bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigen-Induced Proliferation of Guinea Pig Lymphocytes in Vitro: Obligatory Role of Macrophages in the Recognition of Antigen by Immune T-Lymphocytes
Authors: Waldron James A. , Horn Robert G. , Rosenthal Alan S.
Published: December 2022

Highly purified lymphocytes were obtained from the lymph nodes of CFA-immune guinea pigs. Cultures of these lymphocytes did not exhibit proliferative responses upon stimulation with purified protein derivative (PPD), although they were quite responsive to stimulation with phytohemagglutinin. Addition of macrophages to these cultures resulted in marked enhancement of the PPD-induced lymphoproliferative response, but did not affect lymphocyte responsiveness to phytohemagglutinin. The ability of the purified immune lymphocytes to interact directly with soluble PPD was examined by incubating the cells for 1 hr at 37°C with various concentrations of PPD. The lymphocytes were then washed to remove unbound antigen and placed in culture. Addition of macrophages to cultures of antigen-pulsed lymphocytes did not result in the significant DNA synthesis, suggesting that the specifically immune lymphocytes were unable to bind and carry into culture sufficient antigen to allow for their subsequent activation. These results were not due to the induction of tolerance in the lymphocytes, since cultures of antigen-pulsed and untreated lymphocytes were equally responsive to continuous PPD when macrophages were added to the cultures, even when the antigen-pulsed lymphocytes had been exposed to high concentrations of PPD for 1 or 24 hr. In contrast, macrophages which were incubated for 1 hr at 37°C with a given concentration of PPD were as effective as the same concentration of PPD present continuously in macrophage-lymphocyte cultures in the induction of a lymphoproliferative response. Macrophages appear to play an obligatory role in the presentation of antigen to the immunospecific thymus-derived (T)-lymphocyte.
Restricted Clonal Response to DNP in Adult Offspring of Immunized Mice: A Maternal Effect
Authors: Kindred Berenice , Roelants Georges E.
Published: December 2022

Preoccupation with the status of a fetus as a homograft within the mother seems to have led to a virtual overlooking of the mother as the environment of the fetus. The problem of the possible effect on the young of immunization of the mother was studied by comparing adult offspring obtained before and after maternal immunization in their response to nitrophenyl haptens. Maternal immunization has a strong influence on the response to nitrophenyl haptens of subsequently conceived offspring. They show, compared to young obtained before maternal immunization, a more restricted anti-DNP response both in terms of circulating antibody titer, determined by a modified Farr assay, and of the number of DNP-specific clones expressed, determined by isoelectric focusing.

Possible mechanisms for this “maternal effect” and its relevance to studies of families showing antibodies of restricted isoelectric-focuing patterns are discussed.
Metabolic Behavior of C3 in Normal, C4-Deficient (C4D) and Cortisone-Treated Guinea Pigs
Authors: Atkinson John P. , Shin Hyun , Frank Michael M.
Published: December 2022

C3 was isolated from the pseudoglobulin fraction of guinea pig serum by DEAE and CM-cellulose chromatography and Pevikon block electrophoresis. The final product, which was functionally and chemically pure, was radiolabeled with 125I by chloramine-T and used for turnover studies. In normal guinea pigs, metabolic studies revealed the following: fractional catabolic rate (FCR), 7.9 ± 0.4%/hr (S.E.); synthetic rate, 6.2 ± 0.4 mg/kg/hr; and percent of C3 in the plasma pool, 51 ± 2. The corresponding means were not significantly different in C4 deficient animals, suggesting that activation of the classical complement pathway is not essential for significant C3 turnover in the resting state. Guinea pigs were given 20 (low) or 100 (high) mg cortisone acetate/kg. Serum C3 levels fell 27 ± 6% and 52 ± 5%, respectively, over the course of 2 weeks of therapy and stabilized at the lower values. In cortisone-treated animals studied during steady state conditions (after 14 days of continuous therapy), the FCR was 9.2 ± 4%/hr and 12.5 ± 7%/hr for the low and high dose groups, respectively. Synthetic rates and per centage of C3 in the intravascular pool were similar in treated and untreated animals. Two weeks after discontinuing therapy, the serum C3 and FCR had returned to normal. Thus, an increased fractional catabolic rate is primarily responsible for the depressed serum levels of C3 observed in cortisone-treated animals.
Immunologic Functions of Isolated Human Lymphocyte Subpopulations
Authors: Chess Leonard , MacDermott Richard P. , Schlossman Stuart F.
Published: December 2022

Soluble and cell surface antigen-induced triggering of highly purified human T and B cells was studied measuring 3H-thymidine incorporation in vitro. T cells but not B cells from antigen-sensitive donors responded to specific soluble antigen challenge. Nonsensitized cells failed to respond. The addition of sensitized T cells to autologous B cells did not induce B cells to proliferate in response to soluble antigen. In one-way mixed leukocyte cultures, T cells but not B cells incorporated 3H-thymidine in response in allogeneic mitomycin-treated targets. In contrast, both T and B target cells were capable of inducing proliferation of allogeneic T cells.
Studies on the Human T-Lymphocyte Population
Authors: Owen Frances L. , Fanger Michael W.
Published: December 2022

