bluuebunny/crossref_metadata_2025 huggingface dataset:
Isogeneic Lymphocyte Interaction: Responsiveness of Murine Thymocytes to Self Antigens
Authors: Howe Michael L.
Published: December 2022

Previous studies in this laboratory have shown that thymocytes from neonatal CBA/J mice will undergo in vitro blastogenesis in culture with isogeneic spleen cells of adult origin. The present investigation demonstrates that thymocyte reactivity in this isogeneic lymphocyte interaction (ILI) is maximal at 2 to 3 days of age and declines thereafter, whereas the stimulatory capacity of spleen cells is minimal in the neonate but increases with increasing age of the cell donor. Data are also presented which suggest that soluble factors are not responsible for stimulation in the ILI. The results of breeding studies indicate that the capacity of spleen cells to stimulate in this reaction is probably controlled by more than one gene. An evaluation of the capacity of various adult lymphoid tissues to stimulate in the ILI indicates that cells from the spleen and to a lesser extent from lymph nodes are capable of stimulation while those from the thymus and bone marrow are devoid of stimulatory activity. Finally, an examination of several strains of mice indicates that most exhibit weak but positive responses in the ILI.
Delayed Hypersensitivity, as Manifested by Granuloma Formation, Dermal Reactivity, Macrophage Migration Inhibition and Lymphocyte Transformation, Induced and Elicited in Guinea Pigs with Soluble Antigens of Schistosoma Mansoni Eggs
Authors: Boros Dov L. , Schwartz H. J. , Powell A. E. , Warren K. S.
Published: December 2022

Delayed hypersensitivity (DH) was induced in guinea pigs 3 to 6 days after immunization with microgram quantities of soluble antigens derived from Schistosoma mansoni eggs (SEA). Sensitization was achieved by incorporating SEA into complete Freund's adjuvant, but remarkably, animals could also be sensitized by intraperitoneally injected live eggs, or by direct injection of SEA alone into the footpad. DH was measured by comparing dermal and granulomatous reactivity in vivo and 3H-thymidine incorporation and macrophage migration inhibition (MI) in vitro. SEA could be employed successfully also for the elicitation of the different immune responses. The parameters appeared in parallel, MI preceding the rest by 1 day. Granuloma formation, which occurred within 24 hr in the lungs of sensitized animals, correlated well with other parameters of DH. During the 1st 10 days of immunization, when DH was already fully developed, γ1 or γ2 antibodies could not be detected in the circulation of immunized animals.
The Effect of Irradiation on the Immune Response to the Private H-2 Specificities of the H-2K and H-2D Loci
Authors: McKenzie Ian F. C.
Published: December 2022

The H-2 locus in the mouse gives rise to the most immunogenic antigens in an allogeneic transplantation system (1). The locus can be subdivided into two major regions, H-2K and H-2D, each giving rise to private and public specificities, although other regions may exist (2, 3). Private specificities, with few exceptions, are found only in one H-2 allele, or recombinants derived thereof, there being one private specificity for the H-2K locus and one for the H-2D locus (2) for each allele. The H-2 private specificities are considered to be analogous to the HL-A specificities in man (3). The public specificities, which tend to be weaker, have a broad distribution and are found in many alleles. In a recent study (4) it was apparent that the immunogenicity of single, private antigenic specificities of the H-2K locus was greater than those of the H-2D locus. This was demonstrated by both skin grafting and testing for antibody formation.
The First Component (Cln) of the Complement System of the Nurse Shark (Ginglymostoma
Cirratum)
Authors: Ross Gordon D. , Jensen Joerg A.
Published: December 2022

The sedimentation coefficient of Cln varied inversely with the ionic strength in sucrose gradient ultracentrifugation. Natural antibody contamination was removed from impure Cln preparations by zonal ultracentrifugation in a high ionic strength (µ = 0.70) sucrose gradient, in which the Cln was 13.9S. The purified Cln gave a single precipitin line in Ouchterlony and immunoelectrophoresis with anti-whole nurse shark serum. Rabbit antibody against this preparation agglutinated cell-bound Cln, gave a single precipitin line of identity with purified Cln and whole nurse shark serum, and single precipitin arcs immunoelectrophoretically. At 0°C, 0.04 M EDTA inactivated Cln slowly and did not prevent Cln-site formation. EDTA inactivation was irreversible. In the presence of EDTA, immunoelectrophoresis of purified Cln gave three precipitin arcs. Esterase activity, present in the partially purified (urea-precipitated) Cln preparations, was absent in purified Cln with N-acetyl-L-tyrosine ethyl ester, p-tosyl-L-arginine methyl ester, or N-carbobenzoxy-l-tyrosine p-nitrophenyl ester as substrates.
A Comparison of Three Radioimmunoassay Techniques for the Measurement of Serum IgE
Authors: Polmar Stephen H. , Waldmann Thomas A. , Terry William D.
Published: December 2022

