scholarly journals IMMUNOLOGICAL STUDIES ON PURE CULTURES OF VARIOUS SPIROCHETES

1917 ◽  
Vol 25 (6) ◽  
pp. 765-788 ◽  
Author(s):  
Hideyo Noguchi ◽  
Seinai Akatsu
Keyword(s):  
Complement Fixation ◽  
Virulent Strain ◽  
Immune Serum ◽  
The Other ◽  
Rabbit Serum ◽  
Complete Suppression ◽  
Normal Rabbit Serum ◽  
Slight Degree ◽  
Group Reaction

Experiments were carried out for the study of culture spirochetes in their relation to various immunity reactions in vitro. Several strains of Treponema pallidum and one each of Treponema calligyrum, Spirochata refringens, Treponema microdentium, and Treponema mucosum were used. Tests were made of immune substances responsible for agglutination, complement fixation, spirocheticidosis, and opsonization. In cases of agglutination and complement fixation, cross titrations were made. 1. In the sera derived from rabbits immunized with various spirochetes agglutinins were demonstrated in varying quantities for the homologous antigens. The amounts of agglutinins developed were considerably higher in the pallidum immune sera than in the other groups. There was no parallelism between the amounts of antigens injected and the amounts of agglutinins developed. 2. Cross titrations among different pallidum strains revealed that the agglutiantion is not necessarily strongest when homologous antigens and immune sera are brought together. 3. On the other hand, the reactions between the immune sera and antigens belonging to different species were sufficiently specific to justify the grouping. 4. Certain degrees of group reactions were observed between the pallidum immune sera and the calligyrum, and occasionally very faintly also between the pallidum and the refringes antigens and vice versa. There was a much more pronounced group reaction between the calligyrum and refringes. The immune serum and antigen of the microdentium showed a slight affinity for the mucosum but none for the pallidum, calligyrum, or refringes, while the mucosum immune serum caused a slight agglutination with many members of the other groups. Hence, it appears that the pallidum is more or less related to the calligyrum, while the affinity between the calligyrum and refringes, and possibly also between the calligyrum and mucosum in a much smaller degree, seems close. The microdentium showed the least relation to any other spirochetes. 5. Titration of agglutinins in the sera obtained 3 months after the cessation of immunization revealed that the agglutinin contents were already greatly reduced, having fallen roughly to 0.01 of the original strenght. The rates of disappearance were irregular in different animals and bore no direct relation to the initial titers. Titration made of the immune sera which had been preserved aseptically in a refrigerator (6°C.) during the same period (3 months) indicated that the original strength of these sera was reduced to about one-tenth. The agglutinins for spirochetes disappear from the rabbit's body much more rapidly than they are reduced in the separated sera by deterioration on standing at 6°C. 6. Titration of the immune sera for complement fixation power showed with a few exceptions, in which there was only slight complement binding, that the titers were high enough to indicate the presence of this principle. The anti-pallidum sera possessed higher average titers than the other immune sera tested with correspondingly homologous antigens. The least active were the anti-refrigens sera. 7. Cross titration of anti-pallidum immune sera for complement fixation showed that a given serum with a high titer for its own strain of antigen was also strong with most of the other strains of the pallidum. Instances occurred also in which the titers with heterologous pallidum antigens fell far below those of the homologous. Group reactions between the different spirochetes) such as the pallidum and the calligyrum, the calligyrum and the refringens, and the microdentium and the mucosum, were also indicated. The mucosum and the pallidum showed a slight degree of group reaction. No anti-pallidum serum fixed complement with the microdentium. 8. The immune sera were tested for their spirochetiddal properties in vitro against the correspondingly specific and heterologous varieties with and without the addition of complement. Many of the anti-pallidum sera killed their own strains. Normal rabbit serum exhibited only a slight degree of inhibition. Without complement, the immune sera caused a considerable reduction in the number or density of colonies, but not a complete suppression of growth. Complement alone had no injurious effect upon the pallidum strains. The antisera for the calligyrum, refringens, and mucosum showed feeble spirocheticidal action, while the antisera for microdentium was stronger. A syphilitic rabbit serum tested against a strain of culture pallidum gave a feeble inhibitory effect. 9. Under the influence of immune sera and complement, the spirochetes undergo within a few hours complete disintegration or granular degeneration. Without complement, they are more powerfully agglutinated, but no disintegration occurs, even after 20 hours, and complement alone has no effect. 10. In the presence of homologous immune serum and complement, the culture pallidum may be ingested by the leukocytes, but phagocytosis is slight, possibly on account of the filamentous nature of the organisms. The spirochetes in such a mixture disintegrate within a few hours, disintegration being especially rapid when the immune leukocytes are used. In the absence of immune serum, phagocytosis is not noticeable, while without complement but in the presence of immune serum and leukocytes, some phagocytosis, without subsequent lysis, occurs. A virulent strain of pallidum, obtained from syphilitic orchitis in a rabbit, exposed to agglutination, lysis, and phagocytosis by an immune serum prepared by means of culture pallidum strains, showed only slight agglutination and phagocytosis but rapid immobilization without disintegration in the presence of complement.

