M. Richter

The role of bone marrow in the immune response.

Publisher Summary This chapter summarizes the various experimental findings related to the source and function of various immunocompetent cells to define the functions of these cell types with respect to the induction of the humoral and cell mediated immune responses. The chapter evaluates the role of the bone marrow as a source of and as an influence upon stem cells and immunocompetent cell precursors and presents a scheme of cellular interactions culminating in antibody formation that incorporate the findings of all investigations to date in a variety of animal species. The scheme presents the specific multicellular pathways composed of a number of specialized cells rather than the single cell pathway characterized by a single, pluripotential immunocompetent cell. Furthermore, it also appears to supply the microphages as well as the cells that constitute the cellular infiltrations in cell-mediated immune reactions. A better understanding of the immune processes and the cellular events and interactions that lead to the humoral and cell-mediated immune responses will greatly facilitate the comprehension of the etiology, pathogenesis, and management of the immune deficiency syndromes, autoimmune states, and related diseases and will hasten the evolution of more rational and scientifically based approaches to prevent the graft-versus-host reaction and the graft rejection by the host.

The in vitro blastogenic response of lymphocytes of ragweed-sensitive individuals

Abstract Human peripheral leukocytes of normal and ragweed-allergic individuals were cultured with tritiated thymidine in vitro both in the presence and absence of the allergen and in the presence of autologous or homologous plasma. Cells of allergic individuals uniformly incorporated the labeled thymidine to a much greater degree than did cells of normal individuals. The optimal conditions for the incorporation of radioactive thymidine by allergic cells were 7 days in normal homologous plasma in the presence of 100 μg of allergen and with the isotope added 24 hours prior to termination of culture. Attempts to passively transfer blastogenic activity to cells of normal individuals were unsuccessful.

Studies on ragweed pollen. I. Preliminary fractionation of ragweed pollen: Characterization and allergenicity of the isolated fractions☆

Abstract 1.1. Defatted ragweed pollen was extracted with a number of solvents and the various fractions were analyzed by free and paper electrophoresis and ultracentrifugation. All fractions were compared for their allergenic reactivity by skin tests. 2.2. Extraction of defatted ragweed pollen with different solvents yielded fractions of different composition. 3.3. All the extracts soluble in the different solvents displayed allergenic reactivity, while the water-insoluble ragweed was the only one of the insoluble residues to display reactivity consistently. 4.4. The results of this study suggest that zone electrophoresis is suitable for the further fractionation of the various ragweed pollen fractions.

Studies on the allergens of ragweed pollen: A review

Abstract The results reported by numerous investigators who have attempted over the last forty years to isolate and establish the composition of the allergen(s) in the pollens of short and giant ragweed are conflicting. None of the investigations reviewed has provided a satisfactory proof that the allergen(s) have been separated in a pure form. It would seem that the following two studies represent the most advanced stages in the characterization of allergens in ragweed pollens: The work of Goldfarb and associates 58 has led to the isolation from giant ragweed pollen, of a fraction (Trifidin A) which was shown to be chemically, electrophoretically, and immunologically a single entity. On standing in the cold in distilled water, however, it was degraded into at least three components: a ninhydrin-stainable material, arabinose, and another reducing substance. We 117 have recently isolated, by use of paper electrophoresis, a highly allergenically active fraction from short ragweed pollen (AA 1 -D), which was shown to contain a pigment and peptide(s) composed of eight amino acids: arginine, lysine, glutamic acid, glycine, alanine, hydroxyproline, valine, and norleucine. The pigment could be removed from this fraction without loss of allergenic activity. On the other hand, hydrolysis of the peptide(s) resulted in a complete loss of allergenic activity; therefore, it was concluded that the peptide(s) appeared to be the allergen(s) in short ragweed pollen. Furthermore, it was shown that 10 −7 mg. of fraction AA 1 -D was allergenically as potent as 10 −4 mg. of the whole water-soluble extract of ragweed pollen. In view of the extremely high biologic potency of fraction AA 1 -D, and in view of the fact that allergenically active fractions isolated in previous studies all contained some nitrogen material, it may be suggested that the allergenicity of ragweed pollen is due to peptide(s), which may be complexed with other carriers, such as proteins, carbohydrates, or pigments.

Immunologic studies on milk and egg allergy

Abstract 1.1. Immunologic studies were carried out on sera from 25 patients clinically allergic to milk or egg and 9 normal controls. 2.2. Specific hemagglutinins to milk were demonstrated in half of the sera from milk-sensitive individuals and half of the sera from controls. 3.3. Specific hemagglutinating antibodies to egg were demonstrated in half of the sera from egg-sensitive individuals and half of the sera from controls. 4.4. None of the sera fixed complement. 5.5. In the food-sensitive individuals studied, hemagglutinin titers did not correlate with age, sex, results of skin tests, interval since last clinical reaction, or type of clinical reaction. 6.6. It is concluded that a single random determination of hemagglutinin titer cannot be used in the diagnosis of food sensitivity.

