bluuebunny/crossref_metadata_2025 huggingface dataset:
Bactericidal and Opsonic Properties of C4-Deficient Guinea Pig Serum
Authors: Root Richard K. , Ellman Leonard , Frank Michael M.
Published: December 2022

The bactericidal and opsonic properties of serum obtained from a strain of guinea pigs totally lacking in hemolytically active or antigenically detectable C4 were studied and compared to serum from normal guinea pigs and those heterozygous for C4 deficiency. Bactericidal activity against two strains of E. coli was present in C4-deficiency serum; however, the latent period was prolonged and the rate was slower when compared to that in normal or heterozygous sera. C4-deficient serum bactericidal activity could be restored to normal by mixing with normal serum or by adding partially purified C4 from human serum. Treatment of both C4-deficient and normal serum with heat (56°C for 30 min), 0.04 M EDTA, and zymosan abolished bactericidal activity, whereas 0.03 M hydrazine treatment impaired but did not completely destroy the activity of both sera. C4-deficient serum promoted phagocytosis of S. aureus, type 25 pneumococci, E. coli and C. albicans but kinetic studies employing 14C labeled bacteria indicated that ingestion rates were slower than in normal or heterozygous serum. Prior incubation of pneumococci in serum established that opsonization was the rate-limiting step in phagocytosis of these organisms and that this was significantly diminished from normal in C4-deficient serum. Normal opsonization could be restored by the addition of as few as 10 units of partially purified guinea pig C4. Treatment with heat, EDTA and zymosan abolished opsonization in both normal and C4-deficient sera. Hydrazine treatment diminished the opsonic activity of normal serum against pneumococci but did not significantly affect C4-deficient serum pneumococcal opsonization. These observations are consistent with the operation of an alternate pathway for activation of late complement components necessary for bacterial opsonization and killing by serum, but suggest that participation of C4 presumably through the classic C1,4,2,3–9 sequence might be necessary for maximal rates of activity. In addition, the results of studies using the various complement inhibitors offer support for the participation of heat labile factors, divalent cations and zymosan and hydrazine inhibitable components in the alternate pathway. Finally, both normal and C4-deficient guinea pigs were found to be highly resistant to intraperitoneal administration of high doses of type 25 pneumococci or E. coli used in the in vitro studies. Limited studies of the effects of intravenous challenge with pneumococci likewise revealed no striking impairment in the survival of the C4-deficient animals, suggesting that by utilization of the alternate pathway the host defenses are adequately maintained in vivo.
Ontogenesis of the in Vitro Response of Murine Lymphoid Cells to Cellular Antigens and Phytomitogens
Authors: Howe Michael L. , Manziello Bruce
Published: December 2022

The ontogenesis of cellular immunity in CBA/J mice was studied by using in vitro culture of spleen and thymus cells from donors varying in age from 2 to 70 days. Cell suspensions were assayed for reactivity in the mixed lymphocyte interaction (MLI) and isogeneic lymphocyte interaction (ILI) and for responsiveness to phytohemagglutinin (PHA) and pokeweed mitogen (PWM). The capacity of spleen cells to participate in the MLI and ILI and to respond to PWM appeared at an earlier age than did PHA responsiveness. The refractoriness of neonatal spleen cells to PHA stimulation could not be overcome by varying the number of cells or the quantity of PHA used in each culture. The thymus of neonates was found to have a similar selective deficiency of PHA-reactive cells. Cells responsive to PHA could not be detected in the neonatal thymus even after treatment of cell donors with cortisone acetate, although similar treatment of adult donors dramatically enhanced thymic cell responsiveness to PHA. These studies suggest that PHA responsiveness either requires a separate lymphoid cell population or reflects a later stage of lymphocyte maturation than the other in vitro activities assayed in this investigation.
Cell Interactions in Antibody Formation in Vitro
Authors: Leserman Lee D. , Cosenza Humberto , Roseman Jeffrey M.
Published: December 2022

The plaque-forming cell response in vitro of normal mouse spleen cells to sheep erythrocytes requires “adherent” and “nonadherent” cells. Adherent cells incubated only briefly with the antigen and then washed thoroughly with medium stimulate a full response of nonadherent cells which have not been exposed to antigen and are added to the adherent cells. Adherent cells after exposure to antigen and washing with medium were treated briefly with 0.83% NH4Cl and washed again. Such treatment abolished the response of nonadherent cells added to them. The treatment does not injure adherent cells, since their capacity to stimulate a normal response is fully restored by re-adding antigen to them. This requirement for both adherent cells and antigen is demonstrable for the first 2 days of culture of whole spleen cells, but is not demonstrable for so long when cultures are prepared from spleens of mice previously immunized with sheep erythrocytes. Previous studies demonstrated that a subpopulation, present in the adherent population is required, and we refer to this subpopulation as the third cell type. Nonadherent cells contain both bone marrow and thymus-derived cells required for the response to sheep erythrocytes. We suggest that the third cell type may focus the interaction of the other two cell types, presumably through antigen bound on the membranes of its cell surface.
Antigenic Properties of the Fab Fragment of Human IgM: Antigenic Determinants Common to IgM and IgG
Authors: Chen James P. , Tomasi Thomas B.
Published: December 2022

An antigenic determinant common to human IgM and IgG was revealed by the use of a rabbit antiserum produced against the Fabμ fragment of a monoclonal IgM (λ type). This antiserum (R8), after absorption with λ Bence Jones protein, still cross-reacted with IgM and IgG. Sheep erythrocytes coated with either IgM or IgG of λ type were tested with 13 Waldenström IgM, 12 myeloma IgG and 3 myeloma IgA proteins by hemagglutination inhibition. Three out of five IgM (λ) and two out of seven IgG (λ) were markedly inhibitory at concentrations of 0.2 to 4 μg/ml.

