bluuebunny/crossref_metadata_2025 huggingface dataset:
Suppression of Lymphocyte Activation by a Factor Produced by Mycoplasma Arginini
Authors: Callewaert Denis M. , Kaplan Joseph , Peterson Ward D. , Lightbody James J.
Published: December 2022

Mycoplasma arginini, when grown in broth or in cultures of human lymphoblast cell lines, inhibits activation of lymphocytes by allogeneic cells of mitogens. This inhibition can also be produced by cell free media obtained after the growth of this organism. Inhibition is not species specific and inhibits both B and T cell activation in the mouse. The inhibitory factor is required during the first 3 days of the mixed leukocyte culture, but has no effect on the latter phase of this reaction when thymidine uptake is greatest. The factor is stable to treatment at 60°, but not 90°, for 30 min.
The Major Histocompatibility Complex of the Guinea Pig
Authors: Geczy A. F. , de Weck A. L. , Schwartz B. D. , Shevach E. M.
Published: December 2022

Serologic and genetic studies of the antigens which comprise the guinea pig MHC have demonstrated three distinct but linked genetic regions. Antisera to the B region were raised by cross-immunization of random-bred animals; this region controls antigens B.1, B.2, B.3, and B.4 which behave as alleles at a single locus and which resemble the products of the murine D or K region genes in their tissue distribution and molecular characteristics. Cross-immunization of inbred strain 2 and strain 13 animals, both of which bear the B.1 antigen, leads to sera which identify antigens which resemble the products of the I region of the murine MHC. Specific absorption experiments have demonstrated four distinct I region antigens. In addition to the B and I regions, inbred strain 2, strain 13, and some outbred animals bear an antigen (S.1) which is the product of a third genetic region and which also resembles the murine D or K region gene products in molecular size.

The results of these studies should facilitate the use of the guinea pig as an experimental model for studies of genetic control of the immune response and the function of the histocompatibility-linked Ir genes.
In Vitro Selection and Extended Culture of Antigen-Specific T Lymphocytes
Authors: Ben-Sasson S. Z. , Paul William E. , Shevach Ethan M. , Green Ira
Published: December 2022

Functional selection of antigen-specific T lymphocytes can be achieved by culturing thymus-dependent (T) lymphocytes from immunized guinea pigs on “monolayers” of antigen-pulsed adherent peritoneal exudate cells (PEC) from nonimmune syngeneic donors. Several aspects of the in vitro selection of T lymphocyte-rich peritoneal exudate lymphocytes (PEL) were studied. It was shown that irradiated adherent PEC were equivalent to nonirradiated adherent PEC in supporting selection cultures, indicating that the lymphocytes harvested at the end of the selection culture derive from the immune donors of the PEL and not from the nonimmune donor of the adherent PEC.

The relative importance of specific adherence and specific proliferation for achieving selection was determined by comparing the degree of selection obtained when nonadherent cells were discarded at 24 hr with that noted when the discard step was omitted. It was found that omitting the discard step markedly diminished the degree of selection. On the other hand, blocking proliferation with specific alloantisera after the discard step did not diminish the degree of selection, although it did diminish the cell yield. Thus, specific adherence to antigen-pulsed PEC appeared to be critical in the selection culture procedure.

An estimate of the degree of enrichment obtained by the selection culture procedure was obtained by culturing selected cells in an excess of nonprimed PEL, so that auxiliary cells would not be limiting. Under these conditions, it appeared that selected cells were enriched from 4-to 10-fold in antigen-responsive cells with respect to the initial cell population.
Experimental Allergic Aspermatogenic Orchitis
Authors: Hagopian Arpi , Jackson Jesse J. , Carlo Dennis J. , Limjuco Guadalupe A. , Eylar E. H.
Published: December 2022

Four glycoproteins (GP1, 2, 3, and 4) rich in carbohydrate were isolated from guinea pig testes. GP1, 2, and 4 (one or more) were localized in the sperm acrosome by the indirect immunofluorescence technique. Purification consisted of delipidation with chloroform-methanol (2:1), acid extraction at pH 3.0, precipitation with 85% saturated ammonium sulfate, extraction with 5% trichloroacetic acid, and either gel filtration on agarose or ultrafiltration. The final purification steps were isoelectrofocusing or gel filtration on Sephadex G-75 followed by preparative slab gel electrophoresis at pH 8.6. Each glycoprotein appeared homogeneous by gel electrophoresis at pH 2.7 and 8.6, and by immunoelectrophoresis. The crude glycoprotein fraction from the agarose column was resolved into the three major components, GP1, 2, and 3, distinguished by their isoelectric points (pI 3.9, 4.4, and 5.0, respectively), electrophoretic mobilities at pH 8.6, and reactivities with antiserum in immunoelectrophoresis. GP4, isolated by ultrafiltration and Sephadex G-75 chromatography, was differentiated by the same criteria. Approximately 5 mg each of purified GP1, 3, and 4 and 2 to 3 mg of GP2 were isolated from 1000 g of wet guinea pig testes.

