bluuebunny/crossref_metadata_2025 huggingface dataset:
Immune Hemolysis and the Functional Properties of the Second (C2) and Fourth (C4) Components of Complement
Authors: Loos Michael , Borsos Tibor , Rapp Herbert J.
Published: December 2022

Treatment of guinea pig or human C2 with trypsin in the presence of EAC4 leads to the formation of EAC. The kinetics of EAC generation, the nature of the C2 dose response curve and the nature of the decay curve of SA all indicated that EAC formed by the action of trypsin on C2 are not distinguishable from EAC generated by the interaction of EAC1̄4 and C2. The data support the hypothesis that C1̄ is a trypsin-like enzyme.
Lymphocyte Subpopulations in NZB Mice: Deficit of Thymus-Dependent Lymphocytes
Authors: Stutman Osias
Published: December 2022

The relative and absolute numbers of thymus-dependent lymphocytes (positive for ϑ isoantigen) and of thymus-independent lymphocytes (positive for membrane immunoglobulins or for antigen-antibody-complement receptors) were studied in the lymphoid tissues of NZB and CBA/H mice. The ages of the mice ranged from 100 to 600 days. A progressive decline of ϑ-positive lymphocytes was observed with age in NZB, but no such decay was detected in CBA/H mice, a non-autoimmune strain. The decline of ϑ-positive lymphocytes was observed in NZB mice as early as 200 days of age and appeared even in absence of detectable signs of autoimmunity, although the decline was accentuated by the development of autoimmunity and by progressive age. No such changes were observed for the thymus-independent lymphocytes, and only a moderate increase of these cells was observed with age both in NZB and CBA/H strains.
Host Defense Against Bacterial Endotoxemia-Contribution of The Early and Late Components of Complement to Detoxification
Authors: May Joseph E. , Kane Michael A. , Frank Michael M.
Published: December 2022

The precise mechanisms and tissue sites involved in the in vivo detoxification of endotoxin remain unresolved. Although a number of studies have indicated that circulating plasma is a principal site of endotoxin inactivation (1–3), it was believed that the complement system was not required for detoxification of endotoxin (1). However, a recent report which demonstrated the increased susceptibility of C6 deficient rabbits to the injection of bacterial endotoxin suggests that complement may be important (4). Endotoxin can activate the complement system either by the classical mechanism proceeding through the activation of C1, C4 and C2 to activate C3, or via the alternate or bypass pathway proceeding through another series of proteins, as yet incompletely defined, to enter the complement sequence directly at the C3 step (5–7). In this study a further attempt is made to evaluate the role of complement in the detoxification of endotoxin, with particular reference to the contribution of the early (C1, C4 and C2) and late acting (C3–C9) complement components.
Rat Alloantibody Responses against Strong and Weak Histocompatibility Antigens
Authors: Miller Carol , DeWitt Charles
Published: December 2022

Rat alloantisera were prepared in inbred strains of rats by immunization with lymphoid cells of a donor strain. Six host-donor combinations were incompatible at the major (AgB or H-1) as well as other histocompatibility loci. These sera contained complement dependent cytotoxins after two injections of 1 to 3 × 107 cells. Titrations showed 100% kill at several antibody (ab) concentrations and a typical doubling dilution end point. Six host-donor combinations were incompatible only at non-AgB (non-H-1) loci. These sera contained cytotoxins only after two injections of 4 × 108 cells. Titrations showed end points equal to those of the first group of antisera but no concentration was able to kill 100% of available target cells, nor did increments in antiserum concentration yield any change in the low number of cells killed. These characteristics were not the result of: a) the injection schedule; b) presence of complement inhibitors; c) low response capacity of the host; d) the presence of two lymphocyte subpopulations in the target cell suspension nor e) low ab concentrations. Absorption of the two types of antisera by homologous cells suggests that the non-AgB antisera: a) contain both complement-fixing (CF) and non-CF ab; b) the non-CF concentration is as much as 10-fold higher than CF; c) avidities of non-CF and CF are similar. We infer that immunization with non-AgB antigens preferentially induces formation of non-CF ab which is responsible for the odd titration and is enhancng ab.
Blastogenic Factor from Unstimulated Leukocyte Cultures: Further Evidence for Heterogeneity, and Mechanism of Production
Authors: Kasakura Shinpei
Published: December 2022

Blastogenic factor derived from unstimulated leukocyte cultures was studied. The present studies have shown that the properties of the blastogenic factor which stimulates only allogeneic cells (allogeneic blastogenic factor) are different in some respects from those of the blastogenic factor which stimulated autologous cells as much as allogeneic cells (autologous blastogenic factor). Activity of the allogeneic blastogenic factor was labile to exposure to temperature of 60°C, and was concentrated in the precipitate obtained by ultracentrifugation of cell-free culture medium (CFM) at 100,000 × G. In contrast, activity of the autologous blastogenic factor was resistant to the same temperature, and could not be sedimented by ultracentrifugation. The kinetics of in vitro production of the allogeneic blastogenic factor was also different from those of the autologous blastogenic factor. While activity of the autologous blastogenic factor was almost constant throughout the incubation period, that of the allogeneic blastogenic factor reached a maximum after 4 to 5 days of incubation and thereafter decreased. These findings suggest that there may be separate molecules responsible for the allogeneic and autologous blastogenic activities.