Mild reductive cleavage of IgM did not alter its reactivity. The inhibition of Fabμ fragments appeared to parallel that of the parent IgM molecules. Isolated heavy and light chains from macroglobulins produced negligible reactivity. These studies suggest that the conformational antigenic determinant(s) of the anti-Fabμ serum may involve primarily the interaction of the Fd fragment and λ chains of either IgM or IgG molecules. In addition, by using hemagglutination inhibition technique, other antibody system(s) have been found in R8 which show specificity for the Fabμ fragments of both κ and λ macroglobulins.
Hypersensitivity to Ascaris Antigens
Authors: Hussain Rabia , Strejan Gill , Campbell Dan H.
Published: December 2022

An allergen from Ascaris suum crude extract was isolated by a combination of Sephadex G-200 gel filtration and DEAE-cellulose chromatography. The allergen was detected over a wide range of molecular weights but had the property to stimulate homocytotropic (Hc) antibody production only when present in a molecular weight range of 30,000 to 40,000. The main bulk of the immunogenic fraction was negatively charged at pH 8.6, but the entire fraction displayed an extremely broad range of electrophoretic mobilities as detected by bidimensional immunoelectrophoresis; the allergen contained 11% reducing sugars. Bordetella pertussis-treated rats produced relatively high titers of Hc antibodies when injected with as little as 10 μg of the partially purified allergen.

The allergen (Asc-1) was obtained in a highly purified form by associating Sephadex gel filtration with polyvinyl-chloride preparative electrophoresis. A monospecific rabbit precipitating antiserum against Asc-1 was capable of completely removing the allergenic activity by specific absorption.
Immunohistologic Distribution of the Chicken Immunoglobulins
Authors: Lebacq-Verheyden Anne-Marie , Vaerman J. P. , Heremans J. F.
Published: December 2022

An immunoglobulin distinct from IgG and IgM has been identified in chicken serum, intestinal secretions, and bile (1). Its homology with mammalian IgA was inferred from the antigenic individuality of its heavy chains and its high concentration in two exocrine secretions, viz, intestinal juice and bile. The present communication deals with the distribution of the three immunoglobulins, IgG, IgM and IgA in the spleen and intestinal mucosa of five adult conventional chickens.

Specific antisera gainst γ, µ and α chains (1) were labeled with fluorescein isothiocyanate according to standard procedures (2). Tissue samples were processed to 5-µ sections in a Cryo-Cut microtome (American Optical Corporation), incubated with the labeled antisera and examined under ultraviolet light for immunoglobulin-containing cells.

In the spleen (Fig. 1), plasma cells were predominantly distributed around the germinal centers and most of these cells stained with anti-γ or anti-µ reagents. Only few cells reacted with anti-α.
Structural Differences in Mouse IgA Myeloma Proteins of Different Allotypes
Authors: Warner Noel L. , Marchalonis J. J.
Published: December 2022

Mouse IgA myeloma proteins produced by plasma cell tumors derived from BÀLB/c, NZB and (BALB/c × NZB)F1 hybrid mice were examined electrophoretically under dissociating conditions by using reduced and unreduced samples. In confirmation of previous studies by Abel and Grey (3) our electrophoretic patterns are consistent with the conclusion that no L-H disulfide bonds are present in BALB/c-derived IgA proteins. However, 3 out of 3 NZB derived IgA proteins appeared to possess L-H disulfide bonds, as did 5 of 10 NZB hybrid derived proteins. The presence of L-H disulfide bonds in the hybrid-derived proteins completely correlated with the presence of Ig-2e (NZB type) IgA allotypic antigens.
Valence and Temporal Change in Affinity of Purified 7S and 18S Nurse Shark Anti-2,4 Dinitrophenyl Antibodies
Authors: Voss Edward W. , Sigel M. Michael
Published: December 2022

Purified shark 7S and 18S anti-2,4 dinitrophenyl (DNP) antibodies were purified by immunoadsorption. Comparative equilibrium dialysis studies with the 3H-ε-DNP-l-lysine ligand over a 20-month period indicated that the 7S antibody showed a 100-fold increase in the average intrinsic association constant. The binding affinity of the 18S molecule did not change significantly during the same period. Valences of 1.4 and 8 were measured for the 7S and 18S molecules respectively, purified from the 30-month bleedings.
The Application of an in Vitro Cytotoxicity Test to Studies of the Effects of Drugs on the Cellular Immune Response in Mice
Authors: Fass Leroy , Fefer Alexander
Published: December 2022

