bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of Guinea Pig Mitogenic Factors
Authors: Geczy Carolyn L.
Published: December 2022

Mitogenic factor from BCG-sensitized cells stimulated with antigen (PPD) was found to have a m.w. between 20 and 25,000 daltons and an isoelectric point of about 7.5. The blastogenic activity of this factor was not affected by L-fucose or heating at 56°C for up to 1 hr. Mitogenic factor obtained from supernatants of allogeneic cell mixtures (MLC-MF) on the other hand, had a m.w. at 15 to 18,000 daltons and an isoelectric point of 6.5. The blastogenic activity of MLC-MF was inhibited by 0.1 M L-fucose. The factor was stable at 56°C for 1 hr.

An antibody prepared against MLC-MF inhibited the MLC reaction as well as the activity of MLC-MF on non-committed cells. This antibody, however, did not affect the response of lymphocytes to PHA or PPD and had no suppressive effect on PPD-MF. The antibody was not cytotoxic and its suppressive activity in the MLC response could not be absorbed out by lymphoid cells indicating that it is probably directed against a lymphocyte activation product (MLC-MF) rather than membrane antigens.

The chemical and immunologic differences exhibited by PPD-MF and MLC-MF indicate that these two lymphokines are distinct molecular entities.
Genetic Control of the Immune Response to Phage fd
Authors: Kölsch Eckehart , Falkenberg Frank W.
Published: December 2022

The immune response to bacteriophage fd has been tested in congenic resistant strains of mice. B10 mice are high responders; B10.BR and B10.D2 mice are low responders. B10.A mice with the recombinant H-2a haplotype as well as heterozygous H-2k/d (B10.BR × B10.D2) F1 mice respond with high antibody titers, indicating the existence of two complementing Ir genes. From the results obtained with recombinant and F1 hybrid mice, Irfdα can be localized to the right of the crossover between the IE and IC subregions, Irfdβ to the left of IB, presumably at IA. Complementation is observed with selected pairs of alleles only, suggesting α-β coupled complementation.

In addition to genes in the H-2 complex, there are non-H-2 background genes modulating H-2 regulated anti-fd response.

B10.BR mice lack primary IgM and IgG antibodies, yet are able to mount a secondary IgG anti-fd response, giving evidence for intact memory formation in this low responder strain.

The combined data are most easily interpreted by assuming that both H- and non-H- genes modulate the effective immunogenic dose of antigen.
Mechanism for Induction of Anti-DNA Antibodies by Bacterial Lipopolysaccharides in Mice
Authors: Izui Shozo , Kobayakawa Takatoshi , Zryd Marie-José , Louis Jacques , Lambert Paul-Henri
Published: December 2022

The capacity of various polyclonal B lymphocyte activators (PBA) to induce, in mice, the formation of anti-DNA antibodies was compared with their ability to mediate the release of DNA in circulating blood and to stimulate polyclonal antibody synthesis in vivo.

Anti-DNA antibodies or polyclonal antibody synthesis were induced in mice after the injection of at least 10 µg lipopolysaccaride (LPS) from Salmonella typhimurium, 1 mg dextran sulfate (DS), or 2 mg purified protein derivative of tubercle bacteria RT32 (PPD). Smaller quantities of LPS (0.1 µg) or DS (500 µg) were sufficient to cause the release of DNA in circulating blood, whereas PPD was not able to provoke such a release at any concentration used.

The association of anti-DNA antibodies with polyclonal antibody synthesis in mice injected with various PBA contrasts with the lack of correlation between the formation of anti-DNA antibodies and the release of measurable amounts of DNA in circulating blood. These results strongly suggest that the induction of anti-DNA antibodies by PBA is a consequence of the polyclonal B lymphocyte activation.
The Basic Concepts of Immunity
Authors: Manwaring W. H.
Published: December 2022

There are few immunologists who have not had occasion to resent the skepticism with which many of the theoretical phases of our subject have been received by workers in the older medical sciences. “An esoteric cult based on an hypothesis that was discarded generations ago by normal physiologists,” is a typical criticism, that was made in my presence, several years ago, by Professor Cushny. “An ingenious collection of metaphors,” was the parallel comment by Professor Starling. “A pseudoscience of juggled hypotheses and superficial analogies,” in the opinion of Jacques Loeb.

The obvious answer, of course, is to point to the fact that the criticism applies equally well to many phases of their own subjects. Nevertheless, I have thought it might not be without interest if I took up very briefly certain typical concepts of immunology, with an inquiry as to how far these concepts are in accord with known physiological facts.
A Study of Pollen Immune Serum
Authors: Ramsdell Susan Griffith
Published: December 2022

The use of pollen extracts in the treatment of hay-fever occasions certain demands upon the laboratory supplying this material: The solution should not be too irritating for subcutaneous administration.The solution should be the most potent possible.The solution should be stable in content of the antigenic substances.

