bluuebunny/crossref_metadata_2025 huggingface dataset:
An in vitro Model of Lymphocyte Homing
Authors: Stamper Hugh B. , Woodruff Judith J.
Published: December 2022

An in vitro system was used to study lymphocyte interaction with high-endothelial venules (HEV) in rat lymph nodes. The results show that thoracic duct lymphocytes (TDL) are capable of selective adherence to the endothelium of HEV when overlaid onto glutaraldehyde-fixed sections of lymph nodes. At optimal TDL concentration (30 × 106/ml) and incubation temperature (7°C) binding occurs rapidly and the majority of HEV in each section usually shows adherent lymphocytes, the number attaching to each vessel being proportional to the endothelial area. TDL do not bind to other vascular structures in the node and ordinarily there is little adherence to other cells in the section. The results indicate therefore that TDL have surface membrane structures specific for HEV recognition.

Although TDL adhere to glutaraldehyde-fixed HEV, lymphocytes pretreated with fixative lose their capacity to bind. Also incubation performed at 1°C produces a fairly marked reduction in binding capacity of normal TDL. These observations indicate that endothelium of HEV are passive participants in the in vitro reaction whereas metabolic activity is important on the part of lymphocytes. In addition, when incubation is performed at 24°C, binding is severely reduced and at 37°C few, if any, HEV exhibit adherent TDL. It is suggested that lymphocytes that adhere to HEV are capable of rapid dissociation under physiologic conditions, a property that could facilitate entry of lymphocytes into the lymph node after their interaction with the specialized endothelium in vivo.
Mitogen-Stimulated Lymphoid Cells from Human Newborns Suppress the Proliferation of Maternal Lymphocytes across a Cell-Impermeable Membrane
Authors: Olding Lars B. , Murgita Robert A. , Wigzell Hans
Published: December 2022

The immunologic consequences of the interaction between lymphocytes from human newborns and their mothers were investigated in vitro. Maternal and male baby lymphocytes were cocultured in the presence or absence of PHA, and the resulting cellular proliferation was assessed by a) the fluorescent chromosome technique with the Y chromosome as a marker for dividing male baby cells, and by b) measuring the incorporation of 3H-thymidine in one- and two-way mixed lymphocyte cultures (MLC). In confirmation of previous reports, we found that approximately 90 to 100% of the dividing cells in two-way MLC were of baby origin, indicating that lymphocytes from human newborns effectively inhibit the proliferative capacity of maternal lymphocytes in 6-day MLC. The simultaneous measurement of 3H-thymidine uptake in the co-cultures revealed that the maternal cells with an inhibited proliferative capacity did cause a strong allogeneic stimulation in the newborn cell population. The suppressor activity associated with newborn lymphocytes was completely abrogated by irradiation with 6000 rads, showing a requirement for cell division in the in vitro development of the anti-proliferative effect on maternal cells. In contrast to the suppressive effect mediated by actively dividing newborn lymphocytes, irradiated cells from newborns exerted a strong stimulating effect on maternal lymphocyte proliferation in one-way MLC. Experiments using a double chamber culture system showed that mitogen-stimulated cells from newborns could effectively suppress the proliferation of lymphocytes from recently delivered mothers and nonpregnant adult females across a cell-impermeable membrane. PHA-stimulated lymphocytes from nonrelated adults did not, however, inhibit the division of maternal lymphocytes or lymphocytes from another nonpregnant woman when physically separated in double chamber cultures. Furthermore, mitogen-stimulated cells from one newborn failed to inhibit the proliferation of cells from another newborn. These findings suggest that proliferating newborn cells inhibit maternal lymphocyte proliferation by releasing a low m.w.-soluble suppressive factor(s). The characterization of the inhibitory substance released from mitogen- or alloantigen-activated human newborn suppressor cells is in progress.
BCG-Induced Murine Effector Cells
Authors: Tracey Daniel E. , Wolfe Seth A. , Durdik Jeannine M. , Henney Christopher S.
Published: December 2022

Peritoneal exudate cells harvested from mice after i.p. inoculation of 108 viable Bacillus Calmette-Guérin (BCG) organisms demonstrate vigorous lytic activity in vitro toward a large variety of target cells. Lytic activity was demonstrable within hours of BCG administration, was maximal after 4 to 5 days, and fell rapidly thereafter. Cell lysis observed in a 51Cr-release assay resulted from single events of collision with effector cells and was virtually complete within 4 hr. Effector cells were not found in the thymus or bone marrow, and the lytic activity of spleen and peripheral lymph nodes was only minimal. The effector cells were not induced by i.p. injection of thioglycollate medium or proteose peptone broth. Heat-killed BCG induced less activity than did viable BCG organisms.

The induction of lytic activity appeared to be under genetic control and to represent an amplification of a cell population present in the unstimulated animal. Some strains (CBA, B6C3F1) gave consistently high levels of of reactivity after BCG administration; others (A/J) showed much lower reactivity. Normal cells did not serve as targets for the BCG-induced effector cells, whereas tumor cells and long-term tissue culture cell lines were lysed. Susceptible targets included cells syngeneic, allogeneic, and xenogeneic to the effector cell source.

