogenous polysaccharide antibody levels depends on the following factors: the simple structure of the antigen; intravenous immunization; repeated contact with the identical antigen, as well as genetic factors. Breeding studies with such rabbits suggest that not only the magnitude but also the restriction of the immune response are under genetic control.
Chapter
From the clinician’s point of view, the wider use of transplantation as a remedy for disease is being restrained by a number of factors. Despite the considerable degree of success achieved in its most extensive clinical application — kidney transplantation — transplantation of organs must still be regarded as an experimental procedure (Hume, 1971). The central stumbling block is the rejection, since due to an apparently immutable law of immunity the transplantation of tissues to a genetically, i.e. antigenically dissimilar host inevitably mobilizes defense mechanisms against the foreign antigens. It has been suggested that this law may have a crucial loophole. The canine and, particularly, the porcine liver appear to be immunologically favored, since in unmodified hosts identifiable allograft rejection did not occur or reversed spontaneously, whereas in the same animals skin and kidney were regularly rejected (Calne et al., 1969; Garnier et al., 1970). In human liver transplantation it has become the usual practice to ignore the result of HL-A typing (Starzl, 1971; Calne, 1974). Since other experimenters found that canine, porcine, and rat liver allografts were rejected in the usual way (Porter, 1969; Jerusalem et al., 1971b; Jap, 1971), the above findings may represent a particular modification of the immunologic network of responses, rather than an exception to the principle of the transplantation reaction.
Chapter
This chapter describes the antibody response, beginning with the entrance of antigen into certain cells, to all the cellular events that culminate in the synthesis of circulating antibody. The chapter presents two methods for in vitro study of antibody response—maintenance of function of antibody-forming cells and assays of antibody and antigen. The chapter examines certain general factors that affect all aspects of the antibody response and influence interpretation of experimental data, including the chemical and physical nature and dosage of antigen; the age, strain, and species of experimental animal; the amounts and type of antibody produced; the methods utilized for the assay of antigen and antibody. The chapter describes the mechanism of antibody response. This mechanism involved uptake and fate of antigen and examined cells involved in the initiation of the antibody response. For this, an in vitro uptake of antigen followed by antibody synthesis in vivo is performed.
Ribonucleoprotein (RNP)1 particles isolated by DOC treatment from pancreatic microsomes have a RNA content of 35 to 45 per cent of their dry weight. In the analytical ultracentrifuge about 85 per cent of the material has a sedimentation coefficient of ∼85 S. These particles contain amylase, RNase, and trypsin-activatable proteolytic activities which cannot be washed off or detached by incubation in 0.44 M sucrose. The enzymes are released, however, by incubation in the presence of low concentrations of ATP, PP, or EDTA, and high concentrations of IP and AMP. At the same time, and at the same concentrations, ∼80 per cent of the RNA and ∼25 per cent of the protein of the particles becomes also non-sedimentable. The simultaneous addition of Mg++ to the incubation medium prevents these losses. This finding, together with the observation that all the Mg++ of the particles is released by the same agents, makes it likely that Mg++ holds the particles together, and that its removal by the chelators used causes the particles to disintegrate. These findings are discussed in relation to the molecular structure of the RNP particles.
Article
Gamma globulin formation in vitro by various tissues was studied using the incorporation of C14-L-lysine into a protein precipitable by a specific anti rabbit gamma globulin serum prepared in a sheep. It was demonstrated that the rate of gamma globulin formation in similar numbers of spleen cells is much higher if taken from an immunized rabbit at the height of antibody formation than that in normal spleen cells. Besides spleen, other tissues shown to form gamma globulin in normal adult rabbits were: peripheral lymph nodes, bone marrow, lung, mesenteric lymph nodes, appendix, and thymus. In tissues from newborn rabbits gamma globulin formation could not be demonstrated. Rabbits 1 week old already showed a beginning of significant gamma globulin formation in appendix tissue, followed approximately 3 weeks later by gamma globulin synthesis in spleen and thymus. Histological observations on these tissues were described and correlated with findings on gamma globulin formation. In the discussion an attempt has been made to relate these observations on newborn and immature rabbits to those available in the literature on antibody formation in newborn animals.
