bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunosuppression by Leukemia Viruses
Authors: Koo Gloria C. , Ceglowski Walter S. , Friedman Herman
Published: December 2022

The effect of Friend leukemia virus infection on the ultrastructure of spleens of BALB/c mice was studied. Normal mice immunized with sheep erythrocytes developed a characteristic splenic response, with many mature lymphoid cells and plasma cells appearing within 2 to 4 days after immunization. Large numbers of specific antibody plaque-forming cells (PFC) to sheep erythrocytes appeared concomitantly in the spleens of these mice. In contrast, there was a marked suppression in the number of PFC in spleens of mice infected before immunization. Furthermore, infected mice had significant alterations in their spleens, as studied on the ultra-structural level. Very few cells of the plasmacytic series appeared in the spleens of such mice. Mice infected 8 days before immunization had no detectable plasma cells in their spleens, although occasional plasmablasts were observed. Immature lymphoid cells and blast cells were the predominant cell type. Only a slight to moderate cellular change occurred in spleens of mice infected on the day of immunization or 2 days later. Virus particles were not detected in spleens of mice infected on the same day or 2 days after immunization and examined on the 4th day of the immune response. Moderate numbers of virus particles were apparent in extracellular spaces, and occasionally within immature cells in the spleens of mice infected 3 days before immunization and examined 4 days afterwards. In contrast, spleen cells of mice infected 8 days before immunization had numerous Friend leukemia virus particles. Many virus particles budded from the membrane of typical lymphocytes and blast cells, as well as from some cells in mitosis. The alteration in spleen cell morphology of infected animals correlated with the marked immunodepression accompanied by a decrease in the number of antibody plaque-forming cells to sheep erythrocytes. These results seem compatible with the concept that there is competition between leukemia virus and antigen for common stem cells, which can be diverted either to leukemogenesis or immunogenesis depending on the nature of the stimulus.
Immunosuppression by Leukemia Viruses
Authors: Koo Gloria C. , Ceglowski Walter S. , Higgins Michael , Friedman Herman
Published: December 2022

The effects of infection of mice with Friend leukemia virus on single antibody-forming cells was studied by means of electron microscopy. Individual plaque-forming cells (PFC) from normal immunized mice were examined at the 4th day of the immune response and found to consist almost exclusively of cells of the plasmacytic series. Twenty-four per cent of the cells were proplasmacytes and 56% were plasma cells. The remaining PFC consisted of degenerated cells (20%). Virus-like intracisternal A particles were found in many of these cells. In contrast, many cells (55%) from the center of plaques derived from spleens of mice infected with FLV contained typical murine leukemia virus particles, some of which were budding from the plasma membrane. The ultrastructure of these spleen cells was similar to that of the control animals, i.e., most of the cells were either proplasmacytes (10%), or mature plasma cells (60%). Some cells were degenerated (20%). Two PFC from infected animals were lymphocytic cells. The results of this study indicate that leukemia virus infection and antibody formation are not mutually exclusive processes at the single cell level.
Cellular Aspects of Primary Antibody Response in Vitro
Authors: Grobstein Ruth H.
Published: December 2022

A membrane filter, modified organ culture system has been used in order to study the interrelationships between different cell types during the induction of a primary hemolytic response against sheep erythrocytes. Under these conditions, the normal architecture of the spleen fragments is preserved and a multi-cell transfer leading to antibody synthesis can be readily analyzed. The kinetics of the in vitro response using pre-immunized fragments were studied and compared with the kinetics of the in vivo response which it resembles. The kinetics of the primary response induced in vitro were also followed and analyzed with respect to the participation of the cells. The cells sort themselves into different populations on the filter.
The Production of Tolerance to Human Erythrocytes in the Rat with Cytosine Arabinoside or Cyclophosphamide. II. Previously Immunized Animals
Authors: Gordon Robert O. , Santanelli James P. , Wade Maclyn E.
Published: December 2022

Cytosine arabinoside (Cytarabine, ara-C), an antimetabolite, and cyclophosphamide (Cytoxan), an alkylating agent, have been shown in an earlier study to produce long-lasting, selective immunologic tolerance in the previously unsensitized rat to human O, Rh positive, erythrocytes (1). Tolerance was produced most successfully by repeated complete suppression of the immune response by simultaneous courses of drug and antigen. It was shown, therefore, that tolerance to cellular antigens could consistently be established using an antimetabolite as well as an alkylating agent. Selective elimination of all or most of the clonal cells by repeated courses of drug at the time of antigen stimulation was proposed as the mechanism of tolerance induction.

As is the case with unsensitized animals, the production of tolerance in previously immunized animals has been accomplished more easily with soluble proteins than with cellular antigens (2, 3). This report shows that specific unresponsiveness to a cellular antigen can be achieved in an already sensitized animal by repeated complete suppression of the anamnestic immune response.
Conversion of Cholera Exo-Enterotoxin (Choleragen) to Natural Toxoid (Choleragenoid)
Authors: Finkelstein Richard A. , Peterson Johnny W. , Lospalluto Joseph J.
Published: December 2022

The phenomenon of spontaneous conversion of bacterial toxins to toxoids has been observed since the time of Ehrlich although information regarding the mechanism of the reaction is limited. The toxoiding process, apparently common to many protein toxins, occurs on aging and may be accelerated by chemical treatment such as with formaldehyde. In the latter case, a different molecular species of toxoid may result. For example, spontaneously formed tetanus toxoid has been reported to be a dimer of the toxin, whereas the formaldehyde toxoid can be similar in size to the parent toxin (1–4). In some instances, notably among other clostridia, toxin may be released into the medium in the form of a protoxin that is antigenically similar to the toxin but lacks toxicity. The protoxin is converted into toxin by bacterial proteases (5).