GP1, 2, and 4 induced precipitating antibody in rabbits and goats. GP1 and GP4 induced allergic aspermatogenic orchitis in guinea pigs, an autoimmune disease characterized by infiltration of mononuclear cells around and within the seminiferous tubules, followed by extensive destruction of the germinal epithelium. The course of the disease induced by 2 μg of either GP1 or GP4 was essentially identical in time course and pathology to that induced by whole testicular homogenates or 1 μg of purified acrosomal protein (AP1).
Communications
Authors: Gerber N. Lynn , Steinberg Alfred D.
Published: December 2022

Thymocytes from 1-month-old NZB/W mice were separated at unit gravity and then studied for helper or suppressor effects in a GVH assay. Thymocytes with a settling rate of 4 to 6 mm/hr suppressed but did not help. In contrast, help but no suppression characterized thymocytes settling at 6 to 10 mm/hr. Thus helper and suppressor thymocytes in this GVH assay were physically separated.
Effect of Cold Exposure on the Metabolism of Immunoglobulins in Rabbits
Authors: Sabiston Brian H. , Ste. Rose John E. M.
Published: December 2022

The effect of low environmental temperature on the metabolism of IgG and IgM was examined in unimmunized rabbits. The half-lives of both IgG and IgM were less in animals kept at 4°C for 6 weeks than in animals kept at 22°C. Serum concentrations of IgM and IgG were unaltered by cold exposure but intravascular pool sizes tended to increase as a consequence of an expanded serum volume. Fractional turnover rates of both IgM and IgG were greater in cold-exposed animals. At both 4°C and 22°C, the fractional catabolic rate of IgM was independent of its serum concentration whereas that of IgG was correlated directly with its serum concentration. Absolute turnover of both IgM and IgG was accelerated by cold exposure. It is suggested that increased synthesis of immunoglobulin could account for the higher levels of antibody reportedly found in cold-exposed rabbits.
Acute and Chronic Infection of Human Lymphoblastoid Cell Lines with Measles Virus
Authors: Barry David W. , Sullivan John L. , Lucas Susan J. , Dunlap Ruth C. , Albrecht Paul
Published: December 2022

Several human continuous lymphoblastoid cell lines (LCL) having T or B characteristics were infected with low and high passage strains of measles virus. All of the cell lines were susceptible to one or the other or to both strains of measles virus with the production of typical syncytial giant cells and released cell-free infectious virus into the supernatant medium. There was no consistent pattern of susceptibility of LCL with either T or B characteristics to infection by measles virus. Viral induced cytolysis of the lymphoblastoid cells in many of the lines was marked, but in the LCL that could be maintained over longer periods of time, a state of chronic, less cytolytic and persistent infection could be established. The infection was characterized by the production of moderate amounts of cell-free infectious virus for up to 4½ months after initial infection with little change in the number of viable cells in culture. Long-term low multiplicity of infection (MOI) experiments demonstrated that the cell-free infectious virus was being produced only by a small number of cells, but the majority of cells in culture contained measles antigen that was in a cell-restricted, noninfectious, or defective form. Electron microscopic examination of the chronically infected cells demonstrated that many of them contained aggregates of hollow tubular intranuclear nucleocapsids whose “stripped” appearance was in marked contrast to the larger granular intracytoplasmic nucleocapsids found during earlier stages of infection. It is theorized that the persistent infection of LCL may serve as a model in understanding the immune mechanisms which permit latent and chronic measles infection in man.
Does Freund's Adjuvant Denature Protein Antigens? EPR Studies of Emulsified Hemoglobin
Authors: Berzofsky Jay A. , Schechter Alan N. , Kon Hideo
Published: December 2022

Use of complete Freund's adjuvant for production of antibodies to study protein conformation is valid only if emulsification in adjuvant does not denature protein antigens. Using electron paramagnetic resonance to observe directly the protein in situ in the opaque emulsions, we demonstrate that hemoglobin is not denatured by emulsification or storage in adjuvant for 24 hr at 4°C, conditions comparable to the usual handling of antigens before immunization.
Characterization of the Target Cell Receptor for IgE
Authors: Conrad Daniel H. , Froese Arnold
Published: December 2022

Purified rat peritoneal mast cells (RMC) and cultured rat basophilic leukemia (RBL) cells were surface labeled with 125I by using lactoperoxidase, incubated with unlabeled rat monoclonal IgE and subjected to solubilization by treatment with Nonidet P-40 (NP-40). With both cell types significant amounts of radioiodinated material could be specifically precipitated by a “sandwich” system consisting of rabbit anti-rat ε-chain and goat anti-rabbit Ig. The precipitates were dissociated with sodium dodecyl sulfate (SDS) and urea and subsequently analyzed by SDS-polyacrylamide gel electrophoresis. With RMC three radioactive bands were seen. One corresponded to IgE present on the RMC at the time of isolation. A small band migrating in the region of light chain was seen with both specific (anti-IgE) and control precipitates. It showed no demonstrable relationship to IgE. The major radioactive band corresponded to a m.w. of 62,000. This band was dependent upon the presence of IgE and was not found when non-IgE binding control cells were used. With RBL cells, only the IgE-dependent 62,000 dalton peak was present. Saturation of the IgE receptor sites of the RMC or RBL cells before lactoperoxidase labeling almost totally eliminated this radioactive band, indicating that cell-bound IgE rendered this membrane component inaccessible to the radiolabel. These results strongly suggest that this cellular component is identical, at least in part, with the target cell surface receptor for reaginic antibody. The data also further support the hypothesis that the neoplastic RBL cells have a normal surface receptor for IgE.
In Vitro Generation of Secondary Cell-Mediated Cytotoxic Response Against a Syngeneic Gross Virus-Induced Lymphoma in Rats
Authors: Glaser Moshe , Bonnard Guy D. , Herberman Ronald B.
Published: December 2022

