bluuebunny/crossref_metadata_2025 huggingface dataset:
Hemolysis of Sheep Erythrocytes in Guinea Pig Serum Deficient in the Fourth Component of Complement
Authors: May Joseph E. , Frank Michael M.
Published: December 2022

Sheep erythrocytes sensitized with 20 to 40 times the quantity of IgG or IgM antibody required for lysis via the classical complement pathway undergo lysis in C4-deficient guinea pig serum (C4D) or C2-deficient human serum (C2D). The mechanism of this lysis was analyzed in detail, and was found to have an absolute requirement for a Cl-like protein and components of the alternate complement pathway.

Three independent lines of investigation suggested the requirement of C1 for lysis in the deficient sera: 1) C4D serum depleted of C1 by low ionic strength precipitation sustained a marked loss of lytic activity which was overcome by the use of the complement-cell intermediate EAC1. 2) C2D serum depleted of C1 by use of a monospecific anti-C1q immunoabsorbent sustained marked loss of hemolytic activity. Addition of properdin to such sera did not restore hemolytic activity. These absorbed sera were able to lyse the cellular intermediate EAC1. 3) Mg-EGTA completely inhibited lysis in C2D and C4D serum by this pathway while allowing normal function of the alternate complement pathway. Function of C4 and C2 in this lytic mechanism was further excluded by the absence of C2 consumption in C4D serum incubated with heavily sensitized RBC, the failure of anti-C4 to inhibit the lytic reaction, and the inability to lyse heavily sensitized RBC by the sequential addition of purified C1, C2, and C EDTA. Participation of components of the alternate complement pathway was suggested by: 1) conversion of the C3 proactivator by heavily sensitized RBC, 2) abolition or marked reduction of the cytotoxic capacity of C4D and C2D serum heated at 50°C for 30 min, conditions which did not deplete C1, and 3) abolition of lysis in C4D or C2D treated with 0.03 M hydrazine. This lytic mechanism utilizes the later acting complement components as shown by complete absence of lysis in rabbit serum lacking C6.
Recombination of the Polypeptide Chains of a Waldenström's IgM That Binds IgG
Authors: Schrohenloher Ralph E. , Mestecky Jiri
Published: December 2022

An IgM protein that reacted with human IgG was isolated from the plasma of a patient with Waldenström's macroglobulinemia. The IgG-binding site was localized to the F(ab′)2µ fragment of the IgM molecule and the activity was shown to be specific for structures present in the Fcγ fragment of the IgG. µ-and κ-chain fractions were isolated from the IgM after reductive cleavage. Recombination of equimolar quantities of these fractions through both non-covalent and disulfide bond formation resulted in significant levels of anti-IgG activity, as measured by the agglutination of IgG-coated tanned erythrocytes and precipitation of heat-aggregated IgG. This activity was detected shortly after recombination and increased to a maximum after 2 or 3 weeks of storage at 2°C. Ultracentrifugal studies demonstrated that the active products were heterogenous and sedimented over the range of approximately 16S to 40S. Neither chain was active when carried through the recombination procedure in the absence of the other. Maximum anti-IgG activity was recovered from mixtures containing equimolar or greater quantities of the κ-chains. Also, only insignificant levels of activity were detected upon substitution of µ- or κ-chains from another Waldenström's protein that did not bind IgG. The results demonstrated that active molecules were regenerated from the separated polypeptide chains of the IgM and indicate that both chains contributed to the formation of the active site.
Quantitation of Mouse Mammary Tumor Virus (MTV) Virions by Radioimmunoassay
Authors: Cardiff Robert D.
Published: December 2022

A quantitative radioimmune assay for the mouse mammary tumor virus (MTV) has been developed. The assay used highly purified MTV virions which were enzymatically externally labeled with 125I. The radioimmune precipitation (RIP) of 125I-MTV with anti-MTV was specifically inhibited with unlabeled MTV. The degree of inhibition of RIP was directly related to the quantity of unlabeled MTV added. The assay reproducibly detected as little as 5 ng of intact virus. The RIP of 125I-MTV with anti-MTV was not inhibited by murine mammary cell extracts, milk, or “C-type” virus particles. Important variables in this system are discussed.
Quantitative Phagocytosis by Neutrophils
Authors: Ward P. A. , Zvaifler N. J.
Published: December 2022

By using the radiolabeled immune complex method for studying phagocytosis by neutrophils, release of the following lysosomal enzymes has been demonstrated: the C5-cleaving enzyme, β glucuronidase, and acid phosphatase. Enzyme release is directly proportional to cell uptake of immune complexes and is suppressed in antigen excess. It has also been shown that human IgM rheumatoid factor, but not IgM lacking in rheumatoid factor activity, inhibits phagocytic uptake of immune complexes and, correspondingly, depresses the release of lysosomal enzymes.
Isolation of Mucin from Human Submaxillary Secretions
Authors: Baig M. Mansoor , Winzler Richard J. , Rennert Owen M.
Published: December 2022