The rapid, vigorous lysis elicited by 4-day BCG-induced peritoneal exudates against a broad range of target cells indicates that the significance of this new cytotoxic phenomenon may be significant with respect to BCG immunotherapy.
Sensitivity of Amplifier T Cells Involved in the Antibody Response to Type III Pneumococcal Polysaccharide to Anti-Lymphocyte Serum
Authors: Markham Richard B. , Stashak Philip W. , Prescott Benjamin , Amsbaugh Diana F. , Baker Phillip J.
Published: December 2022

Amplifier T cells responsible for enhancement of the antibody response to type III pneumococcal polysaccharide have been shown to be resistant to the effects of antilymphocyte serum (ALS) given at the time of immunization, a treatment that eliminates suppressor T cell activity. The resistance of amplifier T cells to ALS can be attributed to the fact that their activity develops after that of suppressor T cells. ALS given 1 or 2 days after immunization does abrogate amplifier T cell activity, independent of the mode by which that activity is elicited. The data emphasize the importance of kinetic considerations in understanding the effects produced by immunologically active agents such as ALS.
Metabolic Requirements for Hormone-Induced Resistance to Antibody-Complement Mediated Killing of Tumor Cells
Authors: Schlager Seymour I. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

Line-1 guinea pig hepatoma cells are susceptible to killing by anti-Forssman IgM antibody plus guinea pig complement (GPC). When these tumor cells are incubated with insulin, epinephrine, hydrocortisone, or prednisolone, the cells show a marked reduction in their susceptibility to antibody-C-mediated killing. If the ability of the cells to synthesize DNA, RNA, and protein is impaired by pretreatment with metabolic inhibitors, x-irradiation, or culture in nutrient-deficient media, the hormones are no longer effective in rendering the cells resistant to killing. If only DNA synthesis is impaired, but not RNA and protein synthesis, the hormones are effective. The inability of cells inhibited in their macromolecular synthesis to be rendered resistant to killing after hormone treatment is not due to an inability of the cells to bind hormone.
Immunocycte Stimulation in Vitro by Nontoxic Bacterial Lipopolysaccharide Derivatives
Authors: Frank Steven , Specter Steven , Nowotny Alois , Friedman Herman
Published: December 2022

Intact lipopolysaccharides (LPS), considered nonspecific enhancers of B cell responses, as well as nontoxic derivatives from Serratia marcescens LPS, were studied with regard to their ability to stimulate in vitro immune responses to a T-dependent antigen, sheep erythrocytes. Intact LPS, at a dose of 10 to 50 µg, consistently enhanced the in vitro anti-SRBC immune response by normal splenocytes. The LPS also increased the background PFC response to SRBC in nonimmunized cultures. A chemically detoxified preparation derived from LPS (Mex B) had no stimulatory activity in vitro. A completely nontoxic, relatively small m.w., polysaccharide-rich preparation (PS), free of detectable lipid and protein, was stimulatory in vitro and at a dose of 10 µg resulted in a 40 to 70% enhancement of the anti-SRBC response. The PS also stimulated an enhanced background response to SRBC as well as several other RBC species in nonimmunized cultures. PS had no mitogenic effect in vitro since addition of this bacterial derivative failed to stimulate thymidine incorporation into mouse splenocytes, as occurred with the intact LPS. The use of nontoxic preparations from gram-negative bacterial LPS for dissecting the stimulatory vs antigenic properties of bacterial products provides a model system for determining the role of a mitogenic stimulus in B cell activation.
Macrophage Activation for Tumor Cytotoxicity: Induction of Tumoricidal Macrophages by Supernatants of PPD-Stimulated Bacillus Calmette-Guérin-Immune Spleen Cell Cultures
Authors: Ruco Luigi P. , Meltzer Monte S.
Published: December 2022

Resident peritoneal macrophages from normal mice were activated for tumor cytotoxicity in vitro by co-cultivation with BCG1-immune spleen cells and PPD and by incubation with supernatants of PPD-stimulated BCG-immune spleen cell cultures (lymphokine supernatants). Lymphokine activation of macrophages occurred in unfractionated PC suspensions as well as in macrophage monolayers depleted of nonadherent PC. Tumor cytotoxicity by lymphokine-activated macrophages was evident by 3 to 4 hr of culture in active supernatants, reached maximal levels by 8 to 12 hr, and was absent by 20 hr. Continued incubation in lymphokines or even re-exposure after washing did not maintain macrophage cytotoxicity. The capacity of normal resident macrophages to be activated by lymphokines in vitro progressively decreased and was absent by 20 hr in culture. This decrease did not necessarily reflect cell death; macrophage viability as estimated by exclusion of trypan blue or by phagocytic responses did not change over the 20-hr culture period. The short lived nature of both macrophage tumoricidal capacity and capacity of precursor cells to be activated by lymphokines may function as negative feedback mechanisms in immune reactions.
Immune Responses of Outbred Guinea Pigs to the Sequential Polymer Poly(L-Tyr-L-Glu-L-Ala-Gly): Association of Response to Strain 13 Histocompatibility Locus
Authors: Lai Chang-Hai , Maurer Paul H. , Shevach Ethan M.
Published: December 2022

Outbred Hartley guinea pigs were immunized with the sequential polymer poly(L-Tyr-L-Glu-L-Ala-Gly) (T-G-A-Gly)n, the random copolymer poly(Glu50, Tyr50) (GT), and a low dose of dinitrophenylated guinea pig albumin (DNP-GPA). Only the guinea pigs that carry strain 13 histocompatibility antigen responded to these three antigens as measured by the antigen-binding assay and delayed skin reaction. The immune response (Ir) genes controlling responses to (T-G-A-Gly)n, GT, or DNP-GPA, were found to be distinctive genes.

Nonresponder inbred strain 2 guinea pigs were able to produce anti(T-G-A-Gly)n antibodies when immunized with (T-G-A-Gly)n and methylated bovine serum albumin (MBSA) complex, although there was no delayed skin reaction and in vitro cultured lymphocytes did not respond.

