bluuebunny/crossref_metadata_2025 huggingface dataset:
Biological Expressions of Lymphocyte Activation
Authors: Rich Robert R. , Pierce Carl W.
Published: December 2022

Mouse spleen lymphocytes activated by a mitogenic concentration of concanavalin A (Con A) secrete a factor(s), termed soluble immune response suppressor (SIRS), which is a potent, non-cytotoxic inhibitor of primary plaque-forming cell (PFC) responses by mouse spleen cells to heterologous erythrocytes in vitro. SIRS activity was detected in supernatant fluids of spleen cell cultures within 6 hr of stimulation with Con A and was maximal from 12 to 24 hr after stimulation. SIRS profoundly suppressed 5-day PFC responses when added to antigen-stimulated spleen cell cultures at initiation, was less effective when added 24 hr after initiation, and had no significant effect when added 48 or more hours after initiation. SIRS had to be present in antigen-stimulated spleen cell cultures only during the initial 24 hr of incubation to cause suppression of PFC responses which by kinetic analysis was not manifest until 120 hr of incubation. Background and antigen-stimulated IgM and IgG PFC responses were suppressed by SIRS.

Exposure to SIRS did not render spleen cells totally incapable of developing PFC responses to heterologous erythrocytes. Instead, antigen-stimulated PFC responses in control cultures and cultures containing SIRS had the same kinetics and were of comparable magnitude during the first 96 hr of incubation. Thereafter, however, PFC responses in cultures containing SIRS abruptly aborted and the SIRS-induced suppression of PFC responses became evident at 120 and 144 hr of incubation. Although the precise mechanism(s) of action of SIRS is unknown, it appears to function during the early, antigen-dependent phase of the immune response in vitro to abort full expression of the PFC response during the later exponential expansion of the clones of antibody-synthesizing cells.
Susceptibility to Cutaneous Anaphylaxis in Inbred Strains of Mice
Authors: de Souza Celso M. , Maia Luiz C. Sauerbronn , Vaz Nelson M.
Published: December 2022

Inbred mouse strains vary widely in their susceptibility to passive cutaneous anaphylaxis. The susceptibility to IgG1-induced reactions does not parallel susceptibility to IgE-induced reactions.
Attempts to Induce Cultures of BALB/c Myeloma and P3-J Burkitt's Lymphoma Cells to Produce Specific Phage Neutralizing Antibody
Authors: Krueger R. G. , Watkins A. C. , Volkman L. E.
Published: December 2022

An attempt was made to determine if the P3-J line of Burkitt's lymphoma cells could synthesize specific antibody in response after exposure to bacteriophage as reported by Kamei and Moore (1, 2). The cells were exposed to various concentrations of phages MS-2 or T2 and cultured for up to 10 days, but we were unable to detect significant neutralizing activity in any experimental situation. We also attempted to induce a BALB/c myeloma tumor with specific antibody activity for phage MS-2. While two in vivo tumors had characteristics of producing neutralizing antibody for MS-2 on adaptation of one of these tumors to growth in cell culture, no activity could be detected. Further, exposure of the cultured tumor cells to MS-2 did not result in the synthesis of neutralizing antibody.
Comparative in Vitro Studies on Effector Cell Diversity in the Cellular Immune Response to Murine Sarcoma Virus (MSV)-Induced Tumors in Mice
Authors: Plata Fernando , Gomard Elisabeth , Leclerc Jean Claude , Levy Jean Paul
Published: December 2022

The cellular immune response to murine sarcoma virus (MSV)-induced tumors in B6 mice was studied in vitro with two techniques: the 51Cr release test (CRT) and the microcytotoxicity assay (MA). The analysis of the effector cell nature in each of these assays showed that only thymus-dependent (T) cells are effector cells in CRT, while both T and non-T cells can be effector cells in MA. Comparative studies of the same lymphoid cell preparations demonstrated that the T cells revealed in MA might not be identical with the T cells whose activity is measured in CRT.
The Characteristics of a Circulating Antigen in Schistosomiasis
Authors: Nash Theodore E. , Prescott Benjamin , Neva Franklin A.
Published: December 2022

Previous work by others has shown that the sera of mice heavily infected with Schistosoma mansoni contained a schistosome-derived antigen. In this study the same antigen was also found in saline extracts of Schistosoma haematobium and Schistosoma japonicum and in the sera of mice infected with the latter parasite. The trichloroacetic acid-soluble fraction of the worm extract insoluble in chloroform was found to contain practically all the antigen. The antigen was anodic, resistant to boiling, Pronase, trypsin, α-amylase, DNase, RNase but was readily destroyed by sodium metaperiodate. The molecular weight was estimated to be greater than 100,000. Analysis of the trichloroacetic acid-soluble chloroform insoluble fraction, although incomplete, revealed less than 1% nitrogen. The only identifiable carbohydrate in this fraction was glucose. The properties of the antigen are characteristic of a polysaccharide.
Immunoglobulins with Multiple Binding Functions
Authors: Varga Janos M. , Lande Saul , Richards Frank F.
Published: December 2022

A rapid method for screening myeloma immunoglobulins for binding activity is described, based on the principle that low energy interactions between immunoglobulins and haptens are relatively frequent and may be used as indicator systems for finding other interactions of higher energy. Amino groups on partially hydrolyzed nylon screen discs are used as initiators for the polymerization of DL-Serine N-carboxyanhydride to polyserine whiskers. Immunoglobulins are coupled by glutaraldehyde to the whiskers, and the discs are incubated with a bank of nineteen radioactive haptens and antigens. Seven out of 10 myeloma immunoglobulins each bound one or more of these radioactive ligands with K0 of between 1 × 102 and 1 × 103 liters/mole. Non-radioactive prospective antigens were tested for their capacity to inhibit the binding of radioactive ligands.

