bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional Development of the Interacting Cells in the Immune Response
Authors: Chiscon Martha O. , Golub Edward S.
Published: December 2022

The functional ontogeny of the interacting T cell and B cell4 populations in the antibody response to sheep erythrocytes (SRBC) was investigated in CBA mice. It was found that T cell function, as determined by the ability of test cells to interact with adult bone marrow cells, was not detectable in fetal or neonate liver or lung. Moreover, even in the thymus this function remained less than 10% of adult values until birth. At birth T cell function increased exponentially, reaching adult levels within 48 hr. Neonatally thymectomized mice reconstituted with thymus cells of various ages confirmed these results. By day 4 after birth, T cell function could be detected in the spleen. B cell function, as determined by the ability of test cells to interact with adult thymus cells, was detected in the neonate spleen from day one. Therefore, both B cell and T cell function was present in the spleen from day 4 after birth. B cell function also was found in the fetal and neonate liver. B cell function of fetal liver was only 1/6 that of adult bone marrow on fetal day 16, quickly surpassed adult bone marrow activity near birth, and then equalled adult bone marrow activity until day 6 after birth. The ontogeny of B cell and T cell function in the antibody response to SRBC appears to take place at separate times in the mouse. B cell function develops first, most probably from the most primitive stem cell. T cell function develops later, after a structural thymus is present. Migration of T cell lymphocytes to other lymphoid tissue shortly after birth is consistent with the data presented.
A New Serum Factor in Normal Rabbits
Authors: Richie E. R. , Grote D. , Mandy W. J.
Published: December 2022

A hexapeptide was specifically cleaved from the C-terminus of the Fd of rabbit (Fab')2 fragments. The amino acid composition and sequence of the peptide was identical to that reported for the hinge peptide. These studies further showed that the hinge peptide could be recovered as a disulfide bonded dimer from d11 but not from d12 (Fab')2 fragments by digestion with CNBr.

The results of this investigation also demonstrated that the isolated peptides were reactive with a specific natural antiglobulin factor in rabbit sera, namely, homoreactant. As such, the peptide represents one of the smallest naturally occurring antigenic determinants isolated from a protein.

The chemical and serological properties of the isolated hinge peptide were confirmed by comparison with a synthetic peptide of determined amino acid sequence.
Development of Tissue Viral Resistance in Chickens Following Administration of Interferon Inducers
Authors: Finkelstein Martin S.
Published: December 2022

Adult chickens were stimulated by various routes with interferon-inducers: poly rI/rC and statolon. After 7 or 24 hr they were sacrificed, their sera and tracheae were titered for interferon, and their tracheae and kidneys were examined for resistance to several different viruses. By this means the interferon-specificity of resistance could be judged, avoiding any non-interferon-mediated effects of the inducers. It was found that although poly rI/rC is a relatively poor interferon-inducer in chickens when given intramuscularly or intravenously it can induce interferon and viral resistance in the trachea when given locally. Statolon is ineffective when given intratracheally, but when given intravenously, induces considerable circulating interferon; this interferon seems to protect the kidney and to cross over into the trachea and protect that organ as well. The presence of high titers of interferon in the serum or trachea was associated with protection of the kidney or trachea respectively. However, the presence of small amounts of interferon or the absence of detectable interferon is not indicative of protection or a lack of protection. Interferon-like protection can be confined to specific organs depending upon the interferon-inducer used and its route of administration. These studies underscore the difficulty of equating the presence or absence of detectable interferon with tissue resistance and suggest that the technique of testing viral resistance of extirpated organs provides a valuable index of the efficacy of interferon inducers administered in vivo.
Inhibition of Antichlamydial IgM Antibody by IgG Antibody in Immunofluorescence Tests
Authors: Juchau S. Vern , Linscott William D. , Schachter Julius , Jawetz Ernest
Published: December 2022

Monkeys inoculated intramuscularly with chlamydial suspensions responded with antibody demonstrable by indirect immunofluorescence. IgM antibody was not detected in the whole sera of five of six monkeys, but antibody activity could be demonstrated in IgM fractions from these sera. Monkeys inoculated intravenously with large numbers (1011) of chlamydial particles had IgM antibody detectable in whole sera as well as in fractions. IgG antibody interfered with the demonstration of IgM antibody by immunofluorescence. It is suggested that similar interference of IgG with IgM antibody may occur in other systems.
Complementarity of H and L Chains from Antihapten Antibodies of Different Classes
Authors: Eden A. , Lamm M. E. , Nussenzweig V.
Published: December 2022

Recombination experiments were performed with H and L chains of guinea pig anti-DNP γ1 and γ2 immunoglobulins. The immunoglobulins were mildly reduced and alkylated and then exposed to guanidine HCl in order to dissociate their polypeptide chains. Recombination was allowed to occur as the guanidine was subsequently dialyzed away. Recombination of H chains with one another was found to be class-specific. On the other hand, the recombination of L chains with H chains was close to random (i.e., independent of class of origin), especially with antibodies obtained from individual animals. The results are consistent with the idea that during the immune response, clones of antibody-forming cells switch classes (CH genes) while retaining the specificity of the combining site (V genes).
Production of the Second (C2) and Fourth (C4) Components of Guinea Pig Complement by Single Peritoneal Cells: Evidence That One Cell May Produce Both Components
Authors: Wyatt Harold Vivian , Colten Harvey R. , Borsos Tibor
Published: December 2022