The in vitro response to (T-G-A-Gly)n of cultured lymphocytes from responder animals was inhibited by strain 2 anti-strain 13 alloantisera but not the strain 13 anti-strain 2 alloantisera.
Structural Studies on Induced Antibodies with Defined Idiotypic Specificities
Authors: Capra J. Donald , Tung Amar S. , Nisonoff Alfred
Published: December 2022

The complete amino acid sequence of the variable regions of light chains derived from anti-p-azophenylarsonate antibodies from A/J mice bearing a cross-reactive idiotype is reported. At least two and probably more than three distinct light chains are associated with this idiotypically characterized antibody. The antibodies have several differences in their “framework” structures but evidence is presented indicating that all three light chain hypervariable regions have a homogeneous sequence. The data are discussed in relation to the various theories of antibody diversity. In addition, the findings support the view that hypervariable regions, idiotypic determinants, and the antibody-combining site involve, to a large extent, the same molecular structures.
In Vitro Immune Response of Human Peripheral Lymphocytes
Authors: Hirano Toshio , Kuritani Taro , Kishimoto Tadamitsu , Yamamura Yuichi
Published: December 2022

Human peripheral lymphocytes (PBL) upon stimulation with PWM proliferate and differentiate to IgM- and IgG-producing cells. The PWM-induced Ig production in B cells was dependent on T cells, and cell-free supernatant (CFS) obtained from PWM-stimulated PBL or T cell-rich fraction replaced T cell helper functions. The active substance(s) in CFS were most likely derived from T cells. The kinetic studies showed that the proliferation of B cells took place in advance of the final differentiation to Ig-producing cells and that T cells or T cell product(s) had to exist at the initiation of cultures in order to give the maximum helper effect. However, the final differentiation of B cells to Ig-producing cells was not dependent on T cells. The helper effect of T cells or T cell product(s) on PWM-induced proliferation and differentiation of B cells was exerted across the MHC barrier. This may make it possible to apply this experimental system to the assessment of quantitative and/or qualitative changes in human helper T cells in several immunologic diseases.
Partial Purification of Antigens from Eggs of Schistosoma Mansoni That Elicit Delayed Hypersensitivity
Authors: Brown Alan P. , Remold Heinz G. , Warren Kenneth S. , David John R.
Published: December 2022

Schistosoma mansoni egg antigens that elicit delayed hypersensitivity in appropriately sensitized guinea pigs were partially characterized by using ion exchange chromatography and preparative electrophoresis. At least three skin-reactive antigens were found, one of which was purified to homogeneity, as analyzed by polyacrylamide gel electrophoresis (PAGE). This antigen was not adsorbed to CM cellulose, migrated cathodal to guinea pig albumin on electro phoresis, and was adsorbed to DEAE cellulose. A second pool of antigenic activity was obtained by adsorption to CM cellulose and subsequent elution. DEAE cellulose chromatography and preparative electrophoresis of this pool indicated the presence of more than one antigen.
The Rate of Membrane Turnover by the Hairy Cells of Leukemic Reticuloendotheliosis: A Kinetic Study of a Receptor for IgM
Authors: Burns Gordon F. , Baker Christopher R. , Cawley John C. , Hayhoe Frank G. J.
Published: December 2022

The kinetics of turnover of a surface receptor for IgM were studied in six cases of hairy cell leukemia and four cases of chronic lymphocytic leukemia and the results were used as an indicator of membrane turnover in the pathologic cells of these two chronic leukemias. Rosette formation, shedding, and reexpression with EA (IgM) by hairy cells, a process which was shown to require protein synthesis, followed a cyclical pattern with a modal peak-to-peak reexpression time of 6 hr. Membrane turnover time in the cases of chronic lymphocytic leukemia was 12 hr or more. This work indicates that a reduced membrane turnover rate is not an essential feature of chronic leukemias.

The more general application of this type of approach to the study of membrane turnover in normal cells and their pathologic counterparts are discussed.
The Role of Antigen-Antibody Complexes in Mediating Immunologic Unresponsiveness in the Chicken
Authors: Morgan Edward L. , Tempelis Constantine H.
Published: December 2022

A significant suppression of an adoptive secondary response to either BSA or sheep red blood cells (SRBC) resulted when chicken spleen cells were incubated with BSA-antibody (Ab) complexes formed in antigen excess (molar ratio Ag1Ab1.1). Serum from 14-day old birds rendered unresponsive at hatching with large doses of BSA also produced suppression. In addition the in vitro response to SRBC was suppressed by the above treatment. Complexes formed at equivalence either had no effect on the subsequent responses of primed cells or produced an enhancement. The fact that both the homologous and heterologous responses were inhibited could indicate that both were suppressed via a common pathway.
On the Mechanism of Memberane Damage by C: Exposure of Hydrophobic Sites on Activated C Proteins
Authors: Shin Moon L. , Paznekas William A. , Abramovitz Aaron S. , Mayer Manfred M.
Published: December 2022

In previous papers we have presented evidence that peptides from C proteins C5b, C7, C8, and C9 become inserted in the lipid bilayer memberanes and form a transmembrane channel. Presumably, this insertion follows exposure of hydrophobic domains by C activation. In the present experiments liposomes were made with 14C-phosphatidyl choline (PC) and Forssman antigen in the bilayer, and with 86Rb+ in the aqueous compartments. When such liposomes were incubated with anti-Forssman antibody (A) and guinea pig serum (GPS) as a source of C, substantially more 14C-PC and 86Rb+ were released than from liposomes treated with A and C4-deficient GPS, or with A and heated C, or with C alone, or with A alone. The specific release of PC was dependent on the dose of C. Prior treatment of GPS with cobra venom factor abolished its capacity to release PC. The release of PC by A and C7-deficient human serum (C7D-HS) was the same as that of GPS alone, i.e., there was no specific release. A and C8D-HS produced much less specific release than A and GPS; addition of purified guinea pig C7 or C8 to C7D-HS or C8D-HS, respectively, restored the PC release to its full extent. Hence, part of the PC removal is mediated by C5b,6,7; the remainder is attributable to C8 and/or C9.
Induction of I Region-Restricted Hapten-Specific Cytotoxic T Lymphocytes
Authors: Wagner Hermann , Starzinski-Powitz Anna , Jung Hartmut , Röllinghoff Martin
Published: December 2022