An IgG fraction of a goat antibody prepared against human thymus cells and absorbed with red blood cells, insolubilized human Ig, and cells from a patient with chronic lymphocytic leukemia (ATG) has been found to react specifically with the human T lymphocyte population (1). This antibody blocked spontaneous rosette formation by human peripheral lymphocytes and sheep red blood cells (SRBC). The F(ab′)2 and Fab′ fragments of ATG were also effective in blocking rosette formation. Moreover, capping of SRBC on lymphocytes was observed when cells were incubated with the F(ab′)2 fragment of ATG, but not with the Fab′ fragment. The data suggest that on human peripheral lymphocytes the receptor for SRBC and a T-specific antigen may be the same or may be linked. A soluble extract of human peripheral lymphocytes contained material capable of reacting with ATG. Fractions of this extract obtained from Sephadex G-200 and G-75 columns and corresponding to molecular weights of approximately 35,000 and 70,000 were capable of blocking ATD's inhibition of rosette formation. The active components appeared to be glycoproteins and migrated with Rf values of 0.96 on acrylamide gel electrophoresis.
Immune Response of Mice to Thy-1.1 Antigen: Genetic Control by Alleles at the Ir-5 Locus Loosely Linked to the H-2 Complex
Authors: Zaleski Marek , Klein Jan
Published: December 2022

The primary response of mice to Thy-1.1 antigen was measured by means of a plaque assay detecting cells which produce antibodies lytic for AKR thymocytes. It was found that F1 hybrids between the low responder strains C57BL/6J, C57BL/10J, or B10.T and the DBA/2J strain produced significantly more plaque-forming cells per spleen than either parental strain. In backcross populations of these three F1 hybrids to C57BL/6J, C57BL/10J, or B10.T parents, 16.1 to 21% of H-2 heterozygotes were low responders as was the H-2 homozygous parent. These low responder H-2 heterozygotes, when mated to the C57BL/6J parent, produced 70.2% low responder H-2 heterozygotes, whereas when mated to the DBA/2J parent they produced only 5.7% low responder H-2 heterozygotes. The data obtained were interpreted as evidence for the existence of the locus designated Ir-5, which participates in the control of the immune response to the Thy-1.1 antigen together with loci belonging to the I region of the H-2 complex designated Ir-Thy-1. From the recombination frequency between the Ir-5 locus, the H-2 complex, and the T locus in the (B10.T × DBA2/J)F1 × B10.T backcross population, the relative position of the Ir-5 locus has been estimated. This locus falls about 19 map units to the right of the H-2 complex.
A Rabbit with the Allotypic Phenotype: a1a2a3 b4b5b6
Authors: Strosberg A. Donny , Hamers-Casterman Cecile , Van Der Loo Wessel , Hamers Raymond
Published: December 2022

A rabbit of allotype a1a3, b4b5 was hyperimmunized with lyophilized Micrococcus lysodeikticus. Two weeks after the appearance of antibodies of restricted heterogeneity, the serum immunoglobulins of this animal reacted with anti-a1, anti-a2, anti-a3, and anti-b4, anti-b5, and anti-b6 allotype antisera. Control experiments showed that the reaction was specific for the allotypic determinants and that cross-reactions could be excluded. Isolated light chains were shown to contain the b4, b5, and b6 allotypic specificities. Serial bleedings indicated that all six allotypic specificities were present in most bleedings, but occasionally one or more of the allotypic specificities could not be detected.
Studies on the Induction and Expression of T Cell-Mediated Immunity. III. Amplification by Papain of the Generation of Cytotoxic Lymphocytes to Allogeneic Cells
Authors: Kedar Eli , Bonavida Benjamin
Published: December 2022

We have demonstrated in a recent study (1) that mild proteolytic treatment of immune cytotoxic lymphocytes (CL)2 leads to a 3- to 5-fold increase in the in vitro cytolytic activity upon target cells bearing the sensitizing antigens. In the present report we describe the potentiation of the generation of CL after treatment of normal lymphoid cells with low concentrations of papain. Thus, BALB/c (H-2d) splenocytes treated with papain and subsequently sensitized in vitro or in vivo against allogeneic C57BL/6 (H-2b) lymphoid cells exhibit an enhanced specific cytotoxic capacity as compared with untreated cells. Furthermore, the induction of CL with papain-treated cells occurs under conditions in which non-treated cells show little or no reactivity.

Materials and Methods. The preparation of spleen cells and enzymatic treatment with papain were described previously (1).
Modulation of Help and Suppression in a Hapten-Carrier System
Authors: Eardley Diane D. , Sercarz Eli E.
Published: December 2022

Provision of β-galactosidase (GZ) under defined conditions of dose and time can either help or suppress a subsequent response to trinitrophenyl (TNP)-GZ in CBA/J mice. The optimal helper effect occurs when 107 spleen cells from mice primed 9 or more days previously with 10 µg GZ are adoptively transferred to irradiated recipients which are then challenged with 10 µg TNP50GZ. Optimum suppression results from the transfer of spleen cells from mice primed 3 days previously with 100 µg GZ and challenge of recipients with TNP150GZ. Both help and suppression are carrier-specific and mediated by T cells. In experiments where helper or suppressor cells were mixed with normal cells, the anti-TNP response was proportional to the number of primed cells transferred.

