scholarly journals IMMUNOLOGICAL STUDIES ON A NEW PREPARATION OF TYPE SPECIFIC POLYSACCHARIDE FROM PNEUMOCOCCUS TYPE I

1936 ◽  
Vol 64 (6) ◽  
pp. 843-854 ◽  
Author(s):  
Bacon F. Chow

A type specific polysaccharide has been isolated from the autolyzed broth of Type I Pneumococcus by a modified Avery and Goebel's method. The newly prepared polysaccharide reacts with the homologous immune rabbit serum which has been completely absorbed with the acetyl polysaccharide of Avery and Goebel. The newly prepared polysaccharide produces passive immunity in mice and rats and possibly in rabbits. The antigenicity is not lost on boiling in acid or alkaline medium, but the precipitative activity is decreased. In conclusion, it has been shown that the polysaccharide from Type I Pneumococcus, as isolated by a slight modification of Avery and Goebel's method, is a more complete antigen.

1943 ◽  
Vol 78 (4) ◽  
pp. 305-313 ◽  
Author(s):  
George Packer Berry ◽  
Howard B. Slavin

Passive immunity, naturally acquired from immune mothers or artificially induced through the administration of immune rabbit serum, conferred on suckling mice of the albino Swiss strain a high degree of resistance against herpetic infection following the intranasal instillation of the virus. Antibodies, which could be readily demonstrated in the blood of 2-week-old mice, were received by the offspring of immune mothers primarily by the mammary route. Naturally acquired immunity declined rapidly when suckling was interrupted. Herpes virus was not recovered from the fetuses of either immune or infected, non-immune mothers.


1949 ◽  
Vol 32 (6) ◽  
pp. 705-724 ◽  
Author(s):  
John H. Northrop ◽  
Walther F. Goebel

1. The immune precipitate formed by antipneumococcus horse serum and the specific polysaccharide is not hydrolyzed by trypsin as is the diphtheria toxin-antitoxin complex, and purified pneumococcus antibody cannot be isolated by the method used for the isolation and crystallization of diphtheria antitoxin. 2. Type I pneumococcus antibody, completely precipitable by Type I polysaccharide, may be obtained from immune horse serum globulin by precipitation of the inert proteins with acid potassium phthalate. 3. The antibody obtained in this way may be fractionated by precipitation with ammonium sulfate into three main parts. One is insoluble in neutral salts but soluble from pH 4.5 to 3.0 and from pH 9.5 to 10.5. This is the largest fraction. A second fraction is soluble in 0.05 to 0.2 saturated ammonium sulfate and the third fraction is soluble in 0.2 saturated ammonium sulfate and precipitated by 0.35 saturated ammonium sulfate. The second fraction can be further separated by precipitation with 0.17 saturated ammonium sulfate to yield a small amount of protein which is soluble in 0.17 saturated ammonium sulfate but insoluble in 0.25 saturated ammonium sulfate. This fraction crystallizes in poorly formed, rounded rosettes. 4. The crystallization does not improve the purity of the antibody and is accompanied by the formation of an insoluble protein as in the case of diphtheria antitoxin. 5. None of the fractions obtained is even approximately homogeneous as determined by solubility measurements. 6. Purified antibody has also been obtained by dissociating the antigen-antibody complex. 7. The protective value of the fractions is quite different; that of the dissociated antibody being the highest and that of the insoluble fraction, the lowest. 8. All the fractions are immunologically specific since they do not precipitate with Type II polysaccharide nor protect against Type II pneumococci. 9. All the fractions give a positive precipitin reaction with antihorse rabbit serum. The dissociated antibody gives the least reaction. 10. Comparison of the various fractions, either by their solubility in salt solution or through immunological reactions, indicates that there are a large number of proteins present in immune horse serum, all of which precipitate with the specific polysaccharide but which have very different protective values, different reactions with antihorse rabbit serum, and different solubility in salt solutions.