In the effort to meet these conditions many formulae have been used; but the need for stabilizing, and especially for standardizing the process has not thus far been satisfactorily solved. Heretofore, the most dependable factor for measuring the strength of a given solution has been the nitrogen content. That this can be accurately correlated with the antigenic content or with the skin reaction has been lately questioned (1), (2), (3), (4). And even if the nitrogen alone could determine the original antigenic value of a solution, it offered no further evidence as to the stability of the extract alone or as to the influence of other factors, such as preservative.
On the Serological Relationship of Acid-Fast Bacteria
Authors: Furth J.
Published: December 2022

Since the early unsuccessful experiments of Koch (1) to differentiate various acid-fast bacilli by means of agglutination tests, numerous investigators have failed in similar attempts, (Defalle (2), Courmont and Potet (3), Fritzsche (4), Twort, Todd and Perkins (5) and others). Search for distinguishing differences among the various acid-fast microörganisms by means of the complement fixation test has revealed a universally strong group reaction. For this reason Bordet (6), Gengou (7), Fritzsche (4), Harris and Lanford (8), Cooke (9), Twort, Todd and Perkins (5) and others found differentiation practically impossible. In fact, some of these investigations increased the prevailing confusion for they seemed to bring acid-fast bacteria into intimate antigenic relationship with otherwise unrelated microörganisms. For example, Fritzsche (4) extends Koch's observations and finds that Actinomyces resemble the acid-fast bacilli. Massol, Calmette (10) states, has demonstrated that diphtheria bacilli bind antibodies produced by the tubercle bacilli and inversely, tubercle bacilli fix antibodies produced by diphtheria bacilli.
Heterologous Corpuscular Anaphylaxis
Authors: Hyde Roscoe R.
Published: December 2022

This paper furnishes proof for the theory of corpuscular anaphylaxis and disposes of the claim that the phenomenon is, in reality, due to small amounts of serum that may have been introduced with the corpuscles in the sensitizing and test doses. Sheep corpuscles sensitize guinea pigs to chicken corpuscles, and since sheep serum does not sensitize guinea pigs to chicken serum the conditions for serum anaphylaxis are excluded.

An unsuspected difference between the rabbit and guinea pig in their anaphylactic behavior toward these cells comes to light. Chicken cells sensitize rabbits to sheep and goat cells, but sheep cells do not sensitize rabbits to chicken cells. The converse is true of the guinea pig. Sheep and goat cells sensitize guinea pigs to chicken cells but chicken cells do not sensitize to sheep and goat cells.

The serum of the sheep and goat give cross reactions in guinea pigs, a relationship that holds for these corpuscles. The corpuscles of the sheep and goat, however, sensitize guinea pigs for chicken corpuscles. This relationship does not hold for sheep and goat sera, for they do not sensitize guinea pigs to chicken serum.

Table 7 shows that anaphylaxis in the rabbit follows the behavior of the hemolytic antibody in its action toward chicken, sheep and goat cells. The phenomenon does not follow the behavior of the agglutinin toward these cells. In the guinea pig anaphylaxis does not follow the behavior of either the agglutinins or hemolysins toward the red cells of the chicken, sheep and goat.

The death obtained in the guinea pigs with the red cells was typical of anaphylactic shock as evidenced by the onset of symptoms, the violent death and autopsy findings.
The Lethal Action of Brain Extracts for the Rabbit and Guinea Pig
Authors: Hyde Roscoe R. , Parsons Elizabeth I.
Published: December 2022

The following rather curious and unsuspected phenomenon was encountered in connection with the intravenous injection of an extract of rabbit brain into the ear vein of the rabbit. This tissue had been thoroughly macerated in physiological salt solution and allowed to stand for two hours at 37°C. The mixture was centrifuged, the supernatant liquid withdrawn and injected in 2 cc. amounts. The rabbits died within from two to three minutes without outward manifestation of violent symptoms such as one sees in anaphylactic shock. The rabbit dies as if all its vital functions were suddenly overwhelmed. Autopsy showed no distinguishing characteristics beyond a greatly engorged liver.

We were inclined at first to believe that death resulted from mechanical obstruction of the blood stream as it was impossible to centrifuge the macerated brain mixture free from finely suspended particles. This assumption was strengthened by the fact that the material when passed through a Berkefeld filter proved non-“toxic” for the rabbit even when injected in 10 cc. amounts.
Intermediate Molecular Weight Eosinophil Chemotactic Factors in Rat Peritoneal Mast Cells: Immunologic Release, Granule Association, and Demonstration of Structural Heterogeneity
Authors: Boswell R. Neal , Austen K. Frank , Goetzl Edward J.
Published: December 2022

Previously uncharacterized eosinophil chemotactic activity of apparent m.w. 1500 to 2500 by Sephadex G-25 gel filtration was present preformed in rat peritoneal mast cells, was associated with the mast cell granules, and was released by challenge of the mast cells with rabbit anti-rat F(ab′)2 antiserum. Further purification of this activity by Dowex-1 chromatography revealed two peaks of eosinophil chemotactic activity eluting at pH 5.4 to 4.4 and pH 3.1 to 2.1, respectively. On Sephadex G-50 gel filtration both activities filtered just behind the α chain of insulin (m.w. 2540). The more acidic material gave a major peak of eosinophil chemotactic activity on high pressure liquid chromatography whereas the less acidic material separated into two equal peaks of eosinophil chemotactic activity of different hydrophobicity. Both the less acidic and the more acidic intermediate m.w. activities required a concentration gradient for expression of eosinophil chemotactic activity and were capable of rendering eosinophils unresponsive to homologous or heterologous chemotactic stimuli. Checkerboard assays of the more acidic factor over a range of concentrations did not reveal chemokinetic activity in the chemotactic dose-range and neither the more acidic nor the less acidic factors enhanced the spontaneous migration of eosinophils when placed on the cell side alone of the chemotactic chamber. The mast cell serves as a source of several eosinophil chemotactic factors differing in size and charge, which could possibly stabilize the chemotactic gradient and sustain an influx of eosinophils to the site of mast cell activation.
Activation of the Alternative Complement Pathway by Pneumococcal Cell Wall Teichoic Acid
Authors: Winkelstein Jerry A. , Tomasz Alexander
Published: December 2022

Previous studies have shown that the cell wall plays a major role in the activation of the alternative pathway by the pneumococcus. The present studies were performed in order to identify the biochemical component of the pneumococcal cell wall responsible for activating the alternative pathway.