Cell-mediated cytotoxic responses against a syngeneic Gross virus-induced lymphoma, (C58NT)D, in W/Fu rats were generated in vitro by using mixed lymphocyte-tumor cell cultures. The source of responding cells was either spleens from normal rats or spleens from rats carrying or having rejected (C58NT)D tumors. Mitomycin C-treated (C58NT)D tumor cells were used as stimulating cells. The secondary anti-tumor cytotoxic response occurred more rapidly and reached higher levels than the primary response, and it was antigen specific. T cells, but not B cells or macrophages, were essential for both the induction and the effector phases of the secondary anti-tumor responses. These data suggest that specific memory T cells persist for long periods of time in the lymphoid organs of (C58NT)D immune rats, which can rapidly become cytotoxic upon re-exposure to antigen.
The Distribution of Terminal Deoxynucleotidyl Transferase (TdT) among Subsets of Thymocytes in the Rat
Authors: Barton Randall , Goldschneider Irving , Bollum F. J.
Published: December 2022

Terminal deoxynucleotidyl transferase (TdT), a unique DNA-polymerizing enzyme, has been shown to be present in a moderately dense subpopulation of rat thymocytes separated on discontinuous Ficoll density gradients. This subpopulation has been characterized by using antigenic and functional markers to identify directly and quantify cortical and medullary thymocytes. The TdT-positive thymocytes are depleted by cortisone administration, lack responsiveness to phytohemagglutinin, concanavalin-A, and histocompatibility alloantigens, bear surface antigens characteristic of cortical thymocytes (bone marrow lymphocyte antigen) and lack surface antigens characteristic of medullary thymocytes (rat-masked thymocyte antigen and histocompatibility antigens). The results indicate that TdT is present exclusively (or in markedly higher concentrations) in a subset of cells which comprises about 65% of cortical thymocytes. Two other major subsets of cortical thymocytes were identified which appeared to be TdT-negative. A minor subset of very low density cortical thymocytes was also defined. These observations have provided insight into the possible pathways of thymocyte ontogeny.
Structure and Function of Immunoglobulin Domains
Authors: Ellerson J. R. , Yasmeen D. , Painter R. H. , Dorrington K. J.
Published: December 2022

Exposure of Fc fragments derived from human IgG1 myeloma proteins to acid pH rendered the region between the Cγ2 and Cγ3 domains transiently susceptible to cleavage by trypsin upon return to neutral pH. Trypsin covalently linked to Sepharose was used and two fragments derived from the Cγ2 region and one from the Cγ3 region were purified by column chromatography. On the basis of amino acid analysis, primary sequence data, antigenic properties, and m.w., one of the Cγ2 fragments was shown to consist of two polypeptide chains of unequal mass joined by the inter-heavy chain disulfide bonds. The larger chain corresponded to a stretch of γ-chain between Thr 223 and Lys 338 (Eu numbering) and the shorter chain to the section between Thr 223 and Lys 248. The other Cγ2-fragment was a disulfide-linked dimer of the Thr 223 to Lys 338 sections of the paired γ-chains. When this latter fragment was reduced under mild conditions it dissociated into monomers indicating that there was little or no noncovalent interactions between the Cγ2 domains. The Cγ3-fragment was shown to be a noncovalent dimer composed of the Glu 345 to Lys 349 sections of the two γ-chains although some heterogeneity was apparent at the amino-terminus. Circular dichroism was used to probe the conformational relationships between the isolated domains and the parent Fc. The spectral properties of Fc could not be fully accounted for on the basis of the spectra observed for the isolated domains which suggested that inter-domain interaction might be significant in Fc.
The Chemoattraction of Neutrophils by Heterologous and Homologous Cytotoxic Sera
Authors: Williams Dianne , Esquenazi Violet , Cirocco Robert , Jensen Joerg A.
Published: December 2022

Neutrophils (PMN) migrated in vitro chemotactically along concentration gradients of heterologous and homologous leukocytotoxic, complement-active sera. The heterologous systems consisted of normal, non-immune nurse shark or dog serum and human PMN; the homologous systems consisted of human or dog immune sera and human or dog PMN, respectively. Chemotaxis was induced 1) by fresh cytotoxic sera, 2) by complement alone if the cytotoxic antibodies were with the responding cells or if the cells were passively sensitized with the antibodies, and 3) by individual and combined complement components if the corresponding intermediate PMN-antibody-complement component complexes were the migrating cells. These observations represent a new mechanism of immune chemotaxis: the chemotactic stimulation is a direct consequence of a gradient-controlled immune reaction occurring at and involving the surface of the chemoattracted cell. This mechanism is distinctly different from that which depends on chemotactic factors produced or released independent of and at a distance from the responding cells.
Preferential Expression of Anti-Azobenzenearsonate Antibodies of Heterozygous a1a3 Rabbits in the a1 Allotype
Authors: Foshay Miriam C. , Zimmerman Sarah E. , Spencer Lenore K. , Haurowitz Felix , Knight Katherine L.
Published: December 2022

The preferential expression of anti-As antibodies in the allotype a1 of heterozygous a1a3 rabbits immunized with As-TMA-BSA has been investigated by means of quantitative methods. The average content of the anti-As antibodies in a1 and a3 allotypes was 84 and 11%, respectively; the analogous values for anti-TMA antibodies were 41 and 56%, and for anti-BSA antibodies they were 54 and 41%. The molar anti-As/anti-TMA ratios in the heterozygous animals were approximately the same as in homozygous a1a1 rabbits sensitized with As-TMA-BSA. The very low yields of anti-As-antibodies of allotype a3 cannot be caused by a lack of genes for the production of anti-As antibodies of allotype a3 because a3a3 homozygotes produce considerable amounts of anti-As antibody of allotype a3. Competition between lymphoid cells having anti-As receptors of different allotype and different affinity for the antigenic p-azobenzenearsonate determinant is discussed as a possible cause for the preferential expression in the a1 allotype.
Sensitization of T Lymphocytes in Vitro by Syngeneic Macrophages Fed with Tumor Antigens
Authors: Treves Abraham J. , Schechter Bilha , Cohen Irun R. , Feldman Michael
Published: December 2022