The blastogenic factor prepared from mixed cultures of intact cells and treated allogeneic cells was also studied. Although the difference was not significant (p = 0.09) when leukocytes were treated with actinomycin D, the CFM was more active for cells from the donor of the treated cells than for cells from the donor of the intact cells in eight out of 12 experiments. The ultracentrifuged precipitate (probably allogeneic blastogenic factor) was significantly more stimulatory toward cells from the donor of the treated cells than from the donor of the intact cells. When leukocytes were treated with Escherichia coli endotoxin, both the CFM and the ultracentrifuged precipitate stimulated cells from the donor of the treated cells more than cells from the donor of the intact cells. Mytomycin C did not have any effect on the blastogenic activity of the CFM and the ultracentrifuged precipitate prepared from these cultures.
Immunoglobulin Synthesis by Human Lymphoid Tissues: Normal Bone Marrow as a Major Site of IgG Production
Authors: McMillan Robert , Longmire Robert L. , Yelenosky Robert , Lang J. Eugene , Heath Victor , Craddock Charles G.
Published: December 2022

The Fab-anti-Fab assay for human IgG quantitation is used to measure in vitro IgG production by lymphocytes from five normal human lymphoid tissues: bone marrow, spleen, blood, lymph node and thymus. Synthesis of IgG was found to proceed at a constant rate over the 10-day culture period; production could be prevented by hypothermia or mitomycin-C. When corrected for total lymphoid cells, the bone marrow was noted to produce greater than 95% of the IgG synthesized by the organs which were evaluated. Secondary antigenic stimulation with smallpox vaccine resulted in accelerated IgG synthesis by blood and splenic lymphocytes; conversely, no response was noted in cultures containing marrow or thymic cells. It is concluded that the bone marrow must be considered as a major site of IgG production under normal circumstances; however, the inability of the bone marrow to respond to antigenic stimulation suggests that the appropriate quantity or quality of cells is lacking in this organ for optimal antigenic recognition and/or processing.
The Effect of Surgical Bursectomy and Irradiation on Rosette-Forming Cells in the Chicken
Authors: Kiszkiss D. F. , Choi Y. S. , Good R. A.
Published: December 2022

Heterologous erythrocytes are a useful antigen in studies of antigen-binding cells since their binding to lymphoid cells can be detected easily as rosettes of red cells around a central lymphocyte (1, 2). Such binding of antigen is believed to be an early stage in the development of both humoral and cellular immune responses. Because the humoral response to heterologous erythrocytes requires a synergism between thymus dependent (T)3 and bone marrow dependent (B) cells in the mouse (3, 4) and between T and bursal-derived cells in the chicken (5), it is important to assess the contribution of each to the population of rosette-forming cells (RFC). In an effort to explore the origin of RFC in the chicken, we have examined the effect of bursectomy and irradiation on the number of these cells, under conditions which, in normal animals, induce considerable RFC.

In the present report, we demonstrate that animals deprived of bursal influence from the embryonic period lack RFC.
Studies in Microbic Heredity
Authors: Mellon Ralph R. , Yost Elizabeth
Published: December 2022

Introduction. In the second paper (1) of this series it has been shown, first, that the fungoid phase of bacterial cultures can be stabilized in a long filamentous as well as in a coccoid stage, which in mycological terminology is the equivalent of saying that the coccoids (zygospores) may, when they germinate, reproduce themselves as large cocci without returning to the antecedent bacillary or filamentous form; second, under the proper conditions the zygospores can give rise to variants which culturally may be quite aberrant but serologically may be identical with the original strain, or on the other hand, quite specific.

Since the first of these propositions has been accomplished repeatedly in the test tube it is conceivable that this coccoid form may multiply as such in diseased conditions in the human organism and be isolated in this form, which obviously might be misleading as to its real nature.
The Results of the Specific Treatment of Late Hay Fever
Authors: Rackemann Francis M. , Graham Lillian B. , Scully Margaret A.
Published: December 2022

Hay fever in New England occurs in at least three groups. The first depends upon a hypersensitiveness to the pollen of trees which causes symptoms in May and early June. The second depends upon a hypersensitiveness to the grasses. Here the symptoms begin late in May or early in June and last through the month of June and into July. In both groups the causes of trouble are multiple since the patient is rarely, if ever, hypersensitive to the pollen of a single tree or of a single grass or even to any simple combination of a few different pollens: Usually skin tests reveal positive reactions to a wide variety of different pollens. For example, in this second group, it is unusual to find a hypersensitiveness to timothy alone. Red top, orchard grass, Bermuda grass, Johnson grass, rye grass, and other grasses, will frequently produce skin reactions at the same time and often of approximately the same intensity.
Upon the Anticomplementary Action of Pancreatic Extracts
Authors: Grove Ella F.
Published: December 2022

The anticomplementary effect of pancreatic extracts, first described by Michaelis and Skwirsky, was ascribed by these investigators to the tryptic ferments. In the present study, it is shown that the ratio of the digestive power of two pancreatic extracts (1:1) may be very different from the ratio of the anti-complementary power of the two extracts (1:100).

This observation indicates that the proteoclastic and the anti-complementary functions of a pancreatic extract reside in two different substances.