Teichoic acids were extracted from purified pneumococcal cell walls by three different procedures, and in each case they were able to activate the alternative pathway at concentrations as low as 3 to 10 µg/ml. In contrast, the residual cell walls, which contained the peptidoglycan, lost their activity in proportion to the amount of teichoic acid that had been extracted. Notably, cell walls extracted with periodate lost over 98% of their teichoic acid and the residual peptidoglycan was unable to activate the alternative pathway at concentrations as high as 1 mg/ml.

When an autolysin defective pneumococcal strain is grown in the presence of penicillin, cell wall polymers are secreted into the medium. Crude preparations of secreted cell wall polymers were found to be able to activate the alternative pathway. The activity was removed by absorption with TEPC-15 myeloma with anti-choline specificity. Since both cell wall teichoic acid and lipoteichoic acid contain choline and are removed by TEPC-15 absorption, the activity in the crude preparations of cell wall polymers could have been due to one or both of these compounds. However, purified lipoteichoic acid was unable to activate the alternative pathway. Thus, the ability of crude preparations of cell wall polymers to activate the alternative pathway was due to the presence of cell wall teichoic acid.

The results of these studies demonstrate that cell wall teichoic acid, rather than peptidoglycan, is responsible for pneumococcal activation of the alternative pathway.
Immune Response to Phosphorylcholine
Authors: Köhler Heinz , Richardson Bruce C. , Smyk Sue
Published: December 2022

Homologous and isologous anti-idiotypic antibodies against the PC-binding BALB/c myeloma protein HOPC-8 were raised in A/He and BALB/c mice, respectively. Isologous and homologous anti-HOPC-8 serum suppressed the response to PC in vitro specifically. The antibodies were purified by using HOPC-8 immunoabsorbent. Purified isologous and homologous anti-idiotypic antibody were labeled with 125I and compared for Ig class composition and idiotype-binding specificity. Both kinds of anti-idiotypic antibodies were predominantly IgG1, were highly specific for the HOPC-8 idiotype, and had a similar affinities for the PC-binding site. These findings demonstrate that anti-HOPC-8 antibodies raised in A/He and BALB/c mice are very similar in their biologic and immunochemical properties.
Infection with Bacillus Calmette-Guérin Activates Murine Thymus-Independent (B) Lymphocytes
Authors: Sultzer Barnet M.
Published: December 2022

Thymus-independent (B) lymphocytes from BCG-infected mice respond with enhanced or depressed levels of proliferation relative to the nonspecific levels of stimulation induced by tuberculin. The type of response obtained depends on how BCG was administered and the source of the lymphocytes. The proliferation of splenic B lymphocytes was depressed when BCG was given i.v. and unchanged when BCG was given subcutaneously, whereas the lymph node B cell response was enhanced regardless of the route of injecting BCG. BCG was found also to be a mitogen for normal B cells in vitro and to stimulate polyclonal activation of B cells in vivo. Finally, evidence is presented which indicates that the depressed splenic B cell response to tuberculin may be due to the activation of suppressor T cells.
Isolation and Characterization of Distinct Antibody Populations from Antisera Directed against Idiotypes of Rabbit Homogeneous Antibodies
Authors: Mudgett M. , Coligan J. E. , Kindt T. J.
Published: December 2022

A hapten, 3-0-α-N-acetylgalactosaminosyl-N-acetylgalactosamine, was found to inhibit the binding of eight idiotypic antisera directed against different homogeneous antistreptococcal Group C polysaccharide antibodies. The sensitivity of the different idiotypic reactions to inhibition by a given concentration of hapten was found to be quite variable, ranging from 20 to 80% inhibition. This result suggested that idiotypic antisera contained two populations of antibodies: those which are reactive with determinants in the antigen-binding site and those directed against nonbindingsite idiotypic determinants. This suggestion was verified by fractionation of idiotypic antisera on affinity chromatography columns consisting of homogeneous antibodies attached to Sepharose. The absorbed idiotypic antibodies were separated into two populations by successive elution with the Group C hapten and ammonium thiocyanate. The antibody fraction obtained by elution with the hapten comprised approximately 33% of the total idiotypic antibody recovered from the column. The hapten-eluted fractions from different idiotypic antisera varied in their sensitivity to hapten inhibition although each could be inhibited to a greater extent than the original antiserum or the ammonium thiocyanateeluted fraction. One of the hapten anti-idiotype fractions appeared to be homogeneous by isoelectric focusing whereas the other hapten and ammonium thiocyanate-eluted fractions contained multiple bands. Kinetic analysis revealed that idiotypic reactions of antibody fractions obtained by hapten elution were competitively inhibited by the hapten, although the mechanism by which the competition occurs was not completely resolved by this analysis.
Immunochemical and Biologic Studies of the Lipopolysaccharide of Bacteroides Melaninogenicus Subspecies Asaccharolyticus
Authors: Mansheim Bernard J. , Onderdonk Andrew B. , Kasper Dennis L.
Published: December 2022