We investigated the interaction between T lymphocytes and macrophages in the in vitro sensitization of lymphocytes against tumor cells. Spleen cells were sensitized in vitro by syngeneic peritoneal macrophages that had been fed with cell-free antigen preparation of syngeneic tumor cells. The sensitized T lymphocytes acquired specific cytotoxic activity in vitro and the capacity to inhibit tumor development in vivo. Allogeneic macrophages, syngeneic fibroblasts, or the antigen preparation by itself were not able to sensitize the lymphocytes against the tumor.
Eosinophils and Immune Mechanisms
Authors: Greene Bruce M. , Colley Daniel G.
Published: December 2022

The lymphoid cell population responsible for production of eosinophil stimulation promoter (ESP), a lymphokine which increases migration of eosinophils, was investigated in murine Schistosoma mansoni infection. Con A challenge induced ESP production, whereas LPS did not. Prior treatment with anti-θC3H alloantiserum plus complement in vitro eliminated ESP production; in vivo treatment with rabbit anti-mouse thymocyte serum consistently reduced ESP production by splenic lymphoid cells, but affected lymph node cell ESP production only after exceptionally large doses. Thymocytes did not produce significant amounts of ESP; nor did lymphoid cells from congenitally athymic mice. Depletion of B lymphocytes and macrophages by nylon fiber adherence eliminated antigen-induced ESP production; this was partially restored by addition of non-immune, 72-hr peritoneal exudate cells. Con A-induced ESP production was not affected by nylon fiber treatment. These results demonstrate that ESP is produced by an ATS-sensitive, peripheralized T lymphocyte population, and suggest a macrophage requirement for antigen-induced production of this lymphokine.
The Effect of Serum Inhibitor on the Antigenic and Mitogenic Responses to E. Coli Bacteria
Authors: Poe Wanda J. , Michael J. Gabriel
Published: December 2022

The effect of serum inhibitory factors on the induction of the in vitro immune response and mitogenic response to Escherichia coli antigens in murine spleen cell cultures was investigated. Both normal and congenitally athymic mice served as spleen cell and serum donors. Cultured CBA/J spleen cells produced anti-bacterial antibody in the presence of mouse serum, but the mitogenic response to the bacterial antigen was suppressed by the addition of mouse serum. The addition of mouse serum from either CBA/J or nude mice to nude spleen cell cultures led to suppression of both the antigenic and mitogenic responses to bacterial antigen. The polyclonal antibody response by CBA/J spleen cell cultures to SRBC induced by bacteria was significantly enhanced by the addition of either CBA/J or nude mouse serum. Conversely, the nude spleen cell polyclonal response to SRBC was significantly inhibited by high concentrations of nude or CBA/J mouse serum. These data indicate that inhibition of the specific E. coli and nonspecific polyclonal antibody responses by mouse serum occurs only in cultures of spleen cells taken from mice lacking a population of T lymphocytes.
Mitogen Induction of Murine C-Type Viruses
Authors: Schumann Gebhard , Moroni Christoph
Published: December 2022

We reported previously in vitro induction of endogenous C-type viruses from normal mouse spleen cells by lipopolysaccharide (LPS) as well as by combination treatment with concanavalin A and 5-bromo-2′-deoxyuridine (Con A/BrdU). To identify the cell types sensitive to virus induction and to study the relationship of mitogenicity to virus induction we have compared T cell populations (BALB/c thymus cells and cortisone-resistant thymus cells), B cell populations (nu/nu spleen cells and lymph node cells), adherent BALB/c peritoneal cells and mixed populations (BALB/c spleen cells, macrophage-depleted BALB/c spleen cells, and lymph node cells). LPS-induction occurred only in B cell-containing populations. In contrast, induction by Con A/BrdU depended on the presence of both T and B cells. In both instances, neither macrophages nor hemopoietic cells appeared to be a major source of virus. Treatment with anti-Ig serum and complement reduced virus induction by LPS/BrdU but not by Con A/BrdU suggesting that different cell populations produce virus after stimulation with these two different mitogens.
The Induction by Complement of a Change in KSCN-Dissociable Red Cell Membrane Lipids
Authors: Giavedoni Elda B. , Dalmasso Agustin P.
Published: December 2022