An in vitro assay for cytotoxic cellular immunity has recently been developed based on the release of 51Cr from labeled target cells killed by specifically sensitized allogeneic lymphocytes. This test was applied to studies of the effects of chemotherapeutic agents on cellular immunity in mice. Adult C57BL/6 mice (H-2b) were inoculated with x-irradiated ascitic lymphoma cells (LSTRA) of BALB/c origin (H-2d) on day 0 and their spleen cells tested for reactivity against labeled LSTRA cells 4 to 10 days later. Each test was performed on quadruplicate samples with the standard error of the mean being less than 2%. In mice immunized with LSTRA cells but not given drugs a peak cytotoxicity of 16% was observed on day 6. Cells from mice treated on day +1 with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), 48 mg/kg cyclophosphamide (CY) or CY 180 mg/kg exhibited a peak reactivity of 41%, 12% and 2%, respectively, while drug administration on day -1 resulted in a peak reactivity of 36%, 33% and 23%, respectively. The results show that this test can be used to study the effects of drugs on the cellular immune response and that the timing of the drug and antigen administration, as well as the dose of drug, are critical. The increased cytotoxic reactivity of lymphocytes from drug-treated immunized mice suggests that although chemotherapeutic agents may be immunosuppressive as assayed in vivo, the activity of a given number of surviving lymphoid cells when tested in vitro and compared with the same number of cells from immunized but non-drug-treated hosts may be increased.
Immunoglobulins on the Surface of Lymphocytes
Authors: Grey Howard M. , Colón Sonia , Campbell Priscilla , Rabellino Enrique
Published: December 2022

The presence of immunoglobulin (Ig) on the surface of lymphocytes that were negative for Ig by immunofluorescence was investigated by combining a quantitative inhibition assay with immunofluorescence. Thymus cells; thymus, spleen and lymph node cells that had been passed through anti-Ig-coated plastic bead columns; and ϑ-positive mouse lymphomas were evaluated by using these two systems. These cell preparations had 0 to 1% of cells with detectable surface immunoglobulin by immunofluorescence. If the assumption is made that the fluorescent-positive cells were relatively homogeneous with respect to the amount of surface Ig, then it could be calculated that there was more Ig present in the various cell populations studied than was accountable by the small percentage of fluorescent-positive cells. Based on these calculations, the amount of Ig present in the different populations was found to be: fluorescent-positive cells: 5 to 14 ng N/106 cells; fluorescent-negative thymus cells: 0.01 ng N/106 cells; fluorescent-negative spleen and lymph node cells 0.07 to 0.1 ng N/106 cells. Three of four ϑ-positive lymphomas also had detectable Ig in amounts 3 to 10 times that found in normal thymus. Routinely there was more IgM found in both fluorescent-positive and -negative normal cells than IgG2.
The Effect of the Immunogenicity of the Heterologous Carrier on the Early Secondary Anti-2,4-Dinitrophenyl (DNP) Response of BALB/c Mice
Authors: Havas H. Francis , Pickard A. R.
Published: December 2022

The effects of different carriers on the anamnestic response to the DNP-ligand were investigated at the level of circulating antibody and of the single antibody-forming cell (PFC) by a modified hemolysin in gel technique with a number of different indicator sheep red blood cells (SRBC).

In all groups of mice it was found that regardless of the primary antigen the highest number of direct and indirect PFC was obtained on rechallenge with 2,4-dinitrophenyl hemocyanin (DNP-Hcy). The greatest difference between the different groups appared in the number of indirect PFC. The number of PFC after second challenge seemed dependent on the immunogenicity rather than the homology of the carrier used for challenge. Spleen weights were proportional to cell counts but generally did not correlate with the PFC response.

The indicator cell played a crucial role in the number of PFC detected: SRBC coupled with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide HCl DNP-bovine serum albumin (ECDI-DNP-BSA) detected the largest number of cells, presumably those secreting both high and low affinity antibody, while both EDCI-DNP-mouse serum albumin and 1-fluoro-2,4-dinitrobenzene (DNP-F) coupled with SRBC detected fewer cells, presumably only those of high affinity.

It was found that circulating antibody titers were higher in DNP-human γ globulin (DNP-HGG) and DNP-Hcy-primed mice when challenged with the homologous rather than the heterologous conjugate. However, no significant differences were observed in the DNP-BSA-primed group between homologous and heterologous challenge. Antibody concentrations were highest in DNP-HGG-primed and challenged mice but differed little in the other groups. There was little correlation between antibody concentration measured by radioimmunoassay, hemagglutination (HA) titers and the number of direct or indirect PFC.

Affinities varied insignificantly among the different groups but the heterogeneity index varied significantly (0.05 τ; p τ; 0.005). The heterogeneity indices were highest after heterologous and lowest after homologous challenge. These differences were statistically significant and are indicative of a more homogeneous antibody population after heterologous challenge.
Sensitization of Guinea Pigs to Herpes Simplex Virus
Authors: Rogers Howell W. , Scott L. Vernon , Patnode Robert A.
Published: December 2022

Guinea pigs were sensitized to herpes simplex virus (HSV) by inoculation with infective virus, inactive virus or a HSV soluble antigen (SA) preparation. Sensitivity was assayed by means of skin tests with SA. The largest skin reactions (erythema and induration) were observed in guinea pigs that had received two sensitizing injections of infective HSV. Inactive HSV was found to be an effective sensitizing material, suggesting that production of cutaneous hypersensitivity to HSV can occur in the absence of viral infection. A single intracutaneous injection of SA also sensitized animals to a subsequent skin test with SA.