Lipopolysaccharide (LPS) was isolated from the outer membrane complex of Bacteroides melaninogenicus by gel chromatography by using sodium deoxycholate (NaD), an endotoxin-disaggregating detergent in the running buffer. Reaggregation occurred after removal of residual NaD. LPS was also prepared by the phenol/water method for comparison. The LPS was composed of loosely bound lipid (62%) and carbohydrate (32%), with less than 5% protein. Glucose, galactose, and glucosamine were the major sugars as detected by gas-liquid chromatography (GLC). Heptose and 2-keto-3-deoxyoctonate (KDO) were found to be absent when using colorimetric tests. Fatty acid analysis by GLC disclosed an unusual pattern; β-OH myristic acid was notably absent. Biologic activity was assessed with the dermal Shwartzman test, the Limulus lysate assay, and the chick embryo lethality test. LPS preparations gave no positive skin reactions in rabbits in doses up to 1 mg, compared to Salmonella typhi 0:901 endotoxin which was positive at 12.5 µg. Limulus lysate gelation and chick embryo lethality occurred at doses 30-fold or more different from Salmonella endotoxin controls. Hemagglutination inhibition (HAI) studies demonstrated the presence of capsular polysaccharide as a contaminant found in the phenol/water extracted LPS. This polysaccharide could be removed partially by further purifying the LPS with ultracentrifugation.

The LPS of B. melaninogenicus lacks two of the core sugars characteristic of aerobic Gram-negative endotoxins, has a unique fatty acid composition, and displays low biologic potency. These findings may explain the rarity of septic shock in patients infected with this organism. The presence of capsular polysaccharide contaminating the phenol/water-extracted LPS may explain partially the serologic heterogeneity which has been described previously in studies of the LPS of this organism.
Responses of Human Platelets to Immunologic Stimuli: Independent Roles for Complement and IgG in Zymosan Activation
Authors: Martin S. Eric , Breckenridge Robert T. , Rosenfeld Stephen I. , Leddy John P.
Published: December 2022

Analysis of the zymosan (Z)-induced human platelet response with washed platelets, fresh agammaglobulinemic plasma, and purified human IgG has identified a previously unrecognized role of IgG in this reaction, in addition to the previously reported requirements for C and fibrinogen. The rate-limiting step involves the assembly of C components on the Z particles and is independent of immunoglobulin. After the heat-labile components (supplied by agammaglobulinemic plasma) are Z bound, Z-specific IgG (apparently antibody) can bind to Z very rapidly, conferring on the previously inactive Z-complement-fibrinogen intermediate the capacity to induce platelet aggregation and serotonin release. The critical function of this Z-bound IgG appears to be in the interaction of the active Z complex with the platelet through Fc receptors, since soluble Fc fragments of each IgG subclass can totally block platelet activation. It is proposed that the human platelet requires a dual, or multi-site, stimulation to become activated during some forms of immunologic injury. These findings may provide some insight into clinical syndromes involving thrombocytopenia and intravascular coagulation during sepsis.
Generation of Cytolytic T Lymphocytes in Vitro
Authors: Ryser Jean-Etienne , Cerottini Jean-Charles , Brunner K. Theodor
Published: December 2022

Supernatants (SN) from secondary (20) mixed leukocyte cultures (MLC) obtained 18 to 24 hr after restimulation of C57BL/6 anti-DBA/2 long-term MLC cells with irradiated spleen cells of the original stimulating strain are shown to stimulate strong cytolytic T lymphocyte (CTL) responses when added to primary (10) long-term MLC cells. Under the conditions used, no stimulating activity was found in SN obtained from MLC containing normal C57BL/6 spleen cells and irradiated DBA/2 spleen cells, or from control cultures of C57BL/6 anti-DBA/2 long-term MLC cells and irradiated spleen cells of the responding strain. Active SN were also produced after restimulation of long-term C57BL/6 anti-DBA/2 MLC cells that had previously undergone six successive cycles of specific restimulation in vitro. Since the seventh restimulation resulted in a proliferative response without formation of CTL, this finding suggests that the cells involved in the production of active SN may be distinct from CTL or their precursors.

The stimulatory activity of 20 MLC SN was particularly evident after SN addition to 10- to 15-day 10 MLC cells, whereas the response of 20-day 10 MLC cells and of alloimmune spleen cells was of a lower magnitude. No significant lytic activity was generated after addition of 20 MLC SN to normal spleen cells. Under optimal conditions, the kinetics of the CTL response induced by 20 MLC SN was similar to that obtained after restimulation with irradiated allogeneic cells. While the same SN was able to stimulate CTL responses in 10 long-term MLC prepared with unrelated strain combinations, the lytic activity thus obtained was specific for the original stimulating alloantigens used during 10 stimulation. The CTL response of long-term MLC cells was unaffected by prior removal of cellular debris followed by trypsin treatment. Taken together, these results suggest that SN produced in 20 MLC contain nonspecific factors that are able to induce 20 CTL responses without the participation of alloantigen.
Cellular Basis of Tolerance to Serum Albumin in Adult Mice
Authors: Zan-Bar Israel , Murphy Donald B. , Strober Samuel
Published: December 2022