During complement lysis of antibody-sensitized sheep erythrocytes (EA) there was a larger loss of membrane phospholipids than during lysis elicited by hypotonic buffer. In addition, membranes prepared from complement-lysed EA had a marked reduction in KSCN (2.4 M)-dissociable membrane cholesterol and phospholipids, as compared to membranes from EA lysed hypotonically. Complement lysis caused a mild reduction in the amount of KSCN-dissociable membrane hexose but no change in the amount of dissociable protein. The impairment in dissociation of membrane lipids was related to the action of C8; it did not occur with membranes from EA that were treated with heat-inactivated (56°C for 30 min) human serum, C4-deficient guinea pig serum, C6-deficient rabbit serum, or the first seven human complement components. EA lysed with limited amounts of complement exhibited a partial impairment in KSCN-dissociable lipids. Membranes from erythrocytes lysed with melittin showed a large increase in dissociation by KSCN of lipids, proteins, and hexoses. Membranes from erythrocytes lysed with lysolecithin or phospholipase C showed, in addition to a reduction in dissociable lipid, a much larger reduction in dissociable hexose than membranes from complement-lysed cells. These profiles of reactivity with 2.4 M KSCN indicate that the membrane pertubations caused by complement may be specific. We conclude that complement lysis is accompanied by a major rearrangement of membrane cholesterol and phospholipid which could be demonstrated in membranes from cells lysed by only one or very few complement lesions. Therefore, it appears that the lesions induce a propagated change in the lipid organization which extends throughout large areas of the membrane. This change might be responsible for the impairment of membrane permeability that follows the action of complement and results in cell destruction.
Secondary IgG Responses to Type III Pueumococcal Polysaccharide
Authors: Braley-Mullen Helen
Published: December 2022

Mice primed with a thymus- (T) dependent form of Type III pneumococcal polysaccharide (S3), i.e., S3 coupled to erythrocytes (S3-RBC) produce S3-specific IgG antibody after secondary challenge with either S3 or S3-RBC. The production of IgG antibody by mice challenged with S3 was shown to be T independent since secondary responses were enhanced when mice were treated with anti-lymphocyte serum (ALS) at the time of secondary challenge with S3 and T-depleted spleen cells responded as well as unfractionated spleen cells to S3 in an adoptive transfer system. Secondary S3-specific IgG responses in mice challenged with S3-RBC were shown to be T dependent by the same criteria. The results obtained by using S3 as the antigen indicate that IgG-producing B cells (B γ cells) can recognize and respond to antigen in the absence of helper T cells. On the other hand, T cells were required for the induction of S3-specific memory B γ cells since mice depleted of T cells by treatment with ALS at the time of priming with S3-RBC failed to produce S3-specific IgG antibody after secondary challenge with either S3 or S3-RBC. Since RBC-specific memory cells were induced in T-deprived mice the results suggest that T cell regulation of IgG antibody production may vary for different antigens.
Inhibition of Stimulation in Murine Mixed Lymphocyte Cultures with an Alloantiserum Directed Against a Shared Ia Determinant
Authors: Schwartz Ronald H. , Fathman C. Garrison , Sachs David H.
Published: December 2022

Pools of high titered alloantisera were raised by immunizing (B10.A/SgSn × A/WySn)F1 mice with C57BL/10Sn(B10) spleen cells. This serum (F1 anti-B10), when added to one-way mixed lymphocyte cultures (MLC), inhibited stimulation of B10.A splenic responders by both B10 and B10.D2/nSn irradiated, splenic stimulators. The B10 stimulation was suppressed approximately 85% whereas the mean suppression of B10.D2 stimulation was approximately 60%. In the former case, the serum contained antibodies reactive with multiple major histocompatibility complex determinants on the stimulator cells. In the latter case, the cytotoxic reactivity of the serum was directed principally against an I region-associated determinant (Ia.8) shared by B10 and B10.D2 and coded for by a gene(s) in the I-A subregion. The magnitude of the suppression of the response to B10.D2 cells (60%) was similar to the reduction in stimulation observed when the Ia.8 difference was eliminated genetically by using (B10 × B10.A)F1 responder cells against irradiated B10.D2 stimulators. Inhibition of MLC by this antiserum was a function of reactivity with stimulator and not responder cells. Although some pools of F1 anti-B10 antiserum produced partial inhibition of the responder cell in a B10.D2 vs B10.Ax MLC combination, the results were inconsistent and not correlated with the anti-Ia.8 cytotoxicity titers. In addition, an F1 anti-B10 antiserum pool, which consistently failed to inhibit the responder cell, nevertheless inhibited both irradiated B10.D2 and (B10.A × B10.D2)F1 cells from stimulating B10.A responder cells. However, this same antiserum did not inhibit stimulation of B10.D2 responder cells by the (B10.A × B10.D2)F1 stimulators. Thus, the binding of antibodies to the non-stimulating antigens on the F1 stimulator cell did not interfere with the capacity of the appropriate stimulating antigens to cause stimulation. All of these results are consistent with the hypothesis that Ia alloantigens are the major stimulating determinants of I region-associated MLC reactions.
Studies on the Terminal Stages of Antibody-Complement-Mediated Killing of a Tumor Cell
Authors: Boyle Michael D. P. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

Transformation of T* to dead cells was prevented by 3′5′ cAMP. The effect of 3′5′ cAMP was dose, time, and temperature dependent. T* washed free of 3′5′ cAMP after short-term incubation proceeded to die to the same extent as control cells. After 3 hr of incubation of T* with 3′5′ cAMP the level of killing was significantly reduced. The 3′5′ cyclic nucleotides of uridine, guanine, cytosine, and thymidine and the 2′3′ cyclic adenosine nucleotide were not effective. It was concluded that prolonged treatment of T* with 3′5′ cAMP either irreversibly blocked the damage-producing process or facilitated the repair of damaged sites.
An in Vitro Assay for T Cell Immunity to Malaria in Mice
Authors: Weinbaum Fredric I. , Evans Charles B. , Tigelaar Robert E.
Published: December 2022

