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A histoautoradiographic study of the localization of antigen and specific antibodies in the rabbit spleen. I. Comparison of the primary and secondary immune response.

The localization of antigen (chicken serum albumin) and specific antibodies in sections from the spleens of rabbits after primary and secondary immunization was studied by means of histoautoradiography. After primary immunization, both antigen and antibodies were demonstrated in the lymphoid follicles consistently between 3 and 42 days after injection. Antigen was detected in addition in the marginal zone on the 5th day, and antibody-forming cells in the marginal zone and periarteriolar lymphocyte sheaths between 3 and 9 days after immunization. After secondary immunization, follicular antigen trapping was detected as early as 2 hours after injection, reaching a peak in the follicles and marginal zone on the 3rd day after immunization. At later periods of time the presence of antigen in the follicles was less prominent than that of specific antibodies. The numbers of antibody-forming cells were 10-100-fold higher in the secondary than in the primary response. The findings are discussed in relation to the role of antigen-antibody complex in the lymphoid follicles in the development of the capacity to produce the secondary type of immune response.

Animals

Kinetics of the reactive cell clones after immunosuppression and induction of tolerance: (1) inhibition of 19 S and 7 S plaque-forming cells in the primary and secondary immune response to sheep red blood cells by cyclophosphamide and 036.5122 (Asta).

The kinetics of the reactive cell clones after primary and secondary immunization with SRBC1) modified by cyclophosphamide and a newly synthesized cyclophosphamide analogue 036.5122 (Asta), have been studied. After primary immunization, both substances caused a severe and dose dependent depletion of 19 S PFC2). The 7 S PFC in the late primary response were only slightly inhibited by cyclophosphamide in low dose ranges, indicating, that sensitization could not be prevented by this substance. In contrast, 0.36.5122 was fully able to suppress 7 S PFC. Thus, treatment with 0.36.5122 after primary immunization can fully prevent the expression of the specific response. Experiments dealing with inhibition of a secondary immune response revealed that both test substances were able to strongly suppress the 19 S as well as the 7 S PFC. In general, 036.5122 demonstrates a higher suppressive potency, and the timing of its application for optimal suppression is less delicate than that of cyclophosphamide. 036.5122 was equally well inhibitory, whether given directly before or after antigenic challenge. The hypothesis is discussed, whether the immunosuppressive effect of 036.5122 given before antigenic challenge in the secondary immune response is due to cytotoxic damage of antigen reactive clones stimulated by persisting antigen.

Animals

Suppressive effect of Newcastle disease virus on the primary and secondary immune responses of hamsters.

The effect of Newcastle disease virus (NDV) on the formation of 19 S haemolytic plaque-forming cells (HPFC) in the secondary immune response and on the formation of 7 S HPFC in both primary and secondary immune responses of hamsters to sheep red blood cells (SRBC) was studied. The 19 S and the 7 S HPFC in the spleens of hamsters injected with SRBC were demonstrated by the direct and indirect Jerne's method, repectively. A single intraperitoneal injection of NDV 5 days prior to primary immunization markedly depressed the number of 7 S HPFC. NDV administered 5 days prior to the second injection of SRBC resulted in a significant decrease in the number of both 19 S and 7 S HPFC. It is asssumed that interferon induced by NDV is responsible for the suppressive effect of the virus on both the primary and secondary immune responses of hamsters to SRBC.

Animals

Vibriolytic IgG immunocyte response of mice after primary and secondary immunization with cholera somatic antigens.