Based upon the timing of maximal response, the mononuclear nature of the cellular infiltrate at 24 hr and non-transferability of reactivity with serum by the local passive transfer technique, the skin reactions observed in SA-sensitized animals are believed to be those of classical delayed hypersensitivity. While the skin reactions seen in these animals were not as large as those observed in animals sensitized with infective or inactivated HSV, they were shown to be specific for SA. Animals sensitized with infective vaccinia virus or L-cell control antigen did not respond to skin tests with SA.
Use of Dextran-Conjugated SRBC for the Detection of MOPC 104E Tumor Cells
Authors: Ghanta Vithal K. , Hamlin N. Marlene , Pretlow Thomas G. , Hiramoto Raymond N.
Published: December 2022

A dextran, fraction S, from Leuconostoc mesenteroides NRRL strain B-1355 was oxidized with sodium periodate and allowed to react with the side chain amino groups of sheep red blood cells (SRBC). The SRBC coated with this oxidized dextran fraction were used for the estimation of the number of mouse plasma tumor cells actively synthesizing IgM antibody in a population. We report optimal conditions for conjugation of this oxidized dextran to SRBC. The coated cells were agglutinated with the antibody produced by the tumor cells and in the presence of complement, lysis occurred. Formation of plaque-forming units in liquid chamber was readily demonstrable. The IgM antibody gave similar results when compared with actively induced IgM hemolysin to SRBC by the quantitative radial hemolysis in gel technique.
Suppression of Graft-vs-Host Reaction by a Spleen Extract
Authors: Garcia-Giralt E. , Morales V. H. , Lasalvia E. , Mathe G.
Published: December 2022

In 1970, we reported the inhibitory effect of an alcohol extract of calf spleen (S4) on the in vitro proliferation of stimulated lymphocytes (1). The active molecule has been found to be a protein, with a molecular weight of 45,000 (2). Its inhibiting activity in vivo has been shown to be tissue specific and not species specific (1), fulfilling, therefore, the main criterion for a chalone (3, 4). The reduction of the number of hemolytic plaque-forming cells in the spleens of mice immunized with sheep red blood cells and the inhibition of in vitro lymphocyte transformation and proliferation induced by phytohemagglutinin (PHA) or histocompatibility antigens in the mixed lymphocyte culture (5, 6) suggested that this extract could exert an immunosuppressive action. Similar results have been obtained in our laboratory by Kiger in 1971 with a similar thymus extract (7) and later with thymus and spleen extracts by Houck et al. (8).
Synthesis of Macrophage Migration Inhibition Factor (MIF) by Immunoglobulin-Producing Lymphocyte Lines and Their Clones
Authors: Tsuchimoto T. , Tubergen D. G. , Bloom A. D.
Published: December 2022

Studies have established the existence of two distinct populations of immunocompetent cells (1). One of these is thymus dependent (TL) and is responsible for cell-mediated immunity. Cells from this population do not contain intracellular immunoglobulin (Ig) and when stimulated in vitro by antigen these cells release substances which have been described as possible effector molecules of cellular immunity (2–7). The second population consists of cells (BL) capable of producing Ig and specific antibody.

Numerous human lymphocyte cell lines (LCL), derived from peripheral blood, have now been established in various laboratories, and many of the LCL are known to synthesize Ig (8, 9) as well as some of the putative effector molecules of cell-mediated immunity (10, 11). We have developed clonal sublines of established human LCL to test the hypothesis that production of the effector molecules of cell-mediated immunity occurs in a population of cells distinct from those capable of Ig synthesis.
Lysis of Rabbit Platelets by F(ab′)2 Antibody Fragments
Authors: Siraganian Reuben P. , Sandberg Ann L. , Osler Abraham G.
Published: December 2022

Immunologic injury to rabbit platelets provides a model system for the study of tissue damage mediated by unrelated immune complexes, complement and divalent cations (1, 2). Since numerous immunobiologic reactions require the Fc antibody fragment, it was of interest to determine whether the Fc portion is also essential for platelet injury. Recent evidence has indicated that the F(ab')2 fragments of guinea pig (3) and rabbit (4) immunoglobulins can activate the complement system via the alternate pathway.

Immunoglobulins were obtained from the sera of rabbits hyperimmunized with human serum albumin (HSA)4 or ferritin. The γ-globulin fraction precipitated at 40% saturated ammonium sulfate served as intact antibody and was presumed to consist primarily of 7S immunoglobulins. Aliquots of these preparations were subjected to pepsin digestion according to the method of Nisonoff (5) followed by chromatography on Sephadex G-150 and concentration of the 5S eluate by pressure ultrafiltration. The antibody concentrations of the intact and pepsin-digested preparations were determined by quantitative precipitin analysis.
Studies on Heterologous Antilymphocyte and Antithymocyte Sera
Authors: Barth Rolf F. , Singla Om
Published: December 2022

The immunosuppressive properties of rabbit and burro anti-mouse lymphocyte sera (ALS) before and after absorption with sheep erythrocytes (SRBC) have been studied. Five out of nine absorbed sera retained their ability to depress splenic plaque-forming cells (PFC) and serum hemagglutinin and hemolysin titers of mice immunized with SRBC. Four sera completely lost their immunosuppressive activity. Seven of the nine ALS had low titer anti-SRBC antibodies, but their complete removal by absorption did not reduce the potency of five of these sera indicating that their immunosuppressive activity was independent of passively transferred heterophile antibodies. The ability of absorbed ALS to prolong mean survival time of A/J to C57BL/6 skin allografts was significantly reduced when compared to that recorded in mice treated with unabsorbed ALS (16.2 vs 23.8 days). These findings are most consistent with the interpretation that immunosuppressive antilymphocytic antibodies had been removed from ALS by absorption with SRBC.
Molecular Restriction of Anti-Hapten Antibody Elicited in Neonatal Rabbits: Antibody Production in Littermates
Authors: Montgomery Paul C. , Williamson Alan R.
Published: December 2022