Previous work in this laboratory has led to the isolation and purification of the cholera exoenterotoxin, designated choleragen, from vigorously aerated and agitated multi-liter fermenter cultures of Vibrio cholerae 569B (6, 7).
Regulation of Homocytotropic Antibody Formation in the Rat
Authors: Tada Tomio , Taniguchi Masaru , Okumura Ko
Published: December 2022

Homocytotropic antibody (HTA) formation, as measured by the passive cutaneous anaphylaxis (PCA) test, was enhanced, although after a delay, and prolonged by 400 R whole body x-irradiation given shortly before through 8 days after immunization of rats with dinitrophenylated Ascaris extracts (DNP-As) plus killed Bordetella pertussis. Delayed enhancement and prolonged production were most striking when x-irradiation was given 1 day before to 2 days after initial immunization. An adjuvant effect of 400 R was produced in rats immunized with DNP-As alone omitting B. pertussis. As evidenced by passive hemagglutinin titers after 2-ME treatment, the production of γG antibody was greatly suppressed by 400 R x-irradiation. Two possible pathways to account for the irradiation effects on HTA formation were postulated, i.e., the enhancement of γM antibody and the suppression of 7S γG antibody resulted in a defective humoral feed-back regulation, and the depletion of certain populations of lymphoid cells, probably of thymic origin, caused cellular regulatory disturbances.
Decreased C3 Levels in a Chronic Virus Infection: Equine Infectious Anemia
Authors: Perryman L. E. , McGuire T. C. , Banks K. L. , Henson J. B.
Published: December 2022

There is increasing evidence that complement-dependent immune reactions are pathogenetically important in virus diseases. One indication of in vivo complementfixing immune reactions is the lowering of plasma C3 levels. A study was made of the circulating plasma C3 levels in 12 horses infected with equine infectious anemia (EIA) virus. Purified equine C3 was prepared from euglobulin precipitates of horse serum using triethylaminoethyl (TEAE) column chromatography with subsequent Pevikon block electrophoresis. Monospecific antisera prepared in rabbits was used in single radial immunodiffusion to quantitate C3 levels. A significant reduction of C3 was detected in 6 of 12 horses with active clinical disease. All horses with reduced C3 levels had C3-coated erythrocytes and deposits of immunoglobulin and C3 in the glomeruli. These findings further indicate that immune processes are active in the pathogenesis of EIA.
Immunochemical Studies on the Double Specificity of Allotype a2 of Rabbit Immunoglobulin G
Authors: Kakinuma Mitsuaki
Published: December 2022

Immunochemical studies of molecules of rabbit IgG of allotype a2 are presented. Of two precipitin lines which formed between IgG of allotype a2 and anti-a2 antisera, the a2′ antigen (the antigen of the outer precipitin line) was found to be sensitive to digestion by papain whereas the a2" antigen (the inner line) was resistant. Both antigens are carried on Fab fragments of IgG but different molecules appear to carry each of the two antigens. The ratio of a2′ to a2" molecules in IgG populations of allotype a2, either homozygous or heterozygous with respect to the a locus, is roughly 1:1. Antigenic families analogous to a2′ and a2″ were also observed in IgG of allotype a1, but not a3. The anti-a3 was unispecific in its reactions with a3 antigens.
Quantitative Studies on the Binding of Antibody to the Surface of HeLa Cells
Authors: Boone Charles W. , Irving Dianne N. , Rubinstein Susan
Published: December 2022

The binding of horse antibody to viable HeLa cells was studied using mixed immune and normal sera labeled with different iodine radioisotopes, a technique which permits precise correction for nonspecific adsorption. The association constant of the reaction, the maximum antibody-binding capacity of the cells, and the antibody content of the immune serum were determined using two types of titration: a constant concentration of cells titrated with immune serum, and a constant concentration of immune serum titrated with cells. The titration data conformed closely to linear equations derived from the mass action law. When applied to antigenantibody reactions which are heterogeneous with respect to binding affinity, these equations have two advantageous properties: 1) the plots remain linear to within a known degree of error which can be made small, and 2) the Ka value obtained from the slope and ordinate intercept of the linear plots represents the weighted mean of the Ka values of each affinity subspecies in the antibody population.
Studies of Immune Mechanisms in Leprosy
Authors: Bullock Ward E. , Fasal Paul
Published: December 2022

Phytohemagglutinin (PHA)- and antigen-induced DNA synthesis was measured in leukocyte cultures from 16 patients with lepromatous leprosy, 12 with tuberculoid leprosy, and 20 normal persons. In cultures containing 20% autologous plasma, the responses of lepromatous leukocytes to PHA, purified protein derivative (PPD), streptolysin O (SLO) and antigens of Mycobacterium leprae were significantly below those of the control group. Simultaneous culture of lepromatous leukocytes washed and suspended in normal homologous plasma (20%) resulted in significantly increased SLO-induced DNA synthesis and a modest increase in response to PPD. The response to PHA and antigens of M. leprae remained unchanged. Except for PPD, the responses of tuberculoid leukocytes in autologous or homologous plasma were not significantly lower than normal. Nine of 14 lepromatous plasmas were found to contain a factor that depressed the response of normal leukocytes to PPD and SLO but not to PHA. The depressor factor is non-dialyzable, stable after prolonged storage at -20°C, and resistant to heating at 56°C; its activity is lost at relatively low dilutions. It is concluded that the in vitro demonstration of impaired cellular immune function in patients with leprosy may reflect a primary cellular defect, the depressive effect of a humoral factor, or both, to varying degrees.
Immune Hemolysis and the Functional Properties of the Second (C2) and Fourth (C4) Components of Complement
Authors: Opferkuch W. , Rapp H. J. , Colten H. R. , Borsos T.
Published: December 2022