NEW is a human IgG myeloma protein whose three dimensional structure at 2.8 Å resolution is completed and whose structure at 1.8 Å resolution is partially completed. An antigen was required to identify the immunoglobulin-binding region. The method led to the finding that 2-methyl, 3-phytyl 1,4 napthoquinone (Vit. K1) bound to Fab fragment of NEW with a K0 of 1.7 × 105 liters/mole.
Studies of Cellular Immunity in Chagas Disease: Effect of Glutaraldehyde-Treated Specific Antigen on Inhibition of Leukocyte Migration
Authors: Lelchuk Rosalia , Patrucco Aida , Manni Jorge A.
Published: December 2022

Delayed hypersensitivity was studied in chronic protozoal infection produced by the Trypanosoma cruzi (Chagas disease). Blood peripheral leukocyte migration inhibition was selected for studying cellular immune response in vitro. Two kinds of antigen preparations were used: a soluble extract of epimastigotes (SAg), and a modification of the antigen after its treatment with glutaraldehyde (MAg). When SAg was added to the medium no significant inhibition of migration was obtained in spite of using a concentration of antigen as high as 200 µg/ml. Similar results were obtained with normal cells. When the MAg was used, specific inhibition was obtained with chronic Chagas leukocytes. The inhibition was significant with a concentration of MAg as low as 5 µg/ml (p < 0.001).
Identification of a Subpopulation of Mouse Lymphoid Cells Required for Interferon Production After Stimulation with Mitogens
Authors: Stobo Jack , Green Ira , Jackson Lula , Baron Samuel
Published: December 2022

The cell population necessary for interferon production after mitogen stimulation was investigated in the mouse. Thymus cells and cortisone-resistant thymus cells failed to produce interferon after mitogen stimulation. When spleen cells, a mixture of B and T cells, were treated with anti-K chain serum and complement and then stimulated with PHA, no diminution of interferon production was observed. However, when spleen cells were treated with anti-ϑ antibody and complement and then stimulated with PHA, interferon production was markedly reduced. These studies suggest that peripheralized T cells are required for interferon production after mitogen stimulation.
Membrane Properties and Functional Activity of Lymphocytes from Cyclophosphamide-Pretreated Rats
Authors: Guttmann Ronald D.
Published: December 2022

In an attempt to explain the effect of drug pretreatment on the reduction of allograft immunogenicity, lymphocytes from normal and cyclophosphamide- (CY) pretreated rats were studied in leukocyte culture with phytohemagglutinin or allogeneic cells. Pretreatment with 250 mg/kg CY was associated with complete unresponsiveness as well as the inability to stimulate allogeneic cells. This was dose and time dependent and not explicable by just cell death in culture or drug carryover. CY-pretreated cells gave cytotoxic antibody titration curves suggestive of reduced but not absent surface histocompatibility antigens. Lymphocytes from normal and CY-pretreated rats did not show significant differences in the ability to form caps when reacted with antiimmunoglobulin antisera or in sensitivity to lysis in hypotonic solutions. Thus the cells from CY-pretreated rats appear to have surface alterations associated with the gene product of the MLR locus, decreased serologically detectable transplant antigens, and no serious defect in membrane fluidity or extensibility. This approach provides a potential monitoring system for donor pretreatment in organ allograft situations.
Discontinuous Antibody Secretion during the Secondary Response to Sheep Erythrocytes in Vitro
Authors: Thomas David W.
Published: December 2022

Antibody-producing lymphocytes provide a unique model for studying cellular differentiation and patterns of gene expression in eukaryotic cells as recently reviewed by Lerner and Dixon (1). The relationship between gene expression for antibody synthesis and the cell cycle has been studied in several established antibody-producing cell lines with varying results. Human lymphoid cell lines consistently show the highest level of antibody synthesis in the late G1, early S (2–5), and possibly G2 (5) stages of the cell cycle and a reduced synthesis around metaphase. Similarly, Byars and Kidson (6) demonstrated that established mouse myeloma cells also exhibit maximal antibody synthesis in late G1 and early S, and Gandini-Attardi et al. (7) suggest that mouse myeloma cells contain more antibody in S than in other stages of the cycle. On the other hand, studies by Cowan and Milstein (8) and Liberti and Baglioni (9), also using mouse myeloma cell lines, indicate that antibody is synthesized at a fairly constant rate throughout the cell cycle.
Production of α1 and α2 Immunoglobulin Heavy Chains during Fetal Life
Authors: Cederqvist Lars L. , Litwin Stephen D.
Published: December 2022

IgA and its subclasses IgA1 and IgA2 were measured in fetal serum and amniotic fluid throughout gestation by hemagglutination inhibition tests. The IgA1/(IgA1 + IgA2) ratio was higher for fetal serum than for amniotic fluid and was significantly higher in the third than in the second trimester in both amniotic fluid and fetal serum. Am2, a genetic marker for human α2 heavy chains, was typed in a number of combinations of maternal and paternal sera, amniotic fluid, and fetal serum. Three families were informative in that Am2 was present in amniotic fluid, was absent from maternal serum, but present in paternal serum. It was concluded that the fetus synthesized IgA2 in vtero.
Chymopapain C: An Immunosuppressive Protease
Authors: Clagett James A. , Tokuda Sei , Engelhard Warren E.
Published: December 2022