scholarly journals ENHANCEMENT OF THE OPSONIZING AND AGGLUTINATING POWERS OF ANTIPNEUMOCOCCUS SERUM BY SPECIFIC PRECIPITATING SERUM

1920 ◽  
Vol 32 (3) ◽  
pp. 283-293 ◽  
Author(s):  
Ida W. Pritchett
Keyword(s):  
Horse Serum ◽  
Test Tube ◽  
Immune Serum ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Type I ◽  
Type Ii ◽  
Normal Horse Serum ◽  
Serum Type

1. No demonstrable antiopsonins are formed in rabbits following the intravenous injection of monovalent pneumococcus horse sera, Types I, II, and III. 2. The serum of rabbits injected with immune pneumococcus horse serum, Type I, II, or III, or with normal horse serum, when mixed in the proportion of 1:4 with Type I or Type II pneumococcus horse serum, can greatly augment, in vitro, the opsonization and agglutination of Type I and Type II pneumococci by the homologous immune horse sera. No similar effect is obtained with Type III serum and pneumococci. 3. The increase in opsonization and agglutination is dependent upon (a) specific sensitization of the pneumococci by the homologous immune serum and (b) the presence of the precipitating serum. In the absence of sensitization, as when a heterologous or normal horse serum is employed, opsonization and agglutination do not occur, even though a precipitating mixture is provided. The substitution of normal rabbit serum for the precipitating rabbit serum gives opsonization and agglutination in dilutions slightly higher than are effected with salt solution only, due possibly to the more favorable medium created for the leucocytes by the addition of 25 per cent of whole rabbit serum. 4. Different methods of combining the immune horse serum, precipitating rabbit serum, and pneumococci yield very similar results, preliminary sensitization of the bacteria before precipitation, or precipitation in the rabbit-horse serum mixture before the addition of the pneumococci for sensitization causing little if any difference in result from that obtained when immune horse serum, precipitating rabbit serum, and pneumococci are all mixed and incubated together. 5. This increased opsonization in the test-tube does not seem to be paralleled by increased protective power, or at any rate such protection is not readily demonstrated.

scholarly journals A SOLUBLE ANTIGEN OF LYMPHOCYTIC CHORIOMENINGITIS

1940 ◽  
Vol 71 (1) ◽  
pp. 43-53 ◽  
Author(s):  
J. E. Smadel ◽  
M. J. Wall ◽  
R. D. Baird
Keyword(s):  
Guinea Pigs ◽  
Complement Fixation ◽  
Lymphocytic Choriomeningitis ◽  
Immune Serum ◽  
Splenic Tissue ◽  
The Other ◽  
Soluble Antigen ◽  
Precipitin Reaction ◽  
Other Hand ◽  
Protein Nature

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.

