bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effect of Prior Transfusion on Marrow Grafts between Histocompatible Canine Siblings
Authors: Storb Rainer , Epstein Robert B. , Rudolph Robert H. , Thomas E. Donnall
Published: December 2022

Twenty-three canine littermate donor-recipient pairs were matched on the basis of histocompatibility typing with four lymphocytotoxic isoantisera. Eighteen of the 23 pairs were also tested in a one-way mixed leukocyte culture (MLC). Recipient dogs were given 1580 R whole body irradiation followed within 4 hr by infusion of marrow from the donor. Group 1 consisted of 10 recipients not given a preceding blood transfusion. Group 2 was made up of 13 dogs given a single transfusion of 100 ml of blood from the donor 10 days before irradiation. Seven of the 10 recipients in group 1 showed prompt and sustained marrow engraftment and survived beyond 70 days. Three died within 22 days of transplantation, two of marrow graft rejection and one of graft-vs-host disease. These three were mismatched with their donors in MLC. Only one of the 13 pretransfused recipients in group 2 lived beyond 70 days. Twelve dogs died within 36 days including eight that were matched by MLC as well as by serologic test. In most of these animals there was evidence of successful marrow engraftment followed by rejection. The results show that, even in histocompatible donor-recipient pairs, a single prior blood transfusion from the donor to the recipient significantly diminishes the chance of long-term survival following the marrow graft (p < 0.01).
Interaction of Lymphocytes and a Radioresistant Cell in PPD-Stimulated Human Leukocyte Cultures
Authors: Schechter Geraldine P. , McFarland William
Published: December 2022

A 5- to 7-day incubation of heavily irradiated (2000 R) human leukocytes yielded a macrophage-rich population which was capable of markedly increasing blastogenic reactivity to PPD (purified protein derivative) of lymphocytes from sensitive donors as measured by incorporation of tritiated thymidine. These radioresistant cells were also demonstrated in some donors to be capable of initiating the blastogenic response after being only briefly exposed to antigen. This activity was demonstrable only when these cells were intact, viable, in direct contact with the lymphocytes and pulsed with antigen sometime within the preceding 72 hr. This ability to initiate blastogenesis in lymphocytes was inhibited if macrophage exposure to antigen occurred at low temperature or in the presence of actinomycin D or cyanide.
Rabbit IgA, IgM and IgG Antihapten Responses Assayed by Cla Fixation and Transfer
Authors: Clough John D. , Zaccari June , Strober Warren
Published: December 2022

The present report describes an adaptation of the C1a fixation transfer test (C1a FTT) for measurement of rabbit antibody in three immunoglobulin classes—IgM, IgG and IgA—without serum fractionation. IgM antibody was measured directly and IgA and IgG antibodies were measured indirectly using specific antisera against rabbit IgG and IgA to permit complement fixation by antibodies in these immunoglobulin classes. The ability of the test to discriminate among antibodies in these classes was demonstrated by immunochemical examination of the anti-antisera and by use of the C1a FTT to show that 1) the effects of the enhancing anti-antisera could be summated, 2) the enhancing ability of the anti-antisera could be specifically inhibited by purified rabbit immunoglobulins, and 3) antibody activity in rabbit serum could be fractionated by DEAE-cellulose gradient chromatography into its immunoglobulin components. The ability of the test to quantitate antibody in each class was examined by assay of serial dilutions of rabbit antiserum by the C1a FTT; the amount of C1a fixed and quantitated was found to be directly proportional to the amount of antibody present. Primary and secondary humoral responses of rabbits to intravenously administered haptenic arsanilic acid were studied using the C1a FTT. In the primary response, IgA antibody reached an early peak together with IgM antibody. IgG antibody had a more gradual rise and peaked later than IgA or IgM; a small second peak of IgA antibody sometimes accompanied the IgG peak. In the secondary response IgA antibody showed an accelerated onset, rising to an earlier but lower peak than in the primary response. The secondary IgM antibody response was similar to that of IgA while IgG antibody reached an earlier, higher peak level than in the primary response.
Quantitation of Canine Immunoglobulins
Authors: Reynolds Herbert Y. , Johnson John S.
Published: December 2022

Quantitative immunoglobulins were measured in canine serum, colostrum, milk and intestinal secretions, and normal mean values were established. Two groups of dogs, pure bred and mongrel, were used because of significantly different serum levels of IgG subclasses IgGa,b and IgGd. The immunoglobulin content of breast secretions diminished during the 1st week post-partum and only IgA persisted in appreciable amounts until weaning. Intestinal fluid was immunoglobulin-poor except for high levels of IgA, which reaffirms the secretory origin of the molecule.
Initiation of Antibody Responses by Different Classes of Lymphocytes
Authors: Strober Samuel
Published: December 2022

Thoracic duct cells from normal and immunized Lewis rats restored the antibody response of x-irradiated hosts to Salmonella typhi flagella and horse spleen ferritin. Elimination of the large dividing lymphocytes by treatment of the cell donors with the mitotic inhibitor, vinblastine, or by in vitro incubation of the thoracic duct cells markedly reduced the responses restored by cells from normal donors. On the other hand, the responses restored by thoracic duct cells from immunized donors were not reduced by elimination of dividing cells and were dramatically enhanced in the case of horse spleen ferritin. These findings suggest that rapidly dividing thoracic duct cells play an important part in the immunologically competent unit for S. typhi flagella and horse spleen ferritin in normal rats but not in immunized rats.
An in Vitro Correlative Assay for the Immunosuppressive Activity of Horse Anti-Rat Lymphocyte Sera: Estimation of Lymphocytophilic Antibody Activity Using 51Cr-Labeled Thymocytes
Authors: Bach Michael K. , Brashler John R. , Perper Robert J.
Published: December 2022

