Non-immunized Group Research Articles

Detection of platelet associated IgG in immune thrombocytopenia: a new assay employing protein A and peroxidase anti-peroxidase (PROA-PAP).

Immune thrombocytopenia is frequently encountered in medical practice and is generally accepted as being caused by an IgG antibody. The capability of detecting platelet-bound IgG as a diagnostic and therapeutic modality is critical for appropriate care and management of patients with idiopathic thrombocytopenic purpura (ITP), as well as other immune thrombocytopenias. We have modified our previous assay (Br J Haematol 37:265, 1977) by employing protein A and PAP as a labeled antibody. Surface bound platelet IgG was quantitated by phase contrast microscopy after incubation with PAP, graded per 100 platelets and expressed as a reactive index (RI). Controls (n=13) had RIs ranging from 0.49 to 0.72 (mean 0.63 +/- 0.02 SE). The nonimmune thrombocytopenic group (n=7) had an RI ranging from 0.58 to 0.72 (mean 0.64 +/- 0.01 SE). In contrast, the immune thrombocytopenic group (n=28) had RIs ranging from 1.04 to 1.75 (mean 1.43 +/- ;0.03 SE). Platelet-associated IgG was evaluated further by absorbing representative sera samples from each group against washed granulocytes, red cells and platelets. Only when sera from the immune thrombocytopenic group were absorbed against platelets did the reactive indices of pre- and postabsorption samples change significantly. These findings suggest that our assay is clinically applicable in detecting platelet-associated IgG in immune thrombocytopenia and has the advantage of being rapid, reproducible and easy to perform in a clinical laboratory.

Effects of Acquired Resistance on Infection with Eimeria falciformis var. pragensis in Mice

Mice immunized with infections of 500, 5,000, or 20,000 oocysts of E. falciformis var. pragensis were reinfected with 20,000 and 100,000 oocysts at 20 and 38 days, respectively, after the initial infection. After the first challenge infection, none of the immunized mice showed clinical signs of coccidiosis; a few mice passed very low numbers of oocysts, and oocyst discharge seemed to correlate negatively with immunizing dose. None of the mice immunized twice passed oocysts after challenge. Mice immunized with three infections were completely immune to challenge for 4 months. The effect of the immune response on the life cycle of the coccidium was determined by histological examination of the intestines of immune and nonimmune mice infected with the parasite. In both the immune and nonimmune groups, sporozoites penetrated absorptive epithelial cells and migrated to crypt epithelial cells during the first 6 to 24 h postinfection. At 48 to 72 h postinfection, the sporozoites developed into mature first-generation schizonts in the nonimmune mice, whereas the developing first-generation schizonts degenerated within the crypt epithelial cells of the immune mice. In nonimmune mice, third-generation merozoites, inoculated intracecally, developed into mature fourth-generation schizonts, whereas in immune mice the developing fourth-generation schizonts degenerated before maturing. The possibility that a cellmediated immune mechanism is responsible for the arrest in schizogony is discussed.

Post-transfusion alloimmunization in patients with sickle cell disease.

The transfusion histories over a 33-month period of 50 patients with sickle cell disease were reviewed to determine the frequency of alloimmunization to red cell antigens following transfusion in these patients. There were 30 females and 20 males, aged 19--49 years. Eighteen (36%) were immunized of which thirteen were females. Five of the patients have formed only one antibody so far, while the other 13 have formed two or more. Thirty-six antibodies were identified: 16 against various Rh antigens, 12 anti-Lewis, 5 anti-Kell and one each of anti-Jka, -Fya and -M. The immunized patients received, on the average, more transfusions although there was a considerable degree of overlap between the immunized and nonimmunized groups. An approach to the hemotherapy of patients with sickle cell disease (SCD) is discussed.

Mouse potency assay for Bordetella bronchiseptica bacterins.