Human submaxillary mucin isolated by precipitation with cetyltrimethylammonium bromide dissolved in 3 M sodium chloride was subjected to gel filtration on Sephadex G-200. This glycoprotein was free of contaminating proteins as determined by gel filtration, disc electrophoresis, and immunologic studies. The molecular weight was between 500,000 and 1,000,000. The amino acid composition was similar to submaxillary mucins from other species; however, the carbohydrate composition of human mucins varied significantly from different individuals. The mucin from secretor, blood group A individuals is completely absorbed by columns of anti-A antibody coupled with Sepharose 4B, indicating that the submaxillary mucin and the blood group substances in saliva are identical.
Systemic Immunity after Local Antigenic Stimulation of the Lymphoid Tissue of the Gastrointestinal Tract
Authors: Rothberg Richard M. , Kraft Sumner C. , Michalek Suzanne M.
Published: December 2022

The distribution of antibody-producing cells (APC) in the lymphoid tissues of rabbits orally immunized to a low concentration of bovine serum albumin (BSA) was compared to that found in intravenously (i.v.) or subcutaneously (s.c.) immunized animals by determining the concentration of direct and indirect plaque-forming cells in the lymphoid tissues. The spleen was the major site of APC in the i.v. immunized animals and APC predominated in the peripheral lymph nodes of the s.c. immunized group. In contrast, a major site of APC in the orally immunized animals was not directly identified. The concentrations of APC in the spleens of the orally immunized animals were similar to those in the s.c. immunized group, and a relative absence of APC was found in the peripheral and mesenteric lymph nodes, appendix, and Peyer's patches from orally immunized animals.

For determining if systemic immunity is acquired in orally immunized animals in the absence of antibody production by systemic lymphoid tissues, rabbits ingesting the antigen for several weeks were given 50 mg BSA i.v. and the concentrations of APC in the systemic lymphoid tissues were determined 3 days later. Plaque-forming cells were not detectable 3 days after an i.v. injection of BSA given to previously nonimmunized animals, although at this time a marked increase in their concentration was detected in the lymphoid tissues of the i.v. challenged-orally immunized animals.

These studies provide additional evidence that after local antigenic stimulation at gastrointestinal mucosal surfaces, the major site of both secretory and circulating antibody production is the gut-associated lymphoid tissues and that the systemic lymphoid tissues acquire the capacity to produce anti-BSA.
Immunoglobulin on the Surface of Human Lymphocytes. The Detection of Kappa Chain V Region Determinants on B Cells and not on T Cells
Authors: Grey Howard M. , Colon Sonia , Solomon Alan , McLaughlin Carla L.
Published: December 2022

Anti-Vκ region antisera and anti-Cκ,λ region antiserum were used to investigate the nature of antigen recognition units of T cells. The results did not support the hypothesis that T cells contain relatively large amounts of V regions with undetectable amounts of C regions which are buried in the membrane but indicated that with specific anti-κI and anti-κIII antisera the detectable V region determinants were confined to the B cell population.
Thymus Dependence of Lymphocyte Trapping
Authors: Zatz Marion M. , Gershon Richard K.
Published: December 2022

Two models were employed to demonstrate the role of the thymus and T cells in antigen-induced lymphocyte trapping: 1) A graft-vs-host response (GVHR) of parental thymocytes to the antigens of lethally irradiated F1 mice; 2) Challenge of T cell-deprived and reconstituted mice with sheep erythrocytes or keyhole limpet hemocyanin. In both experimental situations T cell-dependent augmented localization of 51Cr-labeled cells was demonstrable in the lymphoid tissues. Adult thymectomy and/or irradiation without T cell supplementation diminished, but did not always abrogate, the trapping response. In the GVHR, trapping preceded the peak DNA synthetic response of the parental thymocytes, and correlated with the dose of injected cells.
Immune Complex in the Renal Glomerulus during Normal Pregnancy. A Study in the Guinea Pig and the Mouse
Authors: Tung Kenneth S. K.
Published: December 2022

Normal pregnant guinea pigs and mice accumulate immune complex in their renal glomeruli. Evidence of immune complex included specific immunofluorescence finding of IgG deposition and typical electron dense deposits on the endothelial and mesangial sides of the glomerular basement membrane. In the guinea pig, C3 was found in the glomeruli; and half of the glomeruli exhibited histopathologic changes although obvious functional disturbance was not evidenced. In the mouse, IgG deposition was not accompanied by C3 or significant histologic changes; and the phenomenon was observed in seven of nine inbred strains of mice studied. Immune complex of pregnancy was found a) to increase with successive pregnancy; b) to appear in the second half of pregnancy; c) not to decrease following pregnancy; and d) to be present in pregnancy of syngeneic mating. Deposition of immune complex, on the other hand, was not found in pseudopregnant mice. The possible nature and significance of the immune complex found in normal pregnancy are discussed.
The Association of J Chain with the Fc Region of Human IgM
Authors: Inman Franklin P. , Ricardo Manuel J.
Published: December 2022

Human IgM (λ) was treated with trypsin at 56°C. A large fragment was isolated by gel filtration and anion exchange chromatography. It sedimented at 10.9S, contained Fcµ epitopes, and upon mild reduction it dissociated to a component which sedimented at 3.4S. Although λ epitopes were exposed serologically, the chains could not be detected by polyacrylamide gel electrophoresis. A protein component of the reduced fragment migrated rapidly anodally during polyacrylamide gel electrophoresis, and was identified as J chain serologically. It was concluded the 10.9S fragment was structurally analogous to Fc5µ and that J chain was covalently associated with it.
Measurement of Antibody Release from Single Cells
Authors: Ghanta Vithal K. , McGhee Jerry R. , Hamlin N. Marlene , Hiramoto Raymond N.
Published: December 2022