After infection with a nonlethal strain of murine malaria (17XNL Plasmodium berghei yoelli), BALB/c mice are then resistant to a lethal strain (17XL P. b. yoelli). BALB/c mice were infected with 17XNL, and challenged 3 weeks later, after clearing their parasitemias, with 17XL. Three weeks thereafter, spleen cells from such immune animals were used to define an early peaking T-dependent (anti-ϑ sensitive) antigen-specific proliferative response when incubated in vitro with 17XL infected RBC, or a saline soluble 17XL antigen preparation. T dependent responsiveness of spleen cells from uninoculated control animals to the 17XL antigen preparation was also observed, but demonstrated a much different (delayed) kinetics from that observed with immune cells.
Naturally Soluble Tumor Antigens from Guinea Pig Hepatomas: Isolation and Partial Characterization
Authors: Detrick-Hooks Barbara , Smith Howard G. , Bast Robert C. , Dunkel Virginia C. , Borsos Tibor
Published: December 2022

Naturally soluble tumor antigens were detected in the ascites fluid of guinea pigs bearing an ascites tumor and from exhausted tissue culture media of cultured tumor cells. Two antigenically distinct cell lines of diethylnitrosamine-induced strain-2 guinea pig hepatomas (line-10 and line-1) served as the source of tumor antigens. Tumor antigen activity was detected by four different techniques: immunodiffusion, inhibition of complement-mediated cytotoxicity, inhibition of membrane immunofluorescence, and delayed cutaneous hypersensitivity. With syngeneic tumorspecific antiserum, line-10 guinea pig tumor antigens were detected by immunofluorescence in the concentrated ascites and tissue culture fluids. With a xenogeneic antiserum, demonstrated to be tumor specific, line-10 tumor antigens were detected not only in the concentrated ascites and tissue culture fluids but also in two of the partially purified fractions of these fluids. When the line-10 concentrated ascites and its fraction I were subjected to ultracentrifugation at 300,000 × G for 1 hr, the antigen activity was retained in the supernatant and thus by this criterion the tumor antigens detected in these samples are soluble.

Immunodiffusion data indicate that more than one antigen is present in the line-10 system since three lines of precipitation were detected when line-10 concentrated ascites was reacted with the line-10 tumor-specific antiserum. In contrast to this, the line-10-concentrated tissue culture fluid displayed only one line of precipitation.

Although tumor antigens could not be demonstrated in the other antigenically distinct tumor cell line, line-1, by immunodiffusion or inhibition of membrane immunofluorescence, inhibition of complement-mediated cytotoxicity was able to detect tumor antigens in the line-1 concentrated ascites and tissue culture fluids.
Shared Antigens between Human Malignant Melanoma Cells and Mycobacterium Bovis (BCG)
Authors: Minden Percy , Sharpton Thomas R. , McClatchy J. Kenneth
Published: December 2022

This study was undertaken to investigate the antigenic relationships between human malignant melanoma cells and Mycobacterium bovis (BCG). Rabbits were immunized with sonicates of BCG or with malignant melanoma cells from different patients and the resulting antisera were tested for their capacity to bind radiolabeled soluble extracts prepared from BCG and melanoma cells.

The binding of antibodies to radiolabeled antigens was studied by precipitation of radiolabeled antigen-antibody complexes by anti-rabbit immunoglobulin. Antibodies in sera from rabbits immunized with either BCG (anti-BCG) or melanoma cells (anti-melanoma) bound both the labeled BCG and melanoma antigens. Control antisera, from rabbits immunized with human acute or chronic lymphatic leukemia cells or with normal human spleen cells, did not bind significant amounts of radiolabeled BCG. Antibodies in sera from rabbits immunized with normal spleen cells bound small but significant amounts of radiolabeled melanoma antigens. Binding by anti-BCG and anti-melanoma to the radiolabeled antigens was studied before and after absorption of antisera with cells from human melanoma, leukemia, guinea pig hepatoma, and normal human spleen cells. Inhibition studies using unlabeled BCG extracts also were carried out. The absorption and inhibition studies confirmed that the binding reactions were specific and that antigens from five melanoma patients shared antigenic determinants with BCG.
The Relative Paucity of IgE in Human Milk
Authors: Underdown Brian J. , Knight Allan , Papsin Fred R.
Published: December 2022

The levels of IgE were determined in paired samples of serum and milk whey obtained 3 to 8 days of postpartum, from 16 human lactating mothers who had reported a history of allergy to a variety of common allergens. Two assay procedures were employed to measure total IgE, a double-antibody assay and a commercially available solid phase assay (RIST). In addition, each sample of serum and whey was tested for specific IgE antibodies to a variety of allergens by the RAST test. The levels of total serum IgE were between 30 and 2300 I.U./ml and relatively good agreement was observed for both the double-antibody and RIST methods. In contrast, total IgE levels in milk whey were either undetectable (<3.0 I.U./ml in 14 of 16 subjects) or very low when analyzed by the double-antibody method, but were very high (400 to 1650 I.U./ml when analyzed by the RIST method. However, IgE added to milk whey could be measured by the double-antibody procedure indicating that the low levels detected in milk were not a fault of the double-antibody assay. It was assumed that the RIST test was subject to nonspecific interference by factors in milk whey which caused the determination of high, but incorrect, levels of IgE.