Murine cytotoxic T lymphocytes (CTL) reactive to TNP-conjugated syngeneic target cells do lyse to a moderate but significant extent TNP-conjugated, I region compatible but H-2K or H-2D region incompatible target cells. Antibody inhibition experiments and “cold inhibition” experiments indicate that some CTL clones recognize TNP-conjugated targets in association with syngeneic I region determinants independently of H-2K or H-2D gene products. The data suggest that besides TNP-conjugated H-2K or H-2D gene products, in principle, also TNP-conjugated I region determinants do stimulate TNP-specific CTL precursor cells and act as target antigens of TNP-specific CTL.
Immune Responses during Measles Infection in Immunosuppressed Rhesus Monkeys
Authors: Hicks John T. , Sullivan John L. , Albrecht Paul
Published: December 2022

Rhesus monkeys immunosuppressed with horse anti-human thymocyte γ-globulin (ATG) were infected with measles and simultaneously inoculated with sheep erythrocytes (SRBC), a thymus-dependent antigen, and with pneumococcal polysaccaride type III (SSS-III), a thymus-independent antigen. ATG treatment alone suppressed SRBC antibody production, had no effect on SSS-III antibody production, and effectively eliminated circulating T cells compared to nonsuppressed monkeys. ATG treatment of measles-infected monkeys resulted in delayed virus clearance and delayed antibody production compared to nonsuppressed infected monkeys. After cessation of ATG treatment, measles antibodies and T cells reached normal levels, and measles virus was eliminated. Thus, immune clearance of measles virus is T cell-dependent, but the relative roles of cellular- and humoral-mediated immunity in vivo could not be clearly separated. Also, measles infection was associated with a decreased T cell mitogen responsiveness of circulating lymphocytes but not of lymph node lymphocytes, suggesting an altered circulating pattern of the cells responsible for delayed hypersensitivity. Also, measles infection had no effect on T-dependent antibody production to SRBC.
Activation of T and B Lymphocytes in Vitro
Authors: Armerding Dieter , Eshhar Zelig , Katz David H.
Published: December 2022

Biochemical and physicochemical analyses were made of allogeneic effect factor (AEF) prepared in either serum-containing or serum-free mixed lymphocyte cultures of alloantigen-activated T cells and the appropriate target cell population. Ion-exchange chromatography of Sephadex G-100 prepurified AEF on DEAE-cellulose indicated that the active molecules in AEF are relatively homogeneous and slightly negative in charge. Affinity chromatography of AEF on concanavalin A-Sepharose demonstrated that AEF contains glycoprotein, since the bulk of AEF activity was retained on such columns and could be recovered by elution with methyl-α-d-glucopyranoside. Analysis of AEF by dissociative chromatography in 6 M guanidinium-HCl demonstrated a subunit composition consisting of one heavy component of approximately 40,000 daltons and one light component of approximately 10,000 to 12,000 daltons associated noncovalently. Neither of these subunits, derived from dissociative chromatography of serum-containing AEF, alone was capable of reconstituting responses to sheep erythrocytes of T cell-depleted spleen cell cultures; however, addition of the two components simultaneously to such cultures resulted in recovery of full AEF activity. Dissociative chormatography of serum-free AEF yielded similar fractions, although the light component alone in such preparations exerted substantial AEF-like activity. Nevertheless, synergistic effects in terms of reconstituting activity were clearly observed when appropriate concentrations of heavy and light subunits from serum-free AEF were recombined in the same cultures.
Immunoregulation of Experimental Allergic Encephalomyelitis: Conditions for Induction of Suppressor Cells and Analysis of Mechanism
Authors: Swierkosz James E. , Swanborg Robert H.
Published: December 2022

We determined requirements for the induction of immunoregulatory suppressor cells in experimental allergic encephalomyelitis (EAE) in Lewis rats. Pretreatment of rats with myelin basic protein (BP) in incomplete Freund's adjuvant (IFA) stimulates the proliferation of suppressor cells that localize in lymph nodes and spleen (but not thymus) and exert control over the development of clinical EAE. Dosage studies revealed that 3 × 107 suppressor cells can adoptively transfer suppression to syngeneic recipients. Transferred unresponsiveness wanes within 3 weeks, indicating that the suppressor cells are short-lived lymphocytes, although actively induced unresponsiveness persists for at least 8 weeks, probably as a result of continual proliferation under the influence of antigen. No evidence was obtained to suggest that antigen carry-over or blocking antibody production accounts for adoptive transfer of unresponsiveness. Suppressor cells apparently act at the inductive phase of the immune response since they had no inhibitory effect on adoptive transfer of disease by effector lymph node cells.