Chymopapain C suppresses the immune responses of mice to both thymus-dependent and independent antigens. We have demonstrated a clear-cut dose and time dependence for the action of this enzyme. When these conditions are optimum, the primary immune response to sheep red cells (SRBC) is completely inhibited when measured by direct plaque-forming cells (PFC), indirect plaque-forming cells, and rosette-forming cells (RFC) during a 10-day experimental period. Immunologic memory was altered in suppressed mice as indicated by quality, quantity, and temporal appearance of the PFC after second injection of antigen. Chymopapain C had a profound suppressive effect on the secondary IgM response when the enzyme was given 1 day before the booster SRBC injection; however, the IgG and RFC responses were not significantly suppressed.
Isolation and Characterization of the Proenzyme form of the C1r Subunit of the First Complement Component
Authors: Valet Gunter , Cooper Neil R.
Published: December 2022

The proenzyme form of C1r was isolated by sequential chromatography of the euglobulin fraction of human serum on DE-32 cellulose, TEAE cellulose, and Sephadex G-200. C1r was found to be a proteolytic enzyme with β-globulin mobility, a sedimentation rate of 7.5S, and a molecular weight of approximately 188,000 daltons. Isolated C1r preparations exhibited an affinity for C1q and C1s, forming a reversible complex with C1q and a firm complex with C1s in free solution.

Isolated C1r preparations did not activate proenzyme C1s, the natural substrate of the C1r enzyme, unless first activated by brief treatment with trypsin, a process accompanied by physicochemical changes in the molecule. It is not known whether the mechanism of conversion of C1r from a proenzyme into an active enzyme during C1 activation is also enzymatically mediated or, alternatively, whether conversion results from conformational changes in C1r induced by the attachment of C1, via C1q, to immunoglobulin molecules.
Detection of Specific Surface Antigens on Cells Transformed by Cytomegalovirus with the Techniques of Mixed Hemagglutination and 125I-Labeled Antiglobulin
Authors: Lausch Robert N. , Murasko Donna M. , Albrecht Thomas , Rapp Fred
Published: December 2022

Cytomegalovirus (CMV)-transformed hamster cells were examined for the presence of virus-specific membrane antigens by using rabbit antiserum prepared against CMV-infected human embryonic lung (HEL) cells. After absorption with sheep erythrocytes and heterologous hamster tumor cells the antiserum reacted specifically with the CMV-transformed cells in the mixed hemagglutination test. The absorbed antiserum did not react with hamster cells transformed by herpes simplex virus type 2 or PARA-adenovirus 7 or with untransformed hamster embryo fibroblasts. These results were confirmed with the 125I-labeled antiglobulin technique. Absorption with HEL cells rendered the antiserum specific for HEL cells infected with CMV. This reactivity was lost after absorption with 107 CMV-transformed cells but not with 107 hamster embryo fibroblasts. These results establish that CMV-transformed hamster cells express virus-related membrane antigen(s), and that the same antigen(s) is also found on CMV-infected human cells.
Secondary Cytotoxic Tumor Immune Response Induced in Vitro
Authors: Röllinghoff M.
Published: December 2022

For the investigation of a secondary cell-mediated anti-tumor immune response a syngeneic mouse plasma cell tumor system was used. The primary immunization was performed in vivo, whereas the secondary response was induced in vitro. Evidence is presented that a secondary anti-tumor cell response was characterized by an accelerated and enhanced reaction when compared to a primary anti-tumor response. Furthermore, our findings suggested that the secondary response studied was antigen-specific. The induction of a secondary response did not appear to depend on the persistence of differentiated cytotoxic effector lymphocytes, as measured in a 51Cr release assay. The effector phase of the secondary anti-tumor response, however, seemed to require T lymphocytes. These data appear to be compatible with the view that a secondary anti-tumor cell response is dependent on the presence of a set of antigen-reactive cells, which operationally can be designated in analogy to the secondary B cell response as memory cells.
Polymerized Ragweed Antigen E
Authors: Patterson Roy , Suszko Irena M.
Published: December 2022

The antigenic characteristics of three preparations of ragweed (RW) antigen E were studied in rabbits. The preparations consisted of the monomeric form, (m.w. 37,800); a lower molecular weight polymer (m.w. 0.2 to 4 million) and a higher molecular weight polymer (m.w. 4 to 20 million). Antibody responses of two Ig classes occurred. A circulating antibody response detectable by an ammonium sulfate-binding technique was consistent with an IgG response. A homocytotropic antibody response, probably rabbit IgE, occurred in a majority of animals. The polymerized RW preparations appeared as antigenic as the monomer in induction of the IgG response. The low molecular weight polymer resulted in a lower and later reaginic antibody response than either the monomer or high molecular weight polymeric preparation.
The Role of Humoral Factors in the Initiation of in Vitro Primary Immune Responses
Authors: Watson James , Thoman Marilyn , Ralph Peter , Trenkner Ekkhart
Published: December 2022