The surface markers and size of suppressor cells were determined in adult (BALB/c × C57BL/Ka)F1 mice which were tolerized with a single injection of deaggregated bovine serum albumin (BSA). Suppressor cells from the spleens of tolerized donors were assayed in a cell transfer system in which graded numbers of cells were injected into irradiated syngeneic mice along with limiting numbers of T cells primed to BSA and an excess of B cells primed to DNP-BSA. Adoptive hosts were challenged with DNP-BSA in saline, and the anti-DNP response was measured. Suppressor cells were antigen specific as shown by the inhibitory activity of BSA-tolerant spleen cells on the response to DNP-BSA, but not to DNP-BGG. Suppressor cells were eliminated by in vitro treatment with anti-Thy 1.2, anti-Ly-2.2, anti-I-J subregion antisera and C, but not with anti-Ly-1 or anti-I-A subregion antisera. Neither unprimed nor primed helper T cells were detected in the spleen of tolerized donors after in vitro treatment with anti-Ly-2.2 antisera. Both helper and suppressor T cells from the spleens of primed or tolerized donors, respectively, showed a rapid sedimentation velocity (S > 3.7 mm/hr).
Inherited Deficiency of the Sixth Component of Complement: a Silent or Null Gene
Authors: Glass David , Raum Donald , Balavitch Diane , Kagan Elliott , Rabson Arthur , Schur Peter H. , Alper Chester A.
Published: December 2022

Four families have been studied, some members of which have inherited deficiency of the sixth component of complement. The genetically determined electrophoretic variants of C6 were evaluated in all family members. Seven individuals were found who did not have the variant found in the serum of the parent from whom they inherited the deficiency. It is inferred that the isolated low levels of C6 in these individuals results from the heterozygous state of a normal C6 variant gene and a silent or null C6 gene; the genes determining electrophoretic variants and the low serum levels of C6 are allelic.
Transplantation in Miniature Swine
Authors: Lunney Joan K. , Sachs David H.
Published: December 2022

Immunochemical analyses of radioactively labeled lymphocyte antigens from miniature swine of three different homozygous major histocompatibility (MHC) types, AA, CC, and DD, have been performed. Anti-MHC sera were incubated with lentil lectin purified Nonidet P-40 swine lymphocyte extracts. Antigen-antibody complexes were then precipitated with protein A bearing staphylococci, eluted, and electrophoresed on sodium dodecyl sulfate polyacrylamide gels. Analysis of antigens from AA or DD cells revealed peaks of 42,000, 31,000, 25,000, and 11,000 dalton m.w. Platelet absorption of the anti-MHC sera yielded antibodies that only precipitated the intermediate m.w. molecules and lysed a subpopulation of swine peripheral blood lymphocytes, suggesting that these molecules were the miniature swine analogues of murine Ia antigens. Antibodies eluted from platelets lysed all lymphocyte populations and precipitated only the 42,000 and 11,000 dalton peaks, indicating that these molecules represent the analogue of murine H-2 histocompatibility antigens, containing a heavy chain and putative swine β2-microglobulin.
Immunofluorescent Detection of Human B Cell Alloantigens on S-Ig-Positive Lymphocytes and Epidermal Langerhans Cells
Authors: Stingl Georg , Katz Stephen I. , Abelson Leslie D. , Mann Dean L.
Published: December 2022

Several similarities exist between Langerhans cells (LC), a subpopulation of the mammalian epidermis, and cells of the monocyte-macrophage-histiocyte series, including the expression of common surface marker characteristics (Fc-IgG and C3 receptors). In further analogy, we asked the question of whether these cells also bear the human equivalent of murine and guinea pig Ia antigens, i.e., B cell alloantigens. Since LC constitute only 2 to 4% of isolated epidermal cells, cytotoxicity techniques did not seem feasible for this purpose. We therefore elaborated an indirect immunofluorescence (IF) technique for the detection of B cell alloantigens, using fluorescein-isothiocyanate-labeled staphylococcal protein A (FITC-SPA) as indicator reagent. Alloantisera were obtained from multiparous women of a highly inbred population. In simultaneous experiments we tested purified B cells from seven normal donors against a panel of alloantisera by a microcytotoxicity test and by IF. Data obtained showed that the alloantigens are selectively expressed on SIg-positive, but not on SIg-negative lymphocytes. Furthermore, there was an excellent agreement between both techniques (correlation coefficient: X2 = 31.09, p <0.001). Isolated epidermal cells from those individuals whose B cells were tested for the expression of alloantigens were subjected to the IF procedure described above. None of the antisera which gave negative reactions on purified B cells of the respective individual reacted with its epidermal cells. By contrast, several of the antisera stained a small percentage of epidermal cells of those individuals whose B cells reacted with the very same antisera. By a simultaneously performed rosette assay with IgG-antibody-coated bovine red blood cells, it could be shown that the fluorescent and rosetted cell populations were identical, which means that—by IF techniques—LC are the only epidermal cells expressing B cell alloantigens.
Labeling Characteristics and Separation of Ia Antigen Subunits
Authors: Schwartz Benjamin D. , Vitetta Ellen S. , Cullen Susan E.
Published: December 2022

Experiments with biosynthetic incorporation of 3H-amino acids into murine and guinea pig Ia antigens have indicated that these antigens consist of two polypeptide chains of 33,000 and 25,000 daltons, respectively, occasionally linked by disulfide bonds into a 58,000 dalton molecule. In contrast, studies with lactoperoxidase-catalyzed radioiodination have indicated that these Ia antigens consist of only a single chain of 25,000 daltons. We therefore undertook a study to explore the basis of these discrepant results. Since 3H-tyrosine labeled both chains well, the lack of tyrosine residues in the 33,000 dalton chain could not be the explanation for the lack of radioiodination. However, by partially purifying the Ia antigen preparation with Lens culinaris (lentil) lectin affinity chromatography before immunoprecipitation and by increasing the resolution of analysis by using discontinuous-SDS polyacrylamide gel electrophoresis, it was possible to show that the 33,000 dalton chain was in fact radioiodinated, though still poorly so relative to the 25,000 dalton chain, and that a radioiodinated 58,000 dalton molecule could be detected. These experiments suggest that the 25,000 dalton chain is more exposed to the external cellular environment, and thus more readily iodinated by lactoperoxidase. In addition, the studies indicate that the choice of labeling method, purification procedures, and analytical methods must be taken into account when interpreting experimental results.
Glycolipid Markers of Murine Lymphocyte Subpopulations
Authors: Stein Kathryn E. , Schwarting Gerald A. , Marcus Donald M.
Published: December 2022