1938 ◽  
Vol 67 (4) ◽  
pp. 619-641 ◽  
Author(s):  
Rose G. Miller

1. Inflammation retards the absorption of horse serum globulin and crystalline egg albumin from the peritoneal cavity and subcutaneous tissue, but retardation of the absorption of crystalline egg albumin is less than that of globulin, which is less diffusible. 2. Inflammation retards the absorption of the specific polysaccharide of pneumococcus Type I from the peritoneal cavity; inflammation may accelerate, but does not hinder, the absorption of glucose from the peritoneal cavity. 3. Inflammation retards the spread of trypan blue in the skin, but accelerates absorption from the skin of the more diffusible dye, brom phenol blue. 4. Phenol red is excreted in the urine with equal rapidity after injection into normal and into inflamed subcutaneous tissue or into normal and into inflamed peritoneal cavities. Direct extractions of phenol red from inflamed subcutaneous sites indicate that inflammation accelerates the absorption of the dye from these areas. 5. Inflammation retards the absorption of the indiffusible proteins, carbohydrates and dyes; it tends to accelerate the absorption of the diffusible carbohydrates and dyes.


1941 ◽  
Vol 73 (1) ◽  
pp. 125-140 ◽  
Author(s):  
Henry P. Treffers ◽  
Michael Heidelberger

1. Rabbits were injected with the washed specific precipitate from Type II antipneumococcus horse serum. Antibody in the resulting antiserum was determined by the quantitative agglutinin method using various specific precipitates as antigens. 2. Suspensions of Types I and II antipneumococcus horse specific precipitates, as well as the specific precipitates derived from Type VIII Pn (anti-C portion), and H. influenzae horse antisera were found to remove the same amount of antibody from the immune rabbit serum. 3. Purified antibody solutions prepared by dissociation methods from Types I and II antipneumococcus horse sera were found to remove the same quantity of antibody as did the homologous specific precipitates. 4. Specific precipitates from anti-crystalline egg albumin and anti-diphtheria horse sera were found to remove only a fraction of the antibody. The reasons for this are discussed. 5. A specific precipitate prepared from pepsin-digested Type I anti-pneumococcus horse serum removed all of the antibody to the homologous antigen from the rabbit anti-precipitate serum, but followed a different quantitative course. 6. From the quantitative course of these reactions and from experiments with specific precipitates from anti-Pn rabbit and pig sera it is concluded that the only antigenic specificity demonstrable for the antibodies investigated was that due to their common origin, and that the groupings responsible for their antibody function constitute either a small part of the total protein molecule or else are non-antigenic.


1924 ◽  
Vol 39 (6) ◽  
pp. 777-802 ◽  
Author(s):  
Thomas M. Rivers ◽  
William S. Tillett

1. The intradermal method of inoculating Virus III, a hitherto unknown filterable virus producing lesions in rabbits, gives more reliable results than those obtained by smearing the virus on the scarified skin. 2. Virus III, heated 10 minutes at 55°C., will not produce visible reactions in the skin of rabbits. 3. Virus III passes through Berkefeld N and V filters. 4. The data obtained so far indicate that the best method of preserving Virus III in an active state is to filter the testicular emulsions containing the virus, add glycerol to the filtrate up to 40 per cent of the total volume, seal with vaseline, and store on ice. 5. Viable Virus III produces a definite immunity in rabbits which persists for at least 6 months. The immunity follows intradermal, intratesticular, intravenous, intracerebral, or intranasal inoculations of the virus. 6. A single intradermal injection of Virus III, which has been killed by heat, will not produce a demonstrable immunity in rabbits. 7. No passive immunity to Virus III could be demonstrated in rabbits which had received intravenous injections of 5 to 10 cc. of immune rabbit serum 24 hours previously. 8. Immune rabbit serum neutralizes Virus III either in vitro, or locally in a rabbit's skin when the immune serum and the virus are injected into the same part of the skin at or about the same time. 9. Three strains of the virus under investigation are immunologically identical. 10. Virus III and vaccine virus are immunologically distinct. 11. Virus III and the virus of symptomatic herpes are immunologically distinct. 12. No passive immunity to Virus III could be demonstrated in rabbits which had received intravenous injections of 5 to 10 cc. of serum or whole blood from patients convalescent from varicella. 13. Sera from two normal adults and from fourteen patients convalescent from varicella did not neutralize Virus III in vitro. 14. Rabbits could not be actively immunized against Virus III by injections of whole blood, vesicle fluid, or nasal washings from patients with varicella. 15. Four of twenty sera collected from stock rabbits of different ages, 20 per cent, neutralized Virus III in vitro. The animals whose sera neutralized Virus III failed to show a reaction at the site of intradermal inoculations with the same virus. About 15 per cent of 200 young stock rabbits (1,800 gm.) used in routine transfers were found to be refractory to Virus III, as evidenced by a failure to react to intradermal inoculations of the virus. 16. No susceptibility to Virus III was observed in guinea pigs, mice, or monkeys. 17. A volunteer who had never suffered from varicella experienced no general reaction and only a mild local one following an intradermal inoculation of Virus III. A volunteer who had had chicken-pox in childhood experienced a moderate general action, viz., fever, headache, backache, and general malaise, and also a moderate local reaction, viz., redness, swelling, tenderness, and pain, following an intradermal inoculation of Virus III. 18. The study of the immunological reactions has failed to bring any evidence that the virus under investigation bears an etiologic relationship to varicella.