By use of the late-acting components of rat complement (rat C-EDTA) as a reagent for the lysis of the intermediate EAC142, each molecule of human C4 and of oxidized human C2 and one of every two molecules of human C2 taken up by the appropriate cell intermediates was shown to be hemolytically active. Furthermore, it was found that each molecule of guinea pig C2 in serum can be measured by its hemolytic activity.
Independent Action of Thymus and Bone Marrow Cells During the Secondary Response of Direct Plaque-Forming Cells
Authors: Jehn Ulrich W. , Karlin Lawrence
Published: December 2022

Thymocytes and bone marrow cells from normal mice or from mice primed with SRBC were used to repopulate heavily irradiated mice. The recipients were challenged with SRBC and the PFC formed in their spleens were enumerated. The thymus-marrow synergism characteristic of unprimed cells was not found when primed cells were used in the restoring inoculum. Thymocytes and marrow cells from primed mice were each capable of restoring immunocompetence in the recipients, and in contrast with the corresponding cells from unprimed animals they appeared to react immunologically as independent units.
Antigenic Markers on Cells Leaving Calf Thymus by Way of the Efferent Lymph and Venous Blood
Authors: Williams R. Michael , Chanana A. D. , Cronkite E. P. , Waksman Byron H.
Published: December 2022

A significant proportion of lymphoid cells which leave the calf thymus by the efferent lymph or blood have thymus-specific antigen (BTA) in their cell membrane. These were found to be larger than the majority of BTA-positive cells within the thymus. Similar cells were not observed in arterial blood, thoracic duct lymph or prescapular lymph nodes.
The Response in Rabbits to Prolonged Immunization with Type III Pneumococci
Authors: Kimball John W. , Pappenheimer A. M. , Jaton Jean-Claude
Published: December 2022

Every rabbit in a group of 17 animals immunized with formalinized type III pneumococci produced type-specific antibody of restricted electrophoretic heterogeneity. The critical factor in eliciting this response appears to be repeated antigenic stimulation over a period of many weeks rather than a particular genetic constitution of the animals. Over long periods of time and with continued immunization, restricted components appear and disappear as shown by 1) the appearance and disappearance of antibody populations of a distinct, restricted electrophoretic mobility and 2) the parallel behavior of the corresponding light chains. Two animals that produced extraordinarily high concentrations of antibodies (43 and 68 mg/ml of serum) showed clinical signs strikingly similar to those associated with multiple myeloma in man.
Differential Effects of Local Lymphoid Irradiation on Delayed Hypersensitivity and the Serum Antibody Response in Rats: Antigen Injection before X-Rays
Authors: Eltringham James R. , Weissman Irving L.
Published: December 2022

Several time-dependent dissociations of the cellular (delayed hypersensitivity response, DHR) and humoral (hemolysin and hemagglutinin tests) responses in rats occurred following irradiation of the draining regional lymph nodes (460 R on two consecutive days or 920 R on one day) at various times after local sensitization with antigen (sheep erythrocytes (SRBC) plus Freund's complete adjuvant). The rats were also locally challenged with SRBC alone on day 9. Thus, irradiation on day 1 or 2 following initial sensitization led to augmentation of the serum hemolysin response and a slight inhibition of DHR. Irradiation on day 3 or 4 selectively inhibited the humoral response, whereas irradiation on day 6 through 8 selectively inhibited DHR. Finally, irradiation on day 9 was ineffective in inhibiting or augmenting either response.
Binding of a Polypeptide Antigen to Ribonucleic Acid from Macrophage, HeLa and Escherichia Coli Cells
Authors: Roelants Georges E. , Goodman Joel W. , McDevitt Hugh O.
Published: December 2022

A synthetic branched polypeptide, (T,G)-A-L, formed complexes with RNA which banded at densities lower than that of pure RNA in cesium sulfate gradients after equilibrium centrifugation. In different experiments, the minor bands appeared at densities ranging from 1.56 to 1.63 whereas pure RNA banded at a reproducible density of 1.68. The presence of minor bands was independent of the addition of (T,G)-A-L, but when (T,G)-A-L was added to cell homogenates prior to extraction of RNA, the minute fraction of polypeptide that remained associated with RNA in the gradients was concentrated in the minor bands. The formation of minor bands was independent of the cell type, since they formed to the same extent when peritoneal exudate cells, HeLa cells or E. coli cells were used. Minor bands were not found when RNA was extracted with hot rather than cold phenol. RNA had a pronounced adjuvant effect in priming CSW (responder) mice for immune responses to (T,G)-A-L, but E. coli RNA was at least as effective in this respect as RNA from peritoneal cells. No support was found for the existence of a ribonucleoprotein fraction unique to macrophages.
Nephritogenic Glycoprotein
Authors: Shibata Sehchi , Nagasawa Toshihiko , Miyakawa Yuzo , Naruse Takuji
Published: December 2022