Other mechanisms also may play a role in unresponsiveness to EAE, since rats pretreated i.v. with high dosages of soluble BP were temporarily rendered unresponsive, although suppressor cells could not be detected in these animals.
Opinions
Authors: Schreiner George F. , Unanue Emil R.
Published: December 2022

Consideration of membrane receptors and their modulation has assumed an increasing importance in our current understanding of the immune system. Basic cell mechanisms for establishing or altering membrane organization have a particular relevance for immunobiology with respect to the membrane-dependent interactions between cells or between a cell and soluble ligands. While our understanding of the organization of the cell membrane has come about from analysis of many cells, but most notably the red cell, our knowledge of the re-organization of the membrane upon interaction of a surface protein with ligands has derived to a great extent from the studies made on lymphocytes. One of the most fascinating and interesting of membrane reorganization phenomena concerns the sequence of membrane and cytoplasmic changes that ensue when B lymphocytes are exposed to antibodies to surface Immunoglobulin (Ig). As first observed by Taylor et al. (1), the lymphocyte rapidly and selectively redistributes its ligand-complexed antigen receptors to one pole of the cell, forming a cap.
Determination of e Antigen and Antibody to e by Means of Passive Hemagglutination Method
Authors: Takahashi Kazuaki , Fukuda Masako , Baba Kiyoshi , Imai Mitsunobu , Miyakawa Yuzo , Mayumi Makoto
Published: December 2022

A simple and sensitive method was developed for the detection of e antigen and antibody to e (anti-e) by means of the passive hemagglutination (PHA) method. Sheep erythrocytes were fixed by glutaraldehyde and coated with either e antigen or anti-e which had been specifically purified from plasma of asymptomatic carriers of hepatitis B antigen by affinity column chromatography, and then by gel filtration on Sephadex G-200. A gain in specific activity of more than 2000-fold for e antigen and 100-fold for anti-e was achieved as compared with the original plasma pool which was used as the source for purification. The PHA method was at least 300 times more sensitive than the conventional immunodiffusion method both for e antigen and anti-e assays. When the PHA method was applied for neat serum samples of 80 asymptomatic carriers, e antigen was found in 21.3% and anti-e in 62.5%, at rates much higher than those determined by immunodiffusion for the identical serum samples at 3-fold concentration (16.3 and 30.0%, respectively).
Mediation of CRP-Dependent Phagocytosis through Mouse Macrophage Fc-Receptors
Authors: Mortensen Richard F. , Duszkiewicz Jo Ann
Published: December 2022

The purified acute phase serum constituent, C-reactive protein (CRP), was examined for its ability to interact with mouse macrophage Fc-receptors when serving as an opsonin for C-polysaccharide (CPS)-coated erythrocytes (E.CPS). The ingestion of E.CPS-CRP by macrophage monolayers was dependent on the presence of CRP on the erythrocyte surface and on the treatment of the CRP-sensitized cells with human complement (C). The ingestion of E.CPS-CRP-C was inhibited by exposing the macrophages to either 2-deoxyglucose, a selective inhibitor of Fc-receptor activity, or to aggregated γ-globulin. Inhibition was also brought about by plating macrophages on glass-bound immune complexes of BSA/anti-BSA (IgG) but not on complexes formed with the F(ab′)2 fragment of the anti-BSA. Substrate-bound complexes of CPS-CRP selectively inhibited the uptake of IgG-coated E, EA(IgG); the inhibition was proportional to the concentration of CRP used to form the complex. The opsonin-independent ingestion of latex particles was not altered by these Fc-receptor blocking procedures. These findings coupled with the recent demonstration of a sequence homology between CRP and the CH3 domain of human IgG lead us to propose shared opsonic functions for antibody and CRP.
Subpopulations of Lymphocytes to Produce Various Lymphokines
Authors: Doroszczak Nadia , Yoshida Takeshi , Cohen Stanley
Published: December 2022

Identification of a subpopulation of lymphocytes producing lymphokines was attempted by fractionating the lymph node cells from guinea pigs immune to DNP-BSA by velocity sedimentation at 1 × G. Each of six fractions obtained by this procedure was cultured with or without the presence of antigen, and the culture supernatants that were separated 24 hr later were assayed for various lymphokine activities. Most of the lymphokines, including migration inhibition factor, chemotactic factor for neutrophils, mitogenic factor, and lymphotoxin were generated by the first two fractions of lymphocytes, which represented the largest, most rapidly sedimenting cells. Although the procedure of cell separation does not depend on cell surface properties, the larger cells contained more cells with T cell surface markers and the smaller contained more cells with B cell surface markers. Proliferative response of those lymphocytes measured by 3H-thymidine uptake, however, has shown that the largest two subpopulations responded poorly either to specific antigens or to mitogens (PHA and LPS), and rather that the medium size cells responded most strongly to the both stimulants. These results indicated that the production of some lymphokines confined to certain subpopulations of lymphocytes which are large in size. Further, these cells are readily separable from the medium sized cells that respond strongly to antigenic and mitogenic stimuli with mitogenic responses.
Ontogeny of B Lymphocyte Function
Authors: Sherr David , Szewczuk Myron R. , Siskind Gregory W.
Published: December 2022

Lethally irradiated mice reconstituted with adult T cells and neonatal or fetal B cells produce an anti-DNP response of restricted heterogeneity of affinity when compared with the response of mice reconstituted with T and B cells from adult donors. The capacity to reconstitute adult mice to give a heterogeneous response matures between 7 and 10 days after birth. The maturation of B cells from day-15 fetal or neonatal donors to produce a heterogeneous response was followed in the adult, cell transfer recipient by immunizing them at different times after cell transfer. It was found that B cells both from day-15 fetal mice and from neonatal mice acquire the capacity to produce a heterogeneous response within 3 days in the adult, cell transfer recipient. Thus, the B cell population matures more rapidly in the cell transfer recipient than in the intact donor. The kinetics of maturation in the adult recipient is the same for B cells from day-15 fetal and neonatal donors. The data imply that all information required to produce a fully heterogeneous response is already present in the day-15 fetus. In addition, the data strongly support the hypothesis that a factor in the adult mouse acts to induce this step in the maturation of the B lymphocyte population. Thus, the data seem to be inconsistent with the view that the timing of the occurrence of this differentiation event is precoded in an internal cell clock in the B lymphocyte line. Clearly, B cells from day-15 fetal mice are already capable of differentiating in response to the inducing factor which is present in the adult animal.
Comparative Studies on the Mechanisms of Nonspecific, Con A-Dependent Cytolysis and Specific T Cell-Mediated Cytolysis
Authors: Gately Maurice K. , Martz Eric
Published: December 2022