A potency assay for Bordetella bronchiseptica bacterins has been developed using mice. The immunogenicities of three bacterins, B, C, and D, were evaluated for ability to prevent death in mice as compared with a reference standard bacterin (RSB-A). Bacterins RSB-A, B, and C were evaluated for ability to prevent death in mice as compared with a reference standard bacterin (RSB-A). Bacterins RSB-A, B, and C were evaluated in swine for efficacy against nasal turbinate atrophy. Swine immunized with RSB-A demonstrated 25% gross nasal turbinate atrophy (GNTA), whereas nonimmunized swine had 85% GNTA. Swine vaccinated with bacterins B and C demonstrated 0 and 100% GNTA, respectively, whereas the nonimmunized groups had 64 and 75% GNTA, respectively. RSB-A and bacterins B, C, and D provided average mouse survivals of 94, 88, 49, and 32%, respectively when the mice were given 1/10,000 of a recommended swine-immunizing dose, whereas an average of 88% of the unvaccinated mice died.

Effects of Local Immunization with Glucosyltransferase Fractions from Streptococcus Mutans on Dental Caries in Rats and Hamsters

The effect of local immunization with glucosyltransferase enzymes of Streptococcus mutans on dental caries in conventional rats, hamsters, and gnotobiotic rats was studied. Injection of these animals with crude or defined glucosyltransferase enzyme preparations incorporated into complete Freund's adjuvant consistently resulted in the presence of antibody in saliva demonstrable by functional inhibition of enzymatic activity and binding of radioactive enzyme. Serum antibody was also present. Two experiments have been completed in conventional rats and one in gnotobiotic rats in which the animals were immunized with crude enzyme antigens. The immunized groups of animals always had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. Hamsters immunized with a defined enzyme preparation, containing no more than three antigenic components (two of which were enzyme), also demonstrated significant reductions in mean caries scores. The numbers of lesions were also always lower in immunized animals. In some cases there were reductions in the numbers of S. mutans that could be recovered from the teeth of immunized, infected animals. The reductions in dental caries and lesions were greater on smooth dental surfaces than on occlusal surfaces, which might be explained as interference with adherence phenomena demonstrated by S. mutans. It is proposed that antibody interference affects dental caries caused by this organism.

Acquired resistance against Listeria monocytogenes in red mice and CF1 mice immunized with strains of BCG or Mycobacterium tuberculosis.

Groups of red mice and CF1 mice immunized intravenously with varying doses of a weak or a strong strain of BCG or a strain of Mycobacterium tuberculosis were challenged 3 weeks after immunization with 0.1 or 0.2 ml 10(-2) Listeria monocytogenes injected intravenously, simultaneously with a non-immunized control group. The acquired resistance was determined on the basis of the number of survivors and the survival times of the animals that died spontaneously. In the red mice, the strong BCG strain induced a definitely higher resistance than the weak strain, and the M. tuberculosis strain a slightly higher resistance than the BCG strains. As in red mice, the resistance of CF1 mice was higher in animals immunized with M. tuberculosis than in those immunized with the BCG strains. However, the difference in the survival times of mice immunized with the two strains of BCG was much less than in red mice, and was only clearly significant as regards one of the doses used. The relationship between the virulence of a mycobacterial strain and its ability to induce acquired resistance against an infection with listeria or against an infection with virulent tubercle bacilli is discussed. It is concluded that red mice are more suitable than CF1 mice for evaluation of the protective potency of a BCG strain.

Effects of Local Immunization with Streptococcus mutans on Induction of Salivary Immunoglobulin A Antibody and Experimental Dental Caries in Rats

The effect of local immunization with Streptococcus mutans on dental caries in conventional and gnotobiotic rats was studied. Injection of these animals with S. mutans strain 6715 incorporated into complete Freund adjuvant consistently resulted in the presence of antibody in saliva directed to this organism. This antibody was primarily of the immunoglobulin A class as demonstrated by specific antiglobulin augmentation and gel filtration of antibody activity. Serum antibody was also present. Five experiments have been completed in conventional rats and two in gnotobiotic animals. The immunized group of animals always had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. The numbers of lesions were also always lower in the immunized animals, suggesting a possible interference with the formation of new lesions in immunized animals. The reductions in dental caries and lesions were greater on smooth surfaces than on occlusal surfaces. which might be explained as interference with adherence phenomena demonstrated by S. mutans. It is proposed that salivary immunoglobulin A antibody may be viewed as an ecological determinant in the oral cavity by affecting oral microorganisms and possibly their by-products.