Rats that were given a single injection in the hind footpads of homologous (watersoluble) renal glycoprotein and Freund's complete (or incomplete) adjuvant developed typical proliferative glomerulonephritis. Proteinuria and morphologic changes of glomerulonephritis continued until death of the animals (the longest period of observation was 12.5 months after injection). The glycoprotein was isolated and purified from rat glomerular basement membrane and had previously been reported from our laboratory to induce nephrotoxic antibody. Immunization of rats with as little as 5 µg of the glycoprotein in Freund's complete adjuvant effectively induced typical proliferative glomerulonephritis. The nephritis could be transferred to previously healthy recipients with the eluate obtained from nephritic kidneys.
Antibody Plaque Responses of Mice Given Vibrio cholerae Antigen as Neonates
Authors: Sajid Mohammad A. , McAlack Robert F. , Cerny Jan , Friedman Herman
Published: December 2022

Neonatal mice were given multiple doses of a lipopolysaccharide (LPS) antigen derived from Vibrio cholerae to induce immunologic tolerance. A direct vibriolytic plaque assay was used to enumerate specific antibody-producing cells to these organisms. Marked suppression occurred in mice given low and high doses, whereas intermediate doses often led to an enhanced response. Normal mice had no “background” antibody-forming cells but many plaque-forming cells (PFC) developed in the spleens of antigen-treated mice before challenge immunization.

It was possible to distinguish between antibody-forming cells to the type-specific and the common somatic antigen. After challenge immunization most antibody-forming cells in spleens of control mice had specificity to the type-specific B antigen, whereas mice injected with LPS as neonates showed a suppression of the anti-B response, the number of anti-A PFC being either unaffected or enhanced. Most of the immunosuppression seemed directed to the major type-specific antigen present in the LPS extract. Concomitant stimulation of antibody-producing cells to the common antigen appeared to reflect its presence in smaller “contaminant” amounts. There was little correlation between serum titers and the number of antibody-forming cells.
Stabilization of Mutant Catalase by Complex Formation with Antibody to Normal Catalase
Authors: Feinstein Robert N. , Jaroslow Bernard N. , Howard Judith B. , Faulhaber Joann T.
Published: December 2022

This study concerns a mouse strain which is mutant with respect to blood and tissue catalase. The mutant catalase is abnormally sensitive to heat and extraordinarily sensitive to mild alkalinity. Antisera prepared by immunizing rabbits to the blood of normal mice stabilize the mutant catalases against the effect of heat and alkaline pH. As a result of this stabilization, the antigen-antibody complex exhibits a greater catalatic activity than was originally added, i.e., an apparent activation.
Hypersensitivity to Ascaris Antigens
Authors: Strejan Gill , Campbell Dan H.
Published: December 2022

Rabbit sera containing relatively high titers of anti-Ascaris homocytotropic antibodies were fractionated by gel filtration on Sephadex G-200 and by DEAE-cellulose chromatography. Anti-Ascaris Hc antibodies were recovered from Sephadex G-200 in the ascending portion of the IgG peak, and from DEAE-cellulose in a peak eluted with 0.1 M sodium phosphate buffer, pH 7.5. Absorption of the IgG component from an Hc antibody-rich fraction had no effect on the intensity of the homologous PCA reaction. Incubation of either whole serum or of isolated fractions at 56°C for 2 or 5 hr brought about only partial inactivation of Hc antibodies, indicating that Hc antibodies with different sensitivities to heat may coexist in the same serum and display similar if not identical gel filtration, chromatographic and electrophoretic properties.
Early Accumulation of Antibody Plaque-Forming Cells in Mouse Spleens Lacking a Pre-Existing Immune Background
Authors: Cerny Jan , McAlack Robert F. , Sajid Mohammed A. , Fronton Jackie , Friedman Herman
Published: December 2022

The early cellular events occurring during antibody formation to Vibrio cholerae somatic antigens were studied in the mouse spleen using a direct vibriolytic plaque assay in agar gel with viable bacteria as the indicator. Plaque-forming cells (PFC) with specificity for either the common A antigen or the type specific B antigen were distinguished by use of appropriate indicator strains of bacteria. No background of PFC to either of these antigens was detectable in spleens of normal non-immunized mice. Immunization with a relatively low dose of antigen (5 µg) resulted in appearance of the first PFC 42 to 46 hr later. There were consistently 9 to 10 PFC in the spleens of mice at this time, four of which were of anti-A specificity and five to six of anti-B specificity. Further accumulation of the cells was non-linear. There was a fivefold increase in the number of PFC at 48 hr, followed by a plateau between 48 and 60 hrt. A third increase, again about fivefold, occurred at 62 hr. Use of a higher dose of angien (500 µg) resulted in a longer latent period before appearance of the first PFC. This seemed due to a “masking” of PFC by excess antigen. Furthermore, no “staircase” accumulation was observed; in contrast there was a more rapid increase in the number of PFC, with periodic “waves.” The early differentiation of PFC could be inhibited by administration of various metabolic inhibitors of DNA between 24 and 36 hr after immunization. This suggests that DNA synthesis occurs during the latent period. The inhibitors had less, if any, effect when administered earlier after immunization.
Communications
Authors: Fauci Anthony S. , Johnson John S.
Published: December 2022

Antigenic competition is an immunologic phenomenon whereby the immunization of an animal with two antigens either simultaneously or in close sequence results in a decreased response to one or both antigens (1). Recently, studies of antigenic competition using two chemically defined haptenic determinants (2, 3) showed that when two haptens conjugated to the same or different protein carriers and emulsified in complete Freund's adjuvant (CFA) were simultaneously injected into the same footpads of rabbits, the specific antibody response to one or both haptens was diminished. Since precipitating antibody was determined at 13 and 20 days, responses being measured were predominantly 7S or IgG antibody.