Three radioimmunoassay techniques [double antibody, Sepharose-bound antibody and bromacetylcellulose (BAC)-bound antibody] were compared. These three methods were used to measure the immunoglobulin E levels in sera of patients with schistosomiasis with very high IgE levels, in sera from normal adults, and in sera that were presumed to have very low IgE levels, including sera from immunodeficient patients and from a variety of non-primate species. There was, in general, good agreement among the three methods when the IgE concentration was elevated, as in patients with schistosomiasis and in some normal adults. In contrast, very significant differences in the estimates of the IgE concentration of the same serum were observed with the different techniques when the serum IgE levels were low. IgE was very low or undetectable in sera of many patients with immunodeficiency when studied by the double antibody technique, but these IgE deficiencies were often missed when the Sepharose or BAC techniques were used. For example, the IgE levels were found to be less than 15% of normal for 80% of the patients with ataxia telangiectasia studied by the double antibody technique, but IgE levels appeared to be normal when assayed by the Sepharose and BAC techniques. The IgE levels determined by the Sepharose method in patients with immunodeficiency were intermediate between the lower double antibody and the higher BAC values. Similarly, sera of various non-primate species were found to inhibit significantly the binding of radiolabeled IgE to the insoluble anti-IgE antibody and gave significant appearent IgE concentrations when the BAC or Sepharose techniques were used, but not when the double antibody technique was used. Thus, the BAC and, to a lesser extent, the Sepharose technique appeared to give falsely high IgE estimates when the levels of human IgE in the serum were low. The double antibody radioimmunoassay appears to be the method of choice for the measurement of serum IgE because it has greater sensitivity and reproducibility, and especially because it gives meaningful estimates for IgE in the critically important low IgE range observed with immunologic deficiency.
Effect of Chronic Viral Infection on the Immune System
Authors: Oldstone Michael B. A. , Tishon Antoinette , Chiller Jacques M. , Weigle William O. , Dixon Frank J.
Published: December 2022

Immunoresponsiveness has been compared in mice chronically infected with lymphocytic choriomeningitis (LCM) virus and in noninfected mice. The antigens used were sheep red blood cells, keyhole limpet hemocyanin, human IgG (HuIgG), bovine IgG, chicken IgG, and turkey IgG. LCM-infected mice were found to have normal immune responses to sheep red blood cells and keyhole limpet hemocyanin over wide dose ranges, whereas against mammalian and avian IgG, LCM-infected mice showed depressed immune responsiveness. This depression in immune responses was not associated with differences in antibody avidity or antigen handling but with decreased numbers of antibody-forming cells. Unresponsiveness to HuIgG was difficult to induce with deaggregated antigen in mice chronically infected with LCM virus.
Studies on Tetanus Antitoxins
Authors: Nagel J. , Cohen H.
Published: December 2022

In this paper the antigenic structure of tetanus toxin is described. It was shown that the toxin as well as the toxoid is carrying at least four antigenic determinants. In addition to antigen-[2] already described in a previous paper, two new degraded antigens could be isolated: antigen-[1] by preparative polyacrylamide gel electrophoresis of spontaneously degraded toxin, and antigen-[3] by pH treatment of toxin. All degraded antigens are immunogenic and nontoxic. The presence of a fourth antigenic determinant on tetanus toxin could be demonstrated only after successive absorption of rabbit antitoxic serum with the antigens-[1], -[2] and -[3]. Toxin-neutralizing antibodies remained in the absorbed serum which reacted only with tetanus toxin and tetanus toxoid. Antibodies against the four antigenic determinants are present in antitoxic serum from rabbits and horses. Human antiserum, however, contained antibodies to these determinants, anti-1 antibodies being absent. All four antitoxins could neutralize tetanus toxin in vivo.
The Genetics of the Immune Response to a Synthetic Double-Stranded RNA in a Mutant CBA Mouse Strain
Authors: Scher Irwin , Frantz Molly M. , Steinberg Alfred D.
Published: December 2022

A mutant strain of mice (CBA/HN) was found that did not form antibody to the synthetic double-stranded ribonucleic acid polyinosinic-polycytidylic acid when given with complete Freund's adjuvant. All other strains investigated responded to this antigen. Genetic studies revealed that the inability to respond to this antigen was secondary to a gene on the X chromosome of the CBA/HN strain. Antibody formation could be induced in this strain by giving the antigen with methylated bovine γ-globulin. Immune responsiveness was transferred to nonresponding animals via lymphoid cells. The possible mechanisms of this defect are discussed.
Age-Dependent Changes in Sensitivity to Antigen in the Mouse
Authors: Rector E. S. , Carter B. G.
Published: December 2022

The maturation of antibody responsiveness was studied in C57BL/6 mice by comparing the sensitivity to antigen of 12-day-old and 12-week-old animals. Sensitivity was established by constructing dose response curves for the IgM and IgG2a PFC responses. Twelve-day-old animals yielded dose response curves with well defined maxima and relatively narrow band widths (BW). The optimal antigen doses for the two antibody classes studied were different—1.2 × 109 sheep red cells for the IgM response and 3.8 × 109 for the IgG2a response. Adult (12-week-old) animals yielded dose response curves significantly different from those of young mice. Optimal antigen doses were lower in the adult group (2.9 × 108 for the IgM and 1.0 × 109 for the IgG2a PFC response), indicating an increased sensitivity to this antigen. The BW (ratio of the two antigen doses which elicited a response equal to 50% of the maximum) of the adult dose response curves were four times greater than the BW obtained with the 12-day-old group. In addition, a low cross-reactivity was demonstrated between sheep red cells and the lipopolysaccharide antigen prepared from Escherichia coli. The increases which occurred with age in both the sensitivity and BW of the responses are discussed in terms of: 1) a thymus-derived cell deficiency in the young mice; 2) the selection of receptor sites by cross-reacting environmental antigens; and 3) changes in the antigen-processing system. It was concluded that the last of these possibilities formed the basis for the changes in sensitivity which were observed.
On the Nature of the Presumed Receptor for IgE on Mast Cells
Authors: Bach Michael K. , Brashler John R.
Published: December 2022