Antibody plaque-forming cells (FC) to the somatic antigens of Vibrio cholerae were enumerated in the spleen of mice after primary and secondary immunization with a heat-killed vaccine prepared from the vibrios. Immunocytes releasing both high efficiency IgM and low efficiency IgG antibody were readily detected using a direct and facilitated plaque procedure in agar gel. Whereas the peak numbers of IgM-PFC after primary immunization occurred on days 12 to 14, the peak IgG-PFC response developed somewhat later (16-18 days). After a second injection of vaccine larger numbers of both IgM- and IgG-PFE appeared in the mouse spleens, with peak responses for both occurring between days 5 and 8. The largest number of IgG-PFC developed in spleens of mice given a second injection of vaccine 6-8 weeks after primary immunization. The dose of killed vibrios used for priming markedly affected both the magnitude and the class of antibody-forming cells appearing during the secondary response; 1--10 mug vaccine was more effective than higher or lower doses for priming the mice to a heightened secondary response. Furthermore, the antigenic specificity of both the IgM- and IgG-PFC appearing after secondary immunization was directly related to the strain of cholera bacilli used for priming. When mice were immunized with the Ogawa strains of cholera most of the secondary PFC after booster immunization with the serologically distinct Inaba strain was directed towards the common antigen shared by both strains and not to the type specific antigen of the Inaba vibrios. The specificity of the anti-vibrio PFC during both the primary and secondary responses was readily demonstrable by inhibition experiments using sonicated or soluble cholera antigens. Prior incubation of these antigens with test spleen cells in the agar gel effictively inhibited development of the vibriolytic plaques, regardless of antibody class. Similar antigen extracts from toher bacteria had no effect. The immunoglobulin nature of the plaques was also demonstrable by inhibition with low dilutions of rabbit anti-mouse globulin serum incorporated into the agar plates prior to testing; both IgM and IgG plaues were inhibited.

Animals

Suppression of the secondary immune response by specific antibody, when given together with the secondary antigenic stimulus.

It is generally believed that antibody-mediated immunosuppression can be only produced in non-primed individuals, and that this applies both to experimental animals and Rh-negative women at risk. However, in this paper it is reported that the additional injection of 0.2 ml of an antiserum to sheep erythrocytes (SE) together with a secondary antigenic stimulus of 10(8) SE into mice, primarily immunized by a tiny dose of 5 x 10(5) SE 28 days before, was capable of producing effective suppression of the secondary immune response.

Animals

[Comparison of the action of carminomycin and rubomycin on the dynamics of the primary and secondary immune responses].

The effect of rubomycin and carminomycin on the dynamics of the primary and secondary immune response and formation of the immunologic memory to sheep red cells in mice was studied. Differences in the character of the antibiotics effect indicative of the higher selective action of carminomycin on multiplying cells, precursors of the antibody-forming plasmids, were found. Theoretically interesting discrepancies in the effect of the antibiotics on the content of the antibodies in the serum and the antibody-producing cells in the spleen were shown. It was demonstrated that carminomycin had no effect on formation of the immunologic memory inspite of a noticeable decrease in the total number of the spleen nuclear cells and the number of the antibody-forming cells at the moment of immunization under the effect of the antibiotic.

Animals

Passive transfer of systemic tumor immunity with cells generated in vitro by a secondary immune response to a syngeneic rat gross virus-induced lymphoma.

Spleen cells taken from W/Fu rats 4 to 6 weeks after immunization with the syngeneic Gross virus-induced lymphoma, (C58NT)D cells, at a time when they lack detectable activity in a short-term 51Cr release assay, were previously shown to retain the capacity to generate cytotoxic activity upon reexposure to mitomycin C-treated lymphoma (C58NT)D cells in vitro. In the studies presented here, we evaluated whether in vitro sensitization of immune lymphoid cells before systemic transfer to a nonimmune recipient allows for more effective transfer of tumor immunity. The results show that the passive transfer of immune spleen cells after in vitro cocultivation with mitomycin-treated (C58NT)D cells allows for inhibition of growth of a subcutaneous inoculum of lymphoma cells. In contrast, spleen cells obtained 4 to 6 weeks after primary sensitization or after secondary in vivo sensitization did not effectively confer anti-tumor immunity. As few as 5 x 107 in vitro sensitized cells permitted complete inhibition of 106 (C58NT)D cells and also allowed for inhibition of the growth of 107 (C58NT)D-F cells, which was lethal to control animals. Immune cells sensitized with syngeneic thymocytes or normal spleen cells sensitized with (C58NT)D cells in vitro did not confer in vivo anti-tumor immunity. After systemic transfer of in vitro sensitized cells, delayed hypersensitivity occurred at the site of tumor inoculation and tumor growth was suppressed. Specificity of the passive immunity was shown by the failure to inhibit growth of a polyoma virus-induced sarcoma in rats which inhibited growth of the Gross virus-induced lymphoma cells. In vitro sensitized cells were more effective in the transfer of anti-tumor protection after 5 days, as compared to 2 days, of cocultivation with tumor. Results show that in vitro sensitized cells can effectively transfer systemic tumor immunity.

