bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional Heterogeneity in Macrophages Activated by Corynebacterium Parvum: Characterization of Subpopulations with Different Activities in Promoting Immune Responses and Suppressing Tumor Cell Growth
Authors: Lee Kwok-Choy , Berry David
Published: December 2022

Peritoneal cells (PEC) from mice injected i.p. with heat-killed Corynebacterium parvum (CP) showed enhanced immunostimulatory (accessory or A cell) activity as measured by their ability to restore the immune responsiveness of nonadherent spleen cells to sheep erythrocytes (SRBC) and polymeric flagellin (POL) of Salmonella adelaide in vitro. This was true whether the PEC and nonadherent spleen cells were in direct contact or separated by a cell-impermeable membrane which allowed the free passage of soluble mediators. CP-activated PEC also exhibited greatly increased cytostatic activity against the growth of syngeneic tumor cells in vitro. After fractionation of the PEC according to cell size by velocity sedimentation, a separation of A cell activity from anti-tumor activity was observed. Although both these functions were associated with phagocytic cells of the monocyte-macrophage series, the highest A cell activity was found in fractions containing small and medium-sized macrophages, whereas the anti-tumor activity increased with cell size to a maximum with the largest macrophages. Thus, there is a relative increase of suppressive activity over stimulatory activity with an increase in cell size. Cytochemical and morphologic evidence suggests that the A cell-rich fractions contained small and medium-sized macrophages which were derived from newly arrived monocytes, whereas the large tumor-suppressive macrophages were relatively more differentiated.
Tumor Growth and Antibodies after RSV-Challenge in Normal Chickens and in Chickens Congenitally Infected with Avian Leukosis Virus
Authors: Meyers Paul , Qualtiere Louis F.
Published: December 2022

Normal chickens and chickens congenitally infected with an avian leukosis virus (ALV) of antigenic subgroup A were challenged with strains of Rous sarcoma virus (RSV) of two different antigenic subgroups (B and C) and tumor induction and growth as well as humoral antibody to viral envelope antigen (VEA) and tumor-specific surface antigen (TSSA) were measured. There was no effect of congenital ALV infection on RSV tumor incidence or latent period but the growth rate and size of the tumors were much higher in congenitally infected birds as compared to controls. Whereas most tumors in the RSV-challenged normal birds regressed, tumors in ALV-infected birds grew progressively. There were no striking differences in the number of birds in either group in the incidence of anti-TSSA or anti-VEA antibodies nor did the presence of either type of antibody reflect the tumor status of the host.
Accumulation of Platelets at Sites of Antigen-Antibody-Mediated Injury: A Possible Role for IgE Antibody and Mast Cells
Authors: Kravis Thomas C. , Henson Peter M.
Published: December 2022

Circulating 51Cr-labeled platelets accumulate at skin sites in which a reversed passive Arthus reaction has been induced. The accumulation is biphasic in time and is accompanied by an increased vascular permeability. Increased permeability itself, however, will not produce localization of platelets. A similar platelet accumulation was observed upon injection of compound 48/80 or anti-IgE antibody into the skin and this was not altered in rabbits depleted of complement or neutrophils. Activation of skin mast cells and release of a platelet-activating factor (PAF) is suggested as a mechanism for the effect produced by anti-IgE and compound 48/80. The first phase of platelet accumulation in the Arthus reaction was also unaffected in rabbits depleted of enutrophils or complement, which may suggest a role for IgE antibody and mast cells. The second phase of accumulation was diminished in complement-depleted animals and abrogated in rabbits without neutrophils, suggesting a complement and neutrophil-mediated process but which still might be mediated through mast cell activation by neutrophil cationic protein.
Evidence That Con A Induces Cytotoxicity in the Same Subclass of T Cells as Does Alloimmunization
Authors: Falkoff Reuben Maverick , Dutton Richard W.
Published: December 2022