Studies of rates of release of IgM were performed in vitro and in vivo. Short term in vivo studies in which 4.98 × 106 tumor cells/mouse were given i.v. in order to quantitate cellular synthetic function showed that the synthetic rate decreased progressively. Samples taken within 1 hr of injection gave rates of release similar to those in the in vitro studies. Nine in vivo samples taken at 30 min gave a rate of 74.03 ± 18.37 and thirteen in vivo samples taken at 1 hr gave 89.98 ± 9.97 molecules/cell/sec. Seventeen separate in vitro measurements gave 76.30 ± 7.44 molecules/cell/sec. The low rate of release of MOPC tumor IgM as compared to the primary response IgM hemolysin to sheep red blood cells is discussed.
Antigen-Binding Thymic Lymphocytes: Specific Binding of Soluble Antigen Molecules and Quantitation of Surface Receptor Sites
Authors: Engers Howard D. , Unanue Emil R.
Published: December 2022

Lymphocytes binding hemocyanin (KLH) have been studied in the thymus and spleen of unimmunized mice. There is a small population of thymic cells which specifically binds KLH—unrelated antigens do not block the binding. A comparison was made of techniques to detect antigen-binding cells. The presence of an antibody to the cell-bound antigen favored the detection of antigen bound to the T cell surface. Quantitation of antigen bound to spleen and thymus cells was determined immunochemically. Spleen cells bind about five times more antigen than thymic cells. Anti-Ig antibodies did not affect consistently the binding of antigen to thymic cells but reduced significantly that bound to spleen cells. We conclude that thymic cells have lower receptor density than spleen cells (3/4 of which may be B lymphocytes) and that this lower receptor density may explain the lower binding strength of the T cell receptor.
Ontogeny of Mouse Lymphocyte Function
Authors: Mosier Donald E.
Published: December 2022

Thymocytes from BALB/c mice ranging in age from 16 days of gestation to adulthood were stimulated with the phytomitogens phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM), or with syngeneic or allogeneic adult spleen cells in the mixed lymphocyte reaction (MLR). Fetal thymocytes responded well to PHA and to allogeneic cells in the MLR, but poorly to Con A and PWM. Neonatal cells showed a sharp decline in PHA and MLR responsiveness and a concomitant increase in Con A and PWM reactivity. The syngeneic MLR was maximal during this period. Adult thymus cells responded poorly to PHA and not at all to syngeneic cells, but showed much increased responsiveness to Con A and PWM. These results are discussed with regard to a model proposing two different but interacting lines of thymus lymphocyte differentiation.
Subpopulations of Mouse T Lymphocytes
Authors: Moorhead John W. , Claman Henry N.
Published: December 2022

The existence of short-lived peripheral T lymphocytes was directly investigated using an in vitro system. Spleen and lymph node cell suspensions from normal LAF1 mice were pulse-treated for 1 hr in vitro with high specific activity tritiated thymidine (“thymidine suicide”). Thymidine suicide allows for the selective, lethal irradiation of those cells which are actively synthesizing DNA, i.e., proliferating cells in S phase, during the pulse period. To determine the “suicide” (loss or inactivation) of T lymphocytes, the isotope-treated cells were stimulated with phytohemagglutinin (PHA). The results showed that: a) a population of dividing (proliferating) T lymphocytes exists in the spleen; b) these cells are not present in lymph nodes; and c) with respect to the spleen, it is the dividing T lymphocytes which are primarily responsible for the PHA response.
Isolation and Characterization of the Proenzyme Form of the C1s Subunit of the First Complement Component
Authors: Valet Gunter , Cooper Neil R.
Published: December 2022

The proenzyme form of C1s was isolated by sequential chromatography of the euglobulin fraction of human serum on DE-32 cellulose, TEAE cellulose, and Sephadex G-200. C1s was found to be an α-globulin with a molecular weight calculated from S and D, of 86,200 daltons. The amino acid composition was unremarkable for a serum protein. Isolated C1s preparations were unable to cleave natural (C4) or synthetic (ALTEE) C1s substrates unless first activated by incubation with C̄1̄r or trypsin. Activation was accomplished by limited proteolytic cleavage.

Isolated C1q, C1r, and C1s formed a macromolecular complex in the presence of calcium which possessed C1 activity. C1 formed in this manner was in the proenzyme form as evidenced by ability to cleave ALTEE. Reconstituted C1, however, was readily activated on addition of aggregated IgG.
Macrophage Control of Time-Dependent Changes in Antigen Sensitivity of Immune T Lymphocyte Populations
Authors: Cohen Benjamin E. , Paul William E.
Published: December 2022

In the course of the immune response to a hapten-protein conjugate, 2,4-dinitrophenyl (DNP)-guinea pig albumin (GPA), a major increase in the in vitro antigen-sensitivity of peritoneal exudate lymphocyte (L) populations occurs. The predominant cell within these populations is the T lymphocyte. In addition a small percentage of macrophages are present. The response measured, antigen-stimulated DNA synthesis, reflects the response of the T lymphocytes but appears to depend upon the presentation of antigen to the T lymphocytes by macrophages. The increase in antigen sensitivity thus might either reflect an increase in the affinity for antigen of the receptors on T lymphocytes or an increase in the efficiency of macrophages in presenting antigen to T lymphocytes. That the latter is the correct interpretation is indicated by experiments involving the mixing of defined lymphocyte and macrophage populations. In these experiments it was shown that macrophages controlled the pattern of antigen sensitivity (relative responsiveness to low and high concentrations of antigen) while the lymphocytes controlled the magnitude of the response to high antigen concentrations. Furthermore, preincubation of cells from animals immunized 7 days earlier with antiserum from animals immunized 14 to 35 days earlier resulted in an increase in the sensitivity of these cells to low concentrations of antigen. This indicates that macrophage-associated cytophilic antibody determines the antigen-sensitivity of the system by allowing macrophages to bind larger amounts of antigen, when antigen is present at low concentration. This, in turn, results in more efficient stimulation of T lymphocytes.