An in vitro method for determining equine cytophilic antibody activity against rat lymphocytes is presented. Because of its reliance on a radioactive counting end point, the assay is rapid, sensitive and capable of handling large numbers of sera. There is a good correlation between the in vitro activity of these sera and their ability to prolong the survival of skin homografts in rats.
Biologic Activities of Peromyscus 7Sγ1 and 7Sγ2 Globulins
Authors: Coe J. , Peel L. , Smith R. F.
Published: December 2022

Peromyscus 7Sγ1- and 7Sγ2-globulins which contained antibody to hen egg albumin (HEA) were isolated, quantitated, and tested for their ability to mediate passive cutaneous anaphylaxis (PCA) and to fix complement. Peromyscus antibody of the 7Sγ2 class induced a PCA reaction in guinea pigs with as little as 4.9 ABC units of antibody/milliliter. With 40 times as much 7Sγ1 antibody, however, no PCA reaction was produced in guinea pigs. In contrast, only Peromyscus 7Sγ1 anti-HEA induced the PCA reaction in Peromyscus. Complement fixation and hemolysis of HEA-conjugated erythrocytes were found only with antibodies of the 7Sγ2 class.
Streptococcal M Protein Vaccines, Rheumatic Fever and Human Histocompatibility Antigens
Authors: Fox Eugene N. , Peterson Raymond D. A.
Published: December 2022

Within the limits of our techniques, we could not demonstrate any serologic cross-reactions between streptococcal M proteins and human histocompatibility antigens, nor could we detect antibodies to human transplantation antigens in persons during the acute or convalescent phases of rheumatic fever, or subsequent to the administration of streptococcal M protein vaccine.
An Immunologic Marker Technique for the Cendehill Vaccine Strain of Rubella Virus
Authors: Oxford J.S. , Potter C.W.
Published: December 2022

Rabbits were immunized with serial, intramuscular doses of low tissue culture passage“wild” and high passage “attenuated” strains of rubella virus. Rubella hemagglutination-inhibiting antibody was detected in animals immunized with two serial doses of 3 log10 TCD50/ml of low passage strains. However, no antibody was detected in animals immunized with 3 log10 TCD50/ml of the Cendehill vaccine strain of rubella virus. A reisolate virus from a person immunized with Cendehill rubella vaccine also had markedly reduced immunogenic properties in the rabbit. Experiments indicated that the Cendehill vaccine virus had a markedly decreased ability to infect rabbits by the intranasal route. The technique may be a useful marker test for this strain of rubella vaccine. The RA/27/3 and the HPV 77 rubella vaccine strains also had a reduced immunogenicity for rabbits but the results were too variable to be used as a marker test for these two strains.
Immunoprecipitation Analysis of Soluble Complement-Fixing Antigens of Dengue Viruses
Authors: Russell Philip K. , Chiewsilp Dumrong , Brandt Walter E.
Published: December 2022

Soluble complement-fixing antigens were obtained from brains of suckling mice infected with each of the four dengue serotypes. Brain suspensions were partially purified by clarification with protamine sulfate, precipitation of the antigen with ammonium sulfate, and gel filtration on Sephadex G-100. Antigenic relationships of the partially purified antigens were examined by complement-fixation and Ouchterlony type immunoprecipitation. Discrete precipitin lines were formed with soluble complement-fixing antigens and hyperimmune mouse ascitic fluid. An antigenic determinant common to all four dengue serotypes was demonstrated by both complement-fixation and immunoprecipitation. Cross-absorption experiments confirmed the presence of the group antigen. Spur formation indicated the presence of a unique type-specific antigen for each serotype. Immunoprecipitation patterns indicated that both group- and type-specific antigenic determinants were on the same molecule and that separate antibodies were formed to each determinant.
Studies of Maleylated Chains of Human IgM
Authors: Habeeb A. F. S. A. , Schrohenloher Ralph E. , Bennett J. Claude
Published: December 2022

An approach to the isolation and physicochemical study of the chains of IgM is presented. The chains were separated after mild reduction in the absence of guanidine as the maleylated derivatives. The maleylated μ and κ chains were isolated in pure form with their intrachain disulfide bonds intact by gel filtration on Sephadex G-200 in Tris-glycine buffer, pH 8.9. Disulfide bond titration of these modified chains revealed 17 such bonds per 7S IgM subunit (5 interchain + 8 intrachain per 2 μ + 4 intrachain per 2 κ). These findings agreed well with the finding of 18 disulfides per subunit obtained on reduction of 19S IgM for 6 hr in 3M guanidine. The maleylated chains were soluble permitting the determination of the molecular weight from the sedimentation and diffusion coefficients in the absence of denaturing agent. The molecular weight determined from these values for the maleylated μ chain was 80,000 and that for the maleylated κ chain was 23,000. Based on the number of maleyl groups on the respective chains, the molecular weight of unmodified μ chain was 75,000 and that of unmodified κ chain was 22,000.
Genetic Control of the Immune Response in Mice
Authors: Gasser David L.
Published: December 2022