The structurally complex antigen dinitrophenylated bovine γ globulin (DNP-BGG) was used to study the immune response of neonatal rabbits. The results of these studies indicated that neonates frequently produce structurally restricted anti-DNP antibody. The isoelectric spectra of certain of these antibodies resembled myeloma proteins or antibodies produced by cell clones. The binding characteristics of these antibodies were consistent with monoclonality or structural homogeneity. An examination of anti-DNP antibody produced by littermates demonstrated that there was no simple genetic control or antibody heterogeneity early in life. Each neonate possessed the potential for anti-DNP antibody production with a differing capacity to express this potential. The results were consistent with the presence at birth of an extensive set of germ-line variable-region genes. while the element of somatic diversity could not be ruled out, somatic processes probably added little to the extent of antibody diversity present in the neonate.
In Vitro Responses of Rat Lymphocytes Following Adult Thymectomy
Authors: Folch Hugo , Waksman Byron H.
Published: December 2022

Inbred DA rats thymectomized at 8 weeks of age showed a marked decrease in in vitro reactivity of spleen cells to phytohemagglutinin-P (PHA) and concanavalin-A, appearing as early as 3 days and vanishing by 35 to 40 days. This change could not be explained by enlargement of the spleen. However, there was a proliferation of blasts in the splenic marginal zones at 1 to 2 weeks, but not at other times. Peripheral (popliteal, inguinal, and axillary) and mesenteric lymph nodes showed a similar effect. This effect was of lesser magnitude and appeared only 20 days after thymectomy, with recovery at the same time as the spleen, and was not observed in rats thymectomized at 4 weeks of age. Spleen cells after thymectomy showed a relatively inconstant decrease in reactivity to hemiallogeneic cells, while the decrease with both types of lymph node cells was, if anything, greater than the decrease in PHA reactivity. The evidence suggested that these findings could not be simply attributed to loss of a putative thymus-derived lymphocyte subpopulation.
Ontogeny of Bursal Function in Chicken
Authors: Toivanen Paavo , Toivanen Auli , Good Robert A.
Published: December 2022

Cyclophosphamide-treated 4-day-old chicks were injected with cells from yolk sac, liver, bursa of Fabricius, bone marrow, spleen and thymus by using embryonic and 4-day-old donors. Recipients and donors were isogeneic at the major histocompatibility locus. Survival pattern, gain of body weight, production of natural and immune antibodies, and microscopic morphology of bursa and spleen were studied to assess the reconstitution achieved. Of all the cell types used, only bursa cells were capable of a functional and morphologic reconstitution of the bursa-dependent lymphoid system. These findings are interpreted to indicate that in embryonic life and immediately after hatching the bursa alone possesses cells that can home to the bursa of 4-day-old chicks and re-establish its functions. These cells derived from embryonic liver and yolk sac are considered stem cells for humoral immunity. The findings illustrate the significance of the critical developmental relationship between the stem cell and its environment.
Alpha and Gamma Heavy Chain Diseases in Man: Intracellular Origin of the Aberrant Polypeptides
Authors: Buxbaum Joel N. , Preud'Homme Jean-Louis
Published: December 2022

Immunofluorescent and in vitro labeling studies were carried out on cells obtained from patients with α and γ heavy chain diseases. No evidence of post-ribosomal intracellular degradation of a normal size heavy chain was found in either case. Intracellular polymerization of the aberrant peptides appeared to take place in a fashion similar to that seen in cells synthesizing intact heavy chains.
Induction of Temporary IgA Deficiency in Mice Injected with Heterologous Anti-Immunoglobulin Heavy Chain Antisera
Authors: Manning Dean D.
Published: December 2022

Complete temporary suppression of IgA in both serum and the lumen of the gut has been achieved by injecting either anti-µ or anti-α antiserum into neonatal BALB/c mice. A model suggesting possible mechanisms of suppression and eventual recovery is discussed.
H-2 Dependence of Co-Operation Between T and B Cells in Vivo
Authors: Kindred Berenice , Shreffler D. C.
Published: December 2022

In nude mice from a stock which has been backcrossed several times to BALB/c (H-2d), a lasting restoration of immune competence could be induced by H-2 compatible (BALB/c, DBA, B10.D2) thymus cells but not by H-2 incompatible thymus cells. Heterozygous, H-2bH-2d, thymus cells from BALB/c × C57, F1 or from a BALB/c × (BALB/c × C57) backcross produce variable effects; only sometimes was a lasting restoration achieved. This cannot be accounted for by rejection of the injected thymus cells since nude mice do not reject tissue grafts. Therefore the H-2 complex must play an active role in an enduring co-operation between bone marrow-derived (B) cells and thymus-derived (T) cells.
The Unique Reassociation of Human IgA2 Immunoglobulins from Dimer Subunits
Authors: Jerry L. M. , Kunkel H. G.
Published: December 2022

The Am2(+) genetic variant of human IgA2 immunoglobulins lacks disulfide bridges between the heavy and light chains. Under dissociating conditions these proteins separate spontaneously into covalent heavy (H2) and light (L2) chain dimer subunits. In this study covalent heavy (H2) and light (L2) chain dimers were prepared from Am2(+) IgA2 myeloma proteins. These subunits recombined readily to give molecules which closely resembled the native proteins in hydrodynamic, antigenic and optical properties. In contrast, in the case of IgG, IgA1 and Am2(-) IgA2 proteins with intact H-L interchain disulfide bridges, covalent heavy and light chain dimers prepared from these proteins failed to recombine, although monomer heavy and light chains recombined readily. Among human immunoglobulins this property of reversible dissociation and reassociation appears to characterize the Am2(+) genetic variant of IgA2.