The nature of the adherent cell type that is required for the induction of immune responses to erythrocyte antigens in mouse spleen culture has been investigated. Bone marrow-derived mononuclear cells were grown in liquid culture systems in the presence of a colony stimulating factor. These mononuclear cells exhibit adherent, phagocytic, staining, rosette-forming, and cytotoxic properties characteristic of macrophages. However, such macrophage-like cells do not replace spleen adherent cells in the induction of immune responses to erythrocyte antigens in mouse spleen culture. Treatment of macrophage-like cells with normal mouse serum, hyperimmune anti-sheep erythrocyte (SRBC) serum or thymus-derived cells activated to SRBC did not render these cells capable of restoring immune responses in adherent cell-depleted cultures. These findings suggest that the macrophage may not be the adherent cell type required for cooperative events in the induction of antibody synthesis. Adherent cell types may play two roles in the induction of immune responses to SRBC. A phagocytic role for the digestion of particulate antigens such as SRBC may be performed by a macrophage-like cell. Another cell type may be required in delivering part of the inductive stimulus to precursor antibody-forming cells or to thymus-derived helper cells.
Aggregation of Fc′µ Subfragments Produced by Tryptic Hydrolysis of Human IgM at Elevated Temperature
Authors: Chen James P. , Beyer Carol B. , Elakovich Stella D.
Published: December 2022

The proteolysis of Waldenstrom macroglobulins into Fabµ and (Fc)5µ fragments by tryptic digestion at 56°C to 65°C was reexamined. It appeared that “(Fc)5µ fragment” is composed of both Fc′µ subfragments and residual “(Fc)5µ fragment” held together by non-covalent forces.

Sephadex G-200 chromatography resolved a 65°C tryptic digest of IgM into three peaks: GI, (Fc)5µ fragment; GII, Fabµ fragment; and peptides. Gel-filtration of an identical IgM digest through Sepharose 6B resulted in four peaks: BI, BII, BIII and peptides. BIII with m.w. 52,000 according to SDS polyacrylamide gel electrophoresis is Fabµ fragment. GI and BII, having sedimentation coefficients of 9.9S and 0.1S, respectively, were found to be antigenically equivalent according to Fcµ specific antisera. BI is apparently a high molecular weight aggregate of Fc′µ subfragments of 10,000 m.w. This aggregate was antigenically deficient in Fcµ determinants to both GI and BII. SDS gel electrophoresis showed that the appearance of BI subfragment was not dependent upon reduction and alkylation. This suggests that BI is derived from a region of the µ-chain other than the area containing inter-µ-chain disulfide bonds. Reduced-alkylated GI is characterized by major protein bands at m.w. 10,000, 12,500, 16,000, and 43,000 in SDS gel electrophoresis. BII, whose reduced-alkylated form is devoid of the band corresponding to m.w. 10,000, is most likely residual “(Fc)5µ fragment”.

It appears that BI results from further fragmentation of (Fc)5µ fragment with subsequent aggregation. The release of BI is temperature dependent starting at 59°C and is greatest at 65°C. BI contains < 2% hexose and is presumably derived from a carbohydrate poor Cµ domain; BII contains 11% hexose. BI fraction was further purified by passage through a Sepharose 6B column equilibrated with 5 M guanidine HCl. Comparison of preliminary N- and C-terminal sequence data with published IgM sequence data shows BI subfragment to be located in the C-terminal region of Fcµ fragment.
IgD: A Major Immunoglobulin on the Surface of Lymphocytes from Patients with Chronic Lymphatic Leukemia
Authors: Kubo Ralph T. , Grey Howard M. , Pirofsky Bernard
Published: December 2022

Previous studies on the characterization of surface immunoglobulins of lymphocytes from CLL2 patients indicated that the cell-associated immunoglobulin was monoclonal with respect to heavy and light chain class with IgM being by far the most common Ig class observed (1–4). Our earlier study of CLL involved analysis for the presence of IgG, IgA, and IgM. Some cases were observed in which neither heavy nor light chains were detectable and others in which only light chains could be detected (1). A more detailed investigation of one of these patients demonstrated IgD as the major Ig class associated with these cells. We have accordingly evaluated other unselected cases of CLL for the presence of IgD on the lymphocyte surface. The current report details these studies and the relationship of IgD to IgM on the surface of CLL lymphocytes.

Methods. Lymphocytes were obtained from patients with CLL having white blood counts ranging from 20,000 to 200,000 cells/mm3.
Reactions between Certain Strains of Pneumococci and Fc of IgG
Authors: Stephens Christopher G. , Reed William P. , Kronvall Göran , Williams Ralph C.
Published: December 2022

A non-immune reaction is known to occur between the Fc region of certain IgG subclasses and cell wall components of staphylococci (protein A) and streptococci. Pneumococci have been studied for similar activity.

Seventy-six strains of Diplococcus pneumoniae were evaluated for non-immune reactivity with γ-globulin Fc by testing their ability to agglutinate erythrocytes which were coated with sub-agglutinating quantities of anti-erythrocyte antibody. Thirty (39%) of the strains were reactive. All strains of some pneumococcal serotypes were reactive, whereas all strains of other serotypes were negative. The reactivity appeared to be a property of the polysaccharides since particles coated with purified polysaccharides from reactive serotypes also caused agglutination. When the capsule size of pneumococci was varied by using encapsulated and non-encapsulated mutants, augmentation by mouse passage, or varying cultural conditions, reactivity was greater if the capsule was larger. Inhibition experiments showed the reactivity to be in the Fc fragment, and IgG1 was the only highly reactive immunoglobulin. IgG2 and IgA were probably non-reactive; and IgG3, IgG4, and IgM were clearly non-reactive.
The Ability of Chemotactic Factors to Induce Lysosomal Enzyme Release
Authors: Becker Elmer L. , Showell H. J.
Published: December 2022