The production of agglutinating antibodies to the Ea-1a erythrocytic alloantigen in Mus musculus has previously been shown to be controlled by a single Mendelian recessive factor when strain YBR was crossed with BALB or CBA. It has now been determined that when YBR is crossed with C57BL/10, responsiveness is dominant and a strong nongenetic effect occurs. When positive assortative matings were made between YBR-B10 F2 segregants, a significant response to selection occurred, and estimates of the heritability of the response could be obtained. After one additional generation of selection the high responder line included 81% responders while the low responder line included no responders.
Antibody Production by Host Cells in X-Irradiated Recipients of Allogeneic Spleen Cells
Authors: Storb U. , Weiser R. S.
Published: December 2022

Experiments were performed to test the hypothesis that two different cells, a radiosensitive antigen reactive cell and a radioresistant precursor of the antibody producing cell collaborate in antibody synthesis. Twenty-four hours following 500 R x-irradiation, experimental recipient group C mice were injected with normal allogeneic spleen cells and sheep red blood cells (SRBC). Control recipients of group A were irradiated similarly and injected with SRBC alone while control recipients of group B were irradiated similarly and injected simultaneously with SRBC and allogeneic spleen cells from heavily irradiated (1000 R) mice. Five to 10 days after antigen injection, the numbers of plaque-forming cells (PFC) of donor and host origin were determined in the spleens of the three groups. The contribution of donor and host PFC was distinguished by the use of anti-histocompatibility sera. On day 5, the spleens of mice in group C contained 26 times as many PFC as did the spleens of mice in group A. Of these, 80% were donor cells and 20% were of host type. Thus more than five times as many host PFC developed in group C than in group A. At least some of the stimulatory effect on host PFC was independent of the normal donor spleen cells because the spleens of control group B contained twice as many PFC as did those of control group A. On days 6 and 7, the PFC in group C decreased, while those in control group A increased, possibly due to alloimmune reactions between donor graft and recipient. The injection of antidonor H-2 serum into group C on day 3 suppressed the development of PFC of both donor and recipient type. Evidently survival of donor cells beyond day 3 was necessary for the enhanced recipient PFC response. It is concluded that existing data establish the importance of cell interactions in the immune response but do not prove the existence of an antigen-sensitive cell capable of transmitting antigen-specific information to the precursor of the antibody-producing cell.
Degradation of Immunoglobulin G by Lysosomal Acid Proteases
Authors: Fehr Kurt , LoSpalluto Joseph , Ziff Morris
Published: December 2022

Enzyme preparations from bovine spleen, human spleen, and lysosomal fractions of human synovial membrane tissue have been shown to degrade human IgG at pH 3.5. The major proteolytic activity is due to cathepsin D. The products of digestion have been shown to consist of fragments of IgG ranging from a slightly altered IgG with a molecular weight of approximately 135,000 to extensively degraded Fc and partially degraded Fab fragments.
Immunochemical and Biologic Properties of Rat IgE
Authors: Orange Robert P. , Stechschulte Daniel J. , Austen K. Frank
Published: December 2022

Rat IgE identified by its physicochemical and antigenic characteristics was shown to mediate the immunologic release of both histamine and slow-reacting substance of anaphylaxis (SRS-A) in the rat. The antibodies in homocytotropic antiserum which mediated histamine and SRS-A release after a 2-hr latent period were shown to be heat-labile, to elute on DE-52 cellulose chromatography and Sephadex G-200 gel filtration in the fractions known to contain rat IgE, and to sediment as an approximately 8S immunoglobulin on sucrose density gradient ultracentrifugation. Absorption with mono-specific guinea pig anti-IgE strikingly reduced the capacity of homocytotropic antiserum to mediate passive cutaneous anaphylaxis or the intraperitoneal release of histamine and SRS-A, thereby establishing that these biologic activities of homocytotropic antiserum are attributable to an antigenically unique rat immunoglobulin. The release of histamine and SRS-A mediated by rat IgE is dependent on the participation of the mast cell but does not require the circulating polymorphonuclear (PMN) leukocyte or an intact complement system, thus differing from the PMN leukocyte- and complement-dependent pathway to SRS-A release mediated by the IgGa antibodies of hyperimmune serum.
Cross-Reactions Among Group a Streptococci
Authors: Wiley Grove G. , Bruno Pauline N.
Published: December 2022

Antigens in type 43 streptococci were studied, which were responsible for confusing cross-reactions among numerous strains isolated during recent years from patients with streptococcal illness, notably pyodermas, sometimes complicated by acute glomerulonephritis. Bactericidal activity, which was either reciprocal or in one direction only, accompanied precipitin cross-reactions between type 43 and the other cross-protective types 33, 41, 52, 53, 56, Ross, C506 and G1. The experiments reported here showed that the cross-protective antibodies could be separated from the type-specific M antibodies by absorption with a strain which shared cross-protection, leaving only type-specific M antibodies. Absorption with one of these strains removed precipitin antibodies for the rest of the cross-reacting strains as well. This indicates that these reactions depend on the same antigenic determinants, and as far as tested, the cross-protective antibodies were removed also. The precipitin reactions between type 43 and strains P24037A and AD996 depended upon one of two non-protective R antigens, R1, which these strains shared with the type 43 representative strain, C126/21. The second R antigen, R2, has not been found in types other than 43 or independent of R1. In other work, a non-protective antigen was also found on M+ and M- variants of strain C107/24, the type 33 representative strain. This antigen accounted for positive precipitin tests with M- variants and anti-M sera. It has not been found in cocci of other types.
The Evolution of the Immune Response
Authors: Litman G. W. , Frommel D. , Finstad J. , Howell J. , Pollara B. W. , Good R. A.
Published: December 2022