The results point to a wave of suppression as the initial event after immunization, which is succeeded by period in which the helper effect dominates.
Experimental Allergic Orchitis and Aspermatogenesis
Authors: Carlo D. J. , Hagopian A. , Jackson J. J. , Limjuco G. A. , Eylar E. H.
Published: December 2022

Typical experimental allergic orchitis (EAO) and aspermatogenesis were successfully transferred to strain 13 guinea pigs with peritoneal exudate and lymph node cells from male and female donor guinea pigs (lacking detectable antibody) previously sensitized with 9 µg of highly purified GP1 glycoprotein isolated from the sperm acrosome. Attempts to transfer the disease with circulating antibody from hyperimmunized animals were not successful. These studies support a cell-mediated basis for the immunopathologic events in EAO.
Production of Large Amounts of Antibodies in Individual Mice
Authors: Tung Amar S. , Ju Shyr-Te , Sato Shigeru , Nisonoff Alfred
Published: December 2022

A method is described for the elicitation of substantial amounts of anti-hapten and anti-protein antibodies in ascitic fluids of individual mice of various strains. The method utilizes repeated i.p. inoculations of 0.5-mg quantities of antien in complete Freund's adjuvant. A high volume ratio of adjuvant to antigen solution, and the volume used per inoculation, were shown to be critical factors. Asctic fluids developed after three to five inoculations and were subsequently tapped repeatedly over a period of several weeks. All strains tested produced substantial amounts of anti-KLH antibodies, but marked differences were noted with respect to response to a hapten. Titers were enhanced in low responders by priming before the induction of ascites. The method is also useful for the production of “nonspecific” IgG and other serum components.
Protective Effects of Specific Antibodies in Trypanosoma Cruzi Infections
Authors: Krettli Antoniana Ursine , Brener Zigman
Published: December 2022

The action of immune sera from animals with chronic Chagas disease on Trypanosoma cruzi bloodstream forms (Y and CL strains) was investigated. Y strain trypomastigotes were agglutinated by anti-CL or anti-Y immune sera and their infectivity was decreased when inoculated into normal mice. CL strain trypomastigotes, however, remained apparently unchanged, with neither agglutination nor decline of infectivity being observed after incubation with both immune sera. Sera from three patients with chronic Chagas disease clearly agglutinated and two of them decreased the infectivity of Y strain trypomastigotes. Passive transfer of immunity was achieved in mice that received anti-CL or anti-Y immune sera and that were challenged with Y strain parasites. These results suggest that humoral immunity participates in the protective immune response against T. cruzi and that bloodstream trypomastigotes represent a more suitable model for studies of antigenic variation in these infections.
Role of Monosaccharides in the Interaction of Two Lymphocyte Mediators with Their Target Cells
Authors: Rocklin Ross E.
Published: December 2022

The role of monosaccharides on the interaction of two lymphocyte mediators related to the human cell migration inhibition systems with their respective target cells has been investigated. Human MIF activity on blood monocytes was significantly reduced by several 5-methylpentose sugars including l-fucose, l-rhamnose and 6 deoxy-d-glucose. Certain structurally unrelated monosaccharides such as d-galactose, methyl α-d-mannoside and N-acetyl-d-glucosamine had no effect on this system. Incubation of human monocytes with α-l-fucosidase, a glycosidase which cleaves terminal l-fucose moieties, reversibly prevented their response to MIF. These results indicate that α-l-fucosyl residues form an integral part of the receptor for MIF on human monocytes, and furthermore, this interaction may be somewhat nonspecific in that the 5-methylpentose configuration per se is recognized. In contrast, human LIF activity on PMN leukocytes was significantly reduced by N-acetyl-d-galactosamine, but not by 5-methylpentoses, d-glucose, N-acetyl-d-galactosamine, methyl α-d-mannoside or d-galactose. The latter finding suggests that a subterminal sugar may form part of the receptor to LIF on human PMN leukocytes.
Response to Antigenic Determinants of Neisseria Meningitidis Lipopolysaccharide Investigated with A New Radioactive Antigen-Binding Assay
Authors: Bertram Mary Ann , Griffiss J. McLeod , Broud Dennis D.
Published: December 2022

Adaptations of the Farr technique have resulted in a specific and reproducible radioactive antigen-binding assay for antibodies directed against the lipopolysaccharide (LPS) of Neisseria meningitidis. LPS was intrinsically labeled with 14C acetate during 16-hr growth in a modified Frantz media, extracted by hot phenol-H2O, and purified by dialysis, ultracentrifugation, and ethanol precipitation. LPS, which aggregates in aqueous solutions, was maintained in a monomeric form in 3% sodium deoxycholate (NaD) as determined by gel filtration on Sephadex G-75. Since NaD is insoluble in (NH4)2SO4, polyethylene glycol, 20%, was used to precipitate immunoglobulins of all three major classes.

Data obtained with the assay provide evidence that host responses to meningococcal LPS vary both quantitatively and qualitatively. Preliminary investigations of these differences by ultra high resolution chromatography indicate that the binding of unchromatographed or “native” LPS represents the summation of individual binding of antigenically distinct molecules, and that the response to these individual determinants within a given LPS may vary considerably from one host to another.
Carrier Determined Tolerance with Various Subclasses of Murine Myeloma IgG
Authors: Borel Yves , Golan Dov T. , Kilham Lynne , Borel Halina
Published: December 2022

Groups of BALB/c mice were treated with various conjugates of 2,4 dinitrophenyl (DNP) and BALB/c myeloma proteins belonging to the four subclasses of IgG (IgG1, IgG2, IgG2b, IgG3). Immediately thereafter, they were challenged with DNP-keyhole limpet hemocyanin in complete Freund's adjuvant and antibody to the hapten was measured by direct and indirect hemolytic plaque assay. The results show that all subclasses of IgG are effective as tolerance-inducing carriers. However, the ability to induce tolerance is dependent upon the concentration of hapten bound to each myeloma protein. Tolerogenic conjugates suppress both direct and indirect plaque-forming cells in all types of antibodies measured (IgG1, IgG2a, IgG2b, IgGa), whereas, non-tolerogenic conjugate failed to suppress them. The intact molecule of IgG but not its fragments (Fab, F(ab)2′, Fc) appear necessary as tolerance-inducing carriers. It is suggested that the ability to induce tolerance is related to the capacity of the tolerogenic conjugates to cause receptor blockade.
Heterologous Sera: A Target for in Vitro Cell-Mediated Cytotoxicity
Authors: Forni Guido , Green Ira
Published: December 2022