The failure to demonstrate “affinity maturation” of T lymphocyte receptors may be explained in a variety of ways but one important possibility is that there is a more limited heterogeneity in the T cells participating in this response than would be true of a comparable population of B lymphocytes.
Chronic Virus Infection and Immune Responsiveness
Authors: Oldstone Michael B. A. , Tishon Antoinette , Chiller Jacques M.
Published: December 2022

Differences in the immune responsiveness of infected mice against one antigen do not necessarily indicate differences in their total immunologic capacities. Antibodies formed to a variety of antigens in mice infected with lactic dehydrogenase virus (LDV) were compared to those made in uninfected mice. The antigens used were keyhole limpet hemocyanin (KLH), human IgG (HuIgG) and goat red blood cells (GRBC). Against KLH, LDV infected mice made lower immune responses than did uninfected controls, whereas against HuIgG, the immune responses of LDV infected mice were elevated over controls. The formation of antibody to HuIgG was enhanced only soon after primary immunization, was associated with an increased number of direct antibody forming cells in the spleen, and was predominantly of the IgM class. Mice infected with LDV and uninfected mice made similar immune responses to large doses of GRBC, whereas when doses of GRBC were reduced infected mice made lower immune responses.
Membrane Receptors and in Vitro Responsiveness of Lymphocytes in Human Immunodeficiency
Authors: Schiff Richard I. , Buckley Rebecca H. , Gilbertsen Richard B. , Metzgar Richard S.
Published: December 2022

Peripheral blood lymphocytes from 24 patients with well-defined immunodeficiency were studied for membrane characteristics and in vitro responsiveness to various stimuli. The mean percentages of lymphocytes bearing easily detectable surface immunoglobulin (Ig) were similar to those of normals except in patients with infantile x-linked agammaglobulinemia (X Ag) who had a significantly lower percentage of only 1.9% Ig bearing cells (p < 0.001). Despite their marked deficiency in surface Ig bearing cells, patients with X Ag had normal percentages of complement receptor rosettes (EAC, HEAC, or B cell rosettes); percentages of these cells were also normal or elevated in the other patient groups. Percentages of spontaneous sheep erythrocyte rosettes (E or T cell rosettes) were significantly increased in X Ag patients and reduced in patients with ataxia telangiectasia and isolated IgA deficiency. Lymphocyte 3H-thymidine incorporation in response to each of the three mitogens, phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM), was usually normal except in X Ag patients where, in several evaluations, the responses of peripheral blood lymphocytes to PHA were significantly lower than those of normal control cells (p < 0.001). Responses to Con A and PWM were normal or elevated in those same experiments. The selective hyporesponsiveness of X Ag lymphocytes to PHA suggests that these patients may be deficient in a restricted population of T cells. It is possible that this population may be necessary for the full maturation of B cell function. Lymphocytes from all patients, including agammaglobulinemic patients, responded normally to stimulation with one or more antigens.
Letters to the Editor
Authors: Spitler Lynn E.
Published: December 2022

August 28, 1973

Sir: In many manuscripts published recently in the Journal of Immunology and in other journals as well, the authors have used the leukocyte migration inhibition test, and have indicated in the results or discussion that they were measuring production of migration inhibitory factor (MIF). I think that this practice is inaccurate and confusing for the following reasons:

The leukocyte migration inhibition test involves evaluation of migration of peripheral buffy coat leukocytes from capillary tubes or from wells in agarose plates. Although both tests have been shown to correlate with skin test reactivity, the leukocyte migration inhibition test differs from the macrophage migration inhibition test in that the migrating cell population probably represents polymorphonuclear leukocytes rather than macrophages. While it does appear that both tests are mediated by cell free mediators, to my knowledge, there have been no reports on the characterization of the mediator responsible for inhibition of leukocyte migration, and it is not known whether or not this mediator is the same as that responsible for inhibition of macrophage migration.
Origin and Development of Rat Thymus-Dependent (T) Lymphocytes
Authors: Balch Charles M. , Feldman Joseph D.
Published: December 2022

Bone marrow (BM) cells of LBNF1 donor rats, from which detectable thymus-dependent (T) lymphocytes were removed by anti-T lymphocyte antisera and C, were infused into lethally irradiated (IrBM) and into thymectomized lethally irradiated (TxIrBM) BN recipients. The lymphoid tissues of these animals were examined from 6 to 24 days after reconstitution for donor cells (AgB+), donor T cells (AgB+T+), and T cells (host and donor). Donor cells and donor T cells progressively increased in number in all lymphoid tissues of IrBM rats. In TxIrBM rats donor cells increased to a lesser extent and donor T cells decreased 24 days after reconstitution. Spleen and lymph node cells of IrBM and TxIrBM responded little or not at all to PHA and Con A, in vitro. The response was less than expected of the number of T cells present in culture. T lymphocytes can differentiate from bone marrow precursor elements and thymus seems to be a necessary organ for maintenance of T cell levels. The newly differentiated T cells differ from normal T cells in their response to mitogens.
The Hapten-Binding Region of Protein 460
Authors: Jackson Peter , Richards Frank F.
Published: December 2022