Several groups have shown that lymphocyte populations rich in previously differentiated, alloimmune cytolytic T cells nonspecifically lyse syngeneic tumor target cells in the presence of concanavalin A (Con A). We have carried out studies comparing the mechanism of this nonspecific, Con A-dependent cytolysis to that of specific T cell-mediated cytolysis of allogeneic targets.

Nonspecific, Con A-dependent cytolysis was experimentally resolved into three successive steps — adhesion formation, programming for lysis, and killer cell-independent lysis — by use of techniques previously developed for studying these steps in specific T cell-mediated cytolysis. The formation of nonspecific, Con A-dependent adhesions proceeded more slowly and was more temperature sensitive than specific adhesion formation. Mg++ in the absence of appreciable Ca++ was sufficient to support optimal adhesion formation in both systems. The kinetics of programming for lysis at 37°C were similar in the two systems; however nonspecific, Con A-dependent programming for lysis was approximately 2- to 3-fold more temperature sensitive than specific programming for lysis in the temperature range 37 to 22°C. Programming for lysis was strongly Ca++ dependent in both systems. Nonspecific programming for lysis occurred within 20 min after the application of Con A, suggesting that Con A causes the expression of a preformed cytolytic mechanism, and that no Con A-induced differentiation is necessary.

A number of known inhibitors of specific T cell-mediated cytolysis including cytochalasin B, DMSO, lidocaine, dibutyryl cyclic AMP, and colchicine were found also to inhibit nonspecific, Con A-dependent cytolysis. Thus, Con A does not bypass any steps inhibited by these agents. Antibody to target cell H-2 inhibited specific cytolysis but not nonspecific, Con A-dependent cytolysis. Overall, the results are compatible with the hypothesis that cytolysis in these two systems proceeds via similar mechanisms.
Chemotaxigenesis and Complement Fixation by Listeria Monocytogenes Cell Wall Fractions
Authors: Baker Lawrence A. , Campbell Priscilla A. , Hollister J. Roger
Published: December 2022

Data reported here show that a crude cell wall fraction of Listeria monocytogenes (LCWF) and its purified derivative (PF) induce chemotaxis of human polymorphonuclear leukocytes in the presence of normal rabbit serum. In addition, both LCWF and PF can fix rabbit and guinea pig complement. The purified cell wall preparation is more active than LCWF in both chemotaxigenesis and complement fixation, indicating that these activities are due primarily to the nonprotein portion of LCWF. Complement fixation by PF was demonstrated in the presence of ethylene glycol tetraacetic acid (EGTA) and in C4-deficient serum, which is evidence that PF activates complement by the alternative pathway. Therefore, chemotaxigenesis by PF and LCWF seems to involve fixation of complement via the alternative pathway.
Measurement of IgD by a Double Antibody Radioimmunoassay: Demonstration of an Apparent Trimodal Distribution of IgD Levels in Normal Human Sera
Authors: Dunnette Sandra L. , Gleich Gerald J. , Miller R. Drew , Kyle Robert A.
Published: December 2022

A double antibody radioimmunoassay (RIA) was developed to measure IgD in serum and secretions. One IgD myeloma protein was used as radiolabeled antigen and standard with antiserum to a second IgD myeloma protein. The IgD standard and normal sera yielded parallel inhibition curves in the RIA and inhibition was produced by IgD and not by any of the other immunoglobulins. The assay had a lower limit of sensitivity of 0.01 International Unit (I.U.)/ml and modifications increased the sensitivity to 0.0008 I.U./ml. Measurable IgD levels were found in all 112 normal adult sera assayed (geometric mean 13.0 I.U./ml, arithmetic mean 30.1 I.U./ml, median 14.8 I.U./ml, range 0.10 to 202 I.U./ml, 95% of values between 0.19 and 156 I.U./ml. The distribution of IgD in the 112 normal sera appeared trimodal with modes at approximately 0.25 I.U./ml, 5 I.U./ml, and 35 I.U./ml. IgD was measurable in nasal and bronchial washes and human milk, but could not be detected in parotid fluid.
Noncytotoxic IgE-Mediated Release of Histamine and Serotonin from Murine Mastocytoma Cells
Authors: Taurog Joel D. , Mendoza Guillermo R. , Hook William A. , Siraganian Reuben P. , Metzger Henry
Published: December 2022

Cultured murine mastocytoma (AB-CBF1-MCT-1) cells were stimulated to release endogenous or incorporated histamine or serotonin by an IgE-mediated mechanism without loss of viability. Stimulation was achieved by incubation of the cells with rat IgE—anti-IgE, rat IgE—anti-light chain, fluoresceinated rat IgE—anti-fluorescein, IgE-enriched mouse anti-ovalbumin-ovalbumin, or covalently linked dimers of rat IgE, at doses similar to those optimal for normal peritoneal mast cells. Active cell metabolism and Ca++ were required to obtain release. Despite the latter, no dose of the calcium ionophore, A23187, could be found which caused release without concomitant cytotoxicity. Phosphatidylserine did not enhance release.
Evanescent Delayed-Type Hypersensitivity: Mediation by Effector Cells with a Short Life Span
Authors: Askenase Philip W. , Hayden Betty , Gershon Richard K.
Published: December 2022