Release of bioactive ACTH by perifused human placenta at early and late gestation

1993 ◽  
Vol 136 (2) ◽  
pp. 345-353 ◽  
Author(s):  
B. J. Waddell ◽  
P. J. Burton
Keyword(s):  
Human Placenta ◽  
Late Gestation ◽  
Rabbit Serum ◽  
Maximal Response ◽  
Normal Rabbit Serum ◽  
Apparent Difference ◽  
Adrenocortical Cells ◽  
Adult Rats ◽  
Constant Rate

ABSTRACT This study assessed whether bioactive ACTH is released by the human placenta during perifusion in vitro at early and late gestation. Human placental villous fragments from early (8–12 weeks) and late (38–40 weeks) gestation were perifused at a constant rate for 6·5 h. To assess ACTH-like bioactivity released by this tissue, the perifusion effluent was redirected through adjacent chambers containing freshly dispersed adrenocortical cells obtained from adult rats. Baseline secretion of corticosterone by these adrenocortical cells averaged 95±26 (s.e.m.) fmol/min, and this increased at least fivefold (P <0·01, two-way ANOVA) in response to placental effluent at early and late gestation. The magnitude of this increase, expressed as a percentage of the maximal response to a subsequent stimulus with ACTH(1–24), was similar for placentas obtained at early (41 ± 12% of maximal response) and late (42 ± 17%) gestation. Immunoreactive (I)-ACTH was readily detectable in placental effluent from all preparations (5·5±2·3 fmol/min per g tissue), and there was no apparent difference with stage of gestation. To determine whether all of the ACTH-like bioactivity released by the placenta was attributable to I-ACTH, a second series of placental/adrenal perifusions was conducted. In these, I-ACTH was selectively removed from placental effluent by immunoneutralization, and the residual bioactivity measured. Immunoneutralization involved preincubation of placental effluent with ACTH antiserum (1:100), and preincubation with normal rabbit serum (NRS) served as a control. Preincubation with ACTH antiserum, but not with NRS, resulted in a marked reduction in ACTH-like bioactivity present in placental effluent at both early (P <0·01, paired t-test) and late (P <0·05) gestation. This inhibition was significantly more effective (P <0·05, unpaired t-test) at early than at late gestation. Overall, these data establish that the human placenta can release bioactive ACTH-like activity at both early and late gestation, and that much, but not all, of this bioactivity is directly attributable to I-ACTH. These findings clearly demonstrate a potential role for placental ACTH in directly influencing the maternal and/or fetal hypothalamic-pituitary-adrenal axes during human pregnancy. Journal of Endocrinology (1993) 136, 345–353

scholarly journals INFECTION INDUCED BY ORAL ADMINISTRATION OF ATTENUATED POLIOVIRUS TO PERSONS POSSESSING HOMOTYPIC ANTIBODY

1957 ◽  
Vol 106 (1) ◽  
pp. 159-177 ◽  
Author(s):  
Dorothy M. Horstmann ◽  
J. R. Paul ◽  
J. L. Melnick ◽  
Joyce V. Deutsch
Keyword(s):  
Tissue Culture ◽  
Neutralizing Antibodies ◽  
Close Contact ◽  
Virulent Strain ◽  
The Other ◽  
Clear Correlation ◽  
Type Iii ◽  
Virus Dose ◽  
Virus Excretion

Four of five individuals possessing homotypic antibody in titers of 8 to 64 were infected on being fed Type III (Leon KP-34) poliovirus attenuated by Sabin by passage through tissue culture. None of the infected subjects or controls showed any evidence of illness which could be attributed to virus infection. There was no evidence of spread of infection to any of the control adult wardmates of the experimental subjects, although the two groups were in close contact: none of the controls excreted virus, none showed any antibody shift. One control who had no Type III antibodies at the start of the experiment was still antibody-negative on the 63rd day of the experiment. Three of the four individuals who became infected had naturally acquired-Type III antibodies; the other had antibodies induced by formalinized vaccine. Virus excretion in the stool was of short duration (7 to 13 days) in the three with natural antibodies, and lasted at least 6 weeks after feeding in the vaccinated child. Virus in the throat was detected only in the two persons receiving the larger virus dose (107.5 TCD50). In them it was present in small amounts between the 2nd and 6th day after feeding. No virus was detected in the blood of any of the infected individuals. The antibody responses of the four infected individuals were variable. There was no clear correlation with virus dosage, amount of virus excretion in the stools, or presence of virus in the throat. Only the child whose neutralizing antibodies were "Salk" vaccine induced showed a marked CF response. The virus excreted by two of the individuals who became infected, as tested in the 2nd tissue culture passage by monkey inoculation, was slightly more neurotropic than the virus which was ingested. Virus excreted by one of these individuals behaved as a virulent strain when tested by the in vitro plaque virulence test, while that isolated from the other had the characteristics of an attenuated strain in this test.