Localized hemolysis-in-gel (LHG) techniques (4) facilitate the study of antibody production at the cellular level. Merchant and Hraba extended this technique to study antibody production against simple haptens (5, 6). Using the LHG technique, we describe the phenomenon of antigenic competition to haptenic determinants at the cellular level in rabbit lymph nodes.
Nonspecific Arbovirus Hemagglutinin Inhibitors in Turtle Sera
Authors: Yuill Thomas M. , Brandt Walter E. , Buescher Edward L.
Published: December 2022

The hemagglutination-inhibition (HI) test has been used extensively to document the presence of antibodies to various arboviruses in man and animals; its interpretation assumes that the observed HI reaction is due to viral antibody. That this is not always so has been reported for large numbers of hens (50% of 422 tested) and “antibody” for western equine encephalitis (WEE) virus (1), but only for a small number of turtles (1% to 2% of 166) reacting nonspecifically with eastern equine encephalitis (EEE) virus (2). During a period of EEE virus dissemination on the Eastern Shore of Maryland, substantial numbers of turtle sera inhibited EEE but not WEE hemagglutinins. Moreover, seroconversions in some of these turtles led us to believe initially that they might be naturally infected. While subsequent tests of these sera showed no neutralizing antibody, the apparent seroconversions by HI test were unexplained.
Selective Inhibition of Plaque-Forming Cells by Bursa-Specific Antisera
Authors: Potworowski E. F. , Richer G. , Borduas A. , Forget A.
Published: December 2022

The distinct role played in birds by the thymus and the bursa of Fabricius in the ontogenesis of the immune response is now well documented (1). Recently, the characterization of the lymphocytes of these two organs by their antigenic specificities has further emphasized their individuality (2). Their respective functional identity could also be demonstrated by selective immunologic inhibition. Thymus-specific antisera were shown to inhibit the graft-versus-host reaction elicited by buffy coat cells on chicken chorioallantoic membranes, whereas bursa-specific antisera had but little effect on that reaction (3). In the present work, we report that bursa-specific antisera can inhibit hemolytic plaque formation in spleen cell preparations while thymus-specific antisera have virtually no inhibiting effect.

The antisera, prepared by immunizing rabbits with fresh thymic and bursal lymphocytes from young chicks, were rendered organ-specific by cross-absorption. Full details of the immunizing schedule, absorptions of antisera and organ-specificity assay have been reported previously (2, 3).
Immunoglobulin Determinants on Mouse Lymphocytes
Authors: Jones G. , Torrigiani G. , Roitt I. M.
Published: December 2022

Specific antisera to mouse light chains and to each mouse Ig subclass were radiolabeled with 125I and used to detect surface Ig determinants on mouse lymphocytes by autoradiography. Light chain determinants were demonstrated on 34%, 29%, 18% and 4% of the lymphocytes in blood, spleen, lymph node and thymus, respectively. The total percentage of cells possessing heavy chain specificities in blood, spleen and lymph node agreed with that of cells bearing light chain determinants, suggesting that individual lymphocytes are restricted to one Ig subclass. Depletion of thymusdependent lymphocytes by thymectomy and anti-lymphocyte serum produced marked increase in the percentage of cells bearing surface Ig, suggesting that these cells belong to the thymus-independent population of lymphocytes.
Rabbit Platelet Injury by Soluble Antigen and Antibody
Authors: Marney Samuel R. , Des Prez Roger M.
Published: December 2022

Experiments with the sodium and magnesium salts of ethylene diamine tetra-acetic acid (EDTA) and ethylene-glycol-diamino-ethyl-tetra-acetic acid (EGTA) demonstrated that only MgEDTA fails to inhibit amine release from rabbit platelets by soluble antigen and antibody when present in a concentration just exceeding that of ionized calcium. Thus this reaction required ionized calcium in a concentration greater than 10-8 M. Addition of 10 mM EDTA to the reaction mixture interrupted amine release after it had begun. Recalcification so as to neutralize the EDTA caused amine release to resume, suggesting that this process may have a requirement for calcium ion other than for its role in the complement sequence. Incubation of platelets with antigen and antibody in the presence of EDTA caused “sensitization” of the platelets, presumably by attachment of immune complex. Platelets “sensitized” in this fashion released amine after washing and resuspension in intact, nonchelated plasma containing no antigen or antibody. The separation of platelet injury by soluble antigen and antibody into a sensitization phase and a release phase was not reported previously.
Complement Fixation in Conventional and Regressing Virus-Induced Murine Leukemia
Authors: Rich Marvin A. , Karl Seung , Clymer Rodney
Published: December 2022

Immunization of Swiss mice with conventional strains of murine leukemia virus did not elicit an appreciable humoral complement fixation (CF) response. In contrast, CF antibody was observed in mice infected with a virus that induced a spontaneously regressing leukemia. After infection with regressing Friend virus, (RFV), CF antibody was present in mice in which leukemia had regressed but was absent in leukemic mice. CF antibody elicited by RFV cross-reacted with other strains of murine leukemia virus.
Antibody Weight Estimates at the Nanogram Level
Authors: Revoltella Roberto , Adler Louise T. , Osler Abraham G.
Published: December 2022