AKR murine leukemia virus

[Studies on the in vitro formation of antibodies. III. Induction of primary and secondary immune response in vitro].

Peritoneal cells from normal, unimmunized mice (female NMRI, 28-32 gr) produced in vitro primary and secondary immune response after induction with the bacteriophage T2 6 hours or 7 day resp. after establishing the cultures. We confirmed the induction of a primary and secondary immunological response in vitro in the very same culture by the following data: 1. In vivo the donor animals were not in contact with the antigen used. We found neither the phage nor its host E. coli B in the gut of 97 mice investigated and no humoral antibodies against T2. The kinetics of humoral antibody production in vivo by different doses of T2 also showed that there are no related or identical antigen structures incorporated in our animals. 2. The T2 neutralizing activity in the culture medium after the first induction had the sedimentation constant of 19.7 +/- 2.3 S (n = 9) but the activity found after the second induction sedimented with 8.1 +/- 0.7 S (n = 10). 3. The primary activity was more sensitive to mercaptoethanol than the secondary. 4. Complement was bound by the complex T2 + neutralizing activity.

Animals

Relationship of cellular proliferation and the generation of cytotoxic cells in an in vitro secondary immune response to syngeneic rat lymphoma cells.

The role of cellular proliferation for the generation of cytotoxic activity in an in vitro secondary immune response to syngeneic lymphoma cells was investigated. Spleen cells from W/Fu rats immunized with the syngeneic (C58NT)D tumor proliferate and generate cytotoxic potential for tumor targets after exposure to mitomycin C-treated (C58NT)D cells in vitro. Elimination of proliferating cells by exposure to high specific activity 3H thymidine at approximate intervals impaired the generation of cytotoxic activity. Elimination of cells proliferating to either syngeneic lymphoma or BN rat alloantigens allowed the remaining cells to generate cytotoxic potential to the second set of antigens. Elimination of proliferating cells also abrogated the ability of the in vitro generated cells to adoptively confer anti-tumor protection on nonimmune recipients. These results demonstrate that cellular division is required for the generation of cells which are cytotoxic in vitro and can adoptively confer anti-tumor protection in vivo.

Animals

[Changes in the electric charge of blood lymphocyte populations after secondary immunization with tetanus antitoxin in man].

The electrophoretic mobility of circulating lymphocytes has been studied in normal human subjects after immunization by tetanus toxoid. The mean migration speed was shown to increase, particularly two and three days after secondary immunization. This increase appeared to be due to the elevation of percentage of T cells migrating at 1.20 and 1.35 micrometer. sec.-1v.-1 cm. (active rosettes-forming cells), with a decrease of the percentage of B cells and T lymphocytes migrating at 1.10 micrometer. sec.-1v.-1 cm. The return to the anterior status was observed between day 4 and 8 after immunization.

Electrophoresis

The effect of long-term exposure to cigarette smoke on the height and specificity of the secondary immune response to influenza virus in a murine model system.