Protein 460, a mouse γ A2 myeloma immunoglobulin, binds at least two structurally diverse haptens, menadione thioglycollate and ε-2, 4-dinitrophenyl-l-lysine, at different sites within the combining region. A “free” cysteine sulfhydryl group within the heavy chain is apparently near the menadione-binding site. In the intact protein, this cysteine residue has been alkylated with iodoacetamide (I-14C); the isolated heavy chain digested with cyanogen bromide, trypsin, and thermolysin; and a nonapeptide has been isolated bearing the 14C 5-methyl carboxy amido cysteine residue. The tentative amino acid sequence of this peptide is Val-Leu-Ser-Gly(Glu-Ala) Cys-Arg-Pro.
Serum Factors in Mammary Neoplasia: Enhancement and Antagonism of Spleen Cell Activity in Vitro Detected by Different Methods of Serum Factor Assay
Authors: Blair Phyllis B. , Lane Mary Ann
Published: December 2022

Spleen cells from female mice bearing progressively-growing MTV-induced mammary tumors reduced tumor cell survival in microcytotoxicity tests. Serum from these females contained blocking factors which inhibited spleen cell activity either partially or completely, depending upon the method of serum assay, whenever serum and target cells had not been obtained from the same donor. Serum obtained from target cell donors had a reverse effect when tested by two of three serum assay methods; tumor cell survival was significantly decreased. More than one type of mammary tumor cell antigen (and host response) was detected in these tests.
Characteristics of Secretory Immune System in Human Middle Ear: Implications in Otitis Media
Authors: Ogra Pearay L. , Bernstein Joel M. , Yurchak Anthony M. , Coppola Peter R. , Tomasi Thomas B.
Published: December 2022

The middle ear washings and biopsy specimens of middle ear mucosa from 20 patients with secretory otitis media were examined for the distribution of major classes of immunoglobulin; presence and nature of antibody activity against mumps, measles, rubella and poliovirus; and immunologic localization of immunocompetent lymphoid cells. Appreciable amounts of secretory component, and γG, 7SγA, and 11S secretory γA immunoglobulin were regularly found in the ear fluids. The ratio of γG: γA immunoglobulin in the serum and middle ear fluids was approximately 10:1 and 10:4, respectively. Fluorescent antibody staining of mucosal tissues of middle ear demonstrated characteristic staining for secretory component in the surface epithelium, and the presence of γG and γA containing lymphoid tissue in the submucosa. Tissue culture studies demonstrated in vitro synthesis of γA and γG immunoglobulin and the secretory component in the mucosal tissues of middle ear. Specific antibody activity in the middle ear fluid against the viruses listed above was essentially limited to γA immunoglobulin, although such activity in the serum was predominantly associated with γG immunoglobulin. These data suggest the existence of a distinct secretory immune system in the middle ear. The implications of these observations may be applicable to the mechanism of protection, or pathogenesis of microbial infections in the middle ear.
Identification and Quantitation of Thymus-Derived Lymphocytes in Human Peripheral Blood
Authors: Bobrove Arthur M. , Strober Samuel , Herzenberg Leonard A. , DePamphilis Jean D.
Published: December 2022

A highly specific anti-T cell antiserum (ATCS) was developed by extensively absorbing a goat anti-human thymocyte antiserum with human red cells and buffy coat cells from a patient with chronic lymphocytic leukemia. The ATCS was used in an in vitro cytotoxicity assay for detecting human thymus-derived (T) peripheral blood lymphocytes. A fluorescent cell sorter provided separated populations of normal immunoglobulin (Ig)-bearing and non-Ig-bearing lymphocytes on which to directly test the sensitivity and specificity of the ATCS. The antiserum killed almost 100% of non-Ig-bearing lymphocytes but less than 10% of Ig-bearing (B) lymphocytes and monocytes. Moreover, ATCS was able to abrogate the response to phytohemagglutinin stimulation, a T cell response.

In further studies, Ficoll-purified peripheral blood cells from 12 normal donors were examined for the per cent Ig-bearing cells, monocytes and cells killed by ATCS. A mean of 21% of these cells were B lymphocytes, 17% were monocytes, and 65% were T lymphocytes, thus accounting for 100% of the cells.
Adsorption Properties in Vitro and in Vivo of Antibodies Raised against Rat Brain Blood Vessels
Authors: Day Eugene D. , Rigsbee Lewis C. , Rosenthal J. Thomas , Shih Janet C.
Published: December 2022