Attempts to separate the two functions in a single extract failed.
The Immunological Characteristics of a Water-Insoluble Protein in Type I Antipneumococcus Sera
Authors: Felton Lloyd D. , Bailey G. Howard
Published: December 2022

The fact that it has been found possible to separate the protective substance of antipneumococcus sera by a method (1) which involves little physical or chemical manipulation, instead of precipitation by high concentration of neutral salts, prolonged dialysis, or by specific antigens, led to a study of the immunological characteristics of this precipitated substance. The work was stimulated by the observations of Izar and Caruso (2) and of Izar (3), and the persistent thought of the possibility of antibodies, as evidenced by agglutination, precipitin, complement-fixation reactions, etc., being the manifestation of a single substance. Two views have been presented, the one sponsored by Ehrlich affirming the multiplicity of specific immune substances, and the other, by Bordet, Nicolle (4), and others, maintaining their essential unity. A comprehensive review of this second thought has been given by Zinsser (5) who advocates the essential identity of antibodies.
Studies in Hypersensitiveness
Authors: Levine Philip , Coca Arthur F.
Published: December 2022

The pioneer observations of Noon (1) and Freeman (2) in England, of Clowes (3), Koessler (4), and Cooke (5), in this country, and the subsequent confirmatory experiences of many others with the specific treatment of atopic conditions (hay fever and bronchial asthma) leave no doubt of a certain effectiveness of the method. This method consists in the subcutaneous injection, at intervals of several days, of gradually increasing quantities of the exciting agent.

The method referred to was conceived upon the basis of the procedure of desensitization in anaphylactic hypersensitiveness. However, there are at hand some observations by R. A. Cooke, which make it at least doubtful that the alleviating effect of the specific treatment of atopic hypersensitiveness is due to a desensitization, in the proper sense of that term. Cooke has found that, in individuals who have obtained complete relief from hay fever, as a result of injections of pollen extract, the degree of sensitiveness of the skin and even of the conjunctiva to the extract is the same as it was before the injections were begun.
Studies of the Toxins of the Hemolytic Streptococci Associated with Scarlet Fever
Authors: Kirkbride Mary B. , Wheeler Mary W.
Published: December 2022

Ever since laboratory procedure established the almost constant presence of hemolytic streptococci in the early stages of scarlet fever, researches have been directed toward determining the etiological relationship of these organisms to the disease. It is only recently, however, that a satisfactory evaluation of clinical and laboratory evidence has been made possible through the application of newer methods by Dochez (1, 2, 3, 4, 5,) and the Dicks (6, 7, 8, 9, 10). These studies are so familiar that it is unnecessary to review them here. It is evident, however, that further advances in our conception of the rôle of hemolytic streptococci in scarlet fever and in other infectious processes are largely dependent on a more exact knowledge of streptococcus toxins and their antitoxins. As a basis for such studies, uniform and readily applicable methods for the preparation and standardization of both the toxins and antitoxins must be developed.
The Effects of Specific Metals on the Antigenic Structure of Alkaline Phosphatase from Escherichia Coli
Authors: Reese R. T. , Treffers H. P.
Published: December 2022

Native alkaline phosphatase has been compared with its Zn (II) and Co (II) reconstituted forms by using direct inhibition of enzyme activity by antiserum, quantitative precipitation and serologic analysis with an aqueous polymer phase system. All three of the enzyme forms are differentiable by this combination of methods which allows measurements to be taken at ratios of enzyme and antibody which are in antigen or antibody excess as well as at equivalence. Although difficult to detect by physical techniques, the removal of divalent cations distinctly alters the structure of the enzyme as measured by quantitative precipitin analysis. Data from kinetic studies were interpreted as suggesting that at best, only part of the antibodies bound close to, or directly modified, the enzyme's active site. This was used as supported for the theory that this enzyme must undergo a conformational change in order to catalyze the hydrolysis of substrate.
Purification and Characterization of Coliphage MS2 Hapten Conjugates
Authors: Curtiss Linda K. , Krueger Robert G.
Published: December 2022

Conjugation of the RNA coliphage MS2 to either dinitrophenol (DNP) or picolinimidate (P), and separation from the reaction mixture on Sephadex G-25 results in the generation of at least two phage populations with differing sedimentation rates. These populations also can be separated on the basis of density. One phage population contains intact, infectious phage particles and conjugated hapten. The data presented shows that neither the density nor sedimentation rate change as a result of conjugation, nor is there any detectable alteration in the purified, conjugated particle that makes it any more sensitive to altered physical and chemical conditions. However, distinct differences between conjugated and unconjugated phage can be seen when they are neutralized with anti-MS2 sera.
Predominance of Theta-Positive Lymphocytes in Gut-Associated and Peripheral Lymphoid Tissues of Newborn Mice
Authors: Chanana A. D. , Schaedeli J. , Hess M. W. , Cottier H.
Published: December 2022

Morphologic and isotope-labeling studies on newborn mice have not only suggested a substantial thymocyte production (1) and peripheralization (2), but have also provided direct evidence for the thymic origin of a majority of lymphocytes in mesenteric lymph nodes and Peyer's patches (3, 4). The presence of θ-alloantigen on thymocytes of mice (5) has been utilized to evaluate the relative proportions of thymus-derived cells in lymphoid tissues (6). The values obtained by the use of cytotoxic anti-θ serum (7) are, however, significantly lower than those suggested by morphologic and isotope-labeling studies (3, 4). We have evaluated the relative numbers of θ-positive cells in the lymphoid tissues of newborn mice by indirect immunofluorescent staining (8). The results indicate that most of the lymphocytes in lymph nodes and Peyer's patches of mice are θ-positive.

