Presence Of Immune Serum Research Articles

Growth of Virulent and Attenuated Ectromelia Virus in Cultured Macrophages from Normal and Ectromelia-Immune Mice

Summary Growth of ectromelia virus in cultured mouse peritoneal macrophages is described. Macrophages in vitro are more susceptible to infection by a virulent than by an attenuated strain of ectromelia virus. Macrophages from susceptible and resistant strains of mouse are equally susceptible to infection in vitro with virulent ectromelia virus. Macrophages from immune mice are more readily infected than are normal macrophages. This effect is attributed to the greater phagocytic activity of the macrophages from immune mice. Even in the presence of immune serum, macrophages from immune mice are more readily infected than are macrophages from normal mice.

A Histochemical Study of Phagocytic and Enzymatic Functions of Rabbit Mononuclear and Polymorphonuclear Exudate Cells and Alveolar Macrophages

Summary and Conclusions The effect of particle ingestion on the activity of certain enzymes of mononuclear (MN) and polymorphonuclear (PMN) exudate cells and alveolar macrophages (AM) was evaluated by quantitative and semiquantitative histochemical techniques. The particles were yeast and chicken red blood cells in the presence of immune serum, and polystyrene in the presence of normal serum. The enzymes were acid and alkaline phosphatase, esterase and succinic dehydrogenase—enzymes probably more concerned with metabolic than with digestive phenomena. The period of ingestion usually varied from 10 to 40 min, in tubes attached to a drum rotating at 6 rpm. Phagocytes containing particles were found to have the same amount of histochemically detectable enzymes as adjacent phagocytes on Mylar strips that had not ingested particles. In other words, the ingestion of particles was not associated with an increased content of enzymes. Biochemical studies on the acid phosphatase of frozen and thawed MN and PMN that had ingested various particles confirmed this conclusion. However, manometric studies under similar conditions showed that the phagocytic process increased the respiration of viable cells. This suggests that their metabolic enzymes, although unchanged in amount, were utilized more fully. The effect of particles on the enzymes of phagocytes was also evaluated over longer periods of mutual association. For this purpose rabbit mononuclear exudate cells were allowed to ingest yeast particles and then were cultured along with controls in 98% autologous serum, a method adopted from Dr. Y. T. Change of the National Institutes of Health. Zero- and 48-hr Mylar strip preparations then were made from these cell cultures for quantitative histochemical evaluation. Not all experiments were successful, but in those that were, the 48-hr cell preparations containing yeast showed a significant increase in the number of MN with high esterase and acid phosphatase contents. This seemed to be associated with digestion of the yeast particles. Although other interpretations are possible, we feel that the increased enzyme content of these cells was due to an adaptive response to the intracellular presence of particles. The above findings are consistent with the following concept: a) An initial protoplasmic excitation, brought about by collision with particles and/or their actual ingestion, results in an increase in the oxygen consumption of intact cells. Such phagocytes are activated, in that they ingest the particles more readily and probably have increased motion and pseudopod formation. Their enzymes are not increased in amount, but seem to be utilized nearer their full capacity and may have a different distribution within the cell. b) This excitatory response initiates and merges with protoplasmic adaptation which is a more lasting response on the part of the phagocyte to ingested material and other irritants. It is expressed by an increase in certain enzymes, and specific and nonspecific changes in the phagocytic, bactericidal and digestive powers of the cell. These two phases of phagocytic activation may be crucial to the host's defense against infectious disease.

The Laboratory Application of Anhydrous Salmonella pullorum Antigen

ALMONELLA PULLORUM was first described by Rettger,4 and its definite relation to the disease was established by him in 1900 and 1901.5 In an effort to eradicate this economically important malady, Rettger and Stoneburn7 attempted to identify carrier hens by bacteriological examination of yolks. This procedure was important in demonstrating that infected hens lay infected eggs, but it was found to be too laborious and uncertain to be of much practical value in identifying infected breeders. Over the years many methods have been described and advocated for the determination of pullorum infection. Aside from the technique of Ward and Gallagher,12 involving the principle of an allergic reaction in specifically infected subjects most methods are based on the specific agglutination of bacterial cells in the presence of immune serum. A macroscopic tube agglutination was described by Jones3 in 1913. In 1914, Rettger,6 after preliminary trials, endorsed this technique, which has since, with some modification, been the official method for identifying pullorum in many areas. In 1927, Runnells8 and his associates devised the so-called rapid method, an antigen-antibody reaction conducted on a clean glass slide. Bunyea et al.1 announced a simplified agglutination test similar to Runnells' approach but differing in that whole blood is utilized in place of serum. In 1931, Schaffer et al.9 published a modification of the whole-blood procedure employing colored antigen.