Guinea pig peritoneal cells were examined by a hemolytic plaque assay for their ability to produce C4 and C2. Of cells obtained by peritoneal wash or from exudates induced by starch, 12% were capable of producing C2 and 12% were capable of producing C4. Peritoneal cells induced by oil or casein showed fewer cells capable of producing C4 or C2. With a plaque assay specifically designed to detect only those peritoneal cells that were producing both C4 and C2, we found one hemolytic plaque (produced by a single cell) per 1000 peritoneal cells. Cells capable of producing both C4 and C2 were identified as large mononuclear cells. The production of C4 and C2 by these cells was a temperature dependent process.
Ultrastructure and Immunofluorescence of Mouse Spleen Cells Obtained by Discontinuous Albumin Gradient Centrifugation
Authors: Waldo Elaine Daimont , Zucker-Franklin Dorothea
Published: December 2022

The ultrastructural, fluorescent and phagocytic properties of mouse spleen cells separated by differential centrifugation in discontinuous albumin gradients were studied. Four bands were consistently obtained. The most dense, and most homogeneous band (D) consisted chiefly of small lymphocytes. Large lymphocytes, monocytes and cells undergoing mitoses occurred chiefly in the least dense layer. All fractions contained about similar numbers of plasma cells. Approximately 50% of the cells in band D had surface immunoglobulin whereas only very few cells in the less dense layers showed surface immunofluorescence. The observations should prove useful for the interpretation of data obtained from experiments in which cells from albumin gradients have been employed.
Delayed Hypersensitivity in the Marsupial Setonix Brachyurus (Quokka)
Authors: Turner Keven J. , Alpers Michael P. , Wight Margaret
Published: December 2022

The capacity of the quokka (Setonix brachyurus) to develop a delayed hypersensitivity response to 2,4-dinitrofluorobenzene (FDNB) was demonstrated and the optimum conditions for it were determined. They were topical application of FDNB in ethyl alcohol (EtOH) for 10 days, challenge intradermally (i.d.) with FDNB in olive oil and reading after 48 hr. There was concomitant development of precipitating antibody to the chemical hapten.

Lymphocytes from normal quokkas undergo blast transformation in response to phytohemagglutinin (PHA).

In the one thymectomized quokka studied there was complete absence of both delayed hypersensitivity response to FDNB and leucocyte response to PHA. A humoral antibody response, however, was still present.
Heat Labile Opsonins to Pneumococcus
Authors: Winkelstein Jerry A. , Shin Hyun S. , Wood W. Barry
Published: December 2022

Previous studies have shown that the interaction of pneumococci and the heat labile opsonin (HLO) system results in the cleavage of C3 and the fixation of C3b on the surface of the pneumococcus. This process is not a direct interaction, but requires the participation of other factors present in normal guinea pig serum (GPS). The present studies concern the identity of these factors.

One component of the HLO system is guinea pig γ2 immunoglobulin. It is bound to the pneumococcus during opsonization and can be demonstrated to participate functionally in the interaction of the pneumococcus and the HLO system.

Using an in vitro phagocytic test, we also found that C4 deficient GPS is able to support the phagocytosis of pneumococci by polymorphonuclear leukocytes as well as normal GPS. Also, when pneumococci are incubated in C4 deficient GPS, they interact with the HLO system to consume C3 as well as they do when incubated in normal GPS. Therefore, C4 is not required for the opsonization of pneumococci by the HLO system and the cleavage and fixation of C3 probably proceeds via the alternate complement pathway.
The Evolution of the Immune Response
Authors: Finstad C. L. , Litman G. W. , Finstad J. , Good R. A.
Published: December 2022

Erythrocyte agglutinins were isolated and purified from the horseshoe crab, Limulus polyphemus, and the starfish, Asterias forbesi. The purified agglutinins were studied with respect to primary and secondary molecular structure and the nature of component subunits. When the two agglutinins were compared to one another, to other invertebrate agglutinins and to known vertebrate immunoglobulins, each was found to be unique.
Formation of an Antibody with Serologic Specificity for N6-(Δ2-Isopentenyl) Adenosine
Authors: Hacker Bruce , VanVunakis Helen , Levine Lawrence
Published: December 2022

Recent reports have dealt with the demonstration of specific antibodies found in response to immunization of rabbits with conjugates of methylated nucleoside haptens (1, 2). Not surprising were the findings that the methyl group and its position on the purine or pyrimidine ring dominated the degree of complementarity between hapten and antibody. Nevertheless, some recognition by these antibodies of heterologous methylated bases and even normal bases was observed. We have recently produced in rabbits antibodies directed toward N6-(Δ2-isopentenyl) adenosine (iA). As expected, the N6-(Δ2-isopentenyl) group on the nucleoside was immunodominant. These antibodies do react with N6-methyladenosine; yet 1 × 104 more N6-methyladenosine molecules were required for equivalent activity. Even at a 1 × 105 higher concentration, normal bases did not cross-react. The iA in the tRNATyryeast was available for reaction with the antibody of iA.