Materials and Methods. Anti-θ and control anti-θ reagents (anti-θ, control reagent, respectively) were prepared by immunizing AKR mice with C3H mouse thymocytes.
Enhancement of an Anti-Hapten Antibody Response by an Antiserum to the Carrier Protein
Authors: Pincus Carolyn , Miller Gary , Nussenzweig Victor
Published: December 2022

The administration of relatively small doses of antiserum to BSA to mice receiving an injection of DNP 12BSA resulted in a concomitant enhancement of both the anti-BSA and anti-DNP antibody responses. It thus appears that under suitable conditions the “helper effect” may be mimicked by anti-carrier antiserum.
Monoclonal IgA and IgM in the Serum of a Single Patient (SC)
Authors: Seon B.-K. , Yagi Y. , Pressman D.
Published: December 2022

A comparative study was carried out with the light chains of IgA and IgM from a single patient (SC). Both light chains were of κ type and were homogeneous. They were not distinguished by urea disc electrophoresis, amino acid composition, N-terminal amino acid sequence (consequently Vκ subgroup), Inv allotype and peptide map. Both were of VκIII subgroup which is found in very low frequency among human κ chains. It appears that both light chains of SC IgA and SC IgM were synthesized by the same genes (the same variable portion gene and the same constant portion gene).
Heterologous Antiserum to Mouse Thymus-Derived Lymphocytes
Authors: Bron Claude , Sauser Daniel
Published: December 2022

By immunization of rabbits with mouse thymocyte membranes, antisera were obtained which reacted, after proper absorption, with 95 to 100% of mouse thymocytes and 35 to 40% of mouse spleen lymphocytes as shown by immunofluorescence and cytotoxic testing. After absorption of such antisera with mouse thymocytes of different θ-allotypes and mouse spleen lymphocytes pretreated with isologous anti-θ serum and complement, the remaining antigen(s) detected by heterologous antisera appeared to be distinct from the θ-alloantigen but present on the same population of spleen lymphocytes.

The simultaneous use of two in vitro tests whereby both humoral and cellular immune responses can be demonstrated in the same animal with one alloantigen shows that the differentiation antigen detected is specific for thymus derived Tlymphocytes.
Effects of Natural Thymocytotoxic Autoantibody of NZB Mice and of Specifically Prepared Antilymphocyte Serum on the Tissue Distribution of 51Cr-Labeled Lymphocytes
Authors: Shirai Toshikazu , Yoshiki Takashi , Mellors Robert C.
Published: December 2022

This study deals with the effect of natural thymocytotoxic autoantibody (NTA) of NZB mouse serum and of specifically prepared antilymphocyte serum upon the tissue distribution of 51Cr-labeled mouse lymph node cells in syngeneic recipients. By cytotoxicity test both sera were found to be cytotoxic for the same population of lymph node cells, which were believed to be thymus dependent lymphocytes (T-cells). C57BL/6J mice were injected intravenously with syngeneic 51Cr-labeled lymph node cells which had been previously incubated at 4°C for 60 min with either of these sera, in the absence of complement, or with normal control sera. Twenty-four hours after injection, a substantial decrease in migration to lymph nodes and spleen and an increase in migration to liver were observed with 51Cr-labeled cells which had been treated with either the NTA-positive NZB mouse serum or the antilymphocyte serum as compared with control groups. These findings are consistent with the hypothesis that NTA has a causal relation to the observed age decrease of recirculating T-cells in NZB mice and that T-cell depletion is mediated by a continuous process of autosensitization phagocytosis.
Chicken γA: Physicochemical and Immunochemical Characteristics
Authors: Bienenstock John , Perey Daniel Y. E. , Gauldie J. , Underdown B. J.
Published: December 2022

Column chromatography on Sephadex G-200 of ammonium sulfate preparations of chicken bile and duodenal washings revealed γA in a high molecular weight form with elution immediately after the void volume. γA from intestinal secretions was further purified by recycling chromatography on a Bio-Gel A-5M column. A constant S value was not obtained for the γA, values of 11.9 to 16.2S being recorded under various conditions. The sedimentation coefficient was relatively unaltered by mild reductive procedures with 0.1 M β-mercaptoethanol and 0.005 M dithiothreitol. Serum γA was partially purified by ammonium sulfate precipitation and DEAE chromatography. The majority of serum γA was in a high molecular weight form, eluting from a G-200 column mainly at the void volume. A smaller amount of γA eluted in the 7S region. The addition of 125I-labeled human secretory component (SC) to fresh chicken serum and subsequent sucrose gradient ultracentrifugation resulted in a 10 to 17% shift of radioactive counts to the heavier portions of the gradients. Radioautography of these fractions with heavy chain specific antisera revealed binding to γA in 3/3, to γM in 1/3 and to γG in 0/3 experiments. Addition of human SC to biliary γA did not result in binding to the γA molecule. This suggests that chicken γA obtained from secretions may possess its own form of SC.
Increased Responsiveness to Phytohemagglutinin and Graft-vs-Host Reactivity of Thymus Cells from Snell-Bagg Dwarf Mice
Authors: Duquesnoy René J. , Ahrens Richard C.
Published: December 2022

Autosomal recessive pituitary dwarf mice of the Snell-Bagg (SB)2 and Ames strains develop severe immunodeficiency which primarily affects the thymus-dependent system (1–5). The thymus undergoes a progressive involution which initially occurs as a depletion of thymocytes from the cortex accompanied by an increased number of lymphoid cells in the medulla. The next stage of thymus degeneration is a loss of lymphoid cellularity of the medulla, thereby causing complete atrophy (5).