ニューカッスル病ウイルスの血清学的研究

It is well known that interest informations on the antigenic structure of viruses have been obtained with complement fixation. The serological properties of Newcastle disease virus, however, have been mainly studied by neutralization and hemagglutination and available references on its complement fixing properties are limited. For complement fixation many kinds of animals may be able to donate antibodies, but the natural host should be considered to be standard as source of antibodies. As chicken serum, which is of the most susceptible natural host for the virus, fails to fix complement when inactivated by heating at 56°C for 30 minutes, indirect complement fixation has been recommended as an available technique. Recently the possibility of direct complement fixation with chicken serum was suggested by Nitzschke (1956) and Brumfield (1957). The author tried to establish the direct complement fixation in the Newcastle disease virus-chicken serum system and to examine complement fixing properties of the virus and antigenic difference among several strains with the technique.The Sato, Miyadera and W strains of Newcastle disease virus, different in origin and pathogenicity, were chiefly employed. The Sato strain was isolated from an affected bird in the outbreak of the acute type disease in 1930 and the Miyadera strain, of the mild type disease in 1951. The W strain, originating from American living vaccine, is an avirulent one in chickens.Results are summarized as follows.1. Direct complement fixation was successfully performed in the Newcastle disease virus-chicken serum system, using sera in the active phase and the incubation at 37°C for 60 minutes in a water bath.2. Complement was specifically fixed with infected embryonic tissues under the presence of immune serum. The highest fixation was shown in the test with chorioallantoic membranes and no fixation was detected in the test with allantoic fluid.3. Two kinds of antigens, soluble and viral, existed in infected embryonic tissues. The former was prepared by the complete elimination of hemagglutinin from the suspension of chorioallantoic membranes, using centrifugation and absorption with chicken red cells, and the latter, by centrifugal concentration of hemagglutinin from allantoic fluid.Although hemagglutinability was approximately the same in titer, the amount of soluble antigen in chorioallantoic membranes seemed to be different among the strains.4. Antibodies against soluble and viral antigens were produced in chickens hyperimmunized with inactivated virus or infected with active virus. Antibody against viral antigen was usually from 2 to 8 times higher than that against soluble antigen in titer.5. Partial antigenic differences among the strains were revealed by cross hemagglutination inhibition with sera absorbed with heterologous antigens, particularly the Sato strain appeared to be different from the others. Cross complement fixation failed to detect the partial differences among them, even when viral antigen was used. The Miyadera and W antigens, both soluble and viral, showed stronger reaction with the Sato immune serum than that with the homologous. The Sato strain seemed to be more effective to produce antibodies in chickens.

Effects of yellow fever (17D) and West Nile viruses on the reactions of human appendix and conjunctiva cells to several other viruses

Established cell lines from human conjunctiva and appendix were found to support multiplication of the 17D strain of yellow fever. Conjunctiva cells showed no cytopathogenic effect, but appendix cells showed a characteristic degeneration followed by regeneration of a cell sheet of normal appearance, which continued to excrete virus. Both appendix and conjunctiva cells infected with West Nile virus (WNV) showed a similar pattern of degeneration, recovery, and continued excretion of virus. Conjunctiva and appendix cells infected with 17D, and appendix cells infected with WNV, were maintained through successive subcultures for periods of from seven to ten months. Virus was present in all fluids tested during that time. The cytopathogenic effects 17D and WNV on appendix cells were neutralized by homologous immune human or rabbit sera. Infected cell lines however could be maintained in the presence of immune serum without removing the infection. Original 17D virus pool prepared in chick embryo was found to give a higher titer in mice than in appendix cells. After seven passages in appendix cells no significant difference in titer could be found. Appendix cells infected with 17D were found to be less susceptible to certain viruses than uninfected cells maintained in parallel. 17D appendix cells were completely resistant to the cytopathogenic effect of 17D and WNV, partly resistant to herpes simplex, polio 1, and polio 3, but as susceptible to vaccinia virus as normal appendix cells. 17D appendix cultures challenged with large doses of WNV continued to release WNV in the fluid, with an apparent fall in amount of 17D released.