Lysosomal enzyme release form rabbit peritoneal neutrophils induced by a bacterial chemotactic factor is temperature dependent; it requires external Ca2+ but not Mg2+ glycolysis as a source of metabolic energy and free sulfhydryl groups. The release is inhibitable by DFP under circumstances suggesting involvement of an activatable serine esterase. The release is enhanced by cytochalasin B. Low concentrations of colchicine and vinblastine enhance lysosomal enzyme release, as well as chemotaxis induced by the same chemotactic factor; higher concentrations inhibit.
Enzymatic Digestion of the First Component of Human Complement (Clq)
Authors: Knobel Hans R. , Heusser Christoph , Rodrick Mary L. , Isliker Henri
Published: December 2022

Human C1q was shown to be sensitive to digestion by trypsin, chymotrypsin and collagenase, but not by elastase. Treatment with trypsin and chymotrypsin caused a rapid loss of the hemolytic activity of C1q and its ability to fix to immune complexes. Collagenase digestion of C1q resulted in a rapid loss of the hemolytic activity also, whereas the ability to bind to immune complexes decreased slowly and remained at a constant value of 15%. In contrast to trypsin and chymotrypsin, collagenase treatment of 125I-labeled C1q led to well defined fragments, only some of which were labeled. Adsorption of collagenase-digested C1q on ovalbumin-antiovalbumin complexes indicated the larger non-covalently bound peripheral subunits of C1q to be involved in fixation to immune complexes. None of the fragments was able to initiate lysis in the presence of RC1q. Immunologic analysis of the digest revealed at least two different fragments with respect to antigenicity.
Cell-Mediated Immunity in Vitro: A Highly Sensitive Assay for Human Lymphotoxin
Authors: Spofford Bryan T. , Daynes Raymond A. , Granger Gale A.
Published: December 2022

A new in vitro technique for the assay of human lymphotoxin (LT) is described. The method uses monolayer tube cultures of an exquisitely sensitive cell type pretreated with mitomycin-C (MC). The use of MC permits the differentiation of growth inhibition and actual cytotoxicity. In addition, the use of MC-treated target cells considerably increases the sensitivity of the LT assay. The method is very simple, rapid, reproducible, and more sensitive than present assays for LT or any other lymphokine. The utility of this assay lends itself to routine application in research and in clinical situations.
Cellular Bases for Relative Radioresistance of the Antibody-Forming System at Advanced Stages of the Secondary Response to Tetanus Toxoid in Mice
Authors: Grobler Peter , Buerki Heinz , Cottier Hans , Hess Max W. , Stoner Richard D.
Published: December 2022

Nearly normal secondary tetanus antitoxin responses were produced by young adult female mice subjected to 600 rads of 60Co gamma whole-body radiation 4 days after secondary antigenic stimulation via the hind leg footpads. In an attempt to correlate the rapid transition from radiosensitivity of the antibody-forming system on day 3 to relative radioresistance on day 4 after booster, we carried out studies on cytokinetics in regional (popliteal) lymph nodes and on incorporation of 3H-l-histidine into circulating antitoxin. Analyses of tritium radioactivity in antigen-antibody precipitates of pooled sera 2 hr after i.v. injection of the labeled amino acid revealed maximum incorporation into antibody around day 7 after booster in nonirradiated controls, and around day 12, i.e., 8 days after radiation, in experimental mice, respectively. Relative radioresistance of the antibody-forming system on day 4 after booster, as opposed to day 3, was attributed to the finding that plasmacellular proliferation in medullary cords reached a marked and narrow peak on day 3 and had almost subsided to control levels between days 4 and 5. Accordingly, many medullary plasma cells survived and continued to proliferate after exposure to radiation. In contrast, germinal centers, which largely determine the overall maximum DNA synthetic activity in the lymph node and attain their full development in non-irradiated animals only 5 to 6 days after booster, were completely destroyed by radiation within 1 day. Since a) antibody production persisted despite the disappearance of germinal centers; and b) mice irradiated 4 days after booster have previously been shown to lack tertiary responsiveness, the present findings lend strong support to the hypothesis that germinal center cells were involved more in the generation of memory cells than in active antibody secretion. The results do not favor the assumption of a marked contribution of germinoblasts to the antibody-forming plasma cell population elicited by the same stimulus which had triggered germinal center formation.
Induction of Marked Hypergammaglobulinemia with Xenogeneic Tumors in Hamsters: Immunopathologic Studies
Authors: Richman Alan Ve , Defendi Vittorio
Published: December 2022

Human heteroploid cells derived from a benign prostatic adenoma, MA160, can grow progressively in the subcutaneous tissues of 2-day-old nonimmunosuppressed hamsters. The growth of these xenografts is associated with certain immunopathologic features. These features are: A marked increase in the serum of electrophoretically restricted IgG which parallels tumor growth.The appearance of noncytotoxic antibody against membrane antigens of the transplanted cells which is related to the IgG elevation but which constitutes only a very minor part of this elevation; and the fixation of antibody to the tumor cell membrane in situ.The extensive plasma cell proliferation, and the presence of circulating immune complexes (as indicated by the granular deposition of IgG and complement (C3) in the renal glomeruli).The lack of demonstrable biologic activity of the serum (enhancement or cytotoxicity).

This transplantation system has analogies with certain chronic viral infections in animals; and our inability to detect antibody specificity for much of the IgG may be related to the presence of circulating immune complexes. Finally we suggest that this system may provide a model from which to study the evolution and pathogenesis of plasma cell dyscrasias.
The Role of the Classical and Alternate Complement Pathways in Host Defenses Against Cryptococcus Neoformans Infection
Authors: Diamond Richard D. , May Joseph E. , Kane Michael A. , Frank Michael M. , Bennett John E.
Published: December 2022

The relationship between complement and the heat labile factors known to be necessary for phagocytosis of Cryptococcus neoformans by human leukocytes has been examined. C. neoformans or purified cryptococcal polysaccharide consumed 36.2 to 76.3% of late complement components (C3-9) in guinea pig serum. Phagocytic studies with C2-deficient human serum and C4-deficient guinea pig serum permitted evaluation of the roles of the classical (C142) and alternate complement pathways in the phagocytosis of these organisms.