Article
Antigene, seien es pathogene Erreger oder künstliche chemische Verbindungen, veranlassen im Körper die Bildung von Antikörpern großer Spezifität, welche die Ursache der späteren Immunität sind. Die Spezifität ist u. a. in den räumlichen Formen der beteiligten Moleküle begründet. Antigen und Antikörper passen zueinander wie Matrize und Abguß (Faltung der Peptidketten). Über Einzelheiten der Antikörperbildung wird sich erst Entscheidendes sagen lassen, wenn wir mehr über die Biosynthese der Proteine und besonders der Serumglobuline wissen. Die wesentlichen Anschauungen und Theorien neuerer Zeit leiten sich noch von Ehrlichs Idee der Komplementarität ab.
Article
Oпиcывaeтcя мeтoд пepyзии ceлeзeнки кpыcы oбoгaщeннoй киcлopoдoм кpoвью, cвoбoднoй oт лeйкoцитoв и тpoмбoцитoв. Пepyзия пpoвoдитcя пpи пocтoянныч тeмпepaтype и дaвлeнии. Изyчeны ocoбeннocти тoкa кpoви зтoгo пpeпapaтa. Пepyзия ceлeзeнки кpыcы пpoвoдиacь дeнaтypиpoвaнным тeплoм aльбyминoм кpыcы, мeчeным I131. Toт akcy;т, чтo пocлe 20–30-минyтнoгo пepиoдa индyкции мeтaбoлчecкий пpoцecc пpoчoдил c пocтoяннoй cкopocтью и пpoдoлжaлcя втeчeниe чeтыpeч чacoв пepyзии, пoкaзывaeт, чтo peтикyлo-зндoтeлиaльнaя cиcтeмa вce зтo вpeмя нeпpepывнo yнкциoниpoвaлa. Пocлe нeбoльшoгo пepиoдa зaдepжки включeниe aминoкиcлoт, мeчeныч C14, в бeлки плaзмы ocyщecтвлялocБ c пocтoяннoй cкopocтью в тeчeниe нecкoлБкич чacoв. Peзyльтaты, выpaжeнныe в пpoцeнтaч oт paдиoaктивнoй мeтки включeннoй в бeлки плaзмы, дaют знaчeния в I-3% пocлe 4,5-чacoвoй пepyзии бeлкoв плaзмы пpoизвoдиocь в paзныч oпытaч зoнaль-ным элeктpoopeзoм, xpoмaтoгpaиeй нa цeллюoзe и бyмaжным элeктpoopeзoм. Былo пoкaзaнo, чтo мeкa включaeтcя в γ-, β- и α-глoбyлины, a включeниe в aльбyмин пpeнeбpeжимo мaлo. Гpyбaя oцeнкa cкopocти oбщeгo cинтeзa глoбyлинa в oднoм из oпытoв дaeт знaчeниe 9,5 мг/24 чaca, из нич 5,1 мг/24 чaca cooтвeтcтвyют γ-paкtscy;ии. Paдиoввктивнocть, в пopoшкe, пpигoтoвлeннoм из бeлкa пяти пepyзиpoвaнныx ceлeзeнoк, cocтaвлялa 3,5–5,4% oт paдиoaктивнocти ввeдeннoй мeтки. Oбcyждaeтcя пpoблeмa вoзмoжnncy;oй пoтepи бeлкa пpи пepyзии, чтo мoжeт пoвpeдить тoчнocти aнaлизa.