Phagocytosis of Influenza Virus

Summary From data obtained using the fluorescent antibody technique of Coons and Kaplan it appeared that the influenza virus could be phagocytosed by human and rabbit leucocytes in vitro. It is believed that the virus was first adsorbed to the leucocyte following which penetration into the cell occurred. The reaction of phagocytosis was markedly enhanced by the presence of immune serum and by leucocytes from immune animals.

Further studies of the phenomenon of capsular swelling of Micrococcus pyogenes var. aureus in the presence of immune serum.

Further studies of the phenomenon of capsular swelling of Micrococcus pyogenes var. aureus in the presence of immune serum.

Intracellular multiplication of Toxoplasma gondi in adult mammalian macrophages cultivated in vitro.

SummaryToxoplasma gondii has been propagated successfully in macrophage cultures of adult laboratory animals. The pattern of the interaction between the macrophages cultured in vitro from different animal species could be correlated with the susceptibility of the species to the infection with Toxoplasma. Macrophages cultured in vitro from animals actively immunized by injection with living Toxoplasma had an inhibitory effect on the intracellular multiplication of these parasites. This effect was increased by the presence of immune serum which had a high titer of cytoplasm-modifying antibodies.

Persistent antigenic variation of influenza A viruses after incomplete neutralization in ovo with heterologous immune serum.

Antigenic variants of influenza A virus strains emerge on serial passage in ovo in the presence of immune serum against different but related strains. An old laboratory strain (PR8) which had been through hundreds of animal passages was as readily modified by this procedure as recently recovered strains. Such variants apparently can be obtained at will and show antigenic patterns which are reproducible and appear to be predictable in terms of the immune serum used for their selection. Variant strains retain their new antigenic patterns on serial passage in ovo in the absence of immune serum. Limited serial passage in ovo of strains in the absence of immune serum did not result in the emergence of antigenic variants. Similarly, serial passages of strains in ovo in the presence of immune serum against widely different strains, which failed to show significant cross-neutralization, did not lead to the appearance of antigenic variants.

The Complement Fixation Reaction in Toxoplasmic Infection

A frozen and thawed extract of toxoplasma-infected rabbit brain (but not similar extracts of certain other heavily parasitized tissues) yielded an antigen which fixed complement specifically in the presence of immune sera from experimentally infected rhesus monkeys but not rabbits, dogs, and cats. This complement fixing antibody appeared within 1 to 4 weeks after inoculation, but unlike the neutralizing antibody disappeared again in most monkeys, sometimes as early as 2 months. A certain number of human beings who were known or suspected to have experienced toxoplasmic infection also gave positive complement fixation reactions. However, twenty selected individuals whose sera had neutralizing antibodies against toxo-plasma gave negative complement fixation reactions, but in none of these was there evidence of recent or active infection. While a positive complement fixation reaction does not in all cases indicate active or even recent infection, it is believed that this test may have its greatest usefulness in the rapid diagnosis of active toxoplasmosis.

Immunological Studies of Pollinosis

Abstract In 1937 Cooke, Loveless and Stull (1) reported that normal individuals respond to pollen-inoculations by forming immune antibody. This antibody can inhibit the urticarial response of sensitized skin to pollen by interfering with the pollen-reagin reaction. (In the absence of immune antibody, skin inoculated with reagin-bearing serum from a hay-fever patient promptly develops erythema and a wheal when pollen-antigen is introduced.) Subsequent studies (2) showed that the cutaneous response fails to occur in the presence of immune serum because pollen is bound and inactivated by the immune antibody. This antibody was found to differ from reagin not only in its greater avidity for pollen-antigen and in its ability to unite with antigen without irritating surrounding tissue, but also in its rapid disappearance from skin into which it had been injected and in its capacity to resist temperatures that inactivate reagin.