In order to evaluate the immunologic reactivity of dwarf thymus and spleen cells, we have investigated their blastogenic response to phytohemagglutinin (PHA) and their ability to induce a graft-vs-host (GVH) reaction in immature F1 hybrid mice. Both tests determine the immunocompetence of thymus-dependent lymphoid cells.

Four-week-old dwarf and normal SB mice of both sexes were sacrificed by cervical dislocation. Their spleens and thymuses were removed and suspensions of each organ were prepared in cold Eagle's minimal essential medium (MEM) (Grand Island Biological Co., Grand Island, N. Y.).
Greater Sensitivity to Inhibition by Anti-Immunoglobulin of Splenic than of Bone Marrow B-Lymphocytes
Authors: Mond J. J. , Thorbecke G. J.
Published: December 2022

Bone marrow derived (B)3 lymphocytes in lymphoid organs represent the precursors of antibody-producing cells and are known to bear immunoglobulin molecules on their cell surface representative of the antibody ultimately to be produced. It has been suggested that B-cells in bone marrow, although at least partially sensitive (1), are more resistant than splenic B-cells to inactivation by incubation with highly radioactive antigen (2). Similarly, induction of tolerance by injection of ultracentrifuged human γ globulin (HGG) occurs much more rapidly in B-cells of the spleen than of the bone marrow (3). In light of these observations and in view of the fact that Ig receptors on the surface of B-cells most probably mediate their interaction with antigen, it seemed interesting to compare the relative sensitivities of mouse bone marrow and spleen precursors of antibody-forming cells to inactivation by anti-Ig antisera.
Evolutionary Studies on the Expression of Allotype Markers on Rabbit Immunoglobulin Molecules
Authors: Conrad A. H. , Rodkey L. Scott
Published: December 2022

Preparations of immunoglobulin F(ab′)2 molecules from individual cottontail sera were examined by indirect precipitation techniques for the expression of allotypic determinants expressed by the a and b loci in domestic rabbits. The data indicate that cottontail immunoglobulins express some but not all of the determinants specified by the a2, a3, b4 and b5 alleles in the domestic rabbit, but none of the determinants specified by the a1 allele in the domestic rabbit. Inhibition tests to compare the domestic rabbit allotype determinants expressed by individual cottontails suggested that the b4 and b5 markers carried by cottontail rabbits were similar from one positive individual to another, but that the a2 and the a3 determinants may differ from one positive individual to another.
Genetic Control of the in Vitro Responses of Rat Peripheral Blood Lymphocytes to Phytohemagglutinin and Concanavalin A
Authors: Newlin Carol M. , Gasser David L.
Published: December 2022

Incorporation of tritiated thymidine was used to quantitate the in vitro responses of rat peripheral blood lymphocytes to phytohemagglutinin (PHA) and concanavalin A (Con A). By this criterion, cells from Brown Norway (BN) rats respond less vigorously to these phytomitogens than cells from Lewis strain animals. Cells from (Lewis × BN) F1 hybrids respond as well as cells from Lewis donors, so high responsiveness appears to be fully dominant over low responsiveness. Studies on the inheritance of mitogen response levels suggest that a single gene is of primary importance in regulating the level of response to both PHA and Con A. The locus involved is not closely linked to the major histocompatibility antigen locus, AgB.
Studies of Guinea Pig Complement Component C9: Reaction Kinetics and Evidence that Lysis of EAC1–8 Results from a Single Membrane Lesion Caused by One Molecule of C9
Authors: Rommel Fred A. , Mayer Manfred M.
Published: December 2022

The generation of EAC1–9 from EAC1–8 by reaction with C9 proceeds without time lag and the initial reaction velocity is directly proportional to the concentrations of C9 and EAC1–8. These characteristics, as well as the low temperature sensitivity, indicate that the reaction is not enzymatic. The subsequent lysis of EAC1–9 is highly sensitive to temperature, the Q10 being approximately 2. Rabbit anti-guinea pig C9 blocks the reaction of C9 with EAC1–8, but does not interfere with lysis of EAC1–9. Input of 1 to 2 molecules of C9 suffices for lysis of 1 EAC1–8. The rate of lysis of EAC1–9 is proportional to the concentration of C9 per cell. The dose-response curve in the titration of C9 follows a monotonic course concave to the abscissa representing the concentration of C9. It is concluded from these results that lysis of EAC1–8 results from a single membrane lesion caused by 1 molecule of C9.
Functional Heterogeneity of Murine Lymphoid Cells
Authors: Stobo John D. , Paul William E. , Henney Christopher S.
Published: December 2022

Murine thymus-derived (T) cells may be divided into two distinct subgroups on the basis of differences in their relative density of the T differentiation antigen theta (ϑ), differential responsiveness to the T dependent mitogens phytohemagglutinin (PHA) and concanavalin A (Con A) as well as by differences in recirculation patterns and radiation sensitivities. Thus, among splenic lymphocytes one population of T cells bears a relatively high density of ϑ determinants, demonstrates relatively equal responsiveness to PHA and to Con A, resides within the recirculating pool of lymphocytes and is relatively radiation sensitive. Another population of splenic T cells bears a relatively reduced density of ϑ determinants, demonstrates reactivity predominantly to Con A and belongs to a sessile, radiation resistant lymphocyte pool. The ability to delineate T cell subgroups by these characteristics provides a useful means of investigating the functional characteristics of T cell subsets.