Four days after i.v. immunization of mice with optimal low doses of heterologous erythrocytes (2 × 105 RBC), strong delayed-type hypersensitivity (DTH) responses can be elicited in the footpad. At later intervals after immunization, DTH responsiveness is progressively diminished and replaced by 4-hr antibody-dependent reactions. These evanescent T cell-mediated DTH responses, which are progressively replaced by antibody-dependent reactions, resemble Jones-Mote type delayed hypersensitivity responses of humans and guinea pigs. Since higher doses of immunizing antigen activate suppressor mechanisms that inhibit DTH responses, we examined the possibility that the evanescence of DTH in mice immunized with an optimal low dose of antigen might also be due to suppression. Using techniques that could clearly demonstrate the suppression produced by high antigen doses, we failed to find evidence for either humoral or cellular suppression in optimally immunized mice with declining DTH responses. Thus, it appears that the evanescence of DTH produced by optimal low dose immunization with RBC may be due to an intrinsic short life span of the effector cells rather than to the activation of an identifiable shut-off mechanism.
Evidence for Adsorption of Heterospecific Host Immunoglobulin on the Tegument of Schistosoma Mansoni
Authors: Kemp Walter M. , Merritt Stephen C. , Bogucki Mary S. , Rosier James G. , Seed John R.
Published: December 2022

Mice infected with Schistosoma mansoni were immunized against human type B Rh-positive (B+) RBC, bovine serum albumin, or horseradish peroxidase. Adult parasites, recovered by perfusion, extensively washed, and incubated in their respective antigens, selectively bound to their tegumental surfaces only those antigens to which their murine host had been immunized. All controls supported the specificity of those reactions leading to the conclusion that adult S. mansoni in mice have the ability to adsorb heterospecific antibody onto their tegumental surfaces. These surface immunoglobulins were lost within 10 min when complexed with their antigens or within 2.5 hr when incubated at 37°C. Parasites that had lost their tegumental immunoglobulins regained them when incubated in normal mouse or rat anti-human type B Rh-negative (B-) RBC serum. Those parasites that had their surface immunoglobulins reconstituted with rat anti-human B- serum specifically bound human B- RBC, suggesting the possible presence of Fc receptors on adult S. mansoni.
Unusual Prevalence of Epstein-Barr Virus Early Antigen (EBV-EA) Antibodies in Ataxia Telangiectasia
Authors: Joncas Jean , Lapointe Normand , Gervais Francine , Leyritz Michele
Published: December 2022

EBV-EA antibodies are consistently present in the serum of patients with nasopharyngeal carcinoma (1, 2) (NPC)2 and Burkitt's lymphoma (3) (BL) at least in the late stages of the disease. In contrast, only 50 to 70% of patients with infectious mononucleosis (4–6) and less than 10% of normal children and adults have detectable serum EA antibodies (4–6). These antibodies in almost all cases of infectious mononucleosis fall to undetectable levels within a year or two after the disease and in patients with Burkitt's tumor disappear during remission but will rise again if a relapse occurs (4–7). It is of interest, therefore, that a high prevalence of Epstein-Barr virus early antigen (EBV-EA) antibodies was found in patients with ataxia telangiectasia (AT) in whom the incidence of lymphoma is increased (8).

Materials and Methods. Patients. All 16 patients had well documented AT with progressive cerebellar ataxia, oculo cutaneous telangiectases, and pedigrees consistent with autosomal recessive inheritance.
Immune Response to Phosphorylcholine
Authors: Köhler Heinz , Richardson Bruce C. , Rowley Donald A. , Smyk Sue
Published: December 2022

Anti-idiotypic antibody raised against the BALB/c myeloma proteins TEPC-15 or HOPC-8 inhibits induction of the immune response to phosphorylcholine (PC). (Fab′)2 and Fab′ fragments from anti-idiotype serum were prepared and anti-idiotypic antibody was purified by absorption and elution from idiotype or anti-IgG subclass immunoabsorbents. The parent material, the purified anti-idiotypic antibodies and the Fab′ fragments were assayed for: i) binding to the idiotype by using a sensitive radioimmunoassay, and ii) specific suppression of the response to PC in vitro. The correlation of binding to suppression was very similar for anti-idiotype serum and both kinds of purified antibody. However, the Fab′ fragments only bound to the idiotype and did not suppress. These findings indicate that the Fc portion of anti-idiotypic anti-receptor antibody (ARA) is essential for inducing suppression but that ARA-induced suppression is independent of the IgG isotype. It is postulated that idiotype suppression is mediated by the interaction of anti-idiotype with the antigen receptor and the Fc receptor thereby cross-linking both receptors.
Partial Purification of Human Lymphocyte-Activating Factor (LAF) by Ultrafiltration and Electrophoretic Techniques
Authors: Lachman Lawrence B. , Hacker Miles P. , Handschumacher Robert E.
Published: December 2022

Lymphocyte-activating factor (LAF) has been produced by culturing human peripheral blood leukocytes in the presence of lipopolysaccharide and autologous human serum. The major LAF activity, identifiable by Sephadex chromatography (m.w. 13,000) was separated from most serum proteins in the culture medium by ultrafiltration with a hollow fiber device. Sucrose gradient isoelectric focusing of the concentrated ultrafiltrate yielded a single peak of LAF activity with an average isoelectric point of pH 6.8. Semi-preparative polyacrylamide gel electrophoresis of the isoelectric focusing sample resulted in separation of the LAF activity from detectable amounts of serum proteins. The recovered LAF activity was estimated to be purified more than 16,000-fold and is judged to be active in submicrogram amounts.
Biologic and Biochemical Activities of Continuous Macrophage Cell Lines P388D1 and J774.1
Authors: Snyderman Ralph , Pike Marilyn C. , Fischer Dina G. , Koren Hillel S.
Published: December 2022