Highly purified preparations of radiolabeled lamprey immunoglobulin have been employed to study the structure of the immunoglobulin of this primitive vertebrate. The identity of the labeled material with that of the native immunoglobulin has been established. The lamprey immunoglobulin appears to be unique among vertebrate immunoglobulins since it has a unique molecular weight, electrophoretic mobility, and sedimentation velocity. It is also extremely labile and dissociates in solvents in a complex manner. The observations presented show that this molecule lacks counterparts of heavy and light chains and does not possess interchain disulfide linkages between its major subunits. A structural model based on its dissociation behavior is presented and the implications of such a structure in terms of molecular function and phylogenetic development are discussed.
Inhibition of Guinea Pig Complement by Aromatic Amidine and Guanidine Compounds
Authors: Bing David H.
Published: December 2022

The enzymatically active form of the first component of guinea pig complement, C1̄,2 has been postulated to have an anionic binding site (1). The evidence for such a site is based on the ability of guinea pig C1̄ to hydrolyze N-tosyl arginine methyl ester (2). Examination of the competitive enzyme kinetics of C1s̄, the esterase enzyme which is part of the macromolecular complex constituting human C1̄, has provided direct evidence for an anionic binding site (3). This report describes the inhibition of the early acting components of guinea pig complement by aromatic amidine and guanidine compounds in the hemolysis of sheep erythrocytes sensitized with anti-Forssman antibody (EA). The data are consistent with inhibition at the level of C1̄, and further substantiate the evidence for an anionic binding site on guinea pig C1̄.

The compounds employed in this study were α phenylguanidine, p-aminobenzamidine, benzamidine and trimethylphenylammonium.
Pepsin Digestion of Mouse Antibody
Authors: Casey Francis B. , Tokuda Sei
Published: December 2022

In 1960 Nisonoff et al. described a method of producing F(ab′)2 fragments from human and rabbit IgG antibodies (1). The immunoglobulins are mixed with pepsin (3% by weight) in 0.1 M acetate buffer, pH 4.5. The mixture is stirred for 18 hr at 37°C. The enzymatic action is terminated by raising the pH to 7.0 and by precipitating the globulins in 18% (w/v) sodium sulfate. This precipitate contains a relatively homogeneous preparation of 5S F(ab′)2 antibody fragments. Recently, Sinclair (2) reported that when this procedure is followed for mouse γ globulins, the pepsin-digested material is grossly contaminated with intact 7S antibodies. In order to obtain the 5S material it was necessary to subject the pepsin digest to sucrose density gradient ultracentrifugation for 40 hr at 40,000 rpm, at 4°C. We have also encountered difficulties in preparing mouse F(ab′)2 according to the method described by Nisonoff et al.
Plasma Kinin System: Proenzyme Components
Authors: Wuepper Kirk D. , Tucker Ernest S. , Cochrane Charles G.
Published: December 2022

Human lymphoeytes can support the replication of 17-D yellow fever virus when stimulated in vitro with phytohemaggiutinin. Between 7 and 11 days following vaccination with 17-D virus, peripheral lymphocytes lose their ability to support virus replication but regain this capacity by the 25th day post-vaccination.
Immunochemical Studies on the Interaction between Staphylococcal Protein a and γG Globulin
Authors: Kronvall Göran , Messner Ronald P. , Williams Ralph C.
Published: December 2022

Reactivity with staphylococcal protein A is a characteristic of IgG-1, -2 and -4 molecules. Human myeloma globulins of these types either precipitate with protein A or are capable of inhibiting such precipitation. In the present study chemical modifications of human myeloma globulins showed distinct effects on the reactivity with protein A. Carbamylation changed a precipitin reaction to one of inhibition. Acetylation rendered myeloma globulins completely non-reactive with protein A. Quantitative precipitin curves for the reaction between normal human γG or various myeloma globulins and staphylococcal protein A revealed marked quantitative differences with varying amounts of soluble complexes formed. Analytical ultracentrifugation of γG preparations with and without protein A revealed similar differences. No intermediate sized complexes were detected. Our results confirm a previous hypothesis that precipitation in this system is a secondary reaction after the primary combination between protein A and the Fc site on human γG globulin. Ultracentrifugational data also indicated that protein A may be univalent, structurally or functionally, in its reaction with γG.
Studies on Rabbit Lymphocytes in Vitro
Authors: Sell Stewart , Mascari Ruth A. , Hughes Stephanie J.
Published: December 2022

The effect of the divalent and univalent fragments of the immune globulins of burro anti-rabbit lymphocyte serum (BALS), sheep anti-rabbit IgG (SAG) and rabbit anti-rabbit allotypic IgG (RAG) on rabbit lymphocytes in vitro was tested. The divalent (F(ab′)2) fragments of each immune globulin stimulated lymphocytes to transform. The univalent fragments (Fab) of SAG and RAG were also stimulatory, but the univalent fragment of BALS was not stimulatory. A different type of reaction site distribution or stimulatory mechanism for BALS in contrast to SAG and RAG is suggested.
Lymphocytotoxic Antibody Response to Cardiac Allotransplantation in Man
Authors: Mittal Kamal K. , Rossen Roger D. , Butler William T. , Johnson Armead H. , Leachman Robert D. , Nora James J. , Rochelle Donald G. , Brooks David K.
Published: December 2022