Antibodies were raised in three rabbits against isolated rat brain blood vessels in Freund's complete adjuvant, were labeled with 125I, and were analyzed by adsorption techniques in vitro and in vivo. After a booster injection two of the three rabbits displayed hyperimmune reactivity. A correlation was established between the in vitro and in vivo data. By the in vitro sequential adsorption technique five categories of antiblood-vessel antibodies were established: a) reactive with isolated rat brain blood vessels; b) reactive with blood vessels, cultured rat astrocytoma cells, and adult rat brain myelin but not rat kidney embryo fibroblasts; c) reactive with blood vessels, fibroblasts, and myelin, but not astrocytes; d) reactive with blood vessels and myelin, but not fibroblasts or astrocytes; and e) reactive with blood vessels, myelin, astrocytes, and fibroblasts. All these antibodies, when tested for adsorption in vivo in rats, after intravenous injection, were found to localize primarily in kidney and liver tissues and to a lesser degree in spleen and lung tissues. They were not adsorbed by heart, thymus, testes, or brain tissues in detectable quantities. The antibodies that were ordinarily adsorbed by kidney tissue in normal rats were adsorbed by liver tissue in nephrectomized rats. When injected intracerebrally most of the antibodies still did not localize extensively in brain tissue but found their way to kidney and liver tissues as before. The question was therefore raised anew whether an active process might not operate in brain tissue under normal circumstances to prevent antibody build-up even in its vascular bed and whether a breakdown in this mechanism might not result in antibody-mediated brain disease.
Evaluation of T Lymphocyte Effector Function in Immunodeficiency Diseases: Abnormality in Mitogen-Stimulated Interferon in Patients with Selective IgA Deficiency
Authors: Epstein Lois B. , Ammann Arthur J.
Published: December 2022

Interferon production in response to the mitogens phytohemagglutinin and pokeweed was studied as a new index of the competence of thymus-derived (T) lymphocyte effector function in patients with various immunodeficiency states. The results indicated that three of four patients with selective IgA deficiency and normal T cell proliferative response had a depressed T cell interferon response.

Two patients who had received successful thymus transplants had a normal T cell interferon response. Before transplantation one had been diagnosed as severe combined immunodeficiency, and the other, thymic hypoplasia with normal serum immunoglobulin levels. By contrast, a patient with thymic hypoplasia and absent serum IgA, low IgM and normal IgG levels, who had had an unsuccessful thymic graft, had a markedly depressed interferon response. Normal T cell interferon responses were seen in one patient with congenital sex-linked hypogammaglobulinemia and three patients with acquired hypogammaglobulinemia.
A Rabbit IgG Antigenic Marker Detected by 19S Anti-IgG Present in Streptococcal Antisera
Authors: Chiao Jen Wei , Bokisch Viktor A. , Christian Charles L. , Krause Richard M.
Published: December 2022

19S anti-IgG, produced in rabbits immunized with streptococcal vaccines, was used in a hemagglutination-inhibition test with rabbit IgG-coated erythrocytes for detecting an antigenic marker on rabbit IgG. The 19S anti-IgG in a panel of streptococcal antisera had different agglutinating specificities. An antigenic marker on rabbit IgG-Fc was detected in some but not all normal rabbit sera by the 19S anti-IgG in one of the streptococcal antisera. Detection of this antigenic marker in breeders and their offspring suggested that this antigenic marker was inherited. Further studies are required to determine if it is identical with or distinct from one of the known allotypic markers.
A Sequence Restriction in the Variable Region of Immunoglobulin Light Chains from Sharks, Birds, and Mammals
Authors: Stanton Thomas , Sledge Carlos , Capra J. Donald , Woods Roy , Clem William , Hood Leroy
Published: December 2022

A relatively conserved pentapeptide sequence is present in the variable region of normal immunoglobulin λ- and κ-chains from animals as diverse as sharks, birds, and mammals. This same conserved sequence is also present in most myeloma light chains from dog, mouse, and man. The conservation of this pentapeptide sequence over the more than 400 million years since sharks and mammals diverged suggests that this region must carry out a very important, but as yet unknown function. A homologous but less highly conserved stretch of sequence is present in the variable region of immunoglobulin heavy chains.
Cyclic AMP, ATP, and Reversed Anaphylactic Histamine Release from Rat Mast Cells
Authors: Kaliner Michael , Austen K. Frank
Published: December 2022

Reversed anaphylactic histamine release by rabbit (Rab) anti-Rat Fab was independent of serum factors and was established as being noncytolytic in that challenged isolated mast cells still retained incorporated 42K and excluded trypan blue dye after releasing histamine. The release reaction involved the activation of a diisopropyl fluorophosphate (DFP)-sensitive esterase and required energy utilization. Prostaglandin E1 (PGE1) consistently produced a dose-related increase in the mast cell content of cyclic 3′,5′-adenosine monophosphate (cyclic AMP) which at the optimal concentration was 2 to 3 times the average baseline of 4.71 pmoles/106 mast cells (range -1 to 10 pmoles/106 mast cells). Aminophylline and PGE1 alone each inhibited Rab anti-Rat Fab-induced histamine release with concomitant increases in cyclic AMP, and the combination of these agents had a synergistic effect on both parameters. The inhibition of histamine release by the endogenous accumulation of cyclic AMP was reproduced by the exogenous administration of dibutyryl cyclic AMP. The interaction of Rab anti-Rat Fab with isolated mast cells resulted in a depletion of the cellular levels of cyclic AMP, ATP, and calcium in association with histamine release. Although these consequences of membrane perturbation may be unrelated, it seems more likely that they are interrelated and intrinsic to reversed anaphylactic histamine release.
The Role of Iron in Nonspecific Resistance to Infection Induced by Endotoxin
Authors: Elin Ronald J. , Wolff Sheldon M.
Published: December 2022