Partial purification of antigens collected during in vitro cultivation of the larval stages of Taenia pisiformis

Parasitology ◽  
1976 ◽  
Vol 72 (3) ◽  
pp. 269-279 ◽  
Author(s):  
M. D. Rickard ◽  
J. C. Katiyar
Keyword(s):  
Protective Immunity ◽  
Culture Medium ◽  
Serum Proteins ◽  
Skin Tests ◽  
Rabbit Serum ◽  
Partial Purification ◽  
In Vitro Cultivation ◽  
Normal Rabbit Serum ◽  
Skin Reactions

SummaryLarvae of Taenia pisiformis were cultured in vitro in medium containing 2·5, 5 or 20% (v/v) of normal rabbit serum (NRS). Greatest development occurred in 20% NRS, and the potency of antigens collected in medium from each culture tested by intradermal (i/d) skin tests in infected rabbits paralleled the in vitro growth rate of larvae. ‘Culture’ antigens from 5% NRS stimulated good immunity in rabbits to a challenge infection with T. pisiformis eggs, although they were poorly reactive in skin tests.T. pisiformis larvae were also cultured in 10% (v/v) of nitrates of serum reduced to one-half of its volume by passage through 300 000 MW cut oif (XM300F) or 100000 MW cut off (XM100F) ultrafiltration membranes. Larvae cultured using XM300F had growth rates comparable with those cultured in 20% NRS, and the antigens released into the culture medium had equal potency in i/d tests and in stimulating protective immunity in rabbits. Larvae did not develop in XM100F orproduce skin-reactive or protective antigens.Crude ‘culture’ antigen from cultures in 20% NRS was separated into 4 fractions by nitration on Sephadex G200. All of these fractions gave i/d skin reactions in infected rabbits. Fraction 3 (F3) was the most active, but was shown by acrylamide gel electrophoresis and immunoelectrophoresis to be highly contaminated with rabbit serum proteins. F3 was separated into fractions on DEAE-Sephadex A50, and the third fraction from this was as active as the original culture medium in i/d skin tests, but had only 5% of the original protein concentration. Electrophoresis demonstrated few serum contaminants, and 2 indistinct protein bands that were not present in a similar fraction of NRS.Neither Sephadex G200 F3 nor DEAE-Sephadex F3 stimulated protective immunity in rabbits, suggesting that antigens stimulating immunity against the establishment of T. pisiformis in rabbits and those provoking cell-mediated immune reactions may be different.

scholarly journals IMMUNE RESPONSE OF RABBITS TO INJECTION OF PLASMODIUM KNOWLESI

1939 ◽  
Vol 70 (2) ◽  
pp. 131-139 ◽  
Author(s):  
Monroe D. Eaton ◽  
L. T. Coggeshall
Keyword(s):  
Immune Response ◽  
Plasmodium Knowlesi ◽  
Conclusive Evidence ◽  
Red Cells ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Passive Protection ◽  
Normal Monkey ◽  
Protective Antibodies

Specific complement-fixing antibodies are produced in the serum of rabbits in response to injections of living or dead Plasmodium knowlesi. Sera from rabbits receiving injections of either parasitized or normal monkey erythrocytes are parasiticidal in vitro for P. knowlesi. Because absorption of parasiticidal rabbit sera with normal monkey erythrocytes abolishes the parasiticidal effect, it is concluded that the effect is largely due to an antibody to the red cells. Normal rabbit serum is not parasiticidal. Experiments on passive protection in monkey malaria with serum from rabbits which have received intraperitoneal injections of living or dead P. knowlesi yield no conclusive evidence that protective antibodies are formed.