In this report, we demonstrate that T cells from the spleens of nonimmunized mice which are reactive in one-way allogeneic mixed lymphocyte cultures (MLC) belong to a T cell subset distinct from that population of T cells, present in spleens of immune mice, which is capable of lysing allogeneic cells. Thus, the administration of limiting amounts of anti-thymocyte serum (ATS) or sublethal irradiation to normal C57BL/6 mice removes the recirculating, radiation sensitive population of cells. The residual spleen T cell population does not manifest mixed lymphocyte reactivity to (C57BL/6 × BALB/c)F1 spleen cells. On the other hand, the cytolytic activity of spleens from alloimmunized C57BL/6 mice is unaffected by these treatments.

Irradiation or ATS treatment before immunization, however, completely ablates the development of cytolytically active splenic lymphocytes. Thus, although the MLC reactive cells and the cytolytically active cells belong to different splenic T cell populations, the MLC cell appears to be required to generate cytolytic activity. Possible mechanisms to explain this requirement are presented.
Production of Migration Inhibitory Factor by Non-Dividing Lymphocytes
Authors: Rocklin R. E.
Published: December 2022

Lymphocyte proliferation in response to antigen was essentially eliminated by treatment with 5-bromo-2-deoxyuridine and light. The remaining lymphocytes were restimulated with the same antigen and the production of migration inhibitory factor (MIF) was determined. Under these conditions, MIF activity was still present. In addition, the absence of dividing cells did not appear to diminish significantly the amount of MIF produced.
Studies on the Mechanism of Lymphocyte-Mediated Cytolysis
Authors: Plaut Marshall , Lichtenstein Lawrence M. , Henney Christopher S.
Published: December 2022

The effect of cytochalasin B and colchicine on thymus-derived lymphocyte-mediated cytolysis in vitro has been studied.

Cytochalasin B reversibly inhibited (range 0.1 µg/ml to 10 µg/ml) the cytolysis of DBA/2 mastocytoma cells by splenic lymphocytes from allo-immunized C57BL mice. The drug itself was not cytotoxic. No evidence was found that “activation” of lymphocytes occurred in the presence of target cells and cytochalasin B. Inhibition occurred at a very early stage in the cytolytic pathway, possibly by preventing intimate contact between target cell antigens and lymphocyte receptor sites.

Colchicine inhibited cytolysis irreversibly (range 10-7 M to 10-3 M). This inhibitor was found to act exclusively on the effector lymphocyte population, preventing, by a non-cytotoxic mechanism, the expression of cytolytic activity. Colchicine appeared to act later in the cytolytic cycle than cytochalasin B. Supportive evidence of this viewpoint was the finding that reaction mixtures inhibited with cytochalasin B were further suppressed when this drug was removed and replaced with colchicine.

Cytochalasin B and colchicine were neither additive nor synergistic in their combined inhibitory activity. The inhibition observed with mixtures of these drugs was always equivalent to that found with the more potent inhibitor alone.

Among other activities, cytochalasin B has been demonstrated to affect microfilaments. The reversibility with which cytochalasin B inhibits cytolysis and the dose range over which it is active are consistent with the view that its inhibitory activity in this system is microfilament associated. Colchicine shows a great specificity for binding to the dimeric sub-unit of microtubules, and by so doing prevents microtubular aggregation. The present study suggests that this binding is the basis for inhibition of cytolysis. Consequently, the present findings support a role for both microfilaments and microtubules in lymphocyte-mediated cytolysis; the former possibly being necessary for effective lymphocyte-target cell interactions, the latter apparently being involved in a terminal cellular secretory event associated with the lysis of the target cell.
Immunologic Events during Listeria Monocytogenes Infection in Mice: Adjuvanticity and Immunogenicity of Macrophage-Bound Antigens
Authors: Lane Fredrick C. , Unanue Emil R.
Published: December 2022

There was a marked enhancement of the immune response to keyhole limpet hemocyanin administered at the time of inoculation with Listeria monocytogenes. Such enhancement was best seen if the antigen was given close to the time of inoculation with Listeria monocytogenes. Such enhancement was best seen if the antigen was given close to the time of inoculation with the bacteria. Bacterial filtrates also produced an adjuvant effect.

Peritoneal exudate cells harvested a few days after Listeria infection contained large activated macrophages. Such macrophages had a greater uptake of 125I-labeled hemocyanin. Metabolic studies indicated that the percentage of hemocyanin retained on the membrane or released from macrophages was increased in activated cells. Hemocyanin bound to activated macrophages was slightly more immunogenic. Activated macrophages (without antigen) did not transfer adjuvant effects. We conclude that the adjuvant effect was not associated with the presence of activated macrophages.
Isolation and Characterization of the Glycopeptides from a Human Immunoglobulin M
Authors: Hurst Mary M. , Niedermeier William , Zikan Jiri , Bennett J. Claude
Published: December 2022

Five glycopeptides, designated CHO1 through CHO5, have been isolated from the heavy chain of a Waldenström IgM. Their approximate location in the molecule, their amino acid sequence and carbohydrate composition have been determined.

Cleavage of the heavy chain with cyanogen bromide (CNBr) produced eight fragments, four of which were found to contain carbohydrate. CHO1 was found in CNBr-3, located toward the COOH-terminus of Fab (trypsin 60°). CHO2 was isolated from CNBr-4 located at the NH2-terminal end of Fc (trypsin 60°). CHO3 and CHO4 were found in CNBr-5 which is located in the Fc (trypsin 60°) portion of the molecule. CHO5 was isolated from CNBr-7 which is near the COOH-terminus of the heavy chain.