The ability of sialidase and phospholipase C to interfere with the capacity of rat peritoneal cells to release histamine in an antigen-induced, IgE antibody-mediated manner was examined in some detail. Both enzymes cause a loss of histamine from the cells which is dependent on the concentration of enzyme employed. Although the loss of histamine from cells treated with phospholipase C ceases when the enzyme is removed by washing, loss from cells treated with sialidase persists for several hours after the free enzyme has been removed. Superimposed on this loss of histamine from enzyme-treated cells is the ability of both enzymes, under certain circumstances, to inhibit the capacity of the cells to release histamine in an antigen-specific manner after their preparation with IgE antibody. Attempts were made to test the hypothesis that this inhibition is a consequence of the destruction of a mast cell “receptor” for IgE by treatment with the enzymes. Phospholipase C does not appear to act on such a receptor since it was equally inhibitory to nonimmunologic histamine release caused by compound 48/80. Furthermore, treatment with this enzyme inhibited immunologic histamine release equally well when enzyme treatment preceded or followed preparation of the cells with IgE. By contrast, treatment with sialidase only inhibited histamine release when treatment with the enzyme preceded preparation of the cells with IgE, suggesting that it may act on the “receptor.” However, the enzyme failed to inhibit histamine release initiated with IgGa antibody and antigen, a reaction which had previously been presumed to involve the same cell “receptor” as the one involved in the IgE-mediated reaction. Possible explanations for these findings are offered.
Lymphocyte Populations: Separation by Discontinuous Density Gradient Centrifugation
Authors: Yu David Tak Yan , Peter James B. , Paulus Harold E. , Machleder Herbert I.
Published: December 2022

A method for separation of human lymphocytes by discontinuous density gradient is described. The method was simple to carry out and utilized Ficoll-salt solutions which were easily standardized. There was good reproducibility with no toxicity to the cells. On applying this method to human thoracic duct lymphocytes, cells were separated into subpopulations which differed in morphology, basal unstimulated 3H-TdR incorporation and response to stimulation by purified phytohemagglutinin P and pokeweek mitogen.
Effect of the Carrier Protein on Idiotypic Specificities of Antihapten Antibodies Produced in an Inbred Strain of Mice
Authors: Wang Ai-Lan , Nisonoff Alfred
Published: December 2022

Antihapten antibodies formed in A/J mice in response to keyhole limpet hemocyanin (KLH)-p-azophenylarsonate (KLH-Ar) show strong intrastrain idiotypic cross-reactions. These antibodies also share idiotypic specificities with antihapten antibodies produced in response to the same hapten coupled to a different carrier, bovine γ-globulin (BGG). Nearly all specificities present in the antihapten antibody population elicited by KLH-Ar were also found in the anti-BGG-Ar antibodies of most mice, as shown by the capacity of the latter antibody populations to displace most of the labeled immunogen from its antiidiotypic antibodies. On the average, however, significantly larger amounts of anti-BGG-Ar, as compared to anti-KLH-Ar antibodies, were required for inhibition. This demonstrates that populations lacking the cross-reacting idiotype were also stimulated by BGG-Ar. The results indicate that many receptors on lymphocytes that are capable of being stimulated by KLH-Ar can also interact effectively with BGG-Ar. The fact that idiotypic specificity is relatively independent of the carrier protein suggests that it is determined at the level of the bone marrow-derived cell.
Immunologic Studies on Drug Addiction
Authors: Liu Chi-Tan , Adler Frank L.
Published: December 2022

Antisera which react with methadone and with urine from users of this drug have been prepared in rabbits. A simple and rapid hemagglutination-inhibition test which detects methadone in concentrations of 1 ng/ml has been applied to human urine specimens with results that demonstrate the specificity of the test and suggest that use of the drug may remain detectable for at least 1 week. Heroin used simultaneously with methadone does not interfere in the test.
Mechanisms of Passive Sensitization
Authors: Ishizaka Teruko , Soto Carmen S. , Ishizaka Kimishige
Published: December 2022

The cell-bound IgE molecules on basophil granulocytes were enumerated by C1 fixation transfer technique. The average number of IgE molecules per basophil granulocyte was 10,000 to 40,000. The number did not have direct correlation with serum IgE level; however, a rough inverse relationship was observed between the number of cell-bound IgE molecules and the optimal concentration of anti-IgE for maximum histamine release from the cells. The results suggested that the number and affinity of cell-bound IgE antibody molecules probably determine the sensitivity of basophils to allergen.

When leukocytes were incubated with 100 µg to 1 mg/ml of E myeloma protein, the number of basophil-bound IgE molecules significantly increased. The number of receptor sites for IgE molecules per basophils was estimated to be between 30,000 and 100,000. IgE molecules have high affinity for the receptor but the binding of the molecules with the receptor is reversible. The association constant of the binding reaction was estimated to be on the order of 108 to 109. Basophils from all normal individuals studied combined IgE molecules upon incubation with E myeloma protein. After sensitization with a reaginic serum, however, the leukocytes from some of these donors failed to release histamine upon challenge with antigen under the usual experimental condition. These cells previously termed “non-acceptor” cells, released histamine if they were challenged in the presence of D2O, which enhanced biochemical processes after the antigen-antibody reaction. The results indicated that even such “non-acceptor” cells combined IgE antibody upon passive sensitization with reaginic serum and, therefore, that failure of antigen-induced histamine release from these cells was not due to lack of IgE antibody on their surface.
Immune Responses in Vitro
Authors: McIntyre John A. , Pierce Carl W.
Published: December 2022

We have demonstrated that hemolytic clusters which develop in cultures of mouse spleen cells stimulated with heterologous erythrocytes contain plaque-forming cells, but are not clonally restricted to production of antibody of one immunoglobulin class or against one antigenic specificity.
Interactions between Human C5, C6, and C7 and Their Functional Significance in Complement-Dependent Cytolysis
Authors: Arroyave Carlos M. , Müller-Eberhard Hans J.
Published: December 2022

Two types of complexes were observed between C5, C6 and C7. In their native form these three complement proteins enter into reversible interactions which were demonstrated by zone ultracentrifugation. Interactions between C5 and C6 occurred in absence of C7 and between C5 and C7 in absence of C6. Presence of C6, however, enhanced the association of C5 and C7.