Lymphocytotoxic antibodies were found in sera collected prior to transplantation from 3 (14%) of 22 cardiac allograft recipients. Of sera collected after transplantation, multiple samples from 15 of 28 patients (54%) contained lymphocytotoxins. These antibodies were found especially frequently in sera from patients who died of infection or who succumbed after repeated episodes of threatened allograft rejection. The findings of the present study suggest that development of lymphocytotoxic antibodies in cardiac allograft recipients is an ominous sign which, within 1 month after transplantation, may identify individuals who will experience an unfavorable clinical course.
The Mitogenic Effects of Endotoxin and Staphylococcal Enterotoxin B on Mouse Spleen Cells and Human Peripheral Lymphocytes
Authors: Peavy Duane L. , Adler William H. , Smith Richard T.
Published: December 2022

The effects of Salmonella endotoxin and staphylococcal enterotoxin B (SEB) were examined in vitro on mouse and human lymphocyte populations from unsensitized subjects. These substances were found to be extremely stimulatory to mouse spleen cells as measured by 3H-thymidine incorporation. In regards to size, general morphologic features, and staining properties, the cell cultures appear identical to cultures stimulated by phytohemagglutinin (PHA). However, endotoxin failed to stimulate human peripheral lymphocytes while PHA and SEB produced a response over a wide dosage range. Attempts to detect stimulation of human lymphoid cells by endotoxin using various mammalian sera as augmentors were unsuccessful.
Requirement for Cross-Linkage in the Stimulation of Transformation of Rabbit Peripheral Lymphocytes by Antiglobulin Reagents
Authors: Fanger Michael W. , Hart David A. , Wells J. Vivian , Nisonoff Alfred
Published: December 2022

Fab′ fragments of goat anti-rabbit fragment Fab and anti-rabbit fragment Fc were ineffective in the stimulation of transformation of rabbit peripheral lymphocytes. F(ab′)2 fragments had stimulatory activity comparable to that of the intact IgG antibody. Combination of Fab′ fragments of both goat anti-rabbit Fc and antirabbit Fab with the lymphocytes was demonstrated by their capacity to inhibit specifically transformation by the corresponding, intact antibody; in addition, transformation was observed when rabbit lymphocytes, pretreated with univalent fragments of goat anti-rabbit Fc, were subsequently allowed to react with bivalent antibody directed to the univalent fragments. These results indicate that the triggering of transformation by antiglobulin reagents is accomplished through crosslinkage of membrane constituents. Such a mechanism may be relevant to the stimulation of sensitive lymphocytes by antigen and thus may aid in providing a basis for understanding the mechanism of induction and paralysis in the immune response.
Reactivity of Normal Shark Immunoglobulins with Nitrophenyl Ligands
Authors: Leslie Gerrie A. , Clem L. William
Published: December 2022

Normal nurse shark and lemon shark sera contain ∼17S and 7S immunoglobulins with anti-dinitrophenyl activity. Specifically purified 17S “anti-DNP” precipitated in gels with DNP-BSA conjugates, bound to DNP-immunoadsorbents (∼60%) but did not give detectable binding by equilibrium dialysis. The 7S “anti-DNP” did not precipitate in gels, nor did it bind radiolabeled hapten and only about 25% bound to DNP-immunoadsorbents. Of special interest was the observation that specifically purified 17S “anti-DNP” also agglutinated sheep erythrocytes. However, the agglutination of sheep erythrocytes could not be inhibited with DNP-OH. Whole nurse shark serum contains agglutinins to both sheep and chicken erythrocytes. Experiments showed that this was not due to heteroligating antibodies. The possibility is raised that many of the shark immunoglobulin molecules have a dual function, that is, as antibody and as a carrier for certain hormones or other small molecules.
Antibody Formation to Equine Anti-Lymphocytic Globulin (ALG) in Man: Effect on Absorption, Distribution and Effectiveness of the ALG
Authors: Butler William T. , Rossen Roger D. , Reisberg Margaret A. , Mazow Jack B. , Trentin John J. , Judd Kenneth P.
Published: December 2022

Increased rates of plasma clearance of 125I-labeled horse IgG were observed in 16 of 17 patients treated with a prolonged course of anti-lymphocytic globulin (ALG) and in 3 of 5 volunteers who received a single small dose of ALG. Rapid immune elimination of ALG developed within 2 weeks of starting ALG treatment, and was accompanied by (1) the development of circulating anti-horse IgG antibodies which could be measured by passive hemagglutination or immunoadsorbent assays and by (2) the return of established delayed hypersensitivity which initially had been suppressed by the ALG treatment.

Radioactively labeled horse IgG was found in the 7S region of sera of four patients tested before development of rapid immune elimination. In three patients studied after onset of rapid immune elimination of horse IgG, the serum proteinbound radioactivity sedimented more rapidly than 7S in the ultracentrifuge. In one case, the labeled protein was completely removed from the circulation within minutes of injection and broken down into small molecular weight fragments. Absorption of ALG after intramuscular injection was impaired in the presence of anti-horse IgG antibodies.