The carbohydrate composition of CHO1 was one fucose, three mannose, one galactose and two glucosamine residues. CHO2 consisted of one fucose, six to seven mannose, two galactose and three glucosamine residues. CHO3 contained two mannose, one galactose and two glucosamine residues. CHO4 and CHO5 were of simpler composition than the other three oligosaccharides, each containing three to four residues of mannose and one to two residues of glucosamine.
The Effect of in Vivo Irradiation on Macrophage Function
Authors: Schmidtke Jon R. , Dixon Frank J.
Published: December 2022

In vivo irradiation (660 R) of donor mice before removal of their peritoneal macrophages caused activation of these cells as indicated by increases in lysosomal enzymes but did not significantly affect the manner in which these cells handled keyhole limpet hemocyanin (KLH) in comparison to how macrophages from unirradiated donors handled KLH. The parameters measured included uptake, catabolism, retention and amount of membrane-bound KLH. In syngeneic recipients, however, the immunogenicity of equal amounts of KLH associated with activated macrophages from irradiated donors was significantly increased compared to that of KLH associated with macrophages from unirradiated donors. Also, addition of activated non-KLH-containing macrophages to the inoculum of normal KLH-containing macrophages increased the immune response. Infection of mice with lactic dehydrogenase virus also enhanced the immunogenicity of KLH associated with macrophages from these mice compared to control, uninfected mice.
Lymphocyte Surface Effects of Soluble Extracts of Streptococcal Membranes (TGCM)
Authors: Rapaport F. T. , Bachvaroff R. , Markowitz A. S.
Published: December 2022

A water-soluble, dialyzable glycoprotein extract of group A type 12 nephritogenic streptococcal membranes, which has previously shown transplantation antigen-like activity in mice and induced nephritis in rhesus monkeys and in dogs, has the capacity to protect human lymphocytes from the cytotoxic effects of isospecific HL-A typing antisera. This effect is not a consequence of a serologic reaction between HL-A antibodies and streptococcal extract (TGCM). Rather, it is due to the firm binding of TGCM to the lymphocyte surface, in a manner which resists dissociation by vigorous repeated washings. The protective effects of TGCM against HL-A antisera appear thus far to be limited to lymphocytes, as TGCM failed to protect human fibroblasts obtained from donors of known HL-A genotypes from the cytotoxic effects of such antisera. Immunofluorescence studies indicate that TGCM binding sites occur in diffuse fashion on the lymphocyte surface. In addition, the results of absorption-inhibition tests utilizing lymphocytes from donors of known HL-A phenotypes, speciesspecific anti-human heteroantisera and isospecific HL-A antisera point to a close spatial relationship between HL-A, species-specific, and TGCM binding sites on the surface of the human lymphocyte.
Reaginic Antibody Formation in the Mouse
Authors: Ishizaka Kimishige , Okudaira Hirokazu
Published: December 2022

DBA/1 mice were immunized with various doses of dinitrophenyl-ovalbumin (DNP-OA) precipitated with aluminum hydroxide gel, and both the reaginic (IgE) and IgG antibody responses were observed. Mice immunized with a minimal dose of DNP-OA, i.e., 0.02 to 0.05 µg, showed a persistent formation of anti-DNP IgE antibody, while an increase of immunizing dose resulted in only a transitory response. Formation of anti-OA IgE antibodies was persistent when animals were immunized with 0.02 to 1 µg of DNP-OA. Anti-DNP IgG antibody appeared in the serum within 10 days after immunization and reached a maximum by 2 weeks, whereas the anti-OA antibody titer increased slowly. The concentration of anti-DNP IgG antibody was much higher than anti-OA antibody during the first 3 weeks. The results showed that an optimal dose of immunizing antigen for maximal response differed with the immunoglobulin class of antibodies and antigenic determinant, even though the determinants are present on the same antigen molecule.

Priming of mice with suboptimal doses, i.e., 0.02 µg to 0.05 µg, of OA for antibody formation 2 weeks before immunization with DNP-OA either enhanced or “accelerated” both IgE and IgG anti-hapten and anti-carrier antibody responses. The results indicated participation of carrier-specific helper cells in the formation of anti-hapten IgE antibody. When animals were primed with 1 to 10 µg of OA, which was sufficient for induction of both IgG and IgE antibody response, and then immunized with DNP-OA, formation of anti-hapten antibodies was suppressed. The immunization of the primed animals with the hapten-carrier conjugate resulted in secondary response of both IgE and IgG anti-carrier antibodies.
Production of Precipitating Primate Antibodies to Human Membrane Antigen(s) with HL-A Activity by Immunization with Soluble HL-A Antigens
Authors: Metzgar Richard S. , Miller June L.
Published: December 2022

A monkey immunized with papain-solubilized HL-A antigens which had been purified on a Sephadex G-150 column produced antibodies which gave a preciptation reaction with the immunizing material as well as with certain other papain soluble spleen antigen preparations. The precipitating antibodies could not be removed by absorption with human serum but could be removed by absorption with peripheral blood leukocytes from all human donors tested. Purified suspensions of blood lymphocytes, lymph node cells or thymocytes were unable to absorb the precipitating antibody. Supernatants of saline suspensions of buffy coat leukocytes “released” antigen which precipitated with the monkey antiserum and gave reactions of identity by agar precipitation with the immunizing papain-solubilized antigen. Peripheral blood leukocytes from a patient with a leukemoid reaction containing 98% neutrophils were able to release precipitating antigen whereas purified blood and lymph node lymphocytes and thymocytes were not. The papain-solubilized antigen preparations as well as the released antigen preparations contained specific HL-A alloantigenic activity as well as precipitating antigen. Precipitation of the papain soluble antigen preparations as well as released antigen by the monkey antiserum resulted in a loss of the HL-A activity from the supernatant which could be recovered in the immune precipitate. These findings suggest that the monkey precipitating antibodies may be reacting with the HL-A base molecule(s) since they do not appear to interfere or react with site(s) conferring HL-A alloantigenic specificity.
Immunologic Reactions Against Insulin
Authors: Patterson Roy , O'Rourke John , Roberts Mary , Suszko Irena
Published: December 2022