We have studied the induction of cytotoxic activity in murine T cells by the T cell mitogen Con A. Here we report the results of experiments that indicate that this cytotoxicity develops in the same class of T cells potentially activatable to cytotoxicity by immunization with allogeneic cells. Cytotoxic activity does not result from activation of cells of the T-helper class by PHA, and extensive reduction of the proportion of cells of the T-helper class by in vivo treatment with ATS does not comparably reduce the cytotoxicity developed in response to Con A. The Con A-activated cytotoxic cell sediments as a large cell. Furthermore, spleen cell populations previously immunized to alloantigens in vivo develop greatly increased cytotoxicity specific for the alloantigen of the immunizing haplotype after culture with Con A in vitro.
Characterization of the Nonphagocytic Adherent Cell from the Peritoneal Cavity of Normal and BCG-Treated Mice
Authors: Nathan Carl F. , Asofsky Richard , Terry William D.
Published: December 2022

A distinctive subpopulation of nonphagocytic, tightly adherent cells (NPAC) comprised approximately 6% of the adherent peritoneal cells from untreated mice, and about 18% of those from mice previously given BCG i.p. A separation procedure based on adherence and lack of phagocytosis was devised. Isolated NPAC were morphologically intermediate between small lymphocytes and macrophages. They were positive for nonspecific esterase, negative for peroxidase, positive for surface IgM, and negative for surface IgG1, IgG2 and IgA. When capped, their surface IgM regenerated in vitro. NPAC had demonstrable Fc receptors but not EAC receptors. They resisted killing by an anti-macrophage serum, were negative by immunofluorescence with an anti-T cell reagent, and incorporated increased amounts of thymidine in response to LPS but not to PHA. They were more readily killed with anti-Ia serum and complement than macrophages, but less readily than splenic B cells. NPAC appeared to represent a subpopulation of B lymphocytes which contaminates some preparations previously regarded as “macrophages” and which may be responsible for some of the activities previously ascribed to “macrophages”.
Effect of Concanavalin a on the Killing of Tumor Cells by Antibody and Complement
Authors: Boyle Michael D. P. , Langone John J. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

Concanavalin A (Con A) was found to inhibit the killing of antibody-sensitized line-1 tumor cells (TA) by guinea pig complement (GPC) but not by human complement (HuC). Other plant lectins (wheat germ, leucoagglutinin, and pokeweed mitogen) were also tested but Con A was the only lectin found to inhibit antibody-GPC-mediated killing. The inhibitory effect of Con A was observed when the GPC was mixed with Con A or when the antibody-sensitized cells were pretreated with Con A (TA-Con A) before the addition of GPC. The effect could be reversed by treatment of such cells with α-d-methylglucopyranoside or by incubation at 37°C for approximately 2 hr. Con A appeared to act by preventing the binding of the first component of GPC (GPC1) to antibody-sensitized tumor cells. Differences in the binding of the first component of HuC (HuC1) and GPC1 to TA-Con A suggested that a difference in the binding site for HuC1 and GPC1 might exist. There was no difference in the number of GPC1 molecules fixed to antibody-sensitized sheep erythrocytes (EA) or EA treated with Con A in experiments using the same antibody as used with the tumor cells and the same Con A preparation. It would consequently appear that the inhibitory effect of Con A on the binding of GPC1 to TA is not due solely to an interaction of Con A with the antibody.
In Vitro Induction of Polyclonal Killer T Cells with 2-Mercaptoethanol and the Essential Role of Macrophages in This Process
Authors: Igarashi Tsuyoshi , Okada Masaji , Kishimoto Tadamitsu , Yamamura Yuichi
Published: December 2022