Studies using an experimental model of the mouse and Candida albicans as the challenge organism attempted to define better the mechanism of nonspecific resistance to infection induced by bacterial endotoxins. In vitro studies showed a positive correlation between the growth of C. albicans in serum from mice obtained at daily intervals for 10 days after endotoxin or saline injection and the percentage iron saturation of the serum. In vivo studies in which one of four different concentrations of iron as ferric ammonium citrate was injected into control and endotoxin-treated mice at the time of challenge with C. albicans showed that the rate of mortality of the mice was directly related to the concentration of iron injected. The endotoxin-induced nonspecific resistance was negated and reversed by iron administration. These studies demonstrate that changes in iron metabolism induced by bacterial endotoxin are a key factor in the mechanism of resistance to C. albicans infection in mice.
A Quantitative Radioisotopic Assay for Hemolytic Antibody by Employing Technetium-99m-Labeled Erythrocytes
Authors: Barth Rolf F. , Singla Om
Published: December 2022

We have developed a quantitative radioisotopic assay for hemolytic antibody with erythrocytes prelabeled with technetium-99m (99mTc), a high specific activity metastable gamma emitter. There was a linear relationship between the amount of 99mTc initially used for labeling and the amount of radioactivity taken up by and subsequently released from erythrocytes which were subjected to complete hemolysis. Hemolytic antibody titrations were carried out with sheep or mouse erythrocytes which had been labeled with varying amounts of either 99mTc or 51Cr and it was concluded that 1 mCi of 99mTc or 100 µCi of 51Cr was sufficient to label adequately the target cells. The same mean log2 endpoint titers were detected in parallel titrations with either mouse anti-sheep RBC serum or an H-2 alloantiserum although the absolute gamma counts of released radioactivity were up to 15 times greater when 99mTc erythrocytes were employed. In contrast to these data on the titration of hemolytic antibodies, it was not possible to use 99mTc-labeled murine thymocytes or lymphocytes as target cells in assays for cytolytic activity of high titer rabbit anti-mouse lymphocyte sera or an H-2 alloantiserum.

The principal advantages of 99mTc compared to 51Cr are its high specific activity, low spontaneous release, and non-reutilizability. From our studies it can be concluded that 99mTc can be successfully employed for the quantitative determination of hemolytic antibodies and that its favorable physical characteristics and ready availability suggest that it may replace 51Cr as a label of choice for erythrocytes.
Activation of the Classic and Alternate Complement Pathways by Endotoxin
Authors: Fine Douglas P.
Published: December 2022

The ability of bacterial endotoxin (LPS) to activate the complement system was studied in guinea pig serum (GPS). In serum chelated with ethyleneglycol tetraacetic acid (EGTA) 10 mM, which permits alternate complement pathway activation but inhibits classic complement pathway activation, lysis of LPS-coated sheep erythrocytes (E-LPS) was inhibited. Furthermore, E-LPS could not decomplement GPS chelated with EGTA. GPS absorbed twice at 0°C with E-LPS retained normal levels of hemolytic complement (CH50) and reacted normally (as measured by CH50 depression) with both zymosan (which activates the alternate pathway) and LPS. The absorbed GPS did not react with E-LPS, whether measured by percentage of lysis of E-LPS or by CH50 depression after incubation with E-LPS. These results suggest that endotoxin is capable of activating both complement pathways but erythrocyte-associated endotoxin can activate only the classic patheay, requiring a serum factor absorbable by E-LPS at 0°C.
Carrier-Specific Enhancement of the Immune Response Using Antigen-Antibody Complexes
Authors: Terres Geronimo , Habicht Gail S. , Stoner Richard D.
Published: December 2022

Antigen-antibody complexes formed at equivalence and injected into adult mice sensitize the mice so that a markedly enhanced antibody response is obtained following an injection of soluble antigen 3 days after the complex injection. The serologic specificity of the enhanced response is determined by the antigen injected on day 3; thus, rabbit anti-dog serum albumin (DSA) in complex with DSA sensitizes the mouse so that when human serum albumin (HSA) is injected on day 3 an enhanced antibody response to determinants unique to HSA is obtained. The antigen injected on day 3 must be related to the antigen in complex in order to produce an enhanced response; tetanus toxoid given on day 3 following injection of DSA-anti-DSA did not stimulate an anti-tetanus response. A mechanism is proposed in which the day 3 antigen-specific bursal-equivalent (B) cells produce an enhanced antibody response as a result of interaction with an increased number of carrier-specific thymus-derived (T) cells which have been stimulated to proliferate by the antigen-antibody complex; in this scheme, the albumin determinants which stimulate T cells to proliferate differ from those which stimulate antibody formation, i.e., B cells.
The Distribution of Lymphocytes with T- and B-Cell Surface Markers in Human Bone Marrow
Authors: Borella Luis , Sen Luisa
Published: December 2022