Formation of a stable C5–C6 complex was effected by the complement enzyme . This complex () had a sedimentation coefficient of 12 to 14S and exhibited the ability to attach itself firmly to non-sensitized sheep erythrocytes (E) in presence of C7.

activity was also generated by trypsin or plasmin, suggesting the existence of a new pathway of complement activation by non-complement enzymes beginning with C5. produced with trypsin or displayed typical ultrastructural membrane lesions and was susceptible to lysis by C8 and C9.

Regardless of experimental conditions, C5, C6, and C7 were taken up by the surface of cells in equimolar quantities. The affinity between native C5, C6, and C7 displayed in free solution, and the equimolarity of the cell-bound equivalents, support the concept of a firm trimolecular assembly as the topologic base of the membrane attack mechanism of complement.
A New Serum Factor in Normal Rabbits
Authors: Hagen K. , Kulkarni S. S. , Mandy W. J.
Published: December 2022

The serum of b4b4 rabbits gave higher HR titers when tested with b4b4 or b9b9 fragments than when tested with fragments bearing the allelic b5 or b6 markers. Reciprocal results were obtained with serum from b5b5 or b6b6 rabbits. Thus, the heterogeneity noted in the HR titers for rabbits is dependent upon the group b L chain allotypic specificities.

The relationship between the group b allotypes and HR activity suggested that the specific Fab HR site required the proper alignment of the Fd piece with the L chain. This was confirmed by reduction-alkylation experiments which established the necessity for the L-H chain disulfide bond.
Interferon Induction in New Zealand Black Mice by Murine Leukemia Virus
Authors: Siegel Benjamin V. , Brown McKay , Morton Jane I.
Published: December 2022

Rauscher leukemia virus (RLV)2 has been reported to induce little or no interferon in a number of murine systems, such as BALB/c (1) and NMRL (2) strains. In the case of the random-bred CD-1 strain, Glasgow and Friedman (1) observed relatively high serum levels of interferon in adult RLV-infected mice with accompanying resistance to the disease. We have extended these studies to an investigation of the interferon response to RLV in the autoimmune New Zealand Black (NZB) model and for comparative purposes have included the C57BL/6 and BALB/c strains.

The female NZB mice employed represented the seventh and eighth generations in this laboratory from breeding stock (generations 57 and 58) obtained from the Otago University Medical School, Dunedin, New Zealand, and have since been maintained in a closed colony. These NZB mice are vigorous, healthy, and relatively long-lived with 50% survival at 17 months; their microbiologic status has not been assessed.
Differentiation of Lymphoid Cells: The in Vitro Emergence of Immunoglobulin M-Secreting Cells
Authors: Zimmerman Daniel H. , Kern Milton
Published: December 2022

Freshly isolated rabbit popliteal lymph node cells synthesize and secrete principally immunoglobulin G (IgG) together with a relatively small amount of immunoglobulin M (IgM). After incubation of such cells in tissue culture for 48 hr, [3H]IgG production decreased markedly and approached zero, in most experiments. On the other hand, [3H]IgM production increased 5- to 10-fold. Thus, at 48 hr and thereafter τ;90% of the [3H]Ig was accounted for as [3H]IgM. In contrast, lymphoid cells derived from rabbit appendix do not show enhanced [3H]IgM production although such cells predominantly secrete [3H]IgM when freshly isolated and in decreasing amounts during tissue culture incubation. The enhanced [3H]IgM production observed with popliteal lymph node cells is dependent on the presence of fetal calf serum during tissue culture incubation. This requirement cannot be replaced by serum from the same rabbit that was the source of the cells and, indeed, such autologous serum in the presence of fetal calf serum inhibited enhanced [3H]IgM production. Serologic techniques, DEAE-cellulose chromatography, Bio-Gel chromatography, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis were used to distinguish IgM from the other classes of Ig. Fluorodeoxyuridine, an inhibitor of DNA synthesis, abolished the emergence of enhanced [3H]IgM production when added to cells during the latent period, but the inhibitor was without effect on Ig secretion when incubated with cells after enhanced [3H]IgM production had already emerged. The results indicate that the cellular source for the enhanced IgM secretion cannot be the original IgG-secreting cells but may be derived from some of the original IgM secreting cells or, alternatively, may be derived from cells that are quiescent in the freshly isolated population.
Antigen-Macrophage Interaction
Authors: Cohen Benjamin E. , Rosenthal Alan S. , Paul William E.
Published: December 2022

Antigen-stimulated tritiated thymidine (3H-TdR) incorporation by lymphocytes is an in vitro correlate of cellular immunity. The stimulation of this response by hapten-carrier conjugates may be inhibited by free hapten, a finding previously interpreted as evidence for independent hapten-specificity of thymus dependent (T) lymphocyte receptors. We confirm the specific inhibition by dinitrophenyl (DNP)-lysine of DNP-guinea pig albumin (GPA) induced stimulation of lymphoid cell cultures from guinea pigs immunized with DNP-GPA and report a similar specific inhibition caused by DNP-ovalbumin (OVA).