1935 ◽  
Vol 62 (2) ◽  
pp. 281-287 ◽  
Author(s):  
L. A. Barnes ◽  
Eleanor C. Wight

An encapsulated strain of Escherichia coli has been isolated which is hemolytic, pathogenic for mice, and which has served to illustrate further evidence of heterogenetic specificity. The relationship appears to be limited to the serological reactions between the colon organism and Type I antipneumococcic horse serum. Type I antipneumococcic rabbit serum failed to agglutinate the organism and no reactions occurred with Types II and III antipneumococcic horse serums, normal horse serum, and a variety of other immune horse serums. Serum from rabbits immunized with the colon bacillus agglutinated the homologous organism and precipitated its soluble substance, but failed to cause agglutination of Type I pneumococci or to precipitate Type I pneumococcic polysaccharide. The evidence indicates a connection somewhat analogous to that between Type II pneumococcus and Type B Friedländer's bacillus.


1931 ◽  
Vol 54 (6) ◽  
pp. 899-923 ◽  
Author(s):  
Victor Ross

1. Feeding the purified soluble specific substance of Type I pneumococcus protects rats against an intraperitoneal injection of the virulent organism. 2. This increased resistance resembles that obtained when the intact (dead) or dissolved bacteria are fed, as follows: (a) one feeding is sufficient, (b) the interval between the feeding and the appearance of the immunity is the same, (c) the duration is approximately the same, (d) when the immunity is exhausted it can be renewed by a new feeding, (e) the immunizing action is type-specific. 3. The differences between the effects of feeding the purified specific substance and the intact or dissolved organism to rats, appear to be quantitative rather than qualitative, the proportion of animals protected and the height of the immunity being generally, though not always, less in the case of the former. 4. In contrast to the immunizing action which the soluble specific substance possesses when administered to rats, feeding it to mice failed to protect them. Neither were mice definitely immunized by parenteral administration. 5. A sodium glycocholate solution of Pneumococcus Type I lost part of its immunizing activity on standing for 1 year. 6. The failure to immunize mice and the loss of activity of the bile salt solution of pneumococcus, on standing, are discussed in terms of (a) the possible presence of a second cell constituent which is active by mouth, and (b) a possible intramolecular change in the type-specific polysaccharide associated with a loss of immunizing action while retaining the precipitin reaction.


Author(s):  
Veronika Burmeister ◽  
N. Ludvig ◽  
P.C. Jobe

Electron microscopic immunocytochemistry provides an important tool to determine the ultrastructural distribution of various molecules in both normal and pathologic tissues. However, the specific immunostaining may be obscured by artifactual immunoreaction product, misleading the investigator. Previous observations show that shortening the incubation period with the primary antibody from the generally used 12-24 hours to 1 hour substantially reduces the artifactual immunostaining. We now extend this finding by the demonstration of artifact-free ultrastructural localization of the Ca2/calmodulindependent cyclic nucleotide phosphodiesterase (CaM-dependent PDE) immunoreactivity in brain.Anesthetized rats were perfused transcardially with phosphate-buffered saline followed by a fixative containing paraformaldehyde (4%) and glutaraldehyde (0.25%) in PBS. The brains were removed, and 40μm sections were cut with a vibratome. The sections were processed for immunocytochemistry as described by Ludvig et al. Both non-immune rabbit serum and specific CaM-dependent PDE antibodies were used. In both experiments incubations were at one hour and overnight. The immunostained sections were processed for electron microscopic examination.


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