Killer T cells against allogeneic and syngeneic tumor cells were generated in vitro by the addition of 2-mercaptoethanol (2-ME) to the murine spleen cell culture in the absence of any antigenic stimulation. The maximum activity of T cell-mediated cytotoxicity (CMC) induced with 2-ME was observed on day 4 of culture and the induction of CMC was completely inhibited by the addition of inhibitor of DNA synthesis, hydroxyurea, or cytosin arabinoside. CMC induced with 2-ME was specifically inhibited by the addition of unlabeled target cells to the 51Cr-release assay system. These results indicated that killer T cells were generated in the presence of 2-ME as a result of nonspecific polyclonal activation of precursors into cytotoxic effector cells and that they recognized target cells with antigen-specific recognition receptors. Spleen cells deprived of adherent cells showed impaired induction of CMC with 2-ME. The addition of peritoneal exudate macrophages to splenic T cells restored this response. The result indicated that macrophages were essential for the induction of CMC with 2-ME. The possibility that the function of macrophages was mediated by soluble factor(s) released from macrophages was demonstrated by the separate culture of splenic T cells and macrophages in double-chambered, Marbrook-type vessels and by the addition of supernatants from macrophage cultures to splenic T cells. 2-ME and soluble factor(s) released from macrophages seemed to be required for the activation of precursors into killer cells.
Expression of Alloantigens Ly-5 and Ly-6 on Cytotoxic Effector Cells
Authors: Woody James N. , Feldmann Marc , Beverley Peter C. L. , McKenzie Ian F. C.
Published: December 2022

With anti-Ly antisera and complement it has been possible to demonstrate that cytotoxic effector cells generated in vitro against allogeneic cells carry the alloantigens Ly-5 and Ly-6. The studies show that antisera directed against the Ly-6 antigens, together with complement, eliminate essentially all of the killer cells, of the appropriate strain, while killing only 50 to 60% of Thy-1+ cells. Anti-Ly-5 antisera and complement lysed 55 to 65% of Thy-1+ cells and killed a significant portion, but not all, of the cytotoxic cells. The later finding was investigated in more detail since it suggested a degree of heterogeneity within the killer cell subpopulation. However, the data did not support that conclusion, at least for allogeneic killer cells.
Variability of the Lytic Susceptibility of Neisseria Gonorrhoeae to Human Sera
Authors: Tramont Edmund C. , Sadoff Jerald C. , Wilson Clarence
Published: December 2022

The bactericidal activity of human sera for Neisseria gonorrhoeae was studied. Sera were obtained from a group of patients with gonococcal infections who had acute urethritis, acute pelvic inflammatory disease, disseminated gonococcal infection, or who were asymptomatic carriers. The homologous and heterologous strains were tested with these sera. The development of serum bactericidal antibodies was not a universal event. With few exceptions, the susceptibility of a particular strain to human antibody and complement appeared to be largely independent of the particular person from whom the serum was obtained and was due instead to antigenic properties intrinsic to each individual strain. Lipopolysaccharide appeared to be the predominant antigen against which bactericidal antibodies were directed. The principal bactericidal antibody class was IgM. Blocking antibodies were not found to account for the lack of lytic activity. A correlation of bactericidal antibodies with protection from developing gonococcal infection could not be demonstrated in three patients.
IgM-Induced Cell-Mediated Cytotoxicity with Antibody and Effector Cells of Human Origin
Authors: Fuson E. W. , Lamon E. W.
Published: December 2022

In vitro lysis of antibody-sensitized target cells by lymphocytes has been demonstrated in a number of systems (1–6). IgG has been implicated as the major antibody class involved in antibody-dependent cell-mediated cytotoxicity (ADCC)3 with human effector cells (1) and IgM had not been previously reported to be active with one recent exception (3).

In contrast, independent investigations in three antigenic systems have demonstrated murine IgM to be active in ADCC with homologous effector cells (4–6). It should be pointed out that Wåhlin et al. (3) reported IgM- and IgG-induced cytotoxicity by using a sensitive plaque assay with rabbit antibody and human effector cells.4 Most investigations with human effector cells have been with heterologous antibodies for sensitization of the target cells. In order to evaluate the ability of human IgM to induce ADCC in an homologous system, we have used human heterophile antibody, which is entirely IgM (7) to sensitize 51Cr-labeled sheep erythrocytes (SRBC).
Induction of Phenotypic Lymphocyte Differentiation in LPS Unresponsive Mice by an LPS-Induced Serum Factor and by Lipid A-Associated Protein
Authors: Koenig Scott , Hoffmann Michael K. , Thomas Lewis
Published: December 2022