Nonetheless, our results indicate that this hapten-specific inhibition is not due to blockade of T cell receptors specific for hapten-carrier conjugates. Thus we show: 1) DNP-OVA must be present before and during “pulse” exposure to DNP-GPA but has no effect if added after such a pulse. 2) Preincubation of macrophage-rich peritoneal exudate cells with DNP-OVA blocks the capacity of these cells to bind low concentrations of DNP-GPA and then to stimulate immune peritoneal exudate lymphocytes to divide. 3) Preincubation of DNP-GPA immune peritoneal exudate lymphocytes in DNP-OVA has no effect on their capacity to subsequently respond to macrophage-associated DNP-GPA.

We conclude that the capacity of DNP-OVA to inhibit DNP-GPA stimulation of 3H-TdR incorporation by lymphoid cell populations from guinea pigs immune to DNP-GPA is due to competition for a hapten-specific cytophilic antibody on the surface of the macrophage and not to competition for a hapten-specific receptor on T lymphocytes.
Selective Cytotoxicity in a Hapten Substituted Cell Culture Model System
Authors: Philpott Gordon W. , Bower Richard J. , Parker Charles W.
Published: December 2022

Selective complement dependent cytotoxicity was demonstrated in a hapten-substituted cell culture model system. Viable hapten substituted HeLa, Hep-2, and L cells were obtained by treatment with 10 to 300 µg/ml of 2, 4, 6-trinitrophenyl (TNP)-sulfonic acid. Binding studies with 14C-TNP-sulfonic acid showed that from 1 to 9 × 109 TNP groups were bound per cell, representing 1 to 4% of the added TNP. High affinity anti-TNP antibody was purified from the sera of rabbits hyperimmunized with TNP-bovine γ-globulin. The amount of anti-TNP Ab binding to TNP-cells was quantitated and found to be dose dependent in this range of TNP substitution. Even at the highest levels of antibody, only about 1% of TNP groups was bound by antibody. The selectivity of antibody binding to TNP-cells was established by inhibition with the hapten, ε-DNP-lysine. Anti-TNP antisera, and to a lesser extent, specifically purified anti-TNP antibody, caused complement dependent cytotoxicity of TNP-cells but not unsubstituted cells. This model system permits comparative cytotoxic antibody studies with a variety of tumor cells.
β2-Microglobulin in Human Blood Cells
Authors: Evrin Per-Eric , Pertoft Håkan
Published: December 2022

β2-Microglobulin has been solubilized from human blood cells by sonication, deoxycholate treatment, or acidification. The average number of β2-microglobulin molecules per cell, measured after acidification, was 0.54 × 105 for platelets, 18 × 105 for mononuclear cells, and 6.9 × 105 for polymorphonuclear cells. No β2-microglobulin was detected in red blood cells. A relatively large proportion of the protein in platelets and mononuclear cells was located on the cell surface as compared with polymorphonuclear cells, according to experiments performed with 125I-labeled rabbit anti-β2-microglobulin IgG.
Detection of Antibodies and Soluble Antigen-Antibody Complexes by Precipitation with Polyethylene Glycol
Authors: Creighton W. D. , Lambert P. H. , Miescher P. A.
Published: December 2022

Polyethylene glycol (PEG) was used in a modification of the Farr antigen binding test in order to separate free antigen from antibody-bound antigen. Most of the immunoglobulins are precipitated at 20% PEG while smaller antigens like bovine serum albumin or insulin are quite soluble at this concentration of PEG. Such antigens are therefore precipitated in 20% PEG if they are bound to antibodies.

Furthermore, PEG has been used at lower concentrations in order to precipitate soluble antigen-antibody complexes in conditions at which free antigen or free antibody would be soluble. This second method could be applied to larger antigens such as IgG or thyroglobulin.
Cellular Studies of the Genetic Control of Immune Response toward the Loop Region of Lysozyme
Authors: Mozes Edna , Shearer G. M. , Maron Elchanan , Arnon Ruth , Sela Michael
Published: December 2022

Specific antibody response to the “loop” peptide of hen egg-white lysozyme has been shown to be controlled by a single dominant gene which is not linked to the major H-2 histocompatibility locus (1, 2). A study of the cellular aspects of the immune response gene regulating immune potential to loop was made with syngeneic and allogeneic thymus and marrow cell transfers in irradiated recipients immunized with loop-A-L, loop-Pro-L or lysozyme. The allogeneic grafts were controlled for optimal cell interaction between the complementary cells of different H-2 origin by observing the expected responses to Pro-L and to lysozyme. No evidence for suboptimal nor superoptimal responses was detected with cell mixtures from SJL/J and DBA/1 donors. Results obtained from the cell transfer experiments were compatible with a defect reflected exclusively in the marrow cell population of low responders SJL/J mice for loop-Pro-L, and expressed in both the thymus and the marrow populations of relevant lymphoid cells from this strain for loop-A-L.