We have shown previously that purified antibodies to ganglioside GM1 react with peripheral T cells and most thymocytes in several strains of mice, independent of Thy-1 phenotype. GM1 and the Thy-1.2 antigen cap independently on C3H thymocytes, which provides additional evidence that GM1 is not the Thy-1.2 antigen. In C3H and nude mice antibodies to GM1 also react with a population of cells, comprising about 25% of lymphocytes from lymph nodes or spleen, that bear surface immunoglobulin. After removal of immunoglobulin from these cells by digestion with proteolytic enzyme, the GM1+ cells regenerate their surface immunoglobulin during 18 hr in culture, which indicates that these double-labeled cells synthesize their surface immunoglobulin. Protease treatment of lymphocytes reveals receptors for antibodies to GM1 on most cells. These data indicate that T and B cells differ in the accessibility of GM1 to antibody, and not necessarily in their content of GM1.

Purified antibodies to asialo GM1 react with mature T cells in all strains of mice tested. In contrast to anti-GM1, these antibodies do not react with most thymocytes, with immunoglobulin-bearing lymphocytes of C3H or nude mice, nor with pronase-treated B cells.
LPS-or 8Br-Cyclic AMP-Induced Production of T Cell-Activating Factor(s) in Macrophage Tumor Cell Line J774.1
Authors: Okada M. , Kishimoto T. , Igarashi T. , Teranishi T. , Yamamura Y.
Published: December 2022

The macrophage tumor cell line J774.1 replaced the function of normal macrophages in the induction of polyclonal killer T cells with 2-mercaptoethanol. J774.1 does not normally release soluble factor(s) which we have shown to be responsible for the differentiation of T cells to killer T cells. However, stimulation of J774.1 with LPS induced soluble factor(s) for T cell activation. An optimum concentration of LPS for the production of soluble factor(s) was 1 to 10 µg/ml, which completely inhibited growth of the tumor cells. The production of soluble factor(s) was observed within 6 hr after LPS stimulation and reached its maximum level at 24 hr. Incubation of the cell line with 8Br-cyclic AMP and theophylline induced soluble factor(s), suggesting that LPS stimulation induced an increase in intracellular cyclic AMP which leads to the synthesis of soluble factor(s).
Comparison of the Homogeneous, Primary Anti-Dextran B1355 Antibody Raised in BALB/c Mice with Protein 104E
Authors: Mayers G. L. , Bankert R. B. , Pressman D.
Published: December 2022

The primary antibody response to dextran B1355 in BALB/c mice is largely a homogenous IgM antibody. This antibody appears to be similar if not identical to the myeloma protein, protein 104E, for the following reasons: a) isoelectric focusing of the 7S monomers, separately and together in co-isoelectric focusing, give the same pattern, both in the presence and absence of 4M urea; b) the inhibition of lysis of the dextran-coated SRBC by specifically purified anti-dextran antibody and by protein 104E required essentially the same concentration of dextran B1355. This similarity was further demonstrated by plaque assays with dextran-coated SRBC, in which the formation of plaques was inhibited by free dextran. Inhibition of plaques produced by both types of cells required essentially the same concentration of dextran B1355. On these bases, there appears to be no difference in properties (or in structure) between the myeloma protein and the induced antibody.
The Effects of Phagocytosis, Dextran Sulfate, and Cell Damage on PGE1 Sensitivity and PGE1 Production of Macrophages
Authors: Gemsa Diethard , Seitz Michael , Kramer Wiebke , Till Gerd , Resch Klaus
Published: December 2022

During phagocytosis of zymosan or latex particles and after treatment with dextran sulfate, rat peritoneal macrophages responded to PGE1 by an elevated and prolonged production of cyclic AMP. Levels of cyclic AMP were not only augmented at the maximal effective concentrations of PGE1 but also at low, normally ineffective concentrations. Phagocytosis-induced enhancement of PGE1 sensitivity was proportional to the amount of interiorized particles, was independent of opsonizing factors, and did not occur in lymphocytes. In contrast, phagocytosis of silica particles or treatment with carrageenan caused a rapid loss of the response to PGE1, which was followed later by cell damage.

Synthesis and release of PGE1 was induced in macrophages by phagocytosis of zymosan but not of latex particles. Zymosan-induced PGE1 release was rather delayed and occurred at a high rate when the enhanced PGE1 sensitivity during phagocytosis had returned to near normal. Exposure of macrophages to silica particles or carrageenan also caused a high release of PGE1, however, it was associated with progressive enzyme leakage, which was used as a criterion for cell damage.