The effect of long-term exposure to cigarette smoke on the height and specificity of the secondary humoral immune response to influenza was investigated in a murine model system. It was shown that if mice were pre-immunized with a sub-lethal infection of influenza virus and then exposed to cigarette smoke daily for 36 weeks, they were able to mount a secondary immune response of normal height on subsequent challenge with the homologous virus strain. The response however, was less specific than that elicited in control mice, with high titres of cross-reacting antibody by haemagglutination-inhibition to the following strain in the same antigenic series. Recall of antibody to the previous strain in the antigenic series was not observed in either control or smoke-exposed animals. These results serve to correct an earlier discrepancy between the murine system and human studies in which the response to influenza infection in mice was depressed by prolonged exposure to cigarette smoke, whereas in man the response of smokers did not differ significantly from that of non-smokers. This apparent discrepancy had been caused by a lack of previous experience of influenza in the mice, which had therefore mounted a primary response, compared with the secondary response observed in the human studies.

Animals

Suppression of secondary immune response by antilymphocyte serum: time relationship between immunization and administration of antilymphocyte serum.

The effect of antilymphocyte serum (ALS) on the secondary humoral immune response to sheep erythrocytes (SRBC) in rats was studied by the Jerne plaque assay technique. Its effect was also studied on the delayed hypersensitivity (DH) response to SRBC by the foot pad swelling test. ALS(N), which was prepared against lymphocytes from normal rats, had no effect on the secondary humoral and cellular response or on the primary cellular response, when administered postantigenically. ALS(I), which was raised against lymph node cells from SRBC immunized rats produced significant immunosuppression of the secondary response to SRBC when administered either before or after the antigenic injections. In the case of DH, ALS(I) behaved just like ALS(N) having no effect on the secondary response and suppressing the primary only when administered prior to the antigen.

Animals

Restriction of primary responses to the IgG class and dependency of IgM responses on secondary immunization for the copolymers of L-glutamic acid, L-tyrosine, and L-alanine.

Primary responses to the linear polymers of L-glutamic acid, L-tyrosine, and L-alanine are restricted to the IgG class of antibodies. The appearance of specific IgM antibodies against these antigens is dependent upon secondary immunization, in contrast to many classical antigenic systems. The presence of an IgM response was verified by a direct plaque-forming cell assay, the inhibition of direct plaques by an antiserum specific for mouse micron-chain, and the physical separation of IgM and IgG GAT-specific antibodies by gel filtration. Preimmunization of the appropriate nonresponder strain with GAT or GT inhibits both the secondary IgM and IgG responses to GAT-MBSA and GT-MBSA, respectively. The tolerance observed is due to the induction of suppressor cells as demonstrated by cell transfer experiments.

Alanine

[Rate of development of immunologic memory and features of the secondary immune response in mice of high and low-reacting strains].

Immunological memory to sheep red blood cells developed in mice of strain CBA, DBA/2, and hybrids (CBAXXC57BL/6) F1 24 hours after the administration of a low dose of the antigen, and in C57BL/6 mice -- in 48 hours. The level of the secondary immune response in CBA, C57BL/6, and hybrid F1 mice was much greater than in the DBA/2 mice. The maximal production of the antibody-forming cells in the spleen of CBA mice occurred after twofold administration of low antigen doses. In contrast to this, repeated administration of a high antigen dose is required to obtain a marked immune response in adoptive transfer of spleen cells of the C57BL/6 strain.

Animals

Factors influencing the secondary immune response in rabbits to Salmonella typhosa.

The characteristics of the primary response, as well as those of the secondary response, to different doses of Salmonella typhosa when elicited at two different time intervals were analysed. The antibody response demonstrates that the size of the primary dose, and the size of the secondary dose, have significant effect on the subsequent synthesis of the secondary IgM and IgG antibody response to both 'H' and 'O' antigens. The time elapsed between the primary and secondary response tends to decrease the IgG response. Due to the high immunogenicity of the 'H' antigen the maximum IgG levels were reached within the range of doses used. These high levels of antibody inhibited the secondary IgM response by a feedback mechanism. Enhanced or equivalent response of predominant anti 'O' IgM system was recorded.

Animals