The light chains exist within these molecules as disulfide-bonded dimers. Selective cleavage of this inter-light (L-L) chain disulfide bridge was possible by gentle reduction with cysteine, with the inter-heavy (H-H) chain bridges remaining intact.
Porcine Immunoglobulins
Authors: Kaltreider H. Benfer , Johnson John S.
Published: December 2022

Antibody activity was demonstrated in each of six porcine immunoglobulins by the technique of indirect radioimmunodiffusion. Anti-BGG antibody of IgM, IgA and IgG classes were detected in serum simultaneously on the 4th day after primary immunization. The acquisition of antibody activity in the IgG subclasses was variable and no clear pattern was seen. IgA antibody reached peak titer last among the major classes and was the first to decline.
Studies on the Properdin System: Isolation of a Heat-Labile Factor from Guinea Pig Serum Related to a Human Glycine-Rich Beta-Glycoprotein (Gbg or Factor B)
Authors: Brade V. , Cook C. T. , Shin H. S. , Mayer M. M.
Published: December 2022

A heat-labile factor (HLF), which is essential for C3 inactivation by purified cobra venom factor, has been isolated from guinea pig serum. In addition to HLF, other protein(s) are required for inactivation of C3 by the cobra venom factor. HLF has a molecular weight of about 100,000 and is antigenically related to the human glycine-rich beta-glycoprotein (GBG). HLF is believed to be the guinea pig counterpart of factor B of the human properdin system.
Deae-Dextran and Polybrene Cation Enhancement and Dextran Sulfate Anion Inhibition of Immune Cytolysis
Authors: Ebbesen P.
Published: December 2022

The polycations DEAE-dextran and polybrene enhanced the sensitivity of the dye exclusion cytotoxic test performed on mouse spleen cells, whereas the anion dextran sulfate had an inhibitory effect.

Dextran sulfate prevented the enhancing effect of the polycations when admixed before incubation of the cells and reversed the polycation effect when added after the sensitization had taken place.

The in vitro infection of fibroblasts with murine leukemia virus was largely influenced in the same way, indicating a similarity of the membrane processes involved.
Antibodies to a Distinct Antigenic Determinant of Staphylococcal Nuclease
Authors: Sachs David H. , Schechter Alan N. , Eastlake Ann , Anfinsen Christian B.
Published: December 2022

Antibodies to staphylococcal nuclease have been fractionated on immunoabsorbent columns bearing selected polypeptide fragments of the enzyme. By this means a population of antibodies specific for a distinct antigenic determinant in the helix-rich, carboxyl-terminal third of nuclease was prepared. The monospecificity of these antibodies was operationally defined by ultracentrifugal analysis in the presence of nuclease. Although prepared against the intact, native nuclease molecule these antibodies are able to bind to a disordered polypeptide fragment thereof. It is postulated that the native antigenic determinant may be generated in such a fragment by spontaneous folding of the polypeptide chain.
Complement: Effects of C1 Binding Affinity on the Efficiency of the C2 Reaction
Authors: Linscott William D.
Published: December 2022

A variable amount of the C1 will dissociate from IgG-sensitized EAC14 at 30°C, µ = 0.065, depending on the relative amounts of antibody and C1 employed. for this reason, use of 30°C and µ = 0.065 for reacting C2 with IgG-sensitized EAC14 is inefficient, particularly with low IgG concentrations. Decreasing the ionic strength to 0.032 prevents dissociation of weakly-bound C1 during the C2 reaction at 30°C, and thus gives optimal results. Similar but not quite as good results can be obtained by adding 100 to 200 molecules of IgM anti-Forssman antibody/cell. It is suggested that this added IgM may function by capturing C1 which dissociates from IgG-SAC14 at 30°C, µ = 0.065. The resulting IgM-SAC1 could activate C2 molecules which then could bind to the IgG-SAC4, converting them to IgG-SAC42 which in turn could react with C3 and later components.

Excessive amounts of IgM (1000 to 10,000 molecules/cell under the conditions employed here) can reduce the efficiency of the reaction with C2, perhaps by raising the C1 binding affinity above some optimal level and/or by taking control of some of the C4 sites away from the more efficient IgG antibody molecules. Given a relatively constant number of molecules of C1/cell, IgM-sensitized EAC14 show a progressive decrease in C2 reaction efficiency with increasing amounts of IgM/cell over a 100-fold range of SA/SAC1 ratios.
The Kinetics of Antibody Production and Affinity in Balb/c Mice Immune and Partially Tolerant to the 2,4-Dinitrophenyl Determinant
Authors: Pickard Allan R. , Havas H. Francis
Published: December 2022

A study was undertaken of the kinetics of the immune responses to the 2,4-dinitrophenyl (DNP) determinant of immune and partially tolerant adult and neonate BALB/c mice. Neonate and adult mice were pretreated with multiple injections of soluble DNP-bovine serum albumin (DNP-BSA) to induce tolerance. The immune responses of pretreated mice were compared with those of nonpretreated controls following immunization with DNP-hemocyanin (DNP-HCY). The quantity of antibody was measured by radioimmunoassay and the affinity by equilibrium dialysis against ε-DNP-lysine. Maximum serum antibody levels were attained 4 weeks after the first and 1 week after the second immunizing injection of DNP-HCY in all groups. Mice made partially tolerant to the DNP determinant as either neonates or adults responded to primary and secondary immunization with DNP-HCY by producing less DNP-specific antibody which had lower average affinity for DNP-lysine than nontolerized controls. However, in all groups, antibody affinity increased with time so that by the 11th week after primary immunization tolerized mice were producing antibody within the range of affinities seen in controls. The implications of these data are discussed and interpreted in terms of an affinity-dependent selection of cell populations during tolerogenic and immunogenic stimulation.
Reconstitution of the Antibody Response in Vitro of T Cell-Deprived Spleen Cells by Supernatants from Spleen Cell Cultures
Authors: Sjöberg Olof , Andersson Jan , Möller Göran
Published: December 2022