These results are in agreement with the hypothesis (3) that the genetic defect is reflected in the thymus-derived cells, when the defect is at the carrier level, whereas it is expressed exclusively in the bone marrow population when the defect is at the determinant level.
Lymphocytes in Peyer's Patches of the Mouse: Analysis of the Constituent Cells in Terms of Their Capacities to Mediate Functions of Mature T and B Lymphocytes
Authors: Katz David H. , Perey Daniel Y. E.
Published: December 2022

The present studies were designed to assess the functional capacities of Peyer's patch lymphocytes (gut-associated lymphoid tissue [GALT]) in the mouse with a system for adoptive transfer of immunity to hapten-carrier conjugates. These functional studies of GALT have been performed in parallel with studies of spleen lymphocytes from the very same primed donor mice in order to establish a positive baseline of function for comparison of immunocompetence. The results of these experiments demonstrate the presence in GALT of lymphocytes capable of performing a carrier-specific helper cell function which is a well-documented property of mature thymus-derived (T) lymphocytes. In contrast, another function of mature T cells, namely the capacity to mediate a graft vs host (GVH) reaction as assessed by the allogeneic effect, is absent in such GALT lymphocytes. The capacity to function as hapten-specific presursors of antibody-secreting cells, a property of mature bone marrow-derived (B) lymphocytes, is also lacking in GALT lymphocytes. These findings indicate that marked functional differences exist between GALT and recognized peripheral lymphoid tissues and lend support to the notion that such lymphoid aggregates may perform a unique and/or perhaps, in part, a central lymphoid function.
Genetic Control of Thymus-Derived Cell Function
Authors: Willams R. Michael , Moore Michael J. , Benacerraf Baruj
Published: December 2022

Thymus and lymph node cells (LNC) from male Lewis and BN rats of exactly the same age were evaluated for 3HTdR incorporation in culture after stimulation with the mitogens concanavalin A (Con A), phytohemagglutinin (PHA), or Escherichia coli lipopolysaccharide (LPS) over a 1000-fold dose range, and after mixed lymphocyte culture (MLC) with (Lewis × BN)F1 LNC. BN thymus cells stimulated by mitogens or by histocompatibility antigens of hybrid LNC in MLC can incorporate as much 3HTdR as Lewis thymocytes under identical culture conditions. Lewis and BN thymus cells respond identically to PHA or in MLC, and neither type of thymus cell responds to LPS in culture. The only difference observed between lewis and BN thymus cells was that BN cells required 4-fold higher concentrations of Con A to produce a maximal DNA synthetic response. Lewis LNC were superior to BN cells in terms of Con A or PHA stimulated DNA synthetic responses irrespective of mitogen concentration. The strain specific Con A concentration difference noted for thymus cells was also present in cultures of LNC. Lewis LNC responded better than BN LNC under identical MLC conditions with LBNF1 cells. LPS stimulation of DNA synthesis was identical for Lewis and BN LNC. These observations indicate that Lewis and BN thymus dependent (T)-cells differ in at least two parameters: 1) a Con A concentration dependent reactivity irrespective of their location, and 2) a potential for in vitro DNA synthesis by mature, peripheralized T cells from the lymph nodes.
Evidence for Several Cell Populations Active in Antibody Dependent Cellular Cytotoxicity
Authors: Zighelboim Jacob , Bonavida Benjamin , Fahey John L.
Published: December 2022

A sensitive assay system using syngeneic attacker and target cells was developed to characterize the cellular and serum components active in antibody-dependent cellular cytotoxicity (ADCC). Lysis of 51Cr-labeled EL-4 target cells requires only 4-hr incubation in vitro in the presence of normal mouse cells and heat-inactivated anti-EL-4 serum. Cytotoxicity was obtained with high serum dilutions of either xenogeneic (rabbit) or allogeneic (BALB/c) antiserum. The IgG fraction obtained from the rabbit anti-EL-4 serum was also effective in this system. Adherent cells from spleen and peritoneal exudate as well as non-adherent cells from the spleen of normal C57BL/6 mice were effectively cytotoxic in conjunction with the appropriate antibody. These observations indicate that at least two populations of cells may act as effector cells in the ADCC phenomenon.
Quantitative Phagocytosis by Neutrophils
Authors: Ward P. A. , Zvaifler N. J.
Published: December 2022

A new method for the quantitative measurement of phagocytosus by neutrophils, providing reproducibility within 3%, is described. The phagocytizable material consists of radiolabeled immune complexes. Cell uptake is a function of the cell concentration, the amount of immune complexes, the duration of incubation, and the amount of antigen in excess of the point of equivalence. The presence of serum does not enhance phagocytosis.
Introduction of Cellular Immunity to a Chemically Altered Tumor Antigen
Authors: Chao Hsia-Fu , Peiper Stephen C. , Aach Richard D. , Parker Charles W.
Published: December 2022

Carcinoembryonic antigen (CEA) was acetoacetylated with acetoacetic anhydride (Diketene). Guinea pigs immunized with acetoacetylated CEA showed delayed skin hypersensitivity and lymphocyte transformation after stimulation with unmodified CEA, but demonstrated very low levels of circulating anti-CEA antibodies. The dissociation of cellular and humoral immunity could prove useful in the further study of their relative role in immunologic resistance to tumor growth. This method of selectively stimulating cellular immunity may also be of value to the immunotherapy of cancer.
Phagocytic Cells as Effectors in a Cell-Mediated Immunity System
Authors: Dennert Gunther , Lennox Edwin S.
Published: December 2022

Cell-mediated immunity, using a xenogeneic system, i.e., mouse spleen cells attacking chicken erythrocytes (CRBC) is studied.