These observations suggest that phagocytosis-induced enhancement of PGE1 sensitivity combined with a subsequent release of PGE1 may be a sensitive control mechanism that may selectively regulate macrophage function under physiologic conditions. In contrast, the release of large amounts of PGE1 when associated with a loss of PGE1 sensitivity, may lead to an uncontrolled inflammatory response.
The Kinetics of Target Cell Lysis by Cytotoxic T Lymphocytes: A Description by Poisson Statistics
Authors: Chu Gilbert
Published: December 2022

The kinetics of T cell killing are analyzed with the assumption that encounters between killer and target cells occur at random. The application of Poisson statistics leads to a number of theoretical predictions which were then tested experimentally. Good agreement between data and theory was found.
Virus-Induced Immune Complex Disease: Identification of Specific Viral Antigens and Antibodies Deposited in Complexes during Chronic Lymphocytic Choriomeningitis Virus Infection
Authors: Buchmeier Michael J. , Oldstone Michael B. A.
Published: December 2022

Structural proteins of LCMV were identified and their role in the immune complex glomerulonephritis of LCMV carrier mice was examined. Purified LCMV contained three major polypeptides, a single nonglycosylated nucleoprotein with an estimated m.w. of 63,000, and two surface glycoproteins of 54,000 and 35,000. Deposition of nucleoprotein antigen in the glomeruli of LCMV carrier mice of several strains was demonstrated by immunofluorescent staining with a monospecific antibody. In addition, Ig eluted from kidneys of three strains of LCMV carrier mice was shown by immune precipitation to react against all of major viral polypeptides of LCMV. Antibody from normal mice, and from mice with immune complex disease unrelated to LCMV did not show deposition of LCMV antigen in glomeruli, and Ig eluted from the kidneys of these mice did not react against LCMV antigens. Hence, mice infected at birth with LCMV and persistently infected throughout their life make antibodies to all the known structural polypeptides of the virus.
Depression of Mitogen Response in Spleen Cells from Reticuloendotheliosis Virus-Infected Chickens and Their Suppressive Effect on Normal Lymphocyte Response
Authors: Scofield Virginia L. , Bose Henry R.
Published: December 2022

Spleen cells and peripheral blood lymphocytes from chickens infected with reticuloendotheliosis virus (REV) were depressed in their responsiveness to phytohemagglutinin (PHA). When spleen cells from uninfected chickens were co-cultured with spleen cells from chickens infected with REV at 2 weeks of age, the PHA response by the normal cells was completely suppressed. Although spleen cells from chickens infected with REV at 6 or 9 weeks of age were also suppressed in their ability to respond to PHA, they did not suppress the mitogenic response of normal cells in mixed cultures.
Opinion
Authors: Benacerraf Baruj
Published: December 2022

Ever since the discovery of specific immune response (Ir) genes in the I regions of the major histocompatibility complex (MHC) of mammals (1), certain characteristic features of the phenomena they control presented a considerable challenge: H-linked Ir gene control has been observed only for thymus-dependent antigens that are proteins or polypeptides, and is concerned with T cell responses such as the stimulation of delayed-type hypersensitivity (DTH) and specific helper T cells (1, 2).The specificity of Ir gene function is remarkable and distinguishes readily between antigens with different amino acid primary sequences (3–6). This has been demonstrated for synthetic antigens and also for native antigens. It is nevertheless remarkably distinct from the specificity of the T cell responses themselves (7), a matter of considerable concern to me when we initially proposed that the Ir gene product could be the elusive T cell receptor (1).
The C5b-6 Complex: Formation, Isolation, and Inhibition of Its Activity by Lipoprotein and the S-Protein of Human Serum
Authors: Podack Eckhard R. , Kolb William P. , Müller-Eberhard Hans J.
Published: December 2022

C5b-6 complex was generated by activation of C7-depleted serum and purified 1200-fold based on its hemolytic activity. A homogeneous bimolecular complex with an s-rate of 11.5S and a m.w. of 328,000 daltons was obtained. The electrophoretic mobility was -3.4 × 10-5 cm2/volt sec. The subunit composition as revealed by SDS-polyacrylamide electrophoresis without reduction showed two protein bands: C5b (180,000) and C6 (120,000); and with reduction three subunits: C6 (128,000), C5bα (122,000), and C5β (75,000). Immunochemical analysis confirmed complex formation between C5b and C6 and showed the presence of one of the two neoantigens expressed by the SC5b-9 complex. Sixty nanograms of C5b-6 represented 108 hemolytically effective molecules. A hemolytic efficiency of 12 to 20 C5b-6 cell-bound complexes per lytic event was calculated from the uptake of radiolabeled C5b-6. The hemolytic efficiency of C5b-6 is dependent on the cell concentration.

C5b-6-initiated lysis is strongly inhibited by serum low density lipoproteins with an inhibition constant of Ki = 34 µg/ml and by the S-protein with Ki = 39 µg/ml. Lipoproteins and S-protein form a stable, hemolytically inactive complex with nascent C5b-7. The mechanism of C5b-7 inhibition is interpreted as a competition reaction between the target cell membrane and the inhibitors for the transitory binding site in nascent C5b-7.
Lipopolysaccharide Induction of IgM Antibodies to Polyadenylic Acid in Normal Mice
Authors: Fischbach Michael , Roubinian Jirayr R. , Talal Norman
Published: December 2022