These results suggest (1) that antibodies form in allograft recipients against ALG despite simultaneous treatment with large doses of azathioprine and corticosteroids, and (2) that these antibodies interfere with the absorption, distribution and the immunosuppressive effectiveness of ALG.
The Interferon Refractory State: in Vivo and in Vitro Studies of Its Mechanism
Authors: Borden Ernest C. , Murphy Frederick A.
Published: December 2022

Interferon synthesis is followed by a period of refractoriness during which restimulation with inducer fails to elicit an interferon response. This period persisted in mice absolutely for 4 days and partially for 13 days; interferon was initially induced with Newcastle disease virus (NDV) and animals were restimulated with an unrelated viral inducer. The development of refractoriness was found to be a graded function of the amount of inducer given and interferon made. Exhaustion of interferon-producing capabilities did not cause the refractory state; it developed both in vivo and in vitro after doses of NDV which elicited a less than maximal response. Furthermore, after an amount of inducer was given which was insufficient to infect all cells with complete infectious virus, refractoriness developed in cultures. In both mice and L cells refractoriness lagged significantly behind maximal interferon production; it was not discernible for 12 to 24 hr after measurable interferon response. Using this delay in the onset of refractoriness, partial biologic separation of repressive activity from antiviral activity of interferon preparations was achieved. Pretreatment of L cell monolayers with 75 units/ml of interferon preparations, collected 36 hr after inoculation of inducer, repressed interferon synthesis to a decidedly greater degree than those collected at 8 hr. Pretreatment of L cell monolayers with small amounts of interferon, 2 units/ml, enhanced interferon production to NDV. These findings suggest a newly synthesized repressor as the mediator of the interferon refractory state.
Immunofluorescent Localization of Human Immunoglobulin in Tissues from Cardiac Allograft Recipients
Authors: Rossen Roger D. , Butler William T. , Reisberg Margaret A. , Brooks David K. , Leachman Robert D. , Milam John D. , Mittal Kamal K. , Montgomery John R. , Nora James J. , Rochelle Donald G. , Rickaway Ruth
Published: December 2022

Direct immunofluorescence studies of 14 human cardiac allografts revealed extensive deposits of immunoglobulin in the sarcolemma, muscle fibers, and in successive layers within the thickened intima of the coronary arteries. In contrast, original hearts from eight recipients and biopsies of six donor hearts taken before transplantation showed little bound immunoglobulin. Sarcolemma-bound IgG was seen significantly more frequently in hearts which succumbed to early rejection. Sera collected before transplantation from 15 to 17 cardiac allograft recipients contained antibodies reactive with cardiac muscle from unrelated donors by indirect immunofluorescence, but not with kidney tissue or skeletal muscle from the same unrelated donors. Pretransplant sera from eight of nine patients also contained antibodies which reacted with the patient's own heart. These results suggest that human cardiac allografts provoke a strong humoral as well as a cell-mediated rejection response and provide evidence which links deposition of immunoglobulin within the arterial intima to the evolution of the obliterative vascular lesion seen in the majority of the cardiac allografts.
Qualitative and Quantitative Studies of Cytotoxic Immune Cells
Authors: Canty Timothy G. , Wunderlich John R. , Fletcher Freeman
Published: December 2022

A quantitative assay of in vitro cell-mediated cytotoxicity has been used to detect the appearance of cytotoxic cells in the spleens of immunized BALB/c mice. Biometric techniques which utilize dose response curves suggested quantitative and qualitative changes in immune activities of different lymphoid populations. The immune responses to intraperitoneal injections of allogeneic tumor cells were studied as functions of both time and immunizing cell dose. Spleen cell-mediated cytotoxicity was detectable 3 days after immunization. The response was maximal after about 10 days and then fell gradually. The highest response to a single intraperitoneal exposure occurred after injection of 107 EL-4 cells. Higher doses of cells resulted in lower responses. The response to a second immunizing dose of EL-4 cells was quicker and reached a peak at about 7 days. In addition to changes in total immune activity, a qualitative change occurred in cell-mediated immunity during the course of primary and secondary responses. With an increase in total immune activity, cytotoxic activity of the average immune cell appeared to increase. The time course and immunizing dose dependence of cytotoxic antibody levels and cell-mediated responses were similar after primary immunization. Hyperimmunization resulted in increasing cytotoxic antibody titers but decreasing cell-mediated activity. Cellular immunity in the hyperimmune animals appeared to have been stimulated but effectively suppressed. This suppression was reversible by transfer of the hyperimmune spleen cells with antigen to sublethally irradiated hosts.
The Significance of the Mast Cell Response to Bee Venom
Authors: Higginbotham R. D. , Karnella S.
Published: December 2022

Local tissue mast cells were found to rapidly secrete their heparin-containing granules in a dose-related response to subcutaneous injection of relatively small amounts of bee venom. The shed granules formed complexes with cationic protein in vivo and these complexes were subsequently ingested by adjacent mononuclear cells. Addition of heparin to bee venom in vitro resulted in the formation of complexes with reduction in both cytolytic and lethal activities of the venom. It is suggested that mast cell granules may, during transfer from mast cells to phagocytic cells, sequester noxious cationic proteins of bee venom and that dermal mast cells may thus be strategically situated and uniquely adapted to serve as a local means of resistance to bee sting.
The Interaction of Human IgM and Clq
Authors: MacKenzie Malcolm R. , Creevy Nancy , Heh Mona
Published: December 2022

The interaction between human IgM Waldenström's protein and human Clq is demonstrated. One mol of IgM combined with 1 mol of Clq indicating a valence of 5 for C1q. The division of IgM proteins into at least two groups by their affinity for Clq is directly confirmed.
Immunologic Investigations of Meningococcal Disease
Authors: Edwards Earl A.
Published: December 2022