In this system target cell lysis by spleen cells from immunized animals is complement independent, hence cell-mediated. It is specific because trinitrophenyl-(TNP) sensitized spleens lyse TNP-CRBC but not CRBC. CRBC-sensitized thymus-derived (T) lymphocytes cooperate with bone marrow-derived lymphocytes in induction of CRBC plaque-forming cells but cannot lyse CRBC as target. However, anti-CRBC antibody at concentrations too low to be detected by complement dependent lysis do induce cytotoxicity to CRBC in spleen cell suspensions containing T cells as well as in spleen cell suspensions depleted of T cells. Anti-ϑ serum absorbed to remove non-anti-ϑ antibody does not abrogate the cytotoxicity of immune spleen cells. These results indicate that the effector cell in this system is not a ϑ-antigen-bearing T lymphocyte. The effector cell is abundant in spleen and in the peritoneal cavity but not in lymph nodes of normal animals: it is bone marrow-derived, radiation resistant, and sticks to siliconized glass bead columns. Since, in addition, the effector cell can be separated from other spleen cells by treatment with carbonyl iron powder and a magnet, it is, we suggest, a phagocytic cell.
Immune Response of Pathogen-Free Mice Inoculated Intranasally with Mycoplasma Pulmonis
Authors: Cassell Gail H. , Lindsey J. Russell , Baker Henry J.
Published: December 2022

Immunoglobulin and antibody levels in serum and tracheobronchial secretions of pathogen-free mice experimentally infected with Mycoplasma pulmonis were determined using radial diffusion and immunofluorescence. Anti-M. pulmonis antibodies of the IgG1, IgG2, IgM, and IgA classes were detected in both serum and secretions. In serum IgM attained peak levels by 4 weeks, IgG1 and IgG2 by 5 weeks, and IgA by 6 weeks post-inoculation. In secretions IgG1 and IgG2 antibodies were detected first at 2 weeks, IgM at 4 weeks, and IgA at 5 weeks.

Immunofluorescent and histologic studies demonstrated that specific antibodies primarily were formed locally in lung and paratracheal lymph nodes. Varying numbers of cells infiltrating the peribronchial and perivascular spaces during infection were fluorescence-positive for IgM, IgG1, IgG2, and IgA. Cells producing IgM appeared early and were followed by cells producing IgG1 and IgG2. Cells containing IgA increased as early as 2 weeks, became the principal cell type by 3 weeks post-inoculation, and continued to predominate throughout the 8-week observation period. Results of these studies suggest that antibody produced locally may be a major component of acquired immunity to M. pulmonis.
Decreased Response of Cultured New Zealand Mouse Spleen Cells to Sheep Erythrocytes
Authors: McCombs Candace , Hom Joanne , Talal Norman , Mishell Robert I.
Published: December 2022

New Zealand (NZ) mice give excessive immune responses when immunized in vivo with sheep red blood cells (SRBC). Spleen cells of young New Zealand Black and New Zealand Black/White hybrid mice developed low primary responses compared to conventional strains when immunized in vitro. These low responses could be raised to levels achieved by other strains by adding peritoneal cells and 2-mercaptoethanol to the NZ spleen cultures. Studies of the peritoneal cells indicate that the restoring activity is a property of the adherent population and that it is not due to thymus derived lymphocytes.
Antigenicity of Reduced and Alkylated Australia Antigen
Authors: Imai M. , Gotoh A. , Nishioka K. , Kurashina S. , Miyakawa Y. , Mayumi M.
Published: December 2022

The association of Australia antigen (AuAg)2 with human type B hepatitis has been fully established. It has been reported that AuAg, which contains 6.5 M% of cysteine (1), completely lost the ability to react with humoral anti-Au antibodies after reduction and alkylation (2). Recently, Vyas et al. (3) have further reported that the reduced and alkylated AuAg could induce a delayed-type hypersensitivity response in guinea pigs but did not raise humoral antibody, suggesting the significance of the disulfide-bond dependent conformation in stimulating the humoral antibody response.

We have found, however, that a part of the antigenic reactivity of AuAg was relatively resistant to reduction and alkylation, although most of it was sensitive to this procedure. This unexpected finding prompted us to study in detail the hitherto recognized antigenic determinants of AuAg—a, d, y, w and r3—with respect to their susceptibility to the conformational changes induced by this chemical modification.
The Utilization of Properdin in the Alternate Pathway of Complement Activation: Isolation of Properdin Convertase
Authors: Stitzel Ann E. , Spitzer Roger E.
Published: December 2022

Normal human serum contains an α-globulin, designated properdin convertase, which after activation by zymosan will interact with native properdin to generate a C3 reactive enzyme. The activation of properdin convertase as well as its interaction with properdin after activation requires no additional factors. The C3 reactive material which is generated reacts directly with C3 and contains properdin in an altered state; the exact composition of this C3 reactive enzyme, however, is not completely clear.

Removal of properdin convertase from serum aborts the consumption of C3-C9 when zymosan is added. In addition, although C3 proactivator is not required for the action of properdin convertase on properdin or for the subsequent cleavage of C3, it, nevertheless, can be activated by products of the properdin convertase, properdin, C3 reaction. Hence, properdin convertase may be involved as an initiating step in the alternate pathway of complement activation.
Delineation of B and T Lymphoid Cells in the Chicken
Authors: Albini Boris , Wick Georg
Published: December 2022

The percentage of lymphoid cells from chicken bursa of Fabricius, thymus, spleen, blood, cecal tonsils and Harder's glands reacting with an anti-chicken bursa cell serum (ABS) and an anti-chicken thymus cell serum (ATS) raised in turkeys was evaluated in a chronologic study from the day of hatching until 24 weeks of age using indirect immunofluorescence and lymphocytotoxicity tests. The predominant number of bursa cells reacted with ABS, while most thymus cells reacted with ATS. In these central lymphoid organs, the relationship between the two cell populations remained fairly constant over the whole observation period. In the spleen and-even more pronounced-in the lymphoid cell populations of blood, cells reacting with ATS were predominant. In contrast, in Harder's glands a majority of cells reacting with ABS could be detected from hatching until the 23rd week of age. Cecal tonsils contained equal amounts of cells reacting with ABS and ATS at 5 weeks, but a preponderance of ABS reactive cells at 10 weeks of age. These findings implicate very distinct relationships of lymphoid cell populations in different lymphoid organs of the chicken, which may account for the different time course in the acquisition of immunological competence in these various organs and are suggestive of their possible different responsibilities within the immunologic defense system.
Production of Colony-Stimulating Factor in Mitogen-Stimulated Lymphocyte Cultures
Authors: Parker John W. , Metcalf Donald
Published: December 2022