The number of cryptococci ingested per neutrophil (phagocytic index) at 30 min in sera with only alternate pathway function was 40 to 60% of that found in sera with both classical and alternate complement pathways intact. When purified C2 or C4 was added back to the corresponding deficient sera, phagocytic indices increased to ranges obtained by using corresponding normal sera with both complement pathways active. By 2 hr, phagocytic indices for deficient sera equaled phagocytic indices for normal sera. Early classical complement components were therefore required for optimum phagocytic kinetics. The fact that late components of complement were required for phagocytosis was established by the fact that chelation of Ca++ and Mg++ with EDTA inhibited the reaction, as did heating at 56°C and use of serum depleted of the late components of complement by cobra venom factor. None of the sera used in these studies had detectable anti-cryptococcal antibody by an indirect fluorescent antibody technique, but absorption of human sera at 0°C with cryptococci lowered phagocytic indices to about 10% of normal. The loss of activity after absorption was demonstrated to be due to loss of specific antibody which was required to activate the C142 pathway as well as to loss of properdin which was required to activate the alternate pathway. The classical complement pathway appeared to be functioning primarily to activate the alternate pathway, which in turn was responsible for opsonization. Serum depleted of properdin or of the heat labile properdin factor B was markedly deficient in opsonic activity. In the former case addition of properdin restored optimal kinetics. Thus, function of the classical pathway in normal serum did not adequately mediate opsonization.

Unlike serum, spinal fluid did not opsonize cryptococci. With fluorescent anti-human β1C, little or no complement was detectable on the surface of cryptococci obtained from the spinal fluid of four patients with cryptococcal meningitis, but was found on cryptococci after incubation in fresh sera from these patients. However, cerebrospinal fluid from two of three patients with active disease could restore opsonic activity of fresh serum absorbed with cryptococci.
In vitro Studies of Cell-Mediated Immunity in an Acute Viral Infection
Authors: McFarland Henry F.
Published: December 2022

The effect of Sindbis virus-sensitized lymphocytes on acutely infected mouse embryo fibroblasts has been studied in vitro. Sindbis virus-immune lymphocytes are capable of lysing the infected target cells bearing surface viral antigen before viral-induced lysis occurs. The cytotoxicity was virus-specific since lymphocytes sensitized to yellow fever virus were unable to kill the Sindbis virus-infected target cells. The effector cell in this in vitro assay has been shown to be a T cell, and the cytotoxic reaction can be blocked by Sindbis virus-immune serum.

In distribution to lysis of infected cells by T cells, viral clearance is dependent on a population of noncytotoxic, adherent cells, probably macrophages. Further, macrophage function in this assay was virus-specific, since adherent cells from animals immunized to a second virus were unable to reduce significantly virus content. Consequently, the full expression of recovery mechanisms in this model of an acute viral infection most likely depends on a balance between lymphocyte-mediated lysis of infected cells and viral clearance by macrophages and antibody.
The Serum Factor Stimulating Colony Formation In Vitro by Murine Plasmacytoma Cells: Response to Antigens and Mineral Oil
Authors: Metcalf Donald
Published: December 2022

Colony formation in agar-medium by three transplanted mouse plasmacytomas (MOPC-460; MOPC-315; HPC-127) was stimulated by addition of serum from mice injected with endotoxin or a variety of bacterial antigens. This effect was potentiated by the addition of mercaptoethanol. The active serum factor was heat-labile but resisted ether treatment and was non-dialysable. Serum plasmacytoma-stimulating activity was localized electrophoretically as a single peak of β mobility and was separable from the colony-stimulating factor stimulating granulocytic and macrophage colony growth. Injection of mineral oil to BALB/c mice increased the capacity of serum or peritoneal cells to stimulate plasmacytoma colony formation in vitro.
Regulation of T Lymphocyte Function by Immunoregulatory Alphaglobulin (IRA)
Authors: Menzoian James O. , Glasgow Arthur H. , Nimberg Richard D. , Cooperband Sidney R. , Schmid Karl , Saporoschetz Inna , Mannick John A.
Published: December 2022

Immunoregulatory alphaglobulin, IRA, has been shown to suppress T cell-dependent immune responses. The present studies were to determine the effect of IRA on B cell-mediated immune responses. IRA-protein, and a more purified species, IRA peptide were shown to inhibit the T cell-mediated responses of spontaneous rosette formation between human peripheral lymphocytes and sheep red blood cells (SRBC) and the plaque-forming cell response in mice injected with SRBC. The same IRA preparations were ineffective in suppressing rosette formation between human peripheral lymphocytes and EAC-SRBC and the plaque-forming cell response to Escherichia coli lipopolysaccharide, both of which are B cell-mediated, T cell-independent immune responses. Therefore, we believe that IRA is effective in nonspecific suppression of only immune responses dependent on T cell function.
Interaction of Isoantibody and Cytotoxic Lymphocytes with Allogeneic Tumor Cells
Authors: Faanes Ronald B. , Choi Yong Sung
Published: December 2022

Specific isoantibody against DBA/2J mastocytoma cells was purified from immune serum by immunoadsorption without losing its inhibiting activity of lymphocyte-mediated cytotoxicity (LMC). The molecular nature of the isoantibody was studied with 125I-labeled antibody (125I-IS). The Ig nature was verified by specific serology with rabbit anti-mouse Ig, acrylamide gel analysis, and specific binding to mastocytoma cells.