Article
The ability of alveolar macrophages of dogs to bind and metabolize two antigens was studied. The antigens were ragweed antigen E (AgE) and keyhole limpet haemocyanin (KLH). These antigens were chosen because of the availability of dogs with respiratory hypersensitivity to them and because they are of very different molecular weights. It was shown that: (1) antigen processing was identical using macrophages from hypersensitive as compared with normal dogs; (2) in comparison with AgE, much more of the higher molecular weight antigen, KLH, was bound to macrophages and much less degraded during subsequent incubation, leaving a greater proportion of the antigen bound to the cell membrane; (3) respiratory cells which had taken up AgE at 0 degrees C instead of 37 degrees C and were then incubated at 37 degrees C were much more active in catabolizing bound antigen with consequently less membrane bound antigen remaining; (4) increases in catabolism of bound AgE were also found in cells from relatively recently lavaged animals; (5) there was no evidence that cytophilic antibody contributed to the uptake of AgE.
Antibody-forming cells (AFC) produced in the lung-associated lymph nodes after lung immunization enter the blood and accumulate mainly in the immunized lung. In lung interstitial tissues and alveoli, AFC mature into plasma cells and produce specific antibody. In addition to AFC, published data suggest that memory cells are also recruited to and/or are produced in immunized lung lobes, and that these memory cells can respond in the alveoli to secondary antigen challenges. The purpose of this study was to determine if memory cells induced in the lung by multiple antigen exposures could respond in vivo to challenges with low doses of antigen. The degree of inflammation produced by antigen doses that would allow the accumulation of AFC from the blood was evaluated. Beagle dogs were anesthetized, and a fiberoptic bronchoscope used to instill 10(6), 10(7), 10(8), 10(9), or 10(10) sheep red blood cells (SRBC) into individual lung lobes. Of these doses, only 10(9) and 10(10) SRBC significantly increased inflammation (increased number of neutrophils) or vascular permeability (increased total protein). The number of specific IgM AFC and concentration of anti-SRBC IgG antibody were significantly elevated only in lavage fluid from the lung lobes immunized with 10(9) and 10(10) SRBC. Four lung lobes were then given two additional challenges with 10(10) SRBC. In third and fourth challenges, these lung lobes received doses of SRBC of 10(4), 10(5), 10(6), 10(7), 10(8), and 10(9) SRBC. The results showed that significantly increased numbers of specific IgM AFC and concentrations of IgG antibody were found even at the lowest dose of antigen (10(4) SRBC) in the absence of increased inflammation or vascular permeability.(ABSTRACT TRUNCATED AT 250 WORDS)
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This chapter discusses the physical, chemical, and biological aspects of γM antibodies in depth. The chapter describes the physical characteristics, chemical composition, and subunit structure of the typical circular γM pentomers and the relationship of these molecules to low molecular weight γM-like proteins. The characteristics of the interaction of γM antibodies with antigens—for example, the nature and number of antigen combining sites—and the interaction of γM antibodies with the complement system are compared to the corresponding properties of γG antibodies. The biosynthesis and metabolism of γM antibodies and their peculiar role in the immune response are also discussed in the chapter. The chapter presents a brief summary related to the hallmarks of immunoglobulin heterogeneity and the criteria that must be used to judge the restricted heterogeneity or molecular uniformity of an antibody preparation. The chapter discusses human antibodies to certain selected antigens that appear to possess less heterogeneity than normal γ-globulin.
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There is no reason to doubt that textbooks of immunology are correct when they teach that the immune response to antigenic stimulation is a reaction in which all the lymphatic tissues of the body have their share. However, work initiated shortly after the turn of the century and gaining increasing momentum in recent years has brought to light that nearly all individual tissues and organs, when confronted with an antigen, display the capacity to set up their own local immune response which is largely independent of the systemic reaction.