The induction of vascular renal allograft rejection by leukocyte sensitizatin.

Abstract Human renal allograft rejection occurring within the first 2 months after surgery is frequently a vascular phenomenon, characterized by fibrinoid necrosis and endothelial cell proliferation of small vessels and plugging of glomerular capillaries with platelets and fibrin. This contrasts with interstitial edema and mononuclear cell infiltration, and only minimal vascular involvement, usually seen in renal allograft rejection in animals but also seen in man. To investigate this difference, 18 dogs were administered intravenous donor buffy coat prior to transplantation. These were compared to 7 dogs similarly treated except that buffy coat was not administered. Three autografts served as controls. All dogs received azathioprine daily (3 mg. per kilogram) beginning on the day of the operation. Daily assays of renal function, dog survival time, platelet counts, platelet survival times, and repeated biopsy specimens were compared. A marked difference between the immunized and nonimmunized groups was apparent morphologically. In the immunized group, vascular wall necrosis, platelet and fibrin plugging of glomerular capillaries, and endothelial cell proliferation were striking features of the rejection process. The over-all appearance was similar to that often seen in severe early (but not hyperacute) rejection in man. No significant difference in dog survival times or platelet studies was demonstrable between the groups. It is suggested that in man, the morphologic changes seen in early vascular rejection are a result of prior sensitization to transplantation antigen. Drug trials in animal transplantation aimed at studying reversal of the vascular aspects of rejection should be performed on animals exposed preoperatively to donor antigen.

Prevention of Experimental Hematogenous and Retrograde Pyelonephritis by Antibodies Against Enterobacterial Common Antigen

Rabbits were immunized with common antigen (CA) derived from Salmonella typhimurium. Animals with CA hemagglutinin titers of 1-320 to 1-81,920 were injected with 10(7) viable Proteus mirabilis via the retrograde and hematogenous routes. Nonimmune control groups were challenged similarly. From the retrogradely challenged groups sacrificed at 4 weeks, pyelonephritis was found in 89% of the control animals but not in those immunized. Bacteriuria was present in 89% of the controls but in only 61% of the immunized group. Hematogenously challenged, immune animals sacrificed at 6 weeks did not show histological evidence of renal pathology, and only 6% had bacteriuria. Eighty-six per cent of the nonimmune controls showed both pyelonephritis and bacteriuria. The protective effect is specific because of the following. (i) Active and passive CA immunization did not prevent pyelonephritis due to Pseudomonas aeruginosa which does not produce this antigen. (ii) Passive immunization with a CA antiserum conferred protection against a P. mirabilis challenge. (iii) Passive administration of an antiserum from which CA antibodies had been differentially absorbed abolished the protective activity.

Association of Heterospecific Pregnancies and the Presence of Saliva Antibodies

Saliva samples from 442 persons were tested for the presence of anti A and/or anti B saliva antibodies. The material was classified into two groups: 1. The ‘non-immunized’ group comprised group 0 women with group 0 husbands, primiparous women with a group O child, nonparous women, and group O males. 2. The ‘immunized’ group comprised group O women with heterospecific pregnancies or group incompatible husbands. Chance immunization by A and B-like substances from other sources was not considered as both groups would be subjected to these influences. Saliva antibodies were found more frequently in the immunized group than in the non-immunized group. The difference between the two groups was statistically significant. If the figures for males were omitted from the non-sensitized group the difference still remained highly significant. In the compatible group the number of pregnancies had no influence on the presence or absence of saliva antibodies. In the incompatible group there was a statistically significant decrease of saliva antibodies with the increase in the number of pregnancies. Explanations for these findings are discussed.