Isolation and characterization of the iA used for synthesis of the immunizing antigens have been described previously (3).
Enzyme-Linked Immunosorbent Assay, Elisa
Authors: Engvall Eva , Perlmann Peter
Published: December 2022

A sensitive and simple method for the quantitative determination of antibodies is reported. Tubes coated with antigen are incubated with antiserum followed by an enzyme-labeled preparation of anti-immunoglobulin. The enzyme remaining in the tubes after washing provides a measure of the amount of specific antibodies in the serum. Coating of polystyrene tubes with antigen is described, as well as the preparation of specifically purified antibodies against rabbit IgG, and their conjugation to alkaline phosphatase.

When rabbit antisera against human serum albumin or against the dinitrophenyl group (DNP) were incubated in tubes coated with antigen, less than 1 ng/ml of specific antibody could be detected in both systems. Antibodies in unknown sera could be quantitated by comparison with a standard antiserum. The results obtained compared well with those obtained with other methods for antibody determination.

In the DNP system, the specificity of the reaction was assessed by inhibition with hapten. The reaction of immune serum against DNP with DNP-protein, adsorbed to the tubes, was completely inhibited by hapten in solution.
Role of the Cerebrosides and a Galactodiglyceride in the Antigenic Cross-Reaction Between Nerve Tissue and Treponema
Authors: Dupouey P.
Published: December 2022

Various authors have demonstrated an antigenic identity between nervous tissue and treponema. This report shows that this antigenic identity is due to the presence in the treponema extracts of a galactodiglyceride hapten: i.e, 2,3-di-O-acyl-1-O-(β-D-galactopyranosyl)-D-glycerol. This hapten shows a cross-reaction with the cerebrosides; moreover, the nervous tissue contains a lipid which, on deacylation, liberates a O-(β-D-galactosyl)-1-1′-glycerol (11). The part played by each of the constituents as well as the possible consequences of this antigenic community are discussed.
Effect of Haptens on the Reaction of Anti-Idiotype Antibody with a Mouse Anti-Phosphorylcholine Plasmacytoma Protein
Authors: Sher Alan , Cohn Melvin
Published: December 2022

In a radioimmune assay under conditions of excess antigen, the binding of a mouse antiphosphorylcholine plasmacytoma protein by anti-idiotype antibody is partially inhibited by phosphorylcholine and related haptens. The effect of these haptens on the idiotype correlates with the affinity with which they are bound by the plasmacytoma protein.
Reversed in Vitro Anaphylaxis Induced by Anti-IgG: Specificity of the Reaction and Comparison with Antigen-Induced Histamine Release
Authors: Grant J. Andrew , Lichtenstein Lawrence M.
Published: December 2022

Antibodies to human IgE can combine with IgE on human basophils and cause release of intracellular histamine. Previous reports have demonstrated that anti-IgG is capable of eliciting the same response, but the possibility of contamination by antibodies which could also react with IgE remained to be explored. In this study, specifically purified antibodies to: 1) Fc of IgG, 2) Fc of IgE and 3) light chains were used to obtain histamine release from leukocytes of allergic donors. The inhibition of this reaction with purified IgG, IgE and light chains was then determined. It was shown that histamine release by anti-IgG was not due to specificities which recognize the ε chain and was only minimally due to anti-light chain specificities.

Histamine release mediated by allergen and anti-IgG was also inhibited by drugs which increase intracellular cyclic-AMP. Both processes were similarly inhibited. This suggests that anti-IgG-mediated histamine release mimics naturally occurring allergen-mediated release: both are noncytotoxic, secretory processes.
Stimulation of Lymphocyte Transformation by Streptococcal Type M1 Protein: Relationship to HL-A Antigens
Authors: Pellegrino M. A. , Ferrone S. , Safford J. W. , Hirata A. A. , Terasaki P. I. , Reisfeld R. A.
Published: December 2022

M1 protein isolated from group A streptococci, type 1, stimulates human peripheral blood lymphocytes in short-term tissue culture to undergo a dose-dependent increase in DNA synthesis. The degree of this lymphocyte stimulation is not related either to HL-A phenotype, the serum anti-M1 titer of the lymphocyte donor, or the effectiveness of M1 antigen in blocking the cytotoxic activity of HL-A alloantisera. However, both HL-A allo- and heteroantisera are highly effective in specifically inhibiting M1 mitogenic activity. This inhibitory effect clearly depends on anti-HL-A antibody since HL-A alloantisera are rendered ineffective when specifically absorbed with cultured human lymphoid cells. The mechanism by which anti-HL-A antibodies suppress the M1-induced lymphocyte transformation is thought to involve a masking of the M1 protein's mitogenic site such as to prevent interactions with specific lymphocyte receptors required for induction of the proliferative response.
Studies in Experimental Eosinophilia
Authors: Litt Mortimer
Published: December 2022

Puromycin blocked the eosinophil response which is elicited in guinea pig lymph nodes by a single injection of hemocyanin into footpads. The drug was most effective in interfering with the 24-hr response when it was given just before or no later than 4 hr after the antigen. A partial block was noted when the drug preceded the antigen by 2 to 8 hr. Interference with the 15-min vascular response required a higher dose. The secondary type of eosinophil response, which is seen in an immunized animal, was not inhibited by puromycin. The drug was effective when administered into the footpads, but a higher dose was required when it was given by the intraperitoneal route. The results support the notion that the eosinophilia which occurs during the first few minutes of a primary immune response reflects the de novo synthesis of antibody, rather than the presence of preformed antibody.
Studies on the Subunit Structure of Rabbit Secretory IgA
Authors: Underdown Brian J.
Published: December 2022