Article
Peripheral blood lymphocytes from 27 healthy individuals and from 18 patients with a diverse spectrum of defects in humoral immunity were examined for their capacity to undergo terminal differentiation in vitro. Pokeweed mitogen induced cells from normal persons to synthesize and secrete IgM. IgG, and IgA as detected by Immunofluorescence and incorporation of [(14)C]amino acids, Lymphocytes from three boys with X-linked agammaglobulinemia were stimulated to proliferate, but did not synthesize immunoglobulin. Lymphocyte cultures from three of four patients having agammaglobulinemia with B lymphocytes produced different immunoglobulin classes in ratios similar to the in vivo distribution of classes of B lymphocytes, Lymphocytes from a dysgammaglobulinemic boy deficient in serum IgG and IgA, but who had normal numbers of IgM-, IgG-, and IgA-bearing B lymphocytes, could not be stimulated by pokeweed mitogen to make IgG and IgA. Synthesis and secretion of IgA, as well as IgM and IgG, was detected in cell cultures from each of 10 patients with isolated IgA deficiency. The results suggest that deficiencies in immunoglobulin synthesis may reflect either (a) failure to develop B lymphocytes, (b) arrested development of B lymphocytes due to intrinsic metabolic abnormalities, or (c) disturbance of factors extrinsic to the B lymphocyte which are essential for normal induction of plasma cell maturation.
Article
Groups of dogs were immunized with sheep erythrocytes administered either directly into the lower respiratory tract (bronchoalveolar spaces) or intravenously. The hemolytic plaque-forming response (Jerne plaque assay) was studied in various canine lymphoid populations (bronchoalveolar cells, hilar lymph nodes, peripheral lymph nodes, and splenic and peripheral blood leukocytes) as a function of time after immunization and as a function of the dose of antigen administered. Serum hemagglutinating antibody titers against sheep erythrocytes were also measured. Intrapulmonary and intravenous administration of sheep erythrocytes to dogs both result in an immune response, the kinetics of which are identical to those observed in other animal species. At equivalent doses, the intravenous route is more efficient than the intrapulmonary route in generating serum hemagglutinating antibodies and antibody-forming cells. Both routes give rise transiently to circulating antibody-forming cells during the primary response; the distribution in tissues of antibody-forming cells is distinctive and unique, depending on the route of immunization. After i.v. immunization, antibody-forming cells are found predominately in spleen, blood, and bronchoalveolar spaces; after intrapulmonary immunization, they are located predominately in hilar lymph nodes, blood, and bronchoalveolar spaces. The reasons for this pattern of distribution are not known. Both routes of immunization are equally effective in populating bronchoalveolar air spaces with antibody-forming cells, which are predominately IgM-secreting and IgG-secreting cells. IgA-secreting cells were not detected.
Article
In vitro studies of antibody synthesis have great attraction because they offer the hope of isolating and examining separately the variables involved in the interaction of antigen with lymphoid cells, which results in an immunological response. The chapter reviews such studies and critically analyzes the different experimental conditions employed, the types of data obtained, and their interpretation. The mechanisms and problems involved in antibody synthesis appear to be similar to those involved in synthesis of other proteins. Of more peculiarly, immunological interest are attempts to induce and demonstrate specific responses in vitro. One of the simplest changes to measure is increased DNA synthesis and that is applicable to stimulation by cell surface antigens. The phenomenon is clear enough, but despite many ingenious experiments, this chapter shows how little is yet understand about its mechanism. The advantages of an in vitro analysis of the immunological response were clear from the earliest times. In the study of a mechanism involving a complex series of events and the possible interaction of cell populations, it is essential to be able to identify the cells under study, regulate the conditions of their exposure to antigen, and to study their subsequent morphological and biochemical response. In spite of these potential advantages, there was a long induction period during which a great deal of effort was expended and relatively little achieved.