The study of continuous lines of murine cells with macrophage-like properties may provide important information concerning biochemical parameters associated with particular types of macrophage functions. Chemotaxis, phagocytosis, adherence, and antibody-dependent cellular cytotoxicity (ADCC), as well as the activities of several intracellular enzymes, were measured in two such cell lines, P388D1 and J774.1. These measurements were compared to those of “resting” and “stimulated” murine peritoneal macrophages. The data show that the P388D1 and J774.1 cell lines, although displaying many biologic characteristics of normal stimulated or resting macrophages, differ quantitatively from each other and from the normal or stimulated cells. Interestingly enough, the P388D1 cells, although the most cytotoxic in ADCC, displayed virtually no chemotactic responsiveness in vitro. Although the J774.1 cells responded as effectively to the chemotactic stimulus C5a as did the resting peritoneal macrophages, they did not migrate toward lymphocyte-derived chemotactic factor (LDCF). Both cell lines contained levels of acid phosphatase comparable to stimulated peritoneal macrophages; however, the P388D1 cells were markedly deficient in adenosine deaminase when compared to the peritoneal macrophages or to the J774.1 cells.

These studies demonstrate the heterogeneity of biologic activities displayed by various macrophage preparations and illustrate the potential usefulness of continuous cell lines in determining biochemical parameters associated with specific macrophage functions.
Suppressor Cell Activation by Anti-Thy 1.2 in NZB/WF1 Mice
Authors: Klassen Lynell W. , Zarate Alfred , Gelfand Michael C. , Steinberg Alfred D.
Published: December 2022

A single injection of anti-Thy 1.2 serum was used to study the spontaneous loss of suppressor cells in NZB/WF1 mice. When given 1 day before primary allografting with C57BL/6 skin, anti-Thy 1.2 significantly prolonged mean graft survival (MGS) in 5-week-old NZB/NZWF1 graft recipients. However, four times the dose of anti-Thy 1.2 was required to cause graft prolongation when compared with BALB/c recipients. In contrast to its graft-prolonging effect in 5-week NZB/WF1 mice, anti-Thy 1.2 failed to prolong MGS in either 17- or 34-week-old NZB/WF1 animals. Spleen cells from anti-Thy 1.2-treated 5-week-old NZB/WF1 mice could transfer suppression of graft rejection to otherwise untreated 5-week-old syngeneic recipients. This transfer required viable, radioresistant T cells. Although spleen cells from anti-Thy 1.2-treated 5-week-old mice transferred the suppression to 5-week-old recipients, the same cells were not effective if given to 17-week-old recipients. However, spleen cells from 17-week-old-pretreated donor mice were able to prolong MGS when given to the younger 5-week-old syngeneic recipients. The transfer studies of skin allograft prolongation by anti-Thy 1.2-treated spleen cells suggest that the ability to initiate suppressor cell activation is present in both young and older NZB/WF1 mice, whereas suppressor effector mechanisms are deficient in the older mice. In addition to its usefulness in studying the details of suppressor cell action, the studies suggest that anti-Thy 1.2 treatment may be a valuable probe for the presence of suppressor cells and/or their precursors.
Humoral Primary Immune Response in Vitro in a Homologous Mouse System: Replacement of Fetal Calf Serum by a 2-Mercaptoethanol or Macrophage-Activated Fraction of Mouse Serum
Authors: Opitz Hans G. , Opitz Uta , Lemke Hilmar , Flad Hans-D. , Hewlett Guy , Schlumberger H. D.
Published: December 2022

The primary immune response in mouse spleen cell cultures against heterologous red cell antigens is dependent on the medium being supplemented with selected batches of fetal calf serum. Mouse serum itself is not able to support this response. The active immune response-supporting component in fetal calf serum seems to be a distinct factor (s), which has been partially purified by Sephadex G-100 filtration and termed MaSF — 2-mercaptoethanol-activated serum factor. In this report it is demonstrated that MaSF is also present in mouse serum. For functional detection, mouse MaSF has to be separated from higher m.w. inhibitors, and has to be activated by 2-ME. After separation and activation mouse MaSF can support the primary immune response in a completely homologous in vitro culture system. Evidence is presented that MaSF can also be activated by macrophages. It is concluded that macrophages and 2-ME have the same mode of action in the primary immune response in vitro, i.e., induction of lymphocyte competence by activation of a serum factor.
Lymphocyte Cytotoxicity to Influenza Virus-Infected Cells
Authors: Greenberg Stephen B. , Criswell B. Sue , Six Howard R. , Couch Robert B.
Published: December 2022

Peripheral blood lymphocytes (PBL) obtained from humans were cytotoxic for influenza virus-infected target cells. The PBL were shown to have associated influenza virus anti-hemagglutinin antibody (AHAb) detectable only by radioimmunoassay. This antibody could be removed by incubating PBL at 37°C for 30 min. The lymphocyte population that was effective in this system was nonadherent and nonphagocytic cells. PBL gave comparable levels of cytotoxicity when tested by using either a xenogeneic or allogeneic virus-infected target cell.

These results indicate that lymphocyte cytotoxicity to influenza virus-infected cells may be mediated by small quantities of antibody and by lymphocytes that possess characteristics of K cells. No evidence for T cell-mediated cytolysis was found with this xenogeneic system.