Neutralization of Vaccine Virus by Immune Serum

Summary and Discussion Experiments have been performed with a view to gaining information on the protection afforded by immune serum against vaccinal infection of mice, and in order to correlate the protective potency of serum against virus introduced intracerebrally in mice with that introduced intracutaneously in rabbits. It was learned that virus could be titrated satisfactorily by intracerebral inoculation of mice, and that with the strain of virus used the estimates of potency agreed well with those derived from the results of rabbit-inoculation. This was taken to mean that the virulence of this strain of virus for mouse and rabbit was essentially the same. Addition of immune serum to the viral suspensions protected mice from infection. The moderate irregularity of the titration-curve which was found when virus was injected alone was found to be considerably increased, but not to such an extent as to make the use of the “50 per cent point” an invalid one for measuring viral titer. It will be recalled that a similar observation was made when virus was titrated on the rabbit skin in the presence of immune serum, and furthermore that the degree of irregularity of results seemed to be proportional to the concentration of immune serum. The fact that a similar condition obtains in the mouse suggests that it is fundamentally related to the protective activity of immune serum. It was found that on dilution of the immune serum its protective capacity fell off regularly, as was found to be the case when titrations were made in the rabbit. It seemed, however, that the power of serum to protect the intracerebrally injected mouse from fatal infection was greater than its capacity to protect against the development of a cutaneous lesion. In evaluating this observation it must, of course, be borne in mind that we were not in a position to recognize the development of a minimal lesion in the brain. However, fatal infection of mice, and cutaneous lesions in rabbits, appeared to follow the introduction of equivalent amount of the original viral suspension of this strain. Further speculation must await fuller information.

A SOLUBLE ANTIGEN OF LYMPHOCYTIC CHORIOMENINGITIS

The soluble antigen of lymphocytic choriomeningitis which is readily separable from the virus is a relatively stable substance and appears to be of a protein nature. A specific precipitin reaction can be demonstrated when immune serum is added to solutions of antigen which have been freed of certain serologically inactive substances. The complement-fixation and precipitation reactions which occur in the presence of immune serum and non-infectious extracts of splenic tissue obtained from guinea pigs moribund with lymphocytic choriomeningitis seem to be manifestations of union of the same soluble antigen and its antibody. On the other hand, the antisoluble substance antibodies and neutralizing substances appear to be different entities.

A SOLUBLE ANTIGEN OF LYMPHOCYTIC CHORIOMENINGITIS

The virus of lymphocytic choriomeningitis can be sedimented in the ultracentrifuge and washed repeatedly; the virus retains its activity provided that a small amount of normal serum is present in the diluent. A soluble substance capable of fixing complement in the presence of immune serum can be separated from the virus. Washed virus fixes complement poorly. The serologically specific soluble antigen is widely distributed in tissues of infected guinea pigs, mice, and monkeys.

Observations on the Mode of Action of Sulfapyridine on Pneumococcus.

Conclusions1. Sulfapyridine did not affect the capsule of the pneumococcus either in vivo or in vitro in such a way as to be ascertainable by capsule-staining or by affecting the “quellung” reaction in the presence of immune serum. 2. In vivo and in vitro experiments using sulfapyridine in pneumococcal infections or pneumococcal cultures containing leucocytes did not indicate an increase in phagocytosis of the diplococci in the presence of the drug. 3. In vitro there was only slight bacteriostatic effect of 10 mg% sulfa-pyridine in broth cultures of pneumococcus Type I until the culture neared the close of the logarithmic phase of growth. At that time a bactericidal effect was apparent along with increased bacteriostasis and the length of the phase of decline was decreased markedly. 4. In vivo a similar effect was noted. In mice treated with sulfa-pyridine there was a short period of increase in the number of organisms in the peritoneal exudate followed by a rapid decrease in the number of bacteria found ...

Complement-Fixation in Infections with the Virus of Lymphocytic Choriomeningitis

SummaryA soluble specific antigen, which occurs in tissues of animals infected with the virus of lymphocytic choriomeningitis, fixes complement in the presence of immune serum; the virus freed from the soluble antigen fixes complement poorly or not at all. Patients recovered from lymphocytic choriomeningitis develop in their serum antibodies that react with the soluble antigen to fix complement.

IMMUNOLOGICAL STUDIES ON PURE CULTURES OF VARIOUS SPIROCHETES

IMMUNOLOGICAL STUDIES ON PURE CULTURES OF VARIOUS SPIROCHETES