The fate of 125I-IS on the surface of activity metabolizing mastocytoma cells was studied. Almost all 125I-IS are bound to mastocytoma cells, inhibiting T cell-mediated cytotoxicity. At the time of the onset of the “escape” from IS inhibition of LMC, 20 to 30% of bound 125I-IS was released and the surface antigens were regenerated. This phenomenon is also temperature dependent, requiring active metabolism of the target cells.
Isolation of an Enzyme from Streptococcus Sanguis Which Specifically Cleaves IgA
Authors: Plaut Andrew G. , Genco Robert J. , Tomasi Thomas B.
Published: December 2022

IgA protease, a metal-dependent endopeptidase elaborated by bacteria normally found in the human oral cavity and intestinal tract, can be isolated in good yield from culture filtrates of Streptococcus sanguis. The enzyme has pronounced specificity for secretory and serum IgA, which it cleaves to form Fcα and Fabα fragments.
Inactivation of Slow Reacting Substance of Anaphylaxis (SRS-A) by Arylsulfatases
Authors: Orange Robert P. , Murphy Robert C. , Austen K. Frank
Published: December 2022

Two different arylsulfatase preparations inactivated slow reacting substance of anaphylaxis (SRS-A) in a dose-related fashion. The loss of SRS-A activity could not be attributed to β-glucuronidase activity in the arylsulfatase preparations or to nonspecific adsorption to proteins. The arylsulfatase and SRS-A inactivating activities shared similar pH requirements, kinetics, and elution properties on Sephadex G-200 gel filtration. With spark source mass spectrometry, analysis of highly purified preparations of SRS-A revealed an abundance of 32S and 40Ca in the active samples as compared to controls. These findings indicate that SRS-A may contain a sulfate ester group.
Activation of the Alternate Pathway of Human Complement by Rabbit Cells
Authors: Platts-Mills Thomas A. E. , Ishizaka Kimishige
Published: December 2022

The hemolysis of sheep red blood cells (SRBC) occurs via the classical complement pathway and is blocked by ethylene glycol bis-amino tetraacetate (EGTA). By contrast, fresh normal human serum in EGTA buffer was found to cause >90% hemolysis of unsensitized rabbit red blood cells (RaRBC) at a final dilution of 1:15. Absorbing human serum with RaRBC at 0°C will remove only 45% of this hemolytic activity and the same activity is present in human hypogammaglobulinemic serum. When rabbit lymphocytes were incubated with human serum in EGTA buffer, complement fixation occurred on their surface which was demonstrated with radiolabeled antibodies to human C3 or as “blocking” of the complement receptor. With purified complement components it was shown that the EGTA buffer completely blocked C1 but not C4, C2, or the late complement components. These findings show that a rabbit cell surface component can activate the alternate pathway of complement in human serum and suggest that this activation does not involve antibody.
Further Studies on the Inhibition of Allergic Encephalomyelitis by l-Asparaginase
Authors: Khan Amanullah , Levine Seymour
Published: December 2022

l-Asparaginase inhibited the hyperacute form of EAE in rats. The inhibition was inversely proportional to the intensity of the eliciting immunization. Passive transfer experiments proved that at least part of the inhibition was related to effects on the fully immunized lymphoid cells and its interaction with the neural target. The inhibition was caused by specific enzyme activity and not by endotoxin contamination or nonspecific stress.
Comparison of Irradiated and Mitomycin-Treated Mouse Spleen Cells As Stimulating Cells in Mixed Lymphocyte Cultures and in vitro Sensitization
Authors: Hodes Richard J. , Terry William D.
Published: December 2022

Mitomycin-treated and irradiated mouse spleen cells were compared as stimulating cells in allogeneic mixed lymphocyte culture (MLC) and as sensitizing cells in in vitro allogeneic sensitization. C57BL/6 and BALB/C spleen cells were irradiated at doses up to 5000 rads or treated with mitomycin C to inhibit DNA synthesis. Irradiated cells, as well as mitomycin-treated cells, were efficient both in stimulating MLC and in sensitizing in vitro.
Properties of Antisera to Ganglioside GM1 and Asialo GM1
Authors: Naiki Masaharu , Marcus Donald M. , Ledeen Robert
Published: December 2022

The object of this study is to prepare antisera to purified glycosphingolipids for immunocytochemical studies. Rabbits were immunized with ganglioside GM1 complexed either to methylated BSA or to a glycoprotein isolated from human erythrocytes, and to asialo GM1 complexed to the erythrocyte glycoprotein. IgG and IgM antibody fractions were separated by chromatography on Sephadex G-200 and their properties were determined by complement fixation. IgG antibodies to asialo GM1 were highly specific and exhibited little cross-reactivity with GM1 or other glycolipids, but IgM antibodies cross-reacted extensively with gangliosides GM1 and GD1b. Complement fixation of IgG anti-asialo GM1 with asialo GM1 was completely inhibited by about 15 nmoles of ganglio-N-tetraose, the carbohydrate moiety of this glycolipid.