Article
During the last few years isotopes have become one of the most important tools of the biochemists. Isotopically labeled molecules behave, in general, in the same manner as their unlabeled analogues. They allow us, however, to differentiate molecules formed after administration of the isotope from those formed before, and thus enable us to obtain more insight into the site and rate of formation of metabolites. Therefore, we may obtain information on the precursors of metabolites, their turn-over time and their breakdown products. For the same reason, isotopically labeled antigens have been used in immunochemical research. Tracing of isotopically labeled antigens has provided new information on their elimination from the circulation, their deposition in certain tissues or cells, their metabolic fate and their breakdown. In vitro experiments with labeled antigens allow us to measure their incorporation into antigen-antibody complexes. In animals injected with isotopically labeled amino acids, incorporation of these amino acids into antibody molecules can be measured. This enables us to determine the rate of antibody formation. Finally, experiments with isotopically labeled complement give some information on the components of the complement complex which combine with antigen-antibody aggregates. It is clear from this short survey that the application of isotopes enables us to investigate numerous problems which were inaccessible to experimental research prior to the availability of isotopes.
Article
A direct study of the isolated rat liver perfused with oxygenated blood containing amino acids and lysine-ϵ-C14 has yielded facts indicating that the liver synthesizes practically all the plasma fibrinogen, the albumin fraction, and probably more than 80 per cent of the plasma globulin fraction. The response of the isolated perfused liver in protein synthesis is qualitatively and quantitatively analogous to that of the intact animal, notably in (a) the ability to discriminate between natural L-lysine and D-lysine, (b) the per cent of isotopic amino acid converted to CO2, (c) the per cent utilized in liver and plasma protein synthesis. The results obtained with the perfused liver are compared and contrasted with those reported for tissue homogenates, minces, and slices.
Article
1. The thermolability of the specific polysaccharides of Types I, II, and III pneumococcus has been shown by three independent methods: (a) diminution of the viscosity of solutions on heating; (b) decrease in the amount of antibody precipitated from homologous rabbit antisera; and (c) increased tendency (S III) to pass through a collodion membrane. 2. These effects may be explained most simply as a partial depolymerization under the influence of heat. In air, particularly in the presence of broth, oxidation also appears to be involved. 3. Improved and simpler methods of preparation based on these findings, are given for S I, S II, and S III. The resulting products precipitate more anti-S from homologous rabbit antisera than do the earlier preparations. 4. The methyl glycoside of methyl galacturonate has been isolated from the hydrolytic products of S I, and evidence of the ultimate structural unit obtained.
Article
A method for the specific histochemical demonstration of antibody in cells and parts of cells is described. It consists of carrying out a two stage immunological reaction on frozen sections of tissues: (a) allowing reaction between antibody in the tissue and dilute antigen applied in vitro, and (b) the detection of those areas where this antigen has been specifically absorbed by means of a precipitin reaction carried out with fluorescein-labelled antibody. Examination under the fluorescence microscope reveals the yellow-green fluorescence of fluorescein over those areas where a precipitate has formed. A study of the hyperimmune rabbit on the first few days after the last of a series of intravenous antigen injections reveals that antibody against human gamma-globulin or ovalbumin is present in groups of plasma cells in the red pulp of the spleen, the medullary areas of lymph nodes, the submucosa of the ileum, and the portal connective tissue of the liver. Because of extensive non-specific reactions, the bone marrow could not be examined. Small amounts of antibody were occasionally visible in cells in the lymphoid follicles of the spleen and lymph nodes, so that a minor contribution by lymphocytes to antibody synthesis cannot be excluded.
Article
A direct study of the isolated rat liver perfused with oxygenated blood containing amino acids and lysine-epsilon-C(14) has yielded facts indicating that the liver synthesizes practically all the plasma fibrinogen, the albumin fraction, and probably more than 80 per cent of the plasma globulin fraction. The response of the isolated perfused liver in protein synthesis is qualitatively and quantitatively analogous to that of the intact animal, notably in (a) the ability to discriminate between natural L-lysine and D-lysine, (b) the per cent of isotopic amino acid converted to CO(2), (c) the per cent utilized in liver and plasma protein synthesis. The results obtained with the perfused liver are compared and contrasted with those reported for tissue homogenates, minces, and slices.
Rec. Trav. chim. Pays-Bas
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