Studies on Diphtheria-Pertussis-Tetanus Combined Immunization in Children

Summary and Conclusions The immune responses of 239 free-living children who received primary vaccination with 3 monthly doses of alum-precipitated DPT vaccine were analyzed on the basis of latent diphtheria immunity prior to vaccination. It was found that the latently diphtheria-immune group yielded significantly weaker pertussis agglutinin and tetanus antitoxin responses than the diphtheria non-immune group. The responses of the latently diphtheria-immune group showed the following characteristics: The magnitude of pertussis agglutinin formation was suppressed, with a mean titer ratio of 1:3.2 against the diphtheria non-immune group 1 mo after the third dose. During the period of observation, the interfered pertussis agglutinin response curve showed no tendency to cross the normal pertussis agglutinin response curve.Both speed and magnitude of tetanus antitoxin formation were heterologously interfered with, the most striking interference following the second dose (mean titer ratio 1:11.3). The interfered tetanus antitoxin response curve tends to cross the normal tetanus antitoxin response curve at their peaks.Weak but detectable interference of pertussis agglutinin and tetanus antitoxin responses occurs when there pre-exists the slightest degree of latent diphtheria immunity. As the degree of pre-existing diphtheria immunity increases, the interference intensifies. In our studies, a diphtheria antitoxin level of 0.01 u/ml represents that degree of immunity over and under which the intensity of interference differs significantly. A further graded increment of pre-existing diphtheria immunity above this level, however, does not result in proportional enhancement of interference. The significance of these observations is discussed.

THE EPIDEMIOLOGICAL EVALUATION OF A NON-CELLULAR PERTUSSIS ANTIGEN

A brief discussion of the background and current status of pertussis immunization has indicated the need for a more satisfactory immunizing agent. The clinical evaluation of a new non-cellular pertussis antigen is described. Statistical analysis of data from 197 children completely immunized with the non-cellular antigen, 207 completely immunized with whole organism vaccine and 238 non-immunized control children suggest the following conclusions: The epidemiological data relating to the non-cellular antigen indicates its ability to produce a satisfactory level of immunity to pertussis. The significantly higher agglutinin titers and better protection in humans suggest that there is a better response to time non-cellular antigen than to the control vaccine even though mouse protective activities were approximately equal. The non-cellular pertussis antigen caused a febrile reaction of shorter duration than the control vaccine and other less quantitative measures of toxicity indicated that the non-cellular antigen was less toxic than the control vaccine. The frequency of pertussis cases in both immunized groups was significantly lower than in the non-immunized control group. The groups employed in the study were significantly uniform and thus permitted a valid comparison of the results among the various groups.

PREVENTION OF PNEUMOCOCCAL PNEUMONIA BY IMMUNIZATION WITH SPECIFIC CAPSULAR POLYSACCHARIDES

1. Immunization of man with 0.03 to 0.06 mg. of each of the capsular polysaccharides of pneumococcus types I, II, V, and VII, given in a single subcutaneous injection, has been shown to be effective in preventing pneumonia caused by these types but not that due to heterologous types. 2. Immunity appears within a period of 2 weeks following injection of the polysaccharides. Its duration was not determined, although 6 months can be set as a minimum. 3. Immunization of alternate subjects in the population reduced greatly the incidence of pneumonia in the non-immunized. 4. The carrier rate for pneumococcus types I, II, V, and VII was lowered significantly in the immunized group as compared with the controls. It is suggested that an over-all reduction in the incidence of carriers was responsible for the lowered rates for pneumococcal pneumonia in the non-immunized group.

Effect of a Whooping Cough Epidemic upon the Size of the Nonimmune Group in an Urban Community

Effect of a Whooping Cough Epidemic upon the Size of the Nonimmune Group in an Urban Community