During the course of studies in inbred Strain 2 Guinea pigs on in vitro cell-mediated cytotoxicity to viral induced fibrosarcomas, it was observed that fetal calf serum in the medium in which the target cells were grown was responsible in some instances for much of the cytotoxicity observed. This fetal calf serum-specific cytotoxicity seemed mainly mediated by T cells and appeared to be independent on histocompatibility between target and killer cells.
The Development by Cytomegalovirus-Infected Cells of Binding Affinity for Normal Human Immunoglobulin
Authors: Westmoreland Diana , St. Jeor Stephen , Rapp Fred
Published: December 2022

After infection with human cytomegalovirus (CMV), cells develop an affinity for normal human immunoglobulin G (IgG). This was demonstrated using 125iodine-labeled purified IgG. It was further demonstrated that the immunoglobulin molecule binds to CMV-infected cells via its Fc portion, and competition for binding to infected cells occurred between purified preparations of human IgG and the Fc fragment of human IgG.

Whole sera from individuals with or without a high titer of anti-CMV antibody were labeled with 125iodine and it was demonstrated that serum from individuals with no anti-CMV antibody had an affinity for CMV-infected cells which probably reflected binding of IgG via its Fc fragment. The possible significance of these results in immunologic studies of human CMV is considered.
Neonatal Infection with Mouse Thymic Virus: Effects on Cells Regulating the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Morse Herbert C. , Cross Sue S. , Baker Phillip J.
Published: December 2022

Mice infected neonatally with mouse thymic virus (TA) were evaluated at different ages with respect to their ability to give a plaque-forming cell (PFC) response to type III pneumococcal polysaccharide (SSS-III), as well as the degree of amplifier and suppressor thymus-derived (T) cell activity present. B cell activity matured rapidly from 2 to 4 weeks of age and was not affected by TA infection. Amplifier T cell activity matured progressively over the first 8 weeks of life and was transiently suppressed in TA-infected mice at 4 weeks of age. Suppressor T cell activity measured at 2, 4, and 6 weeks of age was unaffected by TA. The findings suggest that TA is highly tropic for T cells and has selective effects on subpopulations of T cells.
Graft-vs-Host Reactions in F1 Mice Induced by Parental Lymphoid Cells: Nature of Recruited F1 Cells
Authors: Romano T. J. , Ponzio N. M. , Thorbecke G. J.
Published: December 2022

Graft versus host (GVH) reactivity of parental lymph node (LN) cells was assayed by measurements of 3H-thymidine incorporation in vivo. Mitomycin (Mit.) treatment of parental cells abolished their proliferative activity but the combination of such Mit.-treated parental cells with F1 LN cells resulted in much higher proliferation than either one population alone. This recruitment into proliferation of F1 cells was prominent on days 3 and 4 after cell injection and amounted to 35 to 51% of the total activity seen after injection of untreated parental cells alone. The F1 cell sensitive to recruitment was resistant to anti-Thy 1.2 treatment, was not removed by carbonyl iron-magnet separation; and was not present in thymus. The parental cell inducing recruitment was, however, sensitive to anti-Thy 1.2. When spleen cells from hapten immune F1 donors were injected together with Mit.-treated parental LN cells and boosted with hapten on another carrier, a typical “allogeneic effect” was observed in the anti-hapten immune response. It was concluded that Mit.-treated parental T cells exerted a mitogenic effect on F1 B cells resulting in extensive recruitment similar to that seen in murine mixed lymphocyte reactions.
The Inhibition of Cutaneous Basophil Hypersensitivity Reactions by a Heterologous Anti-Guinea Pig T Cell Serum
Authors: Stadecker Miguel J. , Leskowitz Sidney
Published: December 2022

Systemic treatment with a heterologous anti-T cell serum of guinea pigs immunized with EA in IFA markedly suppressed CBH reactivity to specific antigen and T cell mitogens, as judged by gross reactivity, histology, and skin histamine. The antiserum produced a marked drop in circulating lymphocytes, mainly at the expense of T cells, as indicated by the ability of surviving lymphocytes to rosette with rabbit RBC.

It was postulated that the suppression of CBH reactivity is due to the depletion of T cells, which would have released a factor chemotactic for basophils. The data therefore provide further evidence that cutaneous reactions rich in basophils are primarily dependent on a population of T cells.
Structural Features and Biologic Properties of Fragments Obtained by Limited Proteolysis of C3
Authors: Conroy Mary Carol , Ozols Juris , Lepow Irwin H.
Published: December 2022

Limited proteolysis of the third component of human complement (C3) was performed by using trypsin and streptococcal proteinase and the digests were analyzed for biologic activity. Incubation of C3 with trypsin for 1 min yielded a peptide with smooth muscle-contracting activity but no chemotactic activity, whereas the digest obtained after 15 min of incubation had only chemotactic activity. The conversion of muscle contracting to chemotactic activity could be correlated with the time course of trypsin hydrolysis. Hydrolysis of C3 with streptococcal proteinase gave a digest demonstrating only chemotactic activity. Each digest was resolved on a Sephadex G-100 column and the fractions containing biologic activities were characterized. The amino acid composition of the trypsin fragments, despite having different biologic activities, was remarkably similar, although differences in NH2-terminal amino acids were demonstrable. The fragments obtained by digestion of C3 with the streptococcal proteinase had an amino acid composition different from the trypsin fragments and displayed marked heterogeneity of NH2-terminal residues. These results suggest that slight alterations in the primary structure of C3 fragments may yield significant changes in their biologic activities and that the structural requirements for chemotactic activity are not confined to a single peptide species.
Suppression of Reaginic Antibody Formation
Authors: Lee Weng Y. , Sehon Alec H.
Published: December 2022