Lipopolysaccharide (LPS) is able to induce autoantibodies reversibly in normal mice. Single or multiple doses of LPS induced a rapid and dose-dependent rise in antibodies to Poly A from 113 ng to 590 ng/ml serum as determined by Millipore filter radioimmunoassay. The response peaked at day 3 and was over by day 21. No response was seen if the LPS was chemically inactivated or injected into genetically nonresponsive mice. Antibodies were specific for Poly A and were not induced by T cell mitogens. Sucrose gradient ultracentrifugation demonstrated only IgM antibody. Neonatal thymectomy altered neither the immunoglobulin class nor quantity of antibody. Neonatal splenectomy did not affect antibody class but reduced the amount produced. No free Poly A could be detected in circulation after LPS stimulation. These findings suggest that normal mice have B lymphocytes capable of limited and reversible autoantibody production.
Binding and Activation of the First Component of Human Complement by the Lipid A Region of Lipopolysaccharides
Authors: Cooper Neil R. , Morrison David C.
Published: December 2022

A number of structurally different LPS preparations differing in content of lipid A and polysaccharide were examined for their ability to bind and activate the first component of human complement, C1. Lipid A-rich LPS preparations as well as isolated lipid A were found to be orders of magnitude more active in binding highly purified precursor C1 as well as C1 than polysaccharide-rich preparations. This interaction was shown to occur through the C1q portion of the C1 macromolecule. Attachment of the lipid A-rich LPS preparations to C1 is a biologically meaningful event as it was demonstrated to lead to activation of proenzyme C1 reconstituted from highly purified C1q and proenzymes C1r and C1s. Further, an unusual dose response profile was obtained for the lipid A-rich LPS preparation from Salmonella minnesota R 595 in that relatively high doses of LPS bound C1 but activation was inhibited. Lower doses, in contrast, bound and also activated C1. Isolated lipid A, by comparison, bound and activated C1 efficiently at high and low dose ranges.

The lipid A region of the LPS molecule was found to be responsible for classical pathway activation. This conclusion derives from several observations. First, isolated lipid A was more active in binding and activating C1 and LPS containing even minor proportions of carbohydrate. Second, polymyxin B, an antibiotic that binds to the lipid A portion of LPS, abrogated the C1-activating ability of the preparations. Third, the presence of LAP, a protein that is associated with lipid A in certain LPS preparations, interfered with C1 binding by LPS. Finally, alteration of lipid A by mild alkaline hydrolysis abrogated its C1-binding capacity. These studies thus show, in a purified system, that the lipid A region of lipid A-rich LPS preparations isolated from rough strains of bacteria can directly and efficiently activate the classical pathway at the C1 step. This reaction proceeds in the absence of antibody. Thus, in this system C1q fulfills a recognition role normally associated with the antibody molecule.
Lymphocytes in the Rabbit Expressing Receptors for the Fc Portion of Rabbit Immunoglobulin
Authors: Birch Ruth E. , Bernier George M. , Fanger Michael W.
Published: December 2022

Nonspecies-specific receptors for the Fc portion of immunoglobulin (RFc) were detected in various rabbit lymphoid tissues by the ability of the RFc-bearing cells to form rosettes with ox red blood cells coated with the IgG fraction of a rabbit antiserum directed against ox red blood cell stroma. The rosette assay was shown to be specific for Fc receptor activity. In lymphocyte preparations containing less than 5% phagocytic cells the average percent of RFc-bearing lymphocytes in the various tissues was as follows: appendix 12%, lymph node 9%, peripheral blood 26%, spleen 31%, and thymus 0%. In double assay experiments, in which the lymphocytes were prestained with an FITC-labeled goat anti-rabbit Fab IgG fraction before rosetting, the RFc-bearing lymphocytes were found to constitute a subpopulation of the surface Ig (sIg)-bearing cells in the tissues examined when normal nonactivated rabbit lymphocytes were studied. After Pronase treatment, under conditions sufficient to remove cell surface Ig, and subsequent incubation to allow resynthesis of sIg, the percentage of sIg-bearing cells was restored to pretreatment levels. Conversely, although the Pronase treatment initially had no effect on RFc, there was a decline in detectable RFc-bearing cells after incubation in both treated and untreated preparations. Even with the loss of RFc+ lymphocytes during the incubation period, the percentage of sIg+ lymphocytes that expressed RFcγ remained fairly constant, suggesting the existence of an RFc+-sIg- subpopulation in the rabbit. Double assays employing FITC-labeled goat anti-rabbit antibody specific for µ and γ determinants were performed to determine if there was any relationship between the class of sIg expressed and the presence of Fc receptor. There was an overlap in IgM- and IgG-bearing lymphocytes, but no apparent overlap between IgM+ RFcγ+ and IgG+ RFcγ+ lymphocytes, suggesting that cells expressing both surface IgM IgG do not possess RFcγ. In the present study cells with receptors for the Fc portion of IgM were not detected in any of the tissues examined under the conditions employed.
Adherent Cell Function in Murine T Lymphocyte Antigen Recognition
Authors: Rosenwasser Lanny J. , Rosenthal Alan S.
Published: December 2022

The data in this report describe a T cell proliferation assay with nylon wool column-purified murine lymph node lymphocytes from animals immunized by footpad injection of antigen in CFA. It was found that the in vitro immune response of sensitized T cells to soluble protein antigens was functionally dependent on the presence of adherent cells, more specifically macrophages, at all concentrations of in vitro antigen challenge. The response was due to T cells in that cytotoxic treatment of the immune lymphocyte cells with anti-Thy 1.2 serum and complement effectively eliminated the antigen-specific DNA synthetic responses. The antigen-specific proliferation of murine lymphocytes depleted of adherent cells could not be reconstituted with either guinea pig macrophages nor murine fibroblasts, indicating the existence of species and cell type specificity. In contrast to previous observations in the guinea pig, soluble products of cultured adherent cells could at least partially replace the function of intact macrophages in the response to antigen.