This study demonstrates that lymphocyte subpopulations are distributed differently in peripheral blood and bone narrow. The proportion of T (thymus-dependent) and B (bone marrow-derived) lymphocytes was simultaneously assayed in the peripheral blood and bone marrow of children with acute lymphocytic leukemia (ALL) in continuous remission for 3 to 8 years who had received no therapy for 1 month to 5 years. T cells were identified by spontaneous rosette formation with sheep erythrocytes (RFL) and B cells by immunofluorescence of surface immunoglobulins. Most bone marrow lymphocytes carried neither B- nor T-cell surface markers. These markers were not detected in 78% of bone marrow lymphocytes and in 39% of peripheral blood lymphocytes. In a majority of these children the proportions of T and B lymphocytes were different in blood and bone marrow. Percentages of RFL and of lymphocytes bearing IgG were greater in blood than in bone marrow in 95% of the patients assayed. In contrast, the proportion of IgM-bearing lymphocytes in the bone marrow of 65% of the children was equal to or greater than in their peripheral blood. In the bone marrow the average ratio of lymphocytes bearing IgG to lymphocytes bearing IgM was 1:5 while it was 1:1 in the peripheral blood. Similar results were obtained when the proportion of T and B lymphocytes were analyzed in a group of nine children in remission who had received no therapy for more than 1 year and who were normal as assayed by several clinical, hematologic and immunologic criteria. Thus, we conclude that a low proportion of T cells and IgG-bearing lymphocytes and a predominance of B lymphocytes with IgM receptors represent the physiologic distribution of lymphocytes in human bone marrow.
Effect of Phytohemagglutinin on Expression of Human Histocompatibility Antigens
Authors: Singal Dharam P. , Villanueva Valentine R. , Naipaul Naresh
Published: December 2022

The phytomitogens, concanavalin A (Con A)2 and phytohemagglutinin (PHA-M) have recently been reported to mask cellular histocompatibility (H-2) antigens on mouse lymphoid cells (1). The antiserum utilized in the masking experiments had antibodies directed against a number of H-2 specificities and was produced in mice after two sequential skin grafts and several (8 to 12) weekly injections of 25 × 106 spleen cells from an allogeneic host in the serum donor (2). It is possible that the phytomitogens may have interfered with the interaction between antibody and the H-2 antigenic sites, perhaps through steric hindrance from a non-H-2 antigen-antibody complex. The present study was designed to study the effects of PHA on the expression of human histocompatibility (HL-A) antigens on peripheral blood lymphocytes with monospecific antisera. We also studied the inhibitory effect of monospecific HL-A antisera on PHA stimulation of lymphocytes.
Immunosuppressive Effects of Anti-lymphocyte Serum in Complement Deficient Mice. II. Evidence That Activity Is Mediated by C3 Independent Antibodies
Authors: Barth Rolf F. , Singla Om , Ward Peter A.
Published: December 2022

The immunosuppressive effects of anti-lymphocyte serum (ALS) have been studied in both normal and C5-deficient mice which were depleted of the third component of complement (C3) by treatment with an inactivator isolated from cobra venom. BALB/c and treated B10·D2 old and new line mice which had been treated with C3 inactivator and either ALS or ALG had a marked reduction in both splenic plaque-forming cells and serum hemagglutinin and hemolysin titers 4 days after immunization with sheep erythrocytes compared to normal serumtreated C3 deficient controls. The effectiveness of ALG was particularly significant since this was devoid of C3. The mean survival time of C57BL/6 skin allografts was significantly prolonged on ALS treated, C3-depleted BALB/c mice compared to controls. These findings indicate that antibody-mediated immune adherence, cytolysis, and phagocytosis that require the participation of C3 are not essential for ALS activity. They support the hypothesis that cytophilic antibodies, which are capable of promoting phagocytosis independent of the complement system, are primarily responsible for the immunosuppressive activity of ALS.
Cellular Proliferation As an Index of Immune Competence
Authors: Asantila Tuula , Vahala Jaakko , Toivanen Paavo
Published: December 2022

Sir: We fully agree with Cunningham and Lafferty (1) that cellular proliferation alone can be an unreliable index of immune competence, particularly when fetal lymphocytes are concerned. Therefore, in our study (2) referred to above, microscopic examination was also employed to show destruction of the tubular epithelium in the rat kidney by human fetal lymphocytes (Fig. 2 in Reference 2). Such a destruction is not obtained with immunoincompetent cells proliferating in a xenogeneic environment (3). For quantitation of the response by the fetal lymphocytes, we preferred cell proliferation indices based on the uptake of a radioactive DNA-precursor. The quantitation revealed an increase in the xenogeneic graft-vs-host competence of fetal lymphocytes with the increasing age of the fetus. These observations clearly indicate that we were not measuring only the spontaneous proliferation of fetal lymphocytes, which has been shown to decrease with the increasing age of the fetus (4).
A Hybrid IgG4-IgG2 Immunoglobulin
Authors: Natvig J. B. , Kunkel H. G.
Published: December 2022

Studies have been described which delineate an unusual myeloma protein as a hybrid molecule consisting of heavy chains that are primarily IgG4 but with the C terminal homology region IgG2. The hybrid character of this protein was demonstrated mainly through the use of a wide variety of antigenic markers which have been localized to specific areas of the heavy chains of the different IgG subclasses. The highly purified myeloma protein contained three IgG4 markers but lacked two others that are found in all other Ig4 proteins. In addition it showed the presence of three IgG2 antigens but lacked five others that are characteristic of this subclass. The IgG4 markers which were present are all found in the CH1 and CH2 homology regions whereas the missing markers are in CH3. The reverse situation applied to the IgG2 antigens. These results indicated that the hybrid protein was made up of IgG4 in the CH1 and CH2 portion of the heavy chain and IgG2 in the CH3 area.

Evidence was obtained for a normal counterpart to this protein in certain Negro sera but not in other populations; the myeloma protein was derived from a Negro. It appeared to represent a genetic variant in this population. The studies on this hybrid protein provided further data for the tentative order of the IgG subclass cistrons in the chromosome. Some indication was obtained for a possible duplication of the IgG2 gene at least in the Negro population.