From the study of the effect of epitope density on the immunogenicity of haptenated ovalbumin (DNP-OA) it was concluded that the lightly haptenated conjugate, DNP0.5-OA, induced, on the one hand, only low titers of anti-DNP hemagglutinating antibody and no reaginic antibodies to the hapten and, on the other, high reaginic and high hemagglutinating antibody responses to the carrier. The conjugate with a slightly higher degree of haptenation, i.e., DNP2.3-OA, induced both reaginic and hemagglutinating antibodies to both the hapten and the carrier. By contrast, the heavily haptenated conjugate, DNP20-OA, elicited reaginic and hemagglutinating antibodies only against the hapten but not against the carrier.

Specific suppression of anti-hapten reaginic antibody formation had been achieved by treatment of mice with a tolerogen consisting of the hapten (DNP) conjugated covalently to isologous γ globulins (MγG). The epitope density of the DNPx-MγG conjugates was shown to play a dominant role in determining whether or not the conjugate was tolerogenic. Thus, lightly haptenated conjugates (DNP0.5-MγG, DNP1.3-MγG or DNP1.9-MγG) were not tolerogenic, moderately haptenated conjugates (DNP4.2-MγG, DNP8-MγG, and DNP14-MγG) were tolerogenic, and heavily haptenated conjugates (DNP32-MγG and DNP53-MγG) were immunogenic, being capable of priming the recipients for the DNP hapten.

Further evidence for the nonimmunogenicity of DNP8-MγG conjugate was inferred from its rate of clearance in tolerized and normal mice. Thus, the half-life of 125I-labeled DNP8-MγG in circulation was not significantly different for normal and tolerized mice; it was 3.7 and 3.5 days, respectively, which is within the range of data reported for clearance of normal MγG. These results suggest that DNP8-MγG was catabolized at a rate similar to that of nonconjugated, isologous MγG. Moreover, there was no significant difference in the localization of DNP8-MγG in identical organs (spleen, thymus, kidney, and liver) of normal and tolerized mice.

All the multivalent DNPx-MγG conjugates were shown to be able to elicit passive cutaneous anaphylaxis (PCA) reaction on i.v. Challenge of rats which had been pre-sensitized i.d. with anti-DNP reaginic antibodies.
Human Lymphocyte Subpopulations: Giant Srbc Rosettes
Authors: Yu David Tak Yan
Published: December 2022

Human thymus-derived lymphocytes have the ability to form rosettes with sheep red blood cells (SRBC) in vitro. In the investigation of rosettes of peripheral blood lymphocytes of 10 normal subjects, the number of SRBC adhering to the lymphocyte in each of 100 rosettes was assessed. The percentage of rosettes with SRBC ≥ 36 per rosette was only 1.2 ± 0.5. These were defined as giant SRBC rosettes. Peripheral blood lymphocytes were stimulated in vitro by four mitogens: sodium periodate, neuraminidase plus galactose oxidase, pokeweed mitogen, and concanavalin A. The lymphocytes were then cultured at 37°C. The giant rosette-forming lymphocytes became significantly increased 4 to 24 hr after stimulation, prior to the appearance of lymphoblasts or increased incorporation of tritiated thymidine. The giant rosettes were not caused by the hemagglutinating properties of pokeweed mitogen and concanavalin A that were adsorbed on the lymphocyte surfaces. This was shown by the fact that, on removal of the receptors by trypsinization, they were regenerated on culture in vitro in the absence of the mitogens. It was concluded that giant SRBC rosettes constituted a marker for some of the activated lymphocytes. Their appearance was independent of the increase in size of the cells or of DNA synthesis. These receptors were intrinsic to lymphocytes and not caused by mitogens adsorbed on their surfaces.
The Humoral Response of Mouse Spleen Cells to Two Types of Sheep Erythrocytes. IV. the in Vivo Response to H and L Srbc and the Genetic Control of the Production of IgM and IgG Antibodies in Discriminator Mice
Authors: McCarthy Margaret M. , Bowman Cecile , Butler Alison , Dutton Richard W.
Published: December 2022

It has been shown previously that the in vitro response of mouse spleen cells to two types of sheep erythrocytes (high-H SRBC; low- L SRBC)2 is under multi-gene control (1). We have concluded that there are common or shared antigen(s) (C) found on both types of SRBC and extra antigen(s) (E) unique to H SRBC. The responses of spleen cell suspensions from various strains of mice to H SRBC have been divided into three categories: non-discriminator (ND), low-discriminator (LD), and high-discriminator (HD) based on the ratios of the number of plaque-forming cells (PFC) seen with H and L SRBC when the response is assayed after 4 days of culture (1). Only LD and HD mice are able to respond to the extra antigens on H SRBC. The in vitro response of discriminator mice to the C antigens is normal in LD and low in HD mice. This concept is illustrated in Table I.
Activation of Complement (C) in Human or Animal Serum by Lymphoblastoid Cell Lines Derived from Patients with Burkitt's Lymphoma (BL) and Infectious Mononucleosis (IM)
Authors: Budzko Delia B. , Lachmann P. J. , McConnell I.
Published: December 2022

Several lymphoblastoid cell lines derived from patients with BL or IM were tested for C3b receptors by the rosette-forming technique using EAC3b. When the cells which possessed C3b receptors (Raji and IM) were incubated in normal human serum (NHS) the C receptor activity was inhibited. This was not found with other C3-rosetting cells such as peripheral blood lymphocytes from normal donors (PBL) or from patients with chronic lymphatic leukemia (CLL) or macroglobulinemia (MG). The blocking of the C receptor did not appear to be caused by native C3 since neither EDTA-plasma nor purified human C3 inhibited rosette formation (RF). Instead, inhibition was accomplished with C3b and with EDTA-plasma supplemented with Mg++ (EGTA-Mg-plasma).