The nature of the intersubunit bonds in rabbit secretory IgA dimer was investigated by employing gel filtration on 2% and 4% agarose columns equilibrated with 6 M guanidine-HCl. Under these conditions, rabbit secretory IgA behaved as a dimer [(H2L2)2] which established that the bonds linking the monomer subunits were covalent in nature. Rabbit secretory component, however, dissociated from the native molecule in 6 M guanidine-HCl, which indicated that the secretory component was bound to the native dimer by non-covalent bonds. Mild reduction and alkylation with dithiothreitol (DTT) in aqueous buffer resulted in dissociation of the rabbit secretory IgA dimer molecule to monomer subunits and lower molecular weight material. Reduction with varying quantities of DTT showed that rabbit secretory IgA dimer was more resistant to reduction than human secretory IgA dimer which in turn was more resistant to reduction than a human IgA dimer myeloma protein.
Macrophage-Bound Antigens
Authors: Seeger Robert C. , Oppenheim Joost J.
Published: December 2022

Macrophage-antigen is quite effective in activating T cells which are involved in mediating delayed hypersensitivity (DH) and in helping B cells produce antibodies. In this study we determined if this property was unique to macrophages by immunizing guinea pigs with antigen bound to macrophages, lymphocytes, thymocytes or hepatoma cells. Cells from unimmunized guinea pigs were incubated in vitro for 60 to 90 min with 125I tetanus toxoid or guinea pig albumin conjugated with orthanilic acid (GPA.OA), washed, and then injected into syngeneic unimmunized guinea pigs.

Macrophages took up 10 to 18 times more antigen than the other cells. To compensate for this difference in uptake, we injected equal doses of antigen rather than cells. Even with injection of fewer macrophages, macrophage-antigen was more effective than lymphocyte-antigen and soluble antigen in inducing DH and in priming for production of antibodies. Thymocyte-antigen and hepatoma antigen were ineffective in inducing immunity. Only macrophage-antigen was capable of activating T cells when guinea pigs were immunized by the intracardiac route or when weakly immunogenic GPA.OA was used.

These observations, with those of the previous paper, suggest that an initial interaction of antigen with macrophages but not with immunocompetent lymphocytes, immature thymocytes or hepatoma cells leads to activation of T cells involved in mediating DH and in helping B cells to form antibodies.
Stabilization of Dissociable IgA2 Proteins by Secretory Component
Authors: Jerry L. M. , Kunkel H. G. , Adams L.
Published: December 2022

The Am2(+) genetic variant of human IgA2 immunoglobulins is potentially unstable because it lacks disulfide bridges between the heavy (H) and light (L) chains. It dissociates spontaneously under denaturing conditions into heavy and light chain dimer subunits (H2, L2), without the need for prior reduction. Isolated secretory component (SC) binds to human Am2(+), IgA2 myeloma proteins in vitro, and stabilizes them so that they will no longer dissociate into subunits in the presence of denaturants. Both human and bovine SC will bind to and stabilize the polymer forms of these myeloma proteins, but only human SC will bind to and stabilize monomeric proteins. The reaction involves the whole IgA2 molecule rather than just its subunits. Moreover, SC will stabilize a pepsin F(ab′)2 fragment. Since stabilization is prevented by iodoacetic acid, covalent disulfide bonding appears to be involved. The ability of SC to stabilize the potentially dissociable Am2(+) genetic variant of IgA2 may enhance its capacity to function in external secretions.
A Specific Mouse B Lymphocyte Antigen (MBLA) Demonstrated by Double Immunofluorescence
Authors: Niederhuber John E. , Britton Sven , Berquist Robert
Published: December 2022

Double immunofluorescent staining was used to demonstrate the presence of a specific antigenic marker (MBLA) on the surface of B lymphocytes in the mouse. The percentages of thymus dependent and thymus independent cells detected by indirect staining of specific antisera-treated cells were consistent with the cytotoxic indices for these sera. The double staining of anti-MBLA-treated cells after incubation with fluorescein-conjugated anti-mouse immunoglobulin serum and rhodamine-conjugated anti-rabbit immunoglobulin serum confirmed the absence of anti-immunoglobulin activity in the anti-MBLA serum.
Subtyping of Hepatitis-Associated Antigen (HB Ag); Simplified Technique with Counterelectrophoresis
Authors: Holland Paul V. , Purcell Robert H. , Smith Holly , Alter Harvey J.
Published: December 2022

A simple and sensitive counterelectrophoresis technique for determining the subtype of hepatitis-associated antigen (HB Ag) in sera is described. Using monovalent antisera, or polyvalent antisera rendered monovalent by appropriate absorption, we detected a group-specific antigenic determinant, a, and two additional specificities, d and y. All HB Ag-containing sera could be subtyped as either ad or ay. The subtype appears to be determined by the viral agent, not the host, and to remain the same in affected individuals.