A method is described for weight estimates of anti-dinitrophenyl antibodies in the nanogram range. The procedure is based on the interaction of the antibody preparation with a relatively large molar excess of the intensely iodinated dinitrophenylated dipeptide, DNP-L-lys,L-tyr,125I. The hapten-antibody complex may be precipitated equally well by 45% saturated ammonium sulfate or a goat antiserum to rabbit γ globulins. Data are given to indicate some of the parameters governing the results, the effect of albumin and the methods of calculation.
Enzymes Produced by a Pseudomonas Species Which Inactivate Inhibitors of Certain Viral Hemagglutinins
Authors: Schmidt Nathalie J. , Gee Pinkie S. , Dennis Juanita , Lennette Edwin H.
Published: December 2022

Elastase and phospholipase C which were purified from cultures of a psychrophylic Pseudomonas sp. were examined for ability to inactivate certain viral hemagglutinin inhibitors present in human sera. Both enzymes were required for inactivation of enterovirus and reovirus inhibitors, but adenovirus inhibitors were inactivated by the elastase alone. The Pseudomonas sp. elastase had much greater inhibitor-destroying activity than did pancreatic elastase or Flavobacterium elastase; the activity of the Pseudomonas sp. phospholipase C was comparable to that of phospholipase C from Clostridium perfringens. Cohn fraction III-0 (containing predominantly β lipoprotein) and fraction IV-1 (containing predominantly α lipoprotein) both inhibited enterovirus hemagglutinins. Immunoelectrophoretic studies showed that the combination of Pseudomonas elastase and phospholipase C produced marked alterations in both the α1 and β lipoproteins in human sera. Kaolin absorption also removed both α1 and β lipoproteins. Both of these treatments inactivate enterovirus HA inhibitors. Treatment of sera with heparin-MnCl2 or dextran sulfate-CaCl2, which precipitate β lipoproteins, removed only a portion of the inhibitory activity for enterovirus hemagglutinins. These findings suggest that enterovirus hemagglutinin inhibitors are present in both α and β lipoproteins of human sera.
Atypical Fluorescence in the Fluorescent Treponemal-Antibody-Absorption (FTA-ABS) Test Related to Deoxyribonucleic Acid (DNA) Antibodies
Authors: Kraus Stephen J. , Haserick John R. , Logan Leslie C. , Bullard Janice C.
Published: December 2022

Two distinct patterns of fluorescent treponemal-antibody-absorption (FTA-ABS) test fluorescence have been observed in sera from certain presumably nonsyphilitic patients with lupus erythematosus (LE); the entire Treponema pallidum antigen fluoresces (homogeneous pattern), or intermittent beads of fluorescence are present along the length of the treponeme (beaded pattern). In LE sera, the factor responsible for the homogeneous pattern was an IgG antibody occurring in titers up to 1:1280. Antibodies that produced the beaded pattern were present in similar titers, but they were distributed among immunoglobulin classes IgG, IgA and IgM. The occurrence of beaded FTA-ABS fluorescence was statistically associated with the presence of anti-nucleoprotein, anti-DNA and LE cell factors. The reaction that results in beaded FTA-ABS fluorescence was inhibited by incubating sera with DNA or nucleoprotein or by treating the T. pallidum antigen with DNAase. Anti-DNA antibody therefore appears to be responsible for the beaded FTA-ABS fluorescence reactions with sera from certain LE patients.
Serum Allotype Loss Due to a Lymphoid Tumor in Chickens
Authors: David C. S. , Fletcher O. J.
Published: December 2022

A rapidly growing transplantable lymphoid tumor in chickens caused the disappearance of selected allotype antigens at the a and c loci, which are presumed to control IgA. Two possible explanations are 1) molecules carrying the allotypes bind to the neoplastic cells, thereby enhancing tumor growth, and 2) the tumor virus infiltrates specific stem cells preventing further differentiation.
Detection of Morphine by Hemagglutination-Inhibition
Authors: Adler Frank L. , Liu Chi-Tan
Published: December 2022

The production of antibodies reactive with morphine and a radioimmunoassay sensitive to 0.5 ng of the drug has been recently described (1). This note confirms these findings and describes a rapid, simple and inexpensive hemagglutination-inhibition (HI) test of equal or greater sensitivity which appears to be adaptable to the screening of large numbers of urine specimens with a minimum of effort and equipment.

The immune serum used in the experiments to be described was obtained from a rabbit 4 weeks after a first injection with 1.0 mg of a conjugate of carboxymethylmorphine (CMM) and bovine serum albumin (BSA) in complete Freund's adjuvant and 1 week after a booster of another 1.0 mg. While the immunizing antigen was kindly donated by Dr. C. W. Parker, preparations of CMM and their conjugates used in the in vitro work and in the immunization of additional rabbits were prepared by us according to Spector and Parker and the references cited in their article (1).
Comparative Ligand-Binding by Purified Nurse Shark Natural Antibodies and Anti-2,4-Dinitrophenyl Antibodies from Other Species
Authors: Voss Edward W. , Rudikoff Stuart , Sigel M. M.
Published: December 2022

A population of 19S IgM specific for the 2,4-dinitrophenyl group was purified from normal nurse shark sera by immune precipitation with high concentrations of dinitrophenylated protein antigens. Hapten-binding properties of the purified antibody were difficult to measure in equilibrium dialysis because of the apparent low affinity of the antibody combining sites. Fluorescence quenching and spectral shift studies revealed significant binding with the bis-α, ε-DNP-l-lysine ligand but not with mono-ε-DNP-l-lysine. Comparative binding studies with these two ligands in the fluorescence quenching assay indicated that the combining sites of purified natural shark antibody resembled low-affinity, antigen-induced antibody in shark and other species. Qmax determinations and fluorometric titrations with antibody subunits indicated that binding of bis-α, ε-DNP-l-lysine was specific for the antibody combining sites.
Cutaneous Basophil Hypersensitivity
Authors: Dvorak Harold F. , Simpson Blanche A. , Bast Robert C. , Leskowitz Sidney
Published: December 2022