Deposition of C3 on membranes, assayed by direct immunofluorescence using F(ab′)2 anti-human C3, was seen on all the cells tested except PBL, CLL, or MG cells, only after incubation with NHS, suggesting that positive cells activated C. Staining was also observed when the prior incubation was performed with EGTA-Mg-plasma but not when purified C3 or EDTA-plasma were used.
Biosynthesis of the Second Component of Complement by Human Monocytes
Authors: Einstein L. P. , Schneeberger E. E. , Colten H. R.
Published: December 2022

A method has been developed for preparation of confluent monolayers of human monocytes from small volumes of blood and for routine maintenance of the pure monocyte cultures for up to 16 weeks in vitro. These cells synthesized and secreted functionally active C2 at maximal rates of approximately 30 molecules/min/cell, after the first 6 days in culture. Occasionally, cultures were maintained for as long as 6.5 months, during which time they continued to produce C2. The rate of C2 production during the 1st week in culture was significantly less than that in cells sampled later, and this change in rate of synthesis was coincident with morphologic evidence of monocyte maturation (increase in cell size, appearance of multinucleated cells, loss of peroxidase-positive granules).

Incubation of monocytes in medium containing 14C-labeled amino acids resulted in apparent incorporation of label into C3 and C4 protein as well as C2. However, no hemolytically active C3 or C4 was detectable.
Subunit Structure and Properties of the Soluble Intermediate Complex C5b-7
Authors: Podack E. R. , Kolb W. P. , Müller-Eberhard H. J.
Published: December 2022

The C5b-9 complex contains, in addition to the five terminal complement components, an unidentified serum protein, termed band V protein. This protein is defined by its position on SDS polyacrylamide gel electrophoresis, a molecular weight of 88,000 and an unusually high carbohydrate content as revealed by periodic acid Schiff stain. To determine at which phase of the assembly of C5b-9 complex band V protein is incorporated, the subunit composition of soluble intermediate complexes was examined. The C5b-7 complex was formed by inulin activation of C8-deficient human serum and was isolated by gel filtration, Pevikon block electrophoresis, and CsC1 isopycinic density gradient ultracentrifugation. The complex has a molecular weight of 668,000 compared to 413,000, which is the sum of the molecular weights of C5b, C6, and C7. It reacts with antisera to C5, C6, and C7, but not with antisera to C8 and C9.
Demonstration of C9 at Complement Induced Membrane “Holes” by Immunoferritin Electron Microscopy
Authors: Rauterberg E. W. , Gebest H. J.
Published: December 2022

Complement-mediated lysis of sheep erythrocytes (SRBC) induces membrane alterations, which appear as electron lucent rings surrounding electron opaque condensations after negative staining in the electron microscope. The molecular structure and function of these “holes” or “lesions” are still unknown.

The present report describes a close association of the complement factor C9 with the “lesion.” SRBC were lysed by incubation with a fresh human serum containing an antibody against SRBC. After several washings in low ionic strength buffer, the ghosts were treated with an antiserum against human C9 produced in rabbits (Behringwerke, Germany). Subsequently the washed membranes were incubated with a ferritin-labeled goat anti-rabbit IgG antibody, mounted on carbon coated electron microscope grids with or without previous negative staining, and examined by direct transmission electron microscopy. The two-dimensional distribution of binding sites for anti-C9 antibodies was determined on unstained membrane preparations. The pattern of distribution was found to be identical with that of the “C-holes.”
The Initiating Factor of the Alternative Pathway and the Assembly of the Properdin (P) Receptor Forming Enzyme
Authors: Schreiber R. D. , Götze O. , Müller-Eberhard H. J.
Published: December 2022

A trace protein of normal human serum (NHS) has been detected by its reaction with antisera to nephritic factor (NF). It appears to be an essential early acting component of the alternative pathway. C4-depleted NHS adsorbed with solid phase anti-NF failed to sustain C3 consumption upon incubation with inulin or zymosan (Z) despite undiminished levels of P, B, and C3. Addition of NF, P̄, or cobra venom factor, however, caused consumption of C3 in the adsorbed serum. Addition of the glycine-HCl eluate (pH 2.2) of the immune adsorbent reconstituted the depleted serum. When C4 and P depleted serum (NHS s̄ C4,P) was incubated with Z for 60 min at 37°, 15 to 25% C3 consumption was consistently observed. The low magnitude C3 consumption was abolished by prior treatment of NHS s̄ C4,P with solid anti-NF. By use of various immunochemically depleted sera and fluoresceinated antisera to C3 and P̄, it was found that C3 could be deposited on Z from NHS s̄ C4,P, but not from serum depleted of B, D̄, or the NF analogue.
Stabilization of C3 and C3b by a New Serum Factor: A Mechanism for Regulation of Complement Action
Authors: Spitzer R. E. , Stitzel A. E. , Kalwinsky D. K. , Urmson J. R.
Published: December 2022