While the frequency of subtypes ad and ay were about the same for patients with acute hepatitis, ad was much more common in patients with chronic hepatitis. Subtype ad was also detected more frequently than ay in blood donor carriers of HB Ag. The major values of subtyping for HB Ag will probably be in the study of the epidemiology of hepatitis B and the development of optimal testing reagents for HB Ag detection.
Simultaneous Induction of Experimental Allergic Encephalomyelitis in Obese Strain (OS) Chickens with Spontaneous, Hereditary Autoimmune Thyroiditis
Authors: Wick Georg , Steiner Renate
Published: December 2022

Experimental allergic encephalomyelitis (EAE) was elicited in Obese strain (OS) chickens, which develop a spontaneous autoimmune thyroiditis, and in normal White Leghorn (NWLH) chickens by immunization with bovine spinal cord and complete Freund's adjuvant alone or Freund's adjuvant and pertussis vaccine. Control groups were similarly immunized with bovine liver. OS chickens developed severe EAE without any influence on the frequency and severity of spontaneous thyroiditis and on the occurrence of circulating thyroglobulin autoantibodies. The use of pertussis vaccine as a coadjuvant led to slightly more severe EAE in both NWLH and OS birds as compared to Freund's adjuvant alone.

Immunization with bovine spinal cord resulted not only in the formation of antibodies to this antigen but also to the appearance of circulating brain autoantibodies in OS and NWLH chickens.

This avian model which simultaneously displays a spontaneous and an experimentally induced autoimmune disease should provide a very useful tool for the study of the role of the bursa- and thymus-dependent part of the immune system in the development of these diseases.
The Clonal Nature of Antibody Formation
Authors: Bosma M. , Davis G.
Published: December 2022

Experiments were designed to test the hypothesis that prospective antibody cell clones can synthesize antibody molecules of only one specificity. This was done with the aid of a cell transfer system as follows: hapten-specific precursor cells of F1-hybrid mice were stimulated with protein carriers bearing random copolymers O-acetyl-D-serine and D-alanine (ser-ala); antibody allotypes served as genetic markers in evaluating the clonal origins of antibodies which were specific for the homopolymer haptens, poly-ser or poly-ala. Our analysis of the results supported the conclusion that different and unrelated antibody precursor cells responded to poly-ser and poly-ala determinants. Apparently the antibody specificity of these precursor cells was genetically restricted before contact with antigen.
The Effects of Aggregated Immunoglobulins on Human Blood Platelets in Relation to Their Complement-Fixing Abilities
Authors: Pfueller Sharron L. , Lüscher Ernst F.
Published: December 2022

Human immunoglobulin (IgG) coupled with bis-diazo-benzidine (BDB-IgG) is capable of causing a release reaction from platelets in a manner similar to heat-aggregated IgG or antigen-antibody complexes. Various organic solvents, e.g., ethanol and acetone, also produce platelet-stimulating complexes. All these aggregated immunoglobulins caused the fixation of complement (C), although not strictly in parallel to their ability to cause the platelet release reaction.

Myeloma proteins from the subclasses IgG1, IgG2 and IgG3, but not IgG4 or IgA, bind C, while all IgG subclasses, including IgG4, stimulate platelets. IgG3 proteins cause a poor release reaction but bind C well.

The release reaction induced by human BDB-IgG is inhibited by monomeric IgG but not by bovine IgG or albumin. Partially purified C1 and purified Clq also inhibit the release reaction. The presence of Ca2+ is not required for any of these effects.

It is concluded that for an aggregated immunoglobulin to stimulate platelets it is not necessary that it activate the complement enzymes in the classical way, but that either it may interact with a cell-bound Clq-like receptor, or the immunoglobulin possesses distinct, but closely situated, sites for platelet and C activation.
The Effects of Aggregated Immunoglobulins on Human Blood Platelets in Relation to Their Complement-Fixing Abilities
Authors: Pfueller Sharron L. , Lüscher Ernst F.
Published: December 2022

When the amino groups of aggregated human immunoglobulin G (IgG) were modified by acetylation or acetimidation, its ability to bind complement and to stimulate the release reaction of platelets disappeared, modification of 40% to 46% of the amino groups by acetimidation being sufficient for marked inhibition. By reaction of 26% of the amino groups with dinitrofluorobenzene, platelet stimulation disappeared completely and complement fixation was reduced by 50%. Modification of 3 to 5 tryptophane residues/molecule caused 40% reduction in both activities. There was some indication that tryptophane residues exposed by conformational changes during aggregation of IgG may be important in endowing the molecule with biologic activity.

Removal of 36% of the IgG hexose caused total loss of platelet stimulating activity, although complement fixation was hardly affected. These results are compatible with the idea that the sites of Clq binding and for platelet interaction have amino acid sequences in common. Studies of the inhibition of the normal release reaction by modified monomeric IgG indicate that tryptophane and carbohydrate, but not the amino groups, are necessary for the initial interaction of IgG with the platelet surface. The amino groups are most probably required for a subsequent step in the release process.
The Lack of Thymic Influence in Regulating the Immune Response to Escherichia Coli 0127 Endotoxin
Authors: Veit Bruce C. , Michael J. Gabriel
Published: December 2022

A possible regulatory effect of thymus-derived lymphocytes on the immune response to a thymus independent antigen, Escherichia coli 0127 somatic antigen in mice, was investigated. Antilymphocyte globulin prepared in rabbits caused a substantial suppression of the antibody response to a thymus dependent antigen (sheep erythrocytes) but exerted no effect on the immune response to the bacterial antigen. Administration of E. coli somatic antigen to mice did not induce an immunosuppressive effect on a subsequently injected unrelated antigen indicating that this thymus independent antigen was incapable of inducing antigenic competition. Likewise, the immune response to the bacterial antigen was not influenced by prior immunization of the mice with sheep erythrocytes. Thus, in these immune reactions in which thymus-derived lymphocytes were affected the response to the bacterial antigen was not modified. These results contradict recent observations which indicated that thymus-derived lymphocytes exert a suppressive effect on the immune response to thymus independent antigens.
Use of the 51Cr Release Test to Demonstrate Patterns of Antibody Response in Humans to Herpesvirus Types 1 and 2
Authors: Smith Jerry W. , Adam Ervin , Melnick Joseph L. , Rawls William E.
Published: December 2022