Both IgG and IgM antibodies to GM1 cross-reacted extensively with asialo GM1 and GD1b, but specific IgG antibodies to GM1 were revealed by absorption with asialo GM1. The specific and cross-reacting IgG antibodies to GM1 were inhibited by the pentaose moiety of this glycoplipid, but these antibodies were much harder to inhibit than IgG anti-asialo GM1. These results demonstrate that it is possible to prepare specific antisera to purified glycolipids with a single administration of antigen, in contrast to previous studies in which repeated injections of ganglioside mixtures were employed.
Immunity to Experimental Cholera
Authors: Pierce Nathaniel F. , Reynolds Herbert Y.
Published: December 2022

The protective role of serum IgG antitoxin was studied in dogs transfused either with whole plasma or purified serum IgG from hyperimmunized dogs. Washings of jejunal Thiry-Vella loops after plasma transfusion yielded IgG antitoxin. Furthermore, dogs transfused with antitoxic plasma were protected against intrajejunal challenge with cholera toxin and this protection could not be diminished by extensive washing of the jejunum prior to toxin challenge. Finally, intact dogs transfused with antitoxic serum IgG were highly protected against orogastric challenge with viable Vibrio cholerae. These results indicate that serum IgG can play a protective role at the surface of the gut mucosa or within its lumen. They also suggest that serum-derived antibody may exert its effect while intimately associated with the mucosa, a position in which it may be protected from rapid destruction by proteolytic enzymes.
The Modulating Effect of Cholera Enterotoxin on the Immune Response
Authors: Chisari Francis V. , Northrup Robert S. , Chen Lincoln C.
Published: December 2022

Cholera enterotoxin (CE) is a potent adjuvant when administered simultaneously with sheep red blood cells (SRBC) and an immunosuppressant when given 12 hr before or after the antigen. Antigen clearance is not modified, but splenic adenyl cyclase activity is significantly enhanced reaching peak levels within 12 hr. Theophylline enhances the modest adjuvant effect of 0.01 µg of CE and inhibits the pronounced adjuvant effect of 0.05 µg. The relationship between adenyl cyclase stimulation and modulation of the immune response is discussed.
Anti-Histone KSAchicken Antibody from Guinea Pig
Authors: Mura Casilda , Huang P. C. , Levy David A.
Published: December 2022

Avian erythrocyte-specific histone KSA was purified from chicken red blood cells by selective extraction and by chromatography on amberlite CG-50 columns. This histone was used in the production of antibody in guinea pigs. By immunoprecipitation and microcomplement-fixation tests, the antibody was shown to be specific for histone KSA; reaction was not seen with histones KAP, ARE, GRK, LAK, and KAS. The observation supports the notion that histone KSA has an independent origin from the other histones.
Antibody Affinity
Authors: Gopalakrishnan P. V. , Karush Fred
Published: December 2022

Rabbit antibody of restricted heterogeneity induced by a lactoside determinant has been specifically purified and further fractionated into functionally homogeneous populations. These preparations have been used to evaluate the association constants for binding to the bacteriophage φX174 to which had been coupled multiple lactoside groups. The constants were obtained from equilibrium neutralization experiments and compared to the association constants for the binding of a monovalent lactosyl-containing ligand. The central finding is that the constant for the binding of the antibody to bacteriophage is greater by a factor of 104 than that for the monovalent ligand. As previously suggested in another system, this effect is attributed to bivalent interaction and serves to establish the generality of the enhancement phenomenon. The use of functionally homogeneous antibody precluded the possibility that selection of high-affinity antibody was responsible for enhancement. The issue of the number of critical sites available for neutralization on the conjugated bacteriophage was clarified by comparison of the observed reactivation kinetics with the behavior predicted from a model system. It was inferred that the number of such sites did not exceed two and could be one. Thus the process of neutralization and reactivation can be adequately described by an analysis based on single-site kinetics.
Cytotoxic Activity of Lymphocytes
Authors: Walker Sharyn M. , Lucas Zoltan J.
Published: December 2022

Antibody to human lymphotoxin was elicited in rabbits immunized with partially purified supernatants of human spleen cells stimulated by phytohemagglutinin; Anti-LT activity, assayed by neutralizing soluble LT toxicity to indicator mouse L cells, is confined to the immunoglobulin fraction of the serum; it cannot be removed by adsorption to L cells, human fibroblasts or lymphocytes, or to human serum. Anti-LT activity is removed by immunoadsorption to Sepharose conjugated with goat antisera to rabbit 7S immunoglobulin; partial recovery of anti-LT activity is effected by elution with high molarity acid buffer. The amounts of antisera necessary to effect neutralization are linearly proportional to LT concentration. Anti-human LT neutralizes the toxicity of supernatants of human lymphocytes activated by other, non-PHA mitogens (antigens to which the lymphocyte donors are sensitive) and allogeneic lymphocytes in mixed leucocyte culture. However, anti-human LT is specific for human LT as it will not neutralize guinea pig or rat LT.
Induction of Lymphocyte-Mediated Cytotoxicity against Allogeneic Tumor Cells by Concanavalin a in Vivo
Authors: Anaclerio Allessandro , Waterfield J. Douglas , Möller Göran
Published: December 2022

The in vivo administration of concanavalin A (Con A) induced a dose and time dependent cytotoxic response in spleen cell populations when tested against allogeneic target cells in vitro. Con A doses between 60 µg and 480 µg induced cytotoxicity, which was maximal 24 hr after injection and thereafter disappeared. Cytotoxicity correlated with the time of maximum DNA synthesis and blast transformation in the spleen. The addition of extra Con A to the in vitro test system did not change the kinetics or intensity of cytotoxicity. The results are discussed in terms of the effect of Con A in the activation of cytotoxic lymphocytes and its possible further role in effecting “killer-target” cell binding.