Studies on patients with renal transplant rejection have revealed the presence of a circulating factor which stabilizes C3 and C3b. Thus, serum containing this material, in a concentration of 10%, will prevent C3 consumption when CoF, preformed C3b, or zymosan is added to normal serum or EAC is mixed with EDTA normal serum. The responsible material has been isolated from a rejected graft by elution, G-150 column chromatography, and polyacrylamide gel separation. It is an α-globulin with an approximate m.w. of 60,000. When the isolated material is added to mixtures of preformed CoF- and C3, EAC and C3 or C3b- and C3, C3 consumption is prevented. The isolated material will also prevent the conversion of factor B as well as C3 when added to a mixture of zymosan and serum. If the purified material is added to preformed EAC cells and then removed, lysis of these cells occurs normally on the addition of C5–C9.
Histocompatibility Requirements for Cellular Cooperation in the Chicken: Generation of Germinal Centers
Authors: Toivanen Auli , Toivanen Paavo
Published: December 2022

Cyclophosphamide-treated newly hatched chicks were transplanted with histocompatible, semiallogeneic and allogeneic combinations of B (bursa) and T (thymus) cells from newly hatched donors. At the age of 5 weeks the birds were studied for an anti-SRBC response and for the generation of germinal centers in the spleen. The results of these experiments are summarized as follows. i) Allogeneic bursal stem cells have the capacity to restore the bursal structures of CY-treated recipients, but not the germinal center or anti-SRBC formation. ii) When allogeneic B cells are combined with T cells histocompatible or semiallogeneic with them, a restoration of the germinal center formation is achieved, but not to the same level as observed in normal birds or in CY-treated birds transplanted with histocompatible or semiallogeneic B cells. iii) Allogeneic B cells, even when complemented with T cells histocompatible with them, fail to restore the antibody production against SRBC; this is achieved only after transplantation of B cells histocompatible or semiallogeneic with the recipient. These findings indicate that germinal center formation is dependent on cooperation of histocompatible or semiallogeneic B and T cells, and furthermore, that an additional factor provided by the host is involved. Studies with transplantation of histocompatible and histoincompatible ‘empty’ splenic stromata revealed that the additional factor is not related to the splenic stroma.
Differential Effects of Alloimmunization on T Cells Mediating Graft-vs-Host Splenomegaly or the Allogeneic Effect in F1 Mice
Authors: Morse Herbert C. , Pasanen Vilho J. , Asofsky Richard
Published: December 2022

Spleen cells from normal BALB/c mice or mice immunized 10 or 30 days previously with C57BL/6 spleen cells were tested for a) their capacity to produce graft-vs-host (GVH) reactions in newborn F1 mice and b) their capacity to produce an allogeneic effect in adult F1 mice immunized with Type III pneumococcal polysaccharide. GVH reactivity of alloimmune spleen cells obtained 10 or 30 days after immunization was significantly increased as compared to the reactivity of normal spleen cells in that a) at comparable cell doses, higher spleen indices were obtained with alloimmune cells than normal cells, and b) alloimmune cells produced severe runting at lower cell doses than normal cells. By comparison, the capacity of alloimmune spleen cells to produce an allogeneic effect was reduced 50% on a per cell basis as compared to normal spleen cells at both 10 and 30 days after immunization. These results give further evidence that T cells producing the allogeneic effect are distinct from the T cell populations which interact to produce GVH splenomegaly.
The Direct Antiviral Cytotoxicity by Bovine Lymphocytes Is Not Restricted by Genetic Incompatibility of Lymphocytes and Target Cells
Authors: Rouse Barry T. , Babiuk Lorne A.
Published: December 2022

Experiments were designed to determine if in the ox a requirement for genetic compatibility between antiviral cytotoxic cells and target cells was needed for cytotoxicity to occur. Unrelated bovine animals were immunized with vaccinia or IBR virus (a herpesvirus), and PBL were collected at various times for measurement of their cytotoxicity against autologous or heterologous uninfected or virus-infected fibroblasts. In all instances, cytotoxicity was expressed against heterologous as well as autologous targets and in most cases there was no evidence of enhanced killing of autologous cells. The cytotoxicity was shown to be direct, presumably T cell mediated, and was not attributable to ADCC. The likelihood of the animals under investigation sharing histocompatibility antigens was considered extremely remote but was not formally excluded.
Human Transfer Factor: Fractionation by Electrofocusing and High Pressure, Reverse Phase Chromatography
Authors: Vandenbark Arthur A. , Burger Denis R. , Dreyer David L. , Daves G. Doyle , Vetto R. Mark
Published: December 2022

The biologically active fraction of human transfer factor (Sephadex G-25, fraction III) has been separated by exclusion chromatography into two component fractions (IIIa and IIIb). Fraction IIIa transferred donor reactivities to PPD or KLH into 9 of 9 recipients. Fraction IIIb did not transfer donor reactivities (0 of 8 transfers) but did produce both antigen-independent and antigen-dependent dermal reactions in recipients.