Antibodies to the surface antigens of cells infected with herpesvirus types 1 or 2 were assayed by the 51Cr release test. Absorption of sera with 107 unlabeled cells infected with herpesvirus type 1 removed all cytolytic activity to herpesvirus type 1 51Cr-labeled cells; however, cytolytic activity to 51Cr-labeled cells infected with herpesvirus type 2 remained in some of the absorbed sera. Sucrose gradient separation of the immunoglobulins revealed that the cytolytic activity was in the IgM fractions in all eight sera collected 3 to 28 days after the onset of illness caused by herpesvirus type 2 and also in the IgG fractions in all six sera obtained 11 to 28 days after the onset of illness. Sera collected 8 months or more after the infection were found to contain cytolytic activity only in the IgG-containing fractions. Cytolytic antibody activity of 20% specific 51Cr release or greater from labeled cells infected with herpesvirus type 2 after absorption with unlabeled type 1 infected cells was found in the convalescent sera of most patients who had no neutralizing antibody to type 1 in their acute phase sera. However, among persons with neutralizing antibodies in their acute phase sera, cytolytic activity to the type 2 cells was absent or at low levels in their convalescent sera after absorption with type 1 unlabeled cells. The data suggest that antibodies reacting only to herpesvirus type 2 infected cells can be detected by the 51Cr release test and that the quantity of the antibody present depends upon whether or not the person had been infected with herpesvirus type 1 before infection with herpesvirus type 2.
Rabbit Precipitin Antibody to Chicken Erythrocyte Histone F2C
Authors: Sotirov N. , Johns E. W.
Published: December 2022

Specific precipitin antibody was induced in rabbits to a pure preparation of the chicken erythrocyte histone F2C. The hyperimmune sera obtained reacted in agarose gel to form a distinct precipitin line with about 1 µg of F2C. No cross-reactivity could be detected with whole histone or histone fractions F1, F2B, F2A1, F2A2 and F3 isolated from calf or chicken thymus. The nonspecific precipitation between all histone fractions and normal serum proteins was completely removed by absorption of the hyperimmune sera with calf or chicken thymus whole histone or the various histone fractions. The specificity of the immune reaction against F2C was also proved by the skin test which was strongly positive in all the rabbits that had developed precipitin antibody. Chicken erythrocyte specific histone F2C was found to be antigenically distinct from any other histone fraction.
Lymphocyte Classes in New Zealand Mice
Authors: Stobo John D. , Talal Norman , Paul William E.
Published: December 2022

The frequency of thymocytes bearing the “T” cell differentiation antigen theta (ϑ) as well as the mean density of ϑ determinants per thymocyte are similar in newborn through 6-week-old NZB and BALB/c mice. Although the capacity of these thymocytes to respond to the mitogen concanavalin A is acquired at a somewhat earlier age in NZB animals, the absolute responsiveness of 3-week-to 1-year-old NZB thymocytes is less than noted for comparably aged BALB/c mice.

Similarly, both the age dependent appearance of ϑ positive splenic lymphocytes as well as the absolute frequency of these cells are similar in newborn through 6-week-old animals of both strains. Spleen cells from 1-year-old NZB mice, however, demonstrate a decreased frequency of these cells when compared to BALB/c animals of similar age. The appearance of phytohemagglutinin and concanavalin A reactivity among spleen cells parallels the appearance of ϑ positive cells, and is similar in both strains.

Thus, no gross abnormalities in the development of either surface “T” markers or in the responsiveness to mitogens which activate “T” cells can be found in newborn through 6-week-old NZB mice. Nonetheless, a defect in either an antigen dependent function of “T” cells or in a “T” cell subset cannot be excluded.
The Humoral Immune Response in Mice after Neutron or X-Irradiation at Different Dose Rates
Authors: Gottlieb Charles F. , Gengozian Nazareth
Published: December 2022

Acute radiation dose rate and radiation quality are two critical, although little studied, variables involved in nonspecific radiation immunosuppression. Mice (C3BF1) were injected with sheep erythrocytes shortly after irradiation, and the humoral immune response measured over a 35-day period. With neutron irradiation, antibody formation was dependent only on the total dose, and not the dose rate. In contrast, the immune response after x-irradiation was dependent on both the dose and dose rate. Maximum dose-rate differences occurred with 500 rads of x-rays. Maximum depression with both neutron and x-irradiation occurred when antigen was given 12 hr after exposure. The exposure-rate effect on immunocompetent cells was directly demonstrated at the cellular level by culturing irradiated spleen cells in lethally irradiated recipients.
The Effect of Antibodies to Complement Components C2, C3 and C5 on the Production and Action of Lymphotoxin
Authors: Gately Maurice K. , Mayer Manfred M.
Published: December 2022