The antibody response of spleen cells against sheep red cells was tested in vitro with the Mishell-Dutton technique. Spleen cells deprived of thymus-derived (T) cells by anti-θ serum treatment gave a low response. Supernatants from cultures of either parental or syngeneic spleen cells could restore the response of T cell-deprived spleen cells against sheep red cells to a certain degree, but supernatants from mixed lymphocyte cultures between two H-2 incompatible parental strains were more efficient in this respect. These supernatants also could stimulate markedly the immune response of spleen cells from athymic “nude” mice. Addition of supernatants to cultures of normal or T cell-depleted cells caused an early appearance of plaque-forming cells by day 2 with a maximal number on day 3 for the T cell-depleted cells. Supernatants harvested on day 1 were more effective than supernatants from day 2 in restoring the immune response, whereas supernatants from days 3 and 4 were inhibitory. This was not due to an excessive production of the restorative factor. Both T cells and adherent cells were necessary for the production of the restorative factor. The active factor in the supernatants could be produced in serum-free medium and was capable of supporting a primary antibody response of spleen cells cultured in normal mouse serum. The findings suggest that T cells may stimulate the antibody response of bone marrow-derived cells by means of a soluble factor.
Naturally Occurring Cell-Mediated Immunity to Purified Glycerol-Teichoic Acid Antigen in Guinea Pigs
Authors: Frederick G. T. , Holmes R. A. , Chorpenning F. W.
Published: December 2022

Previous reports of delayed hypersensitivity in guinea pigs have dealt only with animals artificially immunized with the antigen used to elicit the delayed reaction (1–5). Except for delayed-type isohypersensitivity to serum protein antigens (6), no reports have described a naturally occurring cell-mediated immunity in this species. Although it has been established that delayed hypersensitivity to polysaccharides occurs in a number of mammalian species (7–12), only two investigators have reported using completely defined carbohydrates as the eliciting antigen (10, 11). No previous reports of delayed-type hypersensitivity to teichoic acids in any species have been published, although an immediate reaction to one of these antigens has been demonstrated in humans (13). This report describes the occurrence of delayed hypersensitivity to glycerol-teichoic acid in unimmunized guinea pigs as demonstrated in vivo by a delayed skin reaction and in vitro by the inhibition of macrophage migration.
Observations on the Human Blood Groups
Authors: Landsteiner K. , Witt Dan H.
Published: December 2022

In human sera there exist commonly agglutinins—“cold”-agglutinins—differing from the well-known iso-agglutinins in that their effect is generally much more diminished by increased temperature. These “cold”-agglutinins, apparently related to and in part identical with auto-agglutinins, display a certain specificity (Bialosuknia and Hirszfeld) and under suitable conditions their effects appear as atypical (minor) iso-agglutinin reactions. The sensitiveness to changes in temperature warrants a differentiation of these substances from the common iso-agglutinins, although agglutinins exist which seem to be intermediate.

Very probably there exist other individual differences of human bloods than those disclosed by the ordinary iso-agglutination tests.

Strong anomalous iso-agglutinin reactions occur rarely. Neither these nor the minor reactions mentioned previously or the differences disclosed by the use of animal sera have been studied sufficiently to be arranged in a practicable classification.

The division of human bloods into four groups is still adequate for ordinary purposes.

The iso-agglutinins acting on corpuscles of group II can be separated into two qualitatively different fractions which may be designated as α and α1.

Among the corpuscles of group II (and IV) there are two varieties disinguishable by the different susceptibility to the agglutinin α1.

Some sera of group IV contain agglutinins of the type α1 effective on certain corpuscles of group II.
Experiments on the Sensitivity of the Human Skin to the Toxin of the Bacillus of Shiga-Kruse
Authors: Brokman H. , Przesmycki F.
Published: December 2022

Experiments are presented showing that the negative skin reaction to the products of the Shiga-Kruse bacillus in human beings depends upon the presence of antitoxin in the blood. The antitoxin is specific, and, as in the case of “natural” immunity to diphtheria, it appears spontaneously in the individual, without a previous attack of the disease or artificial immunization. If this presence of dysenteric antitoxin in human beings provides an actual immunity against infection with dysentery, then the way must be open to attempt an active immunity in the same way as is done in respect to diphtheria and scarlet fever. We have actually begun to make this attempt with dysentery anatoxin. These experiments will be the suject of our next communication.
The Effect of a Streptococcus Pyogenes Teichoic Acid on the Immune Response of Mice
Authors: Miller Glenn A. , Jackson Robert W.
Published: December 2022

The immunosuppressive activity of glycerol teichoic acid extracts isolated from Streptococcus pyogenes 1-RP41, a group A beta hemolytic streptococcus, has been investigated by measuring the number of direct plaque-forming cells and indirect plaque-forming cells in isolated spleen cells and serum hemolysin titers in mice treated with the teichoic acid. These extracts caused significant suppression of both the IgM and IgG response which was evident at both the organ level and the circulating antibody level. Inhibition of the immune response seemed to be a function of the poly(glycerolphosphate) backbone of the molecule since inhibition of antibody production was still evident when alanyl esters were removed from the teichoic acid polymer. In addition teichoic acid appeared to be functioning at the early processes of antibody production since treatment with the extract after antigenic challenge did not cause significant suppression.
The Early Response of Immunocompetent Cells as Analyzed by Velocity Sedimentation Separation
Authors: Armstrong W. D. , Kraft N. E.
Published: December 2022