Fractionation by electrofocusing and high pressure reverse phase chromatography revealed that IIIa has a major UV254 absorbing component and that fraction IIIb has at least three major and three minor UV absorbing components. The major component of IIIa was isolated by preparative electrofocusing, had an isoelectric point of 1.6, and transferred KLH reactivity to four of five recipients.
In Vivo Suppression of the Primary Immune Response by a Species of Low Density Serum Lipoprotein
Authors: Curtiss Linda K. , DeHeer David H. , Edgington Thomas S.
Published: December 2022

An apparent subspecies of normal human serum low density lipoprotein (LDL-In) has been identified with suppressive activity for early or facilitating events of human lymphocyte mitogen and allogenic cell stimulation in vitro. This report describes the effects of in vivo administration of LDL-In on the mouse anti-SRBC immune response. Human LDL-In is not species specific and was capable of suppressing the in vivo mouse anti-sheep erythrocyte (SRBC) hemagglutination response by 88% after the administration of 500 to 600 µg LDL-In IV, whereas human serum high density lipoproteins and fibrinogen had no effect. Maximal suppression occurred only when LDL-In was injected 24 to 48 hr before antigen administration. Simultaneous or subsequent injection of LDL-In had no effect. The activity of LDL-In was influenced by antigen dose and maximal at low antigen doses. The number of splenic plaque-forming cells was also reduced indicating a suppression of the clonal expansion of primary B cells to antibody-secreting cells rather than only suppression of antibody synthesis by differentiated B cells and their progeny. These observations suggest the hypothesis that endogenous LDL-In could play an important immunoregulatory role in the maintenance of immune homeostasis and the “natural” suppression of nonproductive lymphocyte proliferation.
On the Biochemical Racial-Index of the Japanese in the Hokuriku District (Northern Part of Middle Japan)
Authors: Furuhata Tanemoto , Kishi Takayoshi
Published: December 2022

In 1919 L. and H. Hirschfeld (30) studied a number of soldiers belonging to the Allied Armies in Macedonia, and showed that agglutinogen A predominates greatly over agglutinogen B in the population of northern Europe, and that agglutinogen B predominates in Asia and Africa.

Finding a remarkable difference in the percentage distribution of the blood groups in the different races, they classified the races in three types examined on the basis of what they called the “Biochemical Racial-Index,” that is the ratio of the percentage of A to the percentage of B or , in the population.

Those races, having a racial-index of more than 2, were called of “European Type,” and those with an index less than 1, of “Asio-African Type,” and those from 2.0 to 1 of “Intermediate Type.” Since the work of Hirschfeld, a large amount of additional information has accumulated in several countries.
Partial Characterization and the Distribution of Chicken Mononuclear Cells Bearing the Fc Receptor
Authors: Duncan Robert L. , McArthur William P.
Published: December 2022

Erythrocyte-antibody (EA) complexes made with chicken antibody (IgY) and sheep erythrocytes (SE) were used to detect antigen-antibody complex-binding cells in the chicken. Nonadherent mononuclear cells, adherent mononuclear cells, and heterophils were found to have a subpopulation of EA complex-binding cells. The binding of the EA complexes was dependent on the Fc portion of IgY since F(ab′)2 anti SE-SE complexes did not form rosettes with chicken mononuclear cells. Cells binding EA complexes were therefore referred to as Fc+ cells.

The percentage of Fc+ cells in the spleen, bursa, and bone marrow varied considerably in birds of different ages. No Fc+ cells were detected in the thymus until birds were 8 weeks old. No significant difference was seen in the percentage of Fc+ cells detected in spleens of bursectomized agammaglobulinemic birds and normal birds the same age. Over 80% of the Fc+ cells in a normal adult spleen stained with a nonspecific esterase stain and less than 5% carried a thymic cell antigen. Most (95%) of the Fc+ cells from bursectomized birds stained with a nonspecific esterase stain.

Rabbit anti-SE (IgG)-SE complexes did not bind to chicken mononuclear cells and the converse, chicken antibody (IgY)-SE complexes did not bind to mouse or rabbit mononuclear cells. A population of chicken mononuclear spleen cells was also identified which had a receptor for activated C in the form of erythrocyte-antibody (IgM)-C (EAC) complex. As with Fc binding, the binding of components of activated C was also homocytotrophic.
Effect of Selective T Cell Priming on Anti-Sheep and Anti-Hapten Humoral Responses
Authors: Trizio Domenico , Cudkowicz Gustavo
Published: December 2022

Subimmunogenic numbers of sheep erythrocytes (2 × 106 SRBC) were injected into young adult (C57BL/6 × DBA/2)F1 and (C57BL/6 × C3H/He)F1 mice 4 days before the transfer of 2 or 1.25 × 106 spleen cells into irradiated syngeneic recipients. This procedure greatly enhanced the anti-sheep or anti-TNP antibody responsiveness of spleen cells to rechallenge with fully immunogenic numbers (5 × 108) of SRBC or TNP-conjugated SRBC, respectively. The anti-sheep response was accelerated, prolonged, and augmented in terms of the numbers of plaque-forming cells (PFC) generated in recipient spleens. Since the augmentation was considerably greater for IgG than for IgM PFC, the value of IgG:IgM ratios became >1. Low-dose priming caused the number of detectable antigen-sensitive units (ASU) to rise in the spleen: on day 4, the augmentation factors were 2.2 for IgM and 7.5 to 8.4 for IgG ASU, but the numbers of PFC generated by each unit remained the same.

Of the cell types constituting splenic ASU (T and B cells, macrophages), peripheral T lymphocytes were the most likely candidates for mediating the effect of priming. The reasons are: a) low doses of antigen preferentially activate T cells (11–15); b) low-dose carrier priming with SRBC augmented both the anti-sheep and the anti-S-TNP responses; c) addition of thymocytes to unprimed spleen cells moderately augmented anti-sheep responses; d) antigens cross-reactive with SRBC at the level of T but not of B cells were effective in priming for anti-S-TNP responses; e) Bordetella pertussis, an adjuvant requiring T cells, was also effective in this system. The possibility that low-dose priming induced quantitative as well as qualitative changes in the T subpopulations required for helper activity (helper and amplifier cells) was discussed.