A study was made on the effects of monospecific rabbit antibodies to guinea pig C2, C3 and C5 on the production and action of guinea pig lymphotoxin. Lymphotoxin was produced by incubating ovalbumin or purified protein derivative (PPD) with lymph node cells from guinea pigs which had been immunized with ovalbumin in complete Freund's adjuvant. Culture fluids from cultures of lymph node cells which had been preincubated with anti-C2, anti-C3 or anti-C5 and then incubated with both antigen and the anti-complement component antiserum were as cytotoxic when assayed on L cell monolayers as were culture fluids from cultures of lymph node cells incubated with antigen alone. Furthermore, anti-complement component antibody activity could be demonstrated in culture fluids containing lymphotoxin produced in the presence of anti-C2, anti-C3 or anti-C5 indicating that the concentrations of antisera used were sufficient to neutralize respectively all the C2, C3 or C5 present in the culture system. It is therefore concluded that the complement system, activated either via the classical pathway or via the “alternate” pathway, plays no role in the lymphotoxin phenomenon.
A Mechanism for the Unresponsive State in BALB/c Mice after Repeated Doses of Sheep Erythrocytes
Authors: Thomson Philip D. , Jutila John W.
Published: December 2022

BALB/c mice were rendered unresponsive to sheep red blood cells (SRBC) by injections of 5 × 108 SRBC every other day from 7 days of age to adulthood. It was shown that unresponsiveness was specific by injection of burro red blood cells which produced normal plaque-forming cell responses in SRBC-unresponsive mice. Neither normal thymus nor bone marrow alone or in combination, nor normal peritoneal exudate cells (80 to 90% macrophages) reconstituted unresponsive mice. However, SRBC-primed peritoneal exudate cells (80 to 90% macrophages) did reconstitute the plaque-forming response of unresponsive mice. Normal BALB/c mice were rendered unresponsive by injection of 32 hemagglutinin units of serum from unresponsive mice. Serum from unresponsive mice was absorbed with IgG2. Serum absorbed in this manner failed to render mice unresponsive when administered passively. It was concluded that unresponsiveness to SRBC involves IgG2 which interacts with the macrophage and, in turn, inhibits antigen-macrophage interaction. Thus this system represents a feedback mechanism which prevents the presentation of antigen to the lymphocyte.
Preparation of Dinitrofluorobenzene-Treated Sheep Erythrocytes for Detection of Anti-DNP Plaque-Forming Cells
Authors: Trump Gary N.
Published: December 2022

A method is described for covalent attachment of dinitrofluorobenzene (DNFB) to sheep red blood cells (RBC). The solubility and ionization characteristics of DNFB are considered in this procedure and relatively large amounts of hapten are reacted with RBC. Unreacted DNFB is partially precipitated and soluble DNFB is carefully removed from DNP-RBC at the end of reaction. Haptenated erythrocytes are extremely sensitive to hemolytic antibody in the direct agarplaque assay as well as in passive hemolysis. Previously reported DNFB techniques were carried out in parallel experiments and results of such comparisons highly recommend the present method for use in detection of direct anti-DNP plaque-forming cells.
Use of the Macrophage Migration Inhibition Test to Monitor Fractionation of Soluble Antigens of Chemically Induced Sarcomas of Inbred Guinea Pigs
Authors: Suter Ludwig , Bloom Barry R. , Wadsworth Elisabeth M. , Oettgen Herbert F.
Published: December 2022

Soluble tumor antigen was prepared from two transplanted sarcomas of strain 13 guinea pigs, induced with 3-methylcholanthrene or 9,10-dimethyl-1,2-benzanthracene, by extraction with isotonic saline followed by ultracentrifugation. Further fractionation by ammonium sulfate precipitation, gel filtration on Sephadex G-150 or Biogel A-5m, and DEAE Sephadex chromatography was performed. Antigenic activity was assayed in vivo by skin tests and in vitro by the macrophage migration inhibition method. Antigenic activity was not restricted to a single species of molecules as defined by size or charge. The major part of the activity was associated with two fractions, one excluded by Sephadex G-150, and the other included in Sephadex G-150. The behavior of the antigen preparations in isopycnic sucrose gradient centrifugation indicated that they were truly soluble and not membrane associated. The active fractions were nontoxic for the skin and for peritoneal cells and stable under the conditions of the macrophage migration test. Their antigenic activity was specific for each of the two tumors. In terms of the amount of antigen required to give a positive reaction, the migration inhibition assay was more sensitive than the skin tests.
C3 Shunt Activation in Human Serum Chelated with EGTA
Authors: Fine Douglas P. , Marney Samuel R. , Colley Daniel G. , Sergent John S. , Des Prez Roger M.
Published: December 2022

Complement (C) fixation via the C3 shunt, as exemplified by zymosan (Z) and Escherichia coli (E. coli), requires ionized magnesium but not ionized calcium and proceeds in the presence of 10 mM ethyleneglycoltetraacetic acid (EGTA). C-fixation via the C142 pathway requires calcium and is inhibited by EGTA. Both pathways of C-activation are inhibited by ethylenediaminetetraacetic acid (EDTA). EGTA serum may be a useful and simple reagent to distinguish between C142 and C3 shunt-mediated complement activation.
Prevalence of Hepatitis B Virus Antigen As Revealed by Direct Radioimmune Assay with 125I-Antibody
Authors: Ling C. M. , Overby L. R.
Published: December 2022

A two-step direct radioimmune test was used to detect hepatitis B virus associated antigen in human serum or plasma. The procedure involved the use of polypropylene tubes coated with hyperimmune guinea pig serum, and 125I-labeled specific hepatitis B virus antigen immune globulin. The technique appeared to be more sensitive than other immunologic assays, including indirect radioimmune methods based on competition of unlabeled antigen with 125I-labeled antigen.