scholarly journals STUDIES ON PNEUMOCOCCUS GROWTH INHIBITION

1924 ◽  
Vol 40 (4) ◽  
pp. 467-485 ◽  
Author(s):  
Oswald H. Robertson ◽  
Richard H. P. Sia
Keyword(s):  
Growth Inhibition ◽  
Protective Action ◽  
Immune Serum ◽  
Rabbit Serum ◽  
In Vitro Test ◽  
High Dilution ◽  
Protective Properties ◽  
Growth Inhibitory ◽  
Vitro Test

Using the method described in an earlier publication for testing the growth-inhibitory and bactericidal activity of serum-leucocyte mixtures for the pneumococcus, a study has been made of the action of antipneumococcus serum added to the serum and leucocytes of the rabbit, which alone lack the power to inhibit pneumococcus growth. It was found that the additiori of relatively small quantities of immune serum to a mixture of rabbit serum and washed rabbit leucocytes conferred on this mixture marked growth-inhibitory and pneumococcidal powers. The effect of the immune serum was found to be quantitative in nature. Beyond a certain point progressive dilution of the serum resulted in a corresponding decrease in the degree of growth inhibition, but this property of the immune serum did not disappear until a high dilution was reached. The leucocytes were also found to exert their effect quantitatively. Within certain limits the number of pneurnococci killed was dependent on the quantity of them present. Tests comparing the action of immune serum in the rabbit serum-leucocyte tubes with their protective action in mice showed a close parallelism between these two effects; that is to say, an immune serum showing marked protective properties for mice was likewise effective in high dilution in the in vitro test, and vice versa.

scholarly journals OBSERVATIONS ON MIXTURES OF ELEMENTARY BODIES OF VACCINIA AND COATED COLLODION PARTICLES BY MEANS OF ULTRACENTRIFUGATION AND ELECTROPHORESIS

1940 ◽  
Vol 72 (5) ◽  
pp. 523-529 ◽  
Author(s):  
J. E. Smadel ◽  
E. G. Pickels ◽  
T. Shedlovsky ◽  
T. M. Rivers
Keyword(s):  
The Other ◽  
Rabbit Serum ◽  
Soluble Antigen ◽  
Normal Rabbit Serum ◽  
Electrical Field ◽  
Heat Stable ◽  
Other Hand ◽  
Single Boundary

It has been shown experimentally that mixtures of two types of particles, namely, elementary bodies of vaccinia and collodion particles coated with protein, sediment with a single boundary in the analytical centrifuge. Such mixtures have been shown to develop one or two boundaries on electrophoresis in the Tiselius apparatus, depending on the type of coating on the surface of the collodion particles. When covered with the heat-stable soluble antigen of vaccinia, collodion particles migrate in the electrical field at the same rate as elementary bodies. On the other hand, if they are coated with a component of normal rabbit serum, they migrate at a different rate. The estimation of purity of preparations of virus by means of data obtained by ultracentrifugation and electrophoresis is discussed.

Studies on the growth of 5-day-old rabbit blastocysts in vitro

Development ◽  
1965 ◽  
Vol 13 (1) ◽  
pp. 83-95
Author(s):  
Joseph C. Daniel
Keyword(s):  
Blastocyst Stage ◽  
Culture Conditions ◽  
Rabbit Serum ◽  
Young Rabbit ◽  
Normal Rabbit Serum ◽  
Excellent Growth ◽  
Growth Promoting ◽  
Initial Medium ◽  
Made In

Rabbit embryos at the early blastocyst stage (5 days post coitum) do not grow well in vitro. The present studies were undertaken for the purpose of defining some of the components of the medium that are optimal for growth of young rabbit blastocysts under culture conditions. Preliminary studies showed that the cell culture medium F10 (Ham, 1963a) supported excellent growth of a variety of rabbit cells in vitro. Rabbit blastocysts would grow, although not at the normal rate, in F10 supplemented with 15 per cent, normal rabbit serum. F10R15 was therefore selected as the initial medium to which additions or alterations were made in an attempt to perfect its blastocyst growth-promoting qualities. It should be clearly understood that throughout this paper the word ‘growth’, used in reference to rabbit blastocysts, means an increase in blastocyst volume without prejudice to the issue of whether or not the total volume or mass of living cells is also increasing.

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