Antibodies against insulin in three cases of insulin resistant diabetes mellitus were studied. These were IgG antibodies and their insulin-binding capacity was determined by a double antibody precipitation technique. These anti-insulin antibodies were associated in one case with lymphadenopathy, responsive to corticosteroids; anti-insulin activity was detected in an in vitro lymph node culture. In the other two cases, insulin antibodies were associated with an acute viral disease and intermittent insulin resistance of long duration. Although the insulin-binding activity as determined by the double antibody system was reproducible with individual serum samples, serum insulin-binding activity varied markedly on different days. This was presumptively because of variable degrees of free antibody associated with insulin administration; variable amounts of insulin-anti-insulin complexes were present in these sera. Cross-reactivities of insulins and certain insulin fragments from different species were compared by an inhibition technique and they demonstrated the importance of the secondary and tertiary structures in the antigenic determinants of insulin.
Immunologic Reactions Against Insulin
Authors: Patterson Roy , Mellies C. J. , Roberts Mary
Published: December 2022

Immediate or anaphylactic type hypersensitivity to insulin may be detected by in vitro analysis of IgE antibodies against insulin antigenic determinants. A solid phase immunoadsorbent prepared with polystyrene tubes and an IgE anti-IgE matrix provides the method of fixation of serum IgE against radiolabeled insulin. The degree of insulin binding by IgE correlates with the clinical allergic state and declines in parallel with the Prausnitz-Küstner titers of patient's sera. A more rapid decline in IgE-binding capacity of insulin occurred after therapeutic insulin desensitization or continuous insulin therapy. The occurrence of insulin allergy and immunologic insulin resistance in the same patients was associated with the presence of both IgE and IgG antibodies against insulin. The immunoadsorbent system for detection of IgE antibodies against insulin was used to evaluate cross-reactivity of heterologous insulins or insulin fragments with bovine insulin and is applicable to other IgE-mediated systems.
Deposition of Renal Tubular Epithelial Antigen Along the Glomerular Capillary Walls of Patients with Membranous Glomerulonephritis
Authors: Naruse T. , Kitamura K. , Miyakawa Y. , Shibata S.
Published: December 2022

Membranous glomerulonephritis (Mb-GN)1 is a primary renal disease in which immunologically characteristic alteration of the glomerular capillary walls, namely the uniform granular deposition of γ- and β1c-globulins along the glomerular capillary walls, is found.

This characteristic immunopathologic finding also has been induced experimentally in animals by the injection of various heterologous antigens and, recently, by the injection of homologous renal tubular epithelial antigen (Tub-Ag) (1). The pathogenesis of these experimental glomerulonephrides, especially the participation of antigen-antibody complexes, has already been well clarified (2, 3).

In human Mb-GN, most investigators are inclined to accept the same immunologic mechanism for the development of the disease. But the antigens which compose the glomerular immune deposits in human idiopathic Mb-GN have not been identified. We have demonstrated with immunofluorescent technique the deposition of Tub-Ag along the glomerular capillary walls of three patients with Mb-GN.

Human Tub-Ag was prepared from autopsied kidneys by the method of Edgington, Glassock, and Dixon (1).
Fluorescent Antibodies to Influenza Virus in Various Immunoglobulin Fractions of Serum after Natural Infection or Vaccination
Authors: Brown Gordon C. , O'Leary Theresa P.
Published: December 2022

Sera from confirmed cases of influenza and from recipients of attenuated or killed influenza virus vaccines were tested for IgM, IgA, and IgG antibodies by the indirect fluorescent antibody technique. Treatment with 2-mercaptoethanol and by sucrose density gradient centrifugation proved that IgM antibodies, as well as IgA and IgG, were located exclusively in the 7S fraction of globulin.

The 7S IgM antibodies rose rapidly after onset of disease in all cases and persisted in decreasing titers for 3 months. The appearance of IgA antibodies was detected in 75% of the cases. These attained lower titers and were of shorter duration. IgG fluorescent antibody (FA) titers increased in all subjects and the levels observed were closely analogous to hemagglutination inhibition titers. The three classes of FA were proven to be independently located in their respective subunits of immunoglobulin and to be specific for either type A or B influenza virus.

IgM antibody increases were measurable in 52% of the vaccinees receiving attenuated viruses administered intranasally but only 12% responded with IgA antibodies. Both classes of antibody were of lower titer and shorter duration than were those after clinical disease. Their development, titer, and decline were independent of the presence or absence of IgG antibodies. In fact, many subjects had IgG antibodies at the time of vaccination.

IgM and IgA antibodies in low titers were detected in 57% of the post-inoculation specimens from persons receiving inactivated vaccine. All had responded with increased titers of antibody in the IgG fraction.

Finally, it was shown that it is possible to detect recent influenza infection or vaccination by the demonstration of IgM or IgA antibodies in serum.