Separation of cells by velocity sedimentation is based primarily on differences in cell size. Immunocompetent cells (ICC) from mice primed for 24 to 48 hr with the polymerized flagellar antigen of Salmonella adelaide (POL) have been separated from nonreactive cells by virtue of their increased sedimentation velocity. In vitro and in vivo POL challenges of the sedimented fractions of ICC have demonstrated a depletion of reactivity in the normally immunocompetent slower sedimenting fractions and an enrichment in the normally unresponsive faster fraction. The modified velocities of ICC have returned to normal 15 weeks after the initial injection of antigen. Changes induced by POL in the sedimentation velocity of certain subpopulations of cells are specific for that antigen. The response of sedimented fractions from normal mice, when cultured with tolerogenic amounts of POL. is uniformly reduced by 75%. The profile of the immune response which remains is the same as that of cells challenged with immunogenic levels of antigen in vitro.
Selective Induction of Delayed Hypersensitivity by a Lipid Conjugated Protein Antigen Which Is Localized in Thymus Dependent Lymphoid Tissue
Authors: Coon John , Hunter Robert
Published: December 2022

Multiple injections of bovine serum albumin (BSA) without adjuvant stimulate production of large amounts of antibody but no detectable delayed type hypersensitivity (DTH) in guinea pigs. In contrast, single or multiple injections of BSA heavily conjugated with a lipid, dodecanoic acid, without adjuvant, stimulated sustained DTH but no detectable antibody production. The DTH produced was specific for native BSA and no detectable immune response was produced against the fatty acid groups. This change in the type of immune response produced was associated with a change in the area of antigen localization in popliteal lymph nodes. Native BSA labeled with 125I was localized in germinal centers in the bone marrow derived cell area but was not detectable in the thymic dependent or thymus derived (T) cell area of the lymph nodes. In contrast, lipid conjugated BSA labeled with 125I was prominently localized in the thymic dependent area of the lymph nodes but was not found in the germinal centers of animals producing pure DTH reactions. These results suggest that the well known ability of lipids to enhance the production of DTH may be in part due to a lipid-mediated localization of antigen within the thymic dependent area of lymph nodes where it is in close association with many T type lymphocytes.
Potentiated Reagin Response: Initiation with Minute Doses of Antigen and Alum Followed by Infection with Nippostrongylus Brasiliensis
Authors: Bloch Kurt J. , Ohman John L. , Waltin Jerieta , Cygan Ralph W.
Published: December 2022

Intraperitoneal injection of 10 µg of egg albumin (EA) or keyhole limpet hemocyanin plus alum elicited a low titer reagin response in most male Sprague-Dawley rats. Infection with Nippostrongylus brasiliensis larvae served to potentiate the reagin response in many animals. The potentiated response yielded impressive reagin titers, at least in rats injected with EA, but was of short duration. The potentiated response appeared to be restricted to antibodies of the IgE class and did not involve IgGa or IgG1 antibodies. Some attempts were made to elucidate the mechanism of the potentiated response.
The First Component of Complement in Serum: Evidence for a Hitherto Unrecognized Factor in C1 Necessary for Internal Activation
Authors: Loos M. , Borsos T. , Rapp H. J.
Published: December 2022

The lack of direct proportionality between the number of SAC14 formed and the concentration of serum C1 was ascribed to the dissociation of C1 molecules into their subcomponents. We have obtained evidence, however, that this non-linear relationship is due to the dissociation from the C1 molecule of a hitherto unrecognized factor necessary for the activation of bound C1. Determination of the hemolytic activity of EAC14 before and after treatment with trypsin demonstrated that EAC14 contained two populations of SAC14, one in which C1 was activable by itself and by trypsin and another in which C1 was activable by trypsin but not by itself. A factor necessary for activation of C1, missing in non-self-activable C1, was designated “X” and the C1 molecules lacking this factor were designated (C1 - X). The finding that the apparent C1 content of whole serum was dependent on EAC4 concentration indicated that the activating factor “X” was bound independently from C1 and (C1 - X).
Antibody-Binding Characteristics at the Cellular Level
Authors: Claflin Latham , Merchant Bruce , Inman John
Published: December 2022

A comparison of the rates and dynamics of the maturation process in specific IgM and IgG immune responses was made at the cellular level in BALB/c mice immunized with an enlarged hapten, N-(2,4-dinitrophenyl)-β-alanylglycylglycine (DnpAGG) coupled to hemocyanin. At intervals after immunization, the relative binding characteristics of IgM and IgG plaque-forming cells (PFC) secreting anti-DnpAGG antibodies were determined by the plaque inhibition technique. With time after immunization, the concentration of free hapten required to cause 50% reduction in both IgM and IgG PFC numbers decreased indicating that an increase in average binding affinity of PFC antibodies occurred. This maturation process was complete for IgM PFC by 9 days after immunization but required about 30 days in the IgG PFC response. A subpopulation analysis showed that maturation of both IgM and IgG responses occurred as the result of a simple selective process involving initial stimulation of a heterogeneous population of DnpAGG-specific PFC followed by preferential retention of that subpopulation of PFC producing antibody of the highest affinity. The data indicated that the IgM response evolved from precursor cells present before specific immunization.