After guinea pigs are sensitized with protein antigens in saline or incomplete Freund's adjuvant (IFA), delayed-onset skin reactions elicited at early intervals have certain distinctive immunogenic requirements and unique histologic features that set them apart from classic delayed hypersensitivity. These reactions have been called “Jones-Mote” or cutaneous basophil hypersensitivity (CBH). In the present study lesions induced with insoluble antigen-antibody complexes in IFA have similar histologic characteristics and could also be regarded as examples of CBH. Delayed reactions elicited in animals sensitized with protein antigens in complete Freund's adjuvant (CFA) included numerous basophils when tested early after sensitization; however, at later intervals, this component became less prominent. Delayed lesions of contact allergy to dinitrofluorobenzene/dinitrochlorobenzene (DNFB/DNCB) similarly contained a prominent basophil component that was not diminished when testing was delayed for several weeks following sensitization. Immunization with DNFB in CFA sensitized animals for contact reactions that were more intense and indurated and contained a somewhat smaller proportion of basophils. CBH was passively transferred with lymph node cells from sensitized animals (but not with serum). Reactions in recipients were weak but histologically characteristic. Neither compound 48/80 nor specific antigen caused basophils to degranulate. Basophils did not accumulate one antigen tested, iodinated human serum albumin.
Immune Paralysis Induced by Pneumococcal Polysaccharide
Authors: Wu William G. , Horve Finn O. , Williams Carole C.
Published: December 2022

The phagocytic and antibody-inductive activity of peritoneal cells (PC) derived from mice immunized or immunologically paralyzed with capsular antigen of Diplococcus pneumoniae type III (SIII) was studied. In vitro exposure of PC from “paralyzed” animals to 32P-labeled bacteria indicated that their rate of phagocytosis was similar to that of controls. Cells from immune animals showed an increased uptake of organisms, this being attributed to cell-associated antibody. When studied by passive cell transfer procedures in normal recipients, PC from “paralyzed” and immune animals conferred protection against challenge with viable organisms. Data obtained from irradiated recipients indicated that PC were necessary for production of antibody by spleen cells. Furthermore, immune paralysis appeared to be disrupted by macrophages through an active process resulting in antibody production by formerly unresponsive cells.
Immune Paralysis Induced by Pneumococcal Polysaccharide
Authors: Williams Carole C. , Wu William G.
Published: December 2022

Passive transfer methods demonstrated that disrupted peritoneal cells (PC) and spleen cells (SpC) from mice immunized or immunologically paralyzed with type III pneumococcal polysaccharide antigen (SIII) could induce specific protection against infection in normal recipients. A part of this response to cell extracts could be ascribed to RNase-sensitive material. Injection of RNA derived from PC of such donors led to antibody production in recipients, and the inducing substance was partially RNase-sensitive. RNA preparations from SpC of “paralyzed” donors also evoked an excellent response in recipients, but did not contain an RNase-sensitive component. SpC of immune animals yielded an RNA which had poor antibody-inducing characteristics. Antigen was detected in the RNA of SpC from “paralyzed” donors, but not in other preparations. SIII was also detected in the blood serum of “paralyzed” donors, approximately 0.1 µg/ml being present 24 days following the injection of 100 µg.
Deficiency of C3 Inactivator in Man
Authors: Abramson Neil , Alper Chester A. , Lachmann Peter J. , Rosen Fred S. , Jandl James H.
Published: December 2022

C3 inactivator was undetectable in the serum of a previously described patient with increased susceptibility to infection, hypercatabolism of C3 and multiple protein deficiencies. His red cells were positive with specific C3-Coombs reagent but became negative after infusion of normal plasma. Incubation of his red cells with normal serum or with purified C3 inactivator rendered them C3-Coombs negative, whereas incubation in his own serum or buffer did not alter the C3-Coombs positivity. C3 deposited on red cells in vitro by treatment of purified C3 with trypsin or with C1s, C4 and C2 became similarly unreactive with C3-Coombs reagent following incubation with C3 inactivator. C3 inactivator, in addition to its effect on cell-bound C3, acted on fluid phase C3b to yield a product of approximately unchanged molecular size but with an electrophoretic mobility similar to that of C3c. C3 inactivator may play a role in conversion of C3b to C3c in vitro and, perhaps, in vitro.
IgA and Other Immunoglobulins from the European Hedgehog
Authors: Vaerman J. P. , Heremans J. F.
Published: December 2022

Hedgehog serum and exocrine secretions contain, besides IgM and two subclasses of IgG, a protein which cross-reacts with an antiserum against human IgA. Hedgehog serum IgA comprises a large proportion of molecules having a molecular size in the range of the 9 to 10S dimer of a 7S unit. The electronegative charge of hedgehog's serum IgA resembles that of human serum IgA as shown by its behavior upon DEAE-chromatography and electrophoresis. IgA was found to be the immunoglobulin having the highest secretion-to-serum concentration ratio in saliva and bile, and also to be the immunoglobulin produced by the majority of plasma cells in the lamina propria of the hedgehog's intestinal mucosa.
Serum Antigens of Cattle
Authors: Blakeslee Dennis , Butler John E. , Stone W. H.
Published: December 2022