Patients with fatal fulminant meningococcemia had readily detectable levels of soluble group-specific meningococcal antigen in their sera. Patients with meningococcal meningitis who recovered did not give a positive test for meningococcal antigen with the test described (counterimmunoelectrophoresis). The test is rapid, sensitive and specific for meningococcal group-specific antigens. Three patients had levels of 1.5 µg/ml to 4.5 µg/ml of meningococcal antigen in their acute-phase blood samples. Such levels may signal the onset of fulminant meningococcemia. An important relationship yet to be determined is the threshold level of circulating meningococcal antigen in the serum and the reversibility of the disease process. Preliminary observations indicate such a level may exist. The rapidity and specificity of the counterimmunoelectrophoresis test for meningococcal antigen should permit its use in studying spinal fluids, as well as serum, for rapid identification of the infecting organism. Eight spinal fluids have been examined to date for Neisseria meningitidis antigen. The results all agreed with the isolation and identification of meningococci by bacteriologic methods.
Study of the Interaction of Concanavalin A with Staphylococcal Teichoic Acids
Authors: Reeder W. J. , Ekstedt R. D.
Published: December 2022

The interaction of Concanavalin A with teichoic acids from S. aureus and S. epieermidis has been studied by gel diffusion and precipitation in a fluid system. It was found that Concanavalin A reacted with alpha-glucosylated but not with beta-glucosylated teichoic acids from S. epidermidis. It was similarly found that Concanavalin A reacted only with the alpha-linked N-acetylglucosamine teichoic acids from strains of S. aureus. These observations were substantiated by quantitative precipitin and hapten inhibition studies using specific antisera to the staphylococcal strains studied. The usefulness of the interaction of Concanavalin A with teichoic acids in studying the immunochemistry of these antigens was demonstrated.
The Antigen Receptor in Delayed-Type Hypersensitivity
Authors: Hill William C.
Published: December 2022

Antigen-specific receptors have been demonstrated on lymphoid cells from guinea pigs sensitized to give delayed-type hypersensitivity reactions. A unique affinity label was used to trace the reactive molecule. After solubilization of the cell membrane, two specifically labeled components of restricted molecular weight were isolated by chromatographic methods.
Virus-Specific Immunologic Depression in Mice Following Combined Immunization and Cyclophosphamide-Induced Immunosuppression
Authors: Weiner Leslie P. , Cole Gerald A. , Nathanson Neal
Published: December 2022

The technique of antigenic stimulation followed by cyclophosphamide-induced immunosuppression was used in an attempt to produce virus-specific immunologic depression which outlasted the acute effects of the drug. A series of experiments indicated that this approach was feasible and that animals so prepared showed an enhanced susceptibility specific for the virus used for immunization-suppression.
The Effect of Ellagic Acid on Delayed Hypersensitivity Reactions in Guinea Pigs
Authors: Schwartz Howard J. , Zimmerman Theodore S.
Published: December 2022

Ellagic acid suppresses the cutaneous reactivity of guinea pigs with active delayed hypersensitivity. The intensity of Arthus reactions is reduced, and both heterologous passive cutaneous anaphylaxis reactions and the vascular component of delayed hypersensitivity lesions are significantly depressed.
The Fifth Component of the Guinea Pig Complement System
Authors: Cook Charles T. , Shin Hyun S. , Mayer Manfred M. , Laudenslayer Karen A.
Published: December 2022

A procedure is described for isolation of guinea pig C5. The product has a sedimentation coefficient of 7.8S and a molecular weight of about 180,000. It is free of the other eight complement components and no substantial non-complement contamination is present, as indicated by immunoelectrophoretic tests.
Observations on a Human Antiglobulin of Intermediate Type
Authors: Gentle T. A. , Fudenberg H. H. , Gold E. R.
Published: December 2022

Antiglobulin Fi. reacts with its own IgG, with some human IgG's of all four subclasses and with rabbit antibody IgG. The determinant is present in the sera of all races tested. It has also been found in the sera of apes and Cercopithecus monkeys, but only in a few high-titer anti-D IgG. It is present in cord serum and serum of aged subjects but is present in very small amounts, if at all, in early fetal sera. It is tentatively assumed that it is present in a basic amino acid sequence of the Fc fragment of the heavy chain of IgG.
Passive Transfer of Delayed Hypersensitivity in Guinea Pigs: Role of Conventional Antibody and Antigen
Authors: Dupuy J. M. , Good R. A.
Published: December 2022

Plasma from irradiated sensitized donors will passively transfer delayed hypersensitivity to DNP-BSA. The transfer is specific and cannot be attributed to transfer of demonstrable γ1 or γ2 antibodies, antigen or super-antigen of DNP-BSA specificity.
Studies on Rat Complement: A Method for Titration of Rat C1, C2, C3, C4, As Well As C5, and the Effect of Rabbit Nephrotoxic Serum on the First Five Components of Complement in Rat Serum
Authors: Miyakawa Yuzo , Sekine Teruaki , Shibata Seiichi , Nishioka Kusuya
Published: December 2022

A method for the titration of the first five components of complement in rat serum is reported. With the guinea pig assay system, rat C4 could not be detected and the titer of rat C2 was low. By contrast, rat C4 could be specifically measured with EDTA rat C and rat C2 could be more efficiently measured with EDTA rat C. Three hours after the injection of rabbit nephrotoxic serum which was proved to fix complement in vitro, the hemolytic titer of complement in rat serum dropped from 106 to 24 and the titer of C1, C4, C2, C3 and C5 lowered to 1/32, 1/8, 1/5, 1/2 and 1/2 of the original respectively.
Specific Antibody Against the First Component of Guinea Pig Complement
Authors: Okuda Tomoko , Tachibana Takehiko
Published: December 2022

The use of antibodies against complement components has provided helpful information concerning the properties and mode of action of complement. Studies with antisera against human C1 sub-components (C1q or C1s) have been reported (1–3), but an antiserum against the first component of guinea pig complement (C1gp) has so far not been obtained in a reliable form.