The addition of phytohemagglutinin, pokeweed mitogen, concanavalin A or bacterial lipopolysaccharide to mouse spleen cultures strongly stimulated production of colony-stimulating factor (CSF), the humoral regulator of granulopoiesis and macrophage formation. Column-filtered spleen cells depleted of macrophages exhibited similar responses. Cultures of macrophage monolayers failed to produce elevated levels of CSF when stimulated by PHA or PWM. The results suggest that activated T- and B-lymphocytes can both produce significant amounts of CSF, providing a possible mechanism for stimulating macrophage formation in chronic immunologic or inflammatory responses.
Characterization and Properties of an Anaphylactic 7S Antibody in Sheep
Authors: Esteves Maria B. , Sant'Anna Osvaldo A. , dos Santos Annes Vera C. , Binaghi Rubén A.
Published: December 2022

Anti-DNP antibody was specifically purified from hyperimmune anti-DNP-BGG sheep antiserum. The antibody was fractionated on DEAE-cellulose. Two components belonging to different immunoglobulin classes were isolated and designated IgS (slow) and IgF (fast) on account of their relative electrophoretic mobility. The molecular size of both antibodies is the same and it is similar to that of human IgG. IgS induces homologous anaphylactic reactions (PCA) and does not fix complement in the classical way. IgF fixes complement and is not anaphylactic for the sheep. The biologic activities of IgS are not destroyed by treatment with 2-mercaptoethanol. The skin sensitization obtained with IgS does not persist more than 2 hr.
Cytotoxic Antibodies and Lymphocytes Induced by Orthotopic Tooth Bud Allografts in Mice
Authors: Riviere George R.
Published: December 2022

Under most circumstances, viable teeth are promptly rejected when transplanted to allogeneic recipients. However, the influence of the perioral environment on the induction of specific immune responses has been little studied. As few as 4 × 104 A/J lymphocytes injected per oris into either C57BL/6 (H-2 plus multiple non-H-2 disparities) or A.BY recipients (H-2 disparity only) stimulated the production of lymphocytoxic alloantibodies. Moreover, 4 × 107 cells administered periorally elicited cytotoxic antibody titers quantitatively similar to those obtained after challenge with similar numbers of cells by the i.p. route. In vitro cytotoxicity was not detected by either means of sensitization among non-H-2 (H-4, H-11) incompatible mice. Tooth buds transplanted orthotopically among juvenile mice differing at the H-2 locus also yielded production of cytotoxic serum antibodies and killer lymphocytes. The antibodies were detected in vitro after 10 days (1 to 2 days past the median survival time of the transplanted teeth), whereas cytotoxic lymphocytes were detectible at 10 to 13 days. Although tooth buds were rejected in an acute manner in the B10.129(21M) to B10.129(10M) non-H-2 disparate combination, they survived indefinitely in the reciprocal direction despite a transient immunologic crisis. Nevertheless, in vitro cytotoxicity was not demonstrable in either strain combination. The implications of the results of these experiments for prolonged or indefinite survival of allogeneic tooth transplants are discussed. The perioral tissue environment as such offers no immunologic privilege to transplanted allogeneic tissue.
Tolerance and Contact Sensitivity to DNFB in Mice. II. Specific in Vitro Stimulation with a Hapten, 2,4-Dinitrobenzene Sulfonic Acid (DNB·SO3Na)
Authors: Phanuphak Praphan , Moorhead John W. , Claman Henry N.
Published: December 2022

A hapten, 2,4-dinitrobenzene sulfonic acidsodium salt (DNB·SO3Na), a tolerogen, was shown to stimulate (in vitro) specifically lymph node or spleen cells from mice contact-sensitized to DNFB. This stimulation could be carried out in the absence of added serum. Cells from normal and from tolerant mice were not stimulated by DNB·SO3Na.
Characterization of the Fv Fragment Isolated from a Human Immunoglobulin M
Authors: Kakimoto Kiichi , Onoue Kaoru
Published: December 2022

A fragment composed of the variable regions of heavy and light chains was isolated from a Waldenström IgM(Hi) protein (κ-type). This fragment was obtained almost quantitatively either by direct peptic digestion of the IgM at pH 4.5, or by peptic digestion of the Fabµ fragment produced by papain digestion of the IgM. The fragment, which was called Fv fragment, had a molecular weight of 30,000. Quantitative and qualitative analyses of the antigenicity indicated that this fragment carried essentially all the antigenicity of the variable region of the Hi protein, except some of that of the variable region of κ chain; but it carried only a trace, if any, of the antigenicity of constant region. The results showed that the variable regions of heavy and light chains of an IgM molecule were isolated in their associated form maintaining the structural integrity as in the native molecule.
Effect of Polyadenine: Polyuridine on Brucellosis in Conventional and Congenitally Athymic Mice
Authors: Cheers Christina , Cone Robert E.
Published: December 2022

The adjuvant poly A:U given immediately after infection with Brucella abortus markedly retarded the growth of this organism in both conventional and congenitally athymic mice. The time of administration of poly A:U was critical, and poly A:U given 2 days after brucella had no effect on the course of infection. The implications of these findings for the mechanism of action of poly A:U are discussed.