Specific IgE antibodies were detected in the serum of 10 of 16 subjects but were not present in milk whey. A comparison of the whey/serum ratios of albumin, IgA, and IgE suggested that little, if any, IgE is selectively synthesized or secreted in the mammary gland.
Motheaten: An Immunodeficient Mouse with Markedly Less Ability to Survive Than the Nude Mouse in a Germfree Environment
Authors: Lutzner Marvin A. , Hansen Carl T.
Published: December 2022

The motheaten mouse is an immunodeficient mutant which has been found to have elevated immunoglobulin levels, to exhibit autoimmunity, and early death. Since survival is not improved by a germfree environment, it appears that some mechanism other than infection results in its early demise.
Relationship of CIF, LT, and PIF Released in vitro by Activated Human Lymphocytes
Authors: Jeffes Edward W. B. , Granger Gale A.
Published: December 2022

Lymphotoxin (LT), and proliferation inhibition factor (PIF) activities found in 5-day supernatants of mitogen-activated human lymphocytes (SAL) were further compared. In agreement with previous results, the activities could not be distinguished functionally. Quantitative differences in the amount of activity detected in the SAL could be accounted for on the basis of target cell differences, concentration of the lymphocyte effector molecules in the supernatant, and the parameter employed to assess cell function. Growth inhibitory activity detected at high supernatant dilutions was completely reversible, whereas the cytotoxic activity detected at low supernatant dilutions was irreversible. When the active medium was fractionated on DEAE, two peaks of inhibitory activity were detected. Depending upon the amount of activity and target cell, both peaks of activity were growth inhibitory or cytotoxic. Since both peaks of material affected HeLa and L-929 cells, the materials were not species specific. Thus, it appears that cloning inhibitor factor, LT, and PIF activities may actually be measures of the same stable materials found in 5-day activated lymphocyte supernatants.
Regulatory Factors Produced by Lymphocytes
Authors: Lee Sheng-Chung , Lucas Zoltan J.
Published: December 2022

Supernatant fluids of mitogen-activated human tonsil lymphocytes contain large amounts of a factor toxic to mouse L cells. This substance, with a m.w. of 80,000 ±5,000 daltons, is called α-lymphotoxin (α-LT), to differentiate it from another toxin elaborated by mitogen-activated human blood lymphocytes, called β-lymphotoxin (β-LT), which differs from α-LT in size (45,000 ±5,000 daltons), antigenicity, and stability. Further purification of α-LT by sequential phosphocellulose and DEAE-cellulose chromatography and polyacrylamide gel electrophoresis (PAGE) identifies a series of cytotoxins differing in ion exchange characteristics and electrophoretic mobilities. The three PAGE fractions (PAGE Ia, Ib and II), recovered in 2, 4.6, and 21% yield from the starting serum-free culture supernatant, represent purifications of 24-, 24- and 1851-fold, respectively. Each cytotoxic fraction has a ribonuclease activity. Comparison of RNase and mouse L cell cytotoxic activities of the three α-LT fractions shows that both activities for all three fractions have a similar temperature stability pattern and that both are similarly inhibited by DNA, single strand forms better than double strands, by glycerol in 5 to 20% concentration, and by protein denaturing reagents. These observations suggest, but do not prove, that mouse L cell toxicity and RNase activity are mediated by the same substance, which appears to occur in multiple or isozymic forms.
Measurement of IgG-Blocking Antibodies: Development and Application of a Radioimmunoassay
Authors: Sobotka Anne K. , Valentine Martin D. , Ishizaka Kimishige , Lichtenstein Lawrence M.
Published: December 2022

A radioimmunoassay for measuring blocking antibodies has been developed. We used the ragweed antigen E system to show that the same blocking antibodies (IgG) measured by inhibition of antigen-induced leukocyte histamine release were precipitated in the binding assay (rs = 0.96, p < 0.001), thus validating a widely applicable technique for measuring blocking antibodies. Binding of phospholipase-A (Phos-A), the major allergen in honey bee venom, was also shown to correlate significantly with inhibition of histamine release.

Hymenoptera (insect) hypersensitivity was used as a model to demonstrate application of the binding assay. Sera obtained from patients undergoing whole body extract therapy contained negligible amounts of specific blocking antibodies. Significantly higher blocking antibody titers to both whole honey bee venom and Phos-A were measured in sera drawn from patients immunized with whole venom. The use of the binding radioimmunoassay should facilitate management of allergic disease processes in which blocking antibodies are thought to be protective.
Low Molecular Weight Immunoglobulins in Rana Catesbeiana Tadpoles
Authors: Green Christopher , Steiner Lisa A.
Published: December 2022

Rana catesbeiana tadpoles formed high and low m. w. antibodies in response to immunization with a bacteriophage. Although the neutralizing activity associated with the low m. w. immunoglobulins was relatively weak, the existence of antibodies in this class was clearly demonstrated by radioimmunoelectrophoresis. Moreover, two antigenically distinct variants of the low m. w. antibodies were detected. These were serologically indistinguishable from the two types of low m. w. immunoglobulin previously isolated from the serum of adult frogs of this species.
Cross-Reactive T-Cell Antigens among Mammalian Species
Authors: Balch C. M. , Dagg M. K. , Cooper M. D.
Published: December 2022

Fluoresceinated heteroantisera prepared against T cells of rats, monkeys, and humans were reacted with thymus and spleen cells from 11 selected species. These reagents recognized cross-reacting T cell antigen(s) among rodent species (mouse, rat, guinea pig, and hamster) and among primate species (monkey and humans). With one exception, the cross-reactivity was restricted to a phylogenetic order. All three antisera required relatively few absorptions to achieve T cell specificity for related species when compared to absorption requirements for the isologous species. Differentiation antigens within a phylogenetic order thus appear to be more homologous than other cell surface constituents on T cells.
B Lymphocytes in the Guinea Pig Thymus Are Specifically Localized in the Central Area
Authors: Baak Jan P. A. , Meijer Chris J. L. M. , Nieuwenhuyzen Kruseman Arie C. , Langevoort Henk L. , Kater Louis
Published: December 2022

Since the central area is an integral part of the guinea pig thymus, the cells in this area were compared with those in the thymic cortex and medulla in cryostat-sections by using methods for demonstration of E-, EA-, EAC-adherence and surface membrane immunoglobulins. In the extra cortical central area (ECCA) 15 to 25% of the lymphocytes showed EAC-adherence and 5 to 10% appeared to bear surface membrane immunoglobulins (SIg). In the lymph sinuses up to 70% of the lymphocytes were EAC- and SIg-positive. A small amount of EAC-adhering cells was present in the medulla of the central area. Cortical lymphocytes were EAC- and SIg-negative.