Cells involved in cell-mediated and transplantation immunity. III. The organ source(s) of the cells in the normal rabbit which mediate a reaction of cellular immunity in vitro

Abstract An in vitro cell culture system has been utilized to demonstrate allogeneic inhibition. Spleen cells obtained from a rabbit immunized with sheep red cells 3–6 months previously (memory rabbit) were uniformly capable of giving rise to plaque-forming cells (PFC) when stimulated with sheep red cells in culture. However, the addition to the culture of lymphoid cells obtained from the circulation or the gut-associated lymphoid organs—the sacculus rotundus, appendix and Peyers patches (SAPP organs)—of a second unrelated rabbit (effector rabbit) resulted in the suppression of the emergence of PFCs from the allogeneic memory spleen cells. The spleen and lymph node cells of the effector rabbit exhibited a significantly lesser degree of PFC-inhibitory activity, whereas the bone marrow and thymus cells exhibited no inhibitory activity. Evidence is presented demonstrating that the inhibition of PFC generation is not mediated by the release of cytotoxic antibodies directed to the memory spleen cells by the allogeneic lymphocytes. The results support the conclusion that the reaction observed represents the in vitro counterpart of the graft-versus-host reaction and strongly imply a SAPP organ origin for the cells mediating this reaction in the rabbit.

The Metabolism of Phytohaemagglutinin during Incubation with Human Peripheral Lymphocytes

The mechanism of action of PHA in in vitro leucocyte cultures was investigated with respect to the fate of the mitogen during the 3‐day blastogenic response. It was demonstrated that incubation of the leucocytes with PHA for only 5 minutes followed by incubation for 72 hours in medium devoid of PHA was sufficient to initiate mitosis and blastogenesis. However, the minimum incubation time with PHA required to induce maximum blastogenesis was 6 hours. Furthermore, it was demonstrated that the mitogen is bound to cells in culture but the experimental findings suggest that this cell‐bound mitogen is not degraded during the 3‐day incubation period.

LACK OF CORRELATION OF ANTIBODY TITERS AS DETERMINED BY THE PRECIPITIN AND HEMAGGLUTINATION TECHNIQUE.

Many different techniques are available today to aid in the detection of an antibody-antigen interaction. These generally fall into the catagories of the complement fixation, precipitation hemagglutination and agar gel techniques. Furthermore, it is a common practice to utilize, for any particular system, the technique which has been demonstrated as being most sensitive to ascertain the presence of the antigen-antibody interaction. It has been demonstrated that the BDB (antigen coupled to red blood cells through bis-diazotized benzidine) hemagglutination reaction is far more sensitive than the conventional precipitin method in detecting small quantities of antibody ( i ) . Furthermore, it has also been shown that only bivalent or precipitating-type antibodies are capable of agglutinating antigensensitized red blood cells (2) . One would therefore expect that a correlation should always exist between antibody titers as determined by the precipitin and by the hemagglutination techniques. During the past few years, however, various discrepancies between the results obtained with these two procedures

Cells involved in cell-mediated and transplantation immunity in the rabbit ☆: IV. The organ localization of the cells capable of migrating in vitro. Inhibition of migration by immunizing antigen

Abstract Rabbits were immunized with ovalbumin emulsified in complete Freunds adjuvant subcutaneously or with ovalbumin (OA) in saline intravenously. They were skin tested at intervals of time in order to determine the optimal sensitization time for the induction of the delayed skin reaction. The rabbits were also sacrificed and cell suspensions were prepared from the following organs: spleen, thymus, bone marrow, lymph nodes (popliteal), appendix, sacculus rotundus, and Peyers patches. Peritoneal exudate cells were also obtained. These cell suspensions were tested for their ability to be inhibited in their migration in vitro by the specific sensitizing antigen. It was observed that the migration of all of the cell suspensions except for the bone marrow and peritoneal exudate cells could be inhibited by OA, but not by BGG, a non-cross-reacting antigen. Inhibition of migration was most marked at 3–4 weeks postsensitization and was negligible by 8–12 weeks, at a time when the delayed skin reaction was as extensive as in the early postsensitization period. Furthermore, the migration of cells of rabbits immunized with OA in saline intravenously was also markedly inhibited. It is concluded that, in the rabbit, different cell pathways are operative in the induction of the delayed skin reaction, on the one hand, and the facilitation of migration inhibition, on the other.

The reversed BDB technique: The agglutination of antibody-coated red blood cells by homologous antigen

Abstract 1.1. The use of immunosorbent-antigen conjugates for the isolation and puri fication of antibodies has been discussed. 2.2. Both the diazotized as well as the nondiazotized polyaminostyrene-antigen conjugates are capable of taking up the specific antibody; however, antibody can be eluted only from the former, and not from the latter, immunosorbent. 3.3. The degree of recovery of the antibodies from an antiserum is directly related to the hemagglutinating titer of the antiserum. Only antisera with hamagglutinating titers of over 20,000 can be used for antibody elution experiments. 4.4. The method of using antibody-sensitized cells for the detection of antigen is described. The agglutination of antibody-sensitized red cells by the homologous antigen is specific as it could only be inhibited by the original antiserum and the eluted antibody but not by the antibody-depleted supernatant or heterologous antiserum. 5.5. Red blood cells conjugated with gamma globulin preparations of these antisera failed to be agglutinated by the homologous antigen.