Two antisera detecting different allotypic specificities, A1 and B1, of cattle immunoglobulins were produced by isoimmunization with purified IgG in Freund's complete adjuvant. The A1 marker was found to be restricted to the Fc portion of IgG2 heavy chains, whereas B1 was a marker of immunoglobulin light chains and was found on IgG, IgA and IgM molecules. Genetic analysis indicated that the alleles controlling A1 and B1, bva1 and bvb1, respectively, were autosomal dominants and that they were inherited independently. The bva1 allele was highly frequent in 11 breeds except Hereford, which showed a low frequency. In contrast, bvb1 was infrequent or absent in all breeds.
Prediction of Immunosuppressive Potency of Antilymphocyte Serum by in Vitro Testing
Authors: Gill P. G. , Venables W. , Ludbrook J.
Published: December 2022

Seventeen batches of rabbit anti-mouse lymphocyte serum (RAMLS) and normal rabbit serum were tested for ability to prolong allografts of CBA skin on BALB/c mice, and the leukoagglutinating, cytotoxic and both direct and indirect macrophage-cytophilic antibody titers of these sera were determined. Simple linear regressions of survival times of individual grafts on titers of each measurement disclosed statistically significant correlation in each case. Multiple linear regression analysis was used to establish the relative value of the four methods of assay in predicting allograft survival. The most efficient predictor was a mathematical model that included only the two cytophilic antibody titers. The predictive value was not improved by including agglutinating and cytotoxic antibody titers. These results established a close association of cytophilic activity with the immunosuppressive potency of RAMLS, and suggested that the roles of cytotoxicity and leukoagglutination per se are minor.
Distribution of Antigen-Ribonucleoprotein Complexes within Rat Peritoneal Exudate Cells
Authors: Bishop David C. , Gottlieb A. Arthur
Published: December 2022

Rat peritoneal macrophages were incubated with 125I-labeled preparations of the linear random copolymer L-Glu60Ala30Tyr10 (L-GAT). Following cell fractionation, the 125I label was distributed between the supernatant and ribosomal fractions in a ratio of 4:1. Over 95% of this ribosome-associated antigen was released by 1 M ammonium chloride. The labeled material released from the ribosomes in this way behaved as a unique macrophage ribonucleoprotein (RNP) species on polyacrylamide gels. This RNP-like material exhibited RNA-rich (ρ = 1.500 g/ml) and copolymerrich (ρ = 1.400 g/ml) components in cesium sulfate density gradients. The distribution of RNP species containing 125I L-GAT between the supernatant and ribosomal fractions of the macrophage was 2:1.
Purification and Structural Studies of Human Clq.
Authors: Yonemasu K. , Stroud R. M.
Published: December 2022

Human C1q has been purified by repeated precipitation at low ionic strength in the presence of chelating agents. The preparation was pure as judged by immunochemical and electrophoretic criteria. The final product had the biologic activities of C1q, but no detectable C1r or C1s activity. The yield of C1q by this method is about 40 to 60%.

Alternatively, C1q can be highly purified in smaller quantities for antigenic stimulation, using a single precipitation step, followed by two identical acrylamide gel steps. Rabbits antisera against this material were monospecific by immunodiffusion and immunoelectrophoresis, and agglutinated EAC1q cells, but not EA.

The molecular weight of C1q was estimated to be 388,000 on acrylamide gels containing SDS. C1q, lyophilized and dried to a constant weight, contained 13% nitrogen and its extinction coefficient, E1%1cm, was 6 to 6.5. Amino acid analyses and carbohydrate analyses have been carried out.
Studies on Human Complement: The Relationship of C3, C5, ß1c and ß1a
Authors: Nelson R. A.
Published: December 2022

Nine proteins of the complement system have been isolated from isolectric focusing columns. The pI values were: C1, 4.99; C4, 6.40; C2, 5.52; C3, 5.75; C5, 4.10; C6, 6.00; C7, 5.59; C8, 5.60 and 6.50; and C9, 4.72. Recovery was between 50% and 100% for each component. Reactivities of the C3 inactivator and of the cobra venom “cofactor” for C3 inactivation were found at pH 5.35. Results with C8 indicated either a double peak of hemolytic reactivity or the presence of a previously undefined inhibitor at pH 6.0.

The protein in fresh serum which precipitates ca. 90% of antibody from four standard antisera coded “anti β1c” was isolated as a sharp peak at pH 6.05 to 6.10 on 11 different occasions over a 2-year period. Another protein in “aged” serum or in preparations of C3 which were subjected to prolonged purification procedures peaked sharply at pH 6.76 and also precipitated most of the antibody in the four sera.
Inhibition of Chemotaxis by Human Leukocytes Following Incubation in Homologous Serum Devoid of Chemotactic Activity
Authors: Miller M. E. , Nilsson U. R.
Published: December 2022

Incubation of rabbit polymorphonuclear leukocytes (PMNS) with chemotactic factors has been shown to lead to a progressive decrease, or “deactivation,” in the chemotactic responsiveness of the PMNS (Ward and Becker). In these studies, we report that human PMNS can be similarly “deactivated” by humoral materials lacking chemotactic activity. Suspensions of human, peripheral blood PMNS (5 × 106 PMNS/ml) were incubated with equal volumes of undiluted homologous sera at 4, 23, or 37°C, or with serum that had first been incubated at 37°C for 10 min followed by 56°C for 20 min, or with heat-inactivated serum (56°C for 60 min), or with serum from which chemotactic activity had first been generated by the addition of pre-formed antigen-antibody complexes. After 30 min incubation, the cell suspensions were washed and tested for chemotactic response. Cell suspensions which had been incubated with materials having significant chemotactic activity had been “deactivated”—they failed to chemotax.