From these results we conclude that in the guinea pig thymus B lymphocytes are specifically localized in the central area.
Studies on the Genetic Control of Antibody Affinity: The Independent Control of Antibody Levels and Affinity in Biozzi Mice
Authors: Katz Fay E. , Steward Michael W.
Published: December 2022

The levels (Abt) and relative affinity (KR) of antibody produced to protein antigens injected in saline have been measured in the 22nd generation of the Biozzi high (Ab/H) and low (Ab/L) antibody-producing mice.

No significant differences in affinity were observed between the two lines of mice (p 0.10) but the levels of antibody (Abt) differed significantly (p 0.0025) when immunized with antigen in saline; however, both Ab/H and Ab/L mice were able to mount a high affinity response to protein antigens injected in Freund's complete adjuvant.

These results substantiate earlier observations that in mice, antibody affinity (KR) and antibody level (Abt) are under independent genetic control.
Cutaneous Basophil Hypersensitivity Uncovered in the Cell Transfer of Classical Tuberculin Hypersensitivity
Authors: Askenase Philip W.
Published: December 2022

Cellular transfer of cutaneous basophil hypersensitivity (CBH) was studied. Guinea pigs immunized for CBH with incomplete Freund's adjuvant (IFA) provided cells which could transfer delayed and basophil-rich reactions in skin tests of recipients. Guinea pigs immunized with complete Freund's adjuvant (CFA) and skin tested at 3 weeks have classical tuberculin-type delayed hypersensitivity reactions (DH), which are characteristically devoid of basophils. However, recipients of cells from donors with DH, surprisingly, were found to have delayed skin reactions containing large basophil infiltrates which were lacking in the donors. Thus, recipients of classical cell transfers of tuberculin-type DH had delayed reactions which resembled CBH. Control experiments verified that the cell transfer of CBH from donors with DH was due to passive transfer with live cells and not transfer of contaminating humoral factors or active sensitization of recipients. It was concluded that cutaneous basophil responses were suppressed in CFA-immunized donors and expressed in cell transfer recipients.

Cells from donors immunized with CFA were enriched for nonadherent and nonimmunoglobulin-bearing lymphocytes by passage through nylon wool columns, and these cells transferred conjugate specific CBH reactions. It was concluded that cells mediating these transfers were probably T cells. The finding of basophils in cell transfers of DH and a variety of other findings suggesting complex regulation of basophil numbers in tissue lead to the conclusion that the term CBH be used to simply describe a basophil-containing skin reaction.
HLA-Related Control of Spontaneous and Antibody-Dependent Cell-Mediated Cytotoxic Activity in Humans
Authors: Santoli Daniela , Trinchieri Giorgio , Zmijewski Chester M. , Koprowski Hilary
Published: December 2022

Normal human lymphocyte preparations were tested for their ability to lyse both antibody-coated and unsensitized human target cell lines. The capacity to induce these two types of cell-mediated cytotoxicity, antibody-dependent (Ab-CMC) and spontaneous (Sp-CMC), was detected by a 51Cr release assay. The reactivity of lymphocytes from individual donors showed a positive and highly significant correlation between Ab-CMC and Sp-CMC, leading to the hypothesis that the same type of effector cell is involved in the two cytotoxic mechanisms. Lymphocytes from male donors were about twice as effective as those from female donors in both systems. Moreover, the effector cells from male donors carrying HLA antigens A3 and B7 displayed a significantly lower reactivity in both Sp-CMC and Ab-CMC when compared with lymphocytes from male donors bearing any other HLA haplotype. The possible significance of the hyporeactivity of lymphocytes from normal subjects with HLA-A3,B7 haplotype in relation to an increased susceptibility to multiple sclerosis (MS) is discussed.
Human Transfer Factor: Effects on Lymphocyte Transformation
Authors: Burger Denis R. , Vandenbark Arthur A. , Finke Patricia , Nolte James E. , Mark Vetto R.
Published: December 2022

Transfer factor preparations from 57 different donors have been compared for effects on mitogen- and antigen-induced lymphocyte transformation. Nine of the preparations were mitogenic when added to cultured lymphocytes although the magnitude of this activity was relatively low. The majority of the preparations (48/57) did not affect PHA-induced lymphocyte transformation although augmentation (6 of 57) and suppression (3 of 57) was observed with some. In addition we observed that most of the preparations tested suppressed ConA stimulation and augmented the PWM response.

When selected preparations were evaluated on antigen-responsive cells, there was a correlation between the magnitude of antigen responsiveness and the magnitude of TF augmentation of antigen-induced lymphocyte transformation (p < 0.005). Cultures that were not responsive to antigen (KLH-negative or BUdR-treated) could not be stimulated by TF from immune donors and antigen. These data suggest that TF preparations contain either stimulatory or inhibitory components and that TF is not capable of activating naive lymphocytes to undergo transformation in response to antigen.