We succeeded in preparing 5S fragment antibody against C1gp that gave a single precipitin line in Ouchterlony gel diffusion, as described in this communication.

Immunization. An immune precipitate was prepared by mixing γ globulin from rabbit antibovine serum albumin (BSA) with BSA at optimal proportions. After partially purified C1gp (4) was allowed to react with the immune precipitate, the complex was washed several times in gl-GVB++ (5). This Ag-Ab-C1 complex in complete Freund's adjuvant was injected into the footpads of a rabbit. One week afterward a second injection was given subcutaneously.
Anti-Antiserum Antibody as a Cause of Double Precipitin Rings in Immunoglobulin Quantitation and Its Relation to Milk Precipitins
Authors: Ammann Arthur J. , Hong Richard
Published: December 2022

During the process of quantitating immunoglobulins by the Mancini technique (1) double precipitin rings were observed in a number of instances despite the use of well-absorbed monospecific antisera (Fig. 1). At first these were attributed to lipemic sera, but double precipitin rings continued to be observed even with the use of agarose and nonlipemic sera. Non-antigen antibody precipitation was also considered as a possibility. Several observations suggested that the double rings might be true antigen-antibody reactions. Buckley and Dees (2) reported that individuals with selective IgA deficiency had an increased incidence of anti-milk antibodies, and Tomasi (3) and Schlegel et al. (4) observed that patients with selective IgA deficiency had antibodies against normal goat serum. We noted that double rings occurred with greatest frequency among patients with selective IgA deficiency who had anti-milk antibodies, and that double rings were present only with certain antisera.
A Quantitative Assay for Cell-Bound Antibody Using a Radiolabeled Coombs-Type Antiserum to a γG Myeloma Protein
Authors: Reif Arnold E.
Published: December 2022

Harder and McKhann have introduced an isotopic antiglobulin technique for measuring antibodies to cell-surface antigens (1). Sparks et al. have refined the technique to detect alloantibodies formed after first-set rejection of skin grafts (2). Further, Ting and Herberman have used it to study the humoral antibody response to a tumor-specific cell surface antigen (3).

Two additional refinements are reported in this paper. The first is the preparation of a Coombs antiserum by immunizing rabbits with a highly purified mouse myeloma protein of γG type, thereby producing highly specific antibodies which cross-react with normal mouse γG globulin (4). The second is the purification of these antibodies by use of an insoluble antigen complex. The resulting purified Coombs antibody is radiolabeled and binds avidly to cell surfaces coated with mouse γG antibody. A very desirable end result, namely direct proportionality between the quantity of mouse antibody incubated with the cells and the amount of radiolabeled Coombs serum bound to the cell surface, has been obtained.
Phosphonate Inhibition of the Accumulation and Retention of K+ by Rabbit Neutrophils in Relation to Chemotaxis
Authors: Becker Elmer L.
Published: December 2022

The finding that the accumulation of K+ by rabbit peritoneal neutrophils is inhibited by p-nitrophenyl ethyl phosphonate esters is confirmed. This inhibition does not increase with time. Moreover, phenyl ethyl phosphonates, which are very poor inactivators of known serine esterases, inhibit K+ accumulation as efficiently as do the corresponding p-nitrophenyl ethyl phosphonates. These findings support the previous conclusion that organophosphorus inhibitors of serine esterases do not inhibit K+ accumulation of rabbit leukocytes by inactivating a serine esterase. The cell-dependent inhibition of chemotaxis of rabbit neutrophils by p-nitrophenyl ethyl phosphonates increases with time. Phenyl ethyl phosphonates do not inhibit chemotaxis but may actually enhance it. These findings indicate that the cell-dependent inhibition of chemotaxis by p-nitrophenyl ethyl phosphonates does not depend on the inhibition of K+ accumulation. They provide further support for the hypothesis that the cell-dependent inhibition of chemotaxis depends on the inhibition of an “activated esterase.” The inhibition by p-nitrophenyl ethyl phosphonates of K+ retention by rabbit neutrophils increases with time. The phenyl ethyl phosphonates are much less active in this regard. Evidence is presented that the inhibition of K+ retention probably cannot account for the cell-dependent inhibition of chemotaxis.
The Evolution of the Immune Response
Authors: Litman G. W. , Frommel D. , Finstad J. , Good R. A.
Published: December 2022

Immunoglobulins were isolated from serum of the bowfin, Amia calva, which had been immunized with Brucella abortus. The isolation procedures involved gel filtration on agarose and Sephadex G-100 and ion exchange on DE52 cellulose columns. Both the high and low molecular weight immunoglobulins were shown to possess a slow β electrophoretic mobility. The most cathodal migrating serum protein was identified as serum transferrin. The high and low molecular weight immunoglobulins showed complete antigenic identity with a variety of antisera. The s20,w values of the two isolated immunoglobulins were 13.6 and 6.3S, respectively. The hexose, hexosamine, fucose, and sialic acid contents of high and low molecular weight immunoglobulins were similar and revealed a relationship to mammalian IgM. Peptide mapping emphasized the homology of primary structure of the two immunoglobulin forms. Conformational studies demonstrated the presence of β-sheet structure which also is seen in human IgM. Solvent perturbation difference spectra in 20% methanol revealed a tyrosine exposure similar to that of human IgM; by contrast, an exposure of tryptophan was noted in the bowfin immunoglobulin and was not demonstrable in mammalian IgM.