Various blood donor populations were tested for antigen by the direct radioimmune test, by complement fixation, agar gel diffusion and counterimmunoelectrophoresis. The increased sensitivity of the radioimmune procedure resulted in the detection of about 3 to 8 times as many positive sera as the above three immunologic methods. The percentages of positive sera by radioimmune assay were: about 1% in volunteer blood donors, about 2% in commercial blood donors and about 40% in hepatitis-implicated patients.
Use of 2-Mercaptoethanol for Distinguishing Between IgM and IgG Antibody-Producing Cells of Mice Immunized with Bovine γ Globulin
Authors: Hosono Masamichi , Muramatsu Shigeru
Published: December 2022

Spleen cells from mice immunized with heat-aggregated bovine γ globulin (BGG) were assayed by the technique of localized hemolysis in gel. Target cells were prepared by the conjugation of BGG to sheep red blood cells by means of chromic chloride. No anti-BGG direct plaques were developed by the addition of guinea pig serum alone throughout the period under examination. Practically all of the plaques were obtained by the indirect method with the aid of anti-mouse immunoglobulin antiserum, and at least two categories of cells which secrete either 2-mercaptoethanol (2ME)-sensitive or 2ME-resistant antibodies were involved. The pretreatment of gel plates with 2ME after cellular antibodies have diffused out may be recommended for distinguishing between IgM and IgG antibody producers against protein antigens. We also discuss why direct plaques do not appear in this experiment.
Production of Antibody in the Canine Tracheal Pouch after Local and Systemic Influenza Immunization
Authors: Lieberman Phillip , Berlin Byron S. , Metzger James , Patterson Roy
Published: December 2022

The canine tracheal pouch model was utilized to compare the secretory and systemic immune response to topical tracheal or subcutaneous immunization with influenza A, swine strain, and influenza B viruses. Serum antibody responses were comparable regardless of the route of administration of virus, but topical tracheal immunization was more effective than subcutaneous immunization for eliciting anti-influenza antibody in tracheal secretions. Both IgG and IgA anti-influenza antibodies were found in tracheal secretions after topical immunization. The IgA response was consistent with the presence of local antibody production. In addition, tracheal secretions contained a nonspecific inhibitor of influenza hemagglutinating activity which reacted to high titer with A2 influenza virus and was susceptible to receptor-destroying enzyme.
Complement Fixation Blocking Activity of Anti-Dengue IgM Antibody
Authors: Scott Robert McNair , Russell Philip K.
Published: December 2022

The serologic tests commonly used for the diagnosis of arbovirus infections are complement-fixation (CF), hemagglutination inhibition (HI), and virus neutralization (N) tests. In group B arbovirus infection such as Japanese encephalitis and dengue, early convalescent sera often have little or no CF activity, although HI and N antibody activity may at times be present in high titer; later in convalescence, CF activity increases, while there may be little change or a drop in HI and N activity (1). This phenomenon has also been observed in experimentally infected guinea pigs (2). In both man and guinea pigs early convalescent serum with high HI and N activity, but lacking CF activity, contains either antiviral IgM antibody alone or both IgM and IgG antibody. Conversely late convalescent sera with high levels of CF reactivity contain IgG antibody but little or no IgM.
Reaginic Antibody Receptors on Murine Mastocytoma Cells
Authors: Minard Patricia , Levy David A.
Published: December 2022

Sensitization for the allergic reaction requires the firm binding of IgE antibody in homologous tissues. Studies with monkey tissues (1) and with human leukocytes (2) show that IgE binds exclusively to mast cells and basophils. Electron micrographs show that IgE is bound to the plasma membrane of the human basophil (3). This localization of IgE suggests that these cells possess a specific membrane receptor for IgE.

A mast cell tumor arose spontaneously in an LAF1 mouse which had been subjected to whole body radiation (4). This mastocytoma was originally a solid tumor and now also grows in an ascitic form. Receptors for homologous immunoglobulins on malignant mouse mast cells have recently been demonstrated by the technique of rosette formation with antibody-coated sheep erythrocytes (5–7). This report presents evidence for the ability of mouse mastocytoma cells to deplete serum of skin sensitizing activity, and suggests that these cells may have receptors for homologous reaginic antibody.
A Simple Automated Method for Complement Estimation in a Continuous Flow System
Authors: Nydegger U. E. , Achermann L. M. , Lambert P. H. , Miescher P. A.
Published: December 2022

The activity of the complement system is routinely evaluated through the quantitation of serum hemolytic activity in presence of sensitized sheep red blood cells (SSRBC).3 Such measurements do not reflect the activity of each complement component but provide a useful screening test for clinical investigation in various immunologic disorders. However, time consumption is still a practical limitation of this hemolytic technique.

Automated methods using continuous flow systems have been described for complement titration (1–4). These methods are based on serial or exponential dilutions of serum with determination of the 50% hemolytic point, and require numerous manipulations for each sample.

We are reporting a similar automated procedure for complement titration in which only one dilution of plasma is assayed in the presence of two SSRBC concentrations. This technique provides a simple, accurate and fast estimation of complement values lying between 20% and 120% of normal.