Spleen cells from C3H/HeJ mice are known to be unresponsive to mitogenic stimulation by LPS. We show here that T and B cell precursors of C3H/HeJ mice are unresponsive to induction of differentiation by LPS. Phenotypic differentiation of C3H/HeJ lymphocytes can be induced with DB-cAMP, Lipid A-associated protein, and with a serum factor induced by LPS in LPS responder strains.
Glucocorticoid Receptors in Purified Subpopulations of Human Peripheral Blood Lymphocytes
Authors: Lippman Marc , Barr Ronald
Published: December 2022

On the premise that the differential effects of glucocorticoids on various aspects of the immune response may be mediated by differences in the glucocorticoid receptors in the effector cells, subpopulations of human peripheral blood lymphocytes were examined for these receptors as well as for glucocorticoid responsiveness. Purified T and non-T lymphocytes, when studied by a sensitive whole cell assay technique, contained equivalent amounts of specific glucocorticoid receptor, which, by binding affinity and specificity measurements, were indistinguishable from each other. Furthermore, under in vitro incubation conditions, macromolecular synthesis in both of these cell populations was inhibited by glucocorticoid at concentrations which saturated the receptor sites. It is concluded that the putative differential effects of glucocorticoids on T and non-T lymphocyte-associated functions are probably not mediated by differences in the glucocorticoid receptors in these cell populations.
A Functional Comparison of Human Fc-Receptor-Bearing Lymphocytes Active in Natural Cytotoxicity and Antibody-Dependent Cellular Cytotoxicity
Authors: Kay H. David , Bonnard Guy D. , West William H. , Herberman Ronald B.
Published: December 2022

Natural (or “spontaneous”) cytotoxicity against the K-562 myeloid cell line and antibody-dependent cellular cytotoxicity (ADCC) against sensitized Chang liver cells were monitored simultaneously in a 4 hr 51Cr release assay, using the same preparations of normal human peripheral blood lymphocytes as effector cells. Both types of cytotoxic activity were expressed by lymphocytes bearing receptors for sheep erythrocytes and for the Fc portion of IgG. Up to 80% of the total natural killer (NK) cell and K cell (ADCC) activities were recovered in E-rosette pellets separated under optimal conditions. Since the ability to form rosettes with sheep erythrocytes is primarily a marker for human T cells, it seems likely that both NK and K cells are in the T cell lineage. Further, both NK and K cell cytolytic activities were severely depressed after brief exposure of lymphocyte suspensions or rosette pellets to ammonium chloride solutions. Both activities returned to normal values after 24-hr incubation at 24°C. Such sensitivity of human lymphocytes to ammonium chloride solutions may explain why other investigators have failed to observe NK or ADCC activity by E-RFC.

When lymphocytes bearing Fc receptors were adsorbed onto plastic surfaces coated with a monolayer of immobilized antigen-antibody complexes made up of TNP and rabbit anti-TNP serum, both NK and ADCC activities were simultaneously depleted. Such depletion was completely blocked when Staphylococcal protein A, which binds selectively to the Fc portion of IgG, was incubated on the antigen-antibody monolayers before adding the lymphocytes. This confirmed that both NK and ADCC activities were mediated by effector lymphocytes bearing Fc receptors.

When protein A at concentrations ranging from 0.01 to 100 µg/ml was added to the 51Cr release assays, ADCC was significantly inhibited, presumably due to binding of protein A to available Fc sites of the sensitizing antibody. However, protein A had no effect on NK reactivity at any of the concentrations tested. When lymphocytes were treated for 30 min at 37°C with purified trypsin or α-chymotrypsin at concentrations from 0.01 to 10 mg/ml, NK activity was significantly inhibited whereas ADCC was unaffected. Other enzymes depressed (pronase, subtilisin) had no effect except at high concentrations (lipase), or stimulated (neuraminidase) both NK and K cell activity similarly, and therefore failed to distinguish between the effector cell types.

Thus, our data indicate that effector cells in NK and ADCC are in overlapping, if not identical, subpopulations of Fc receptor-bearing lymphocytes of probable T cell lineage. The cytolytic mechanisms in NK and ADCC appear to be distinct, since we have been unable to find evidence for an “arming” antibody involved in this NK system.
Suppressor Cells in Tolerance to Hgg: Kinetics and Cross-Suppression in High Dose Tolerance—Absence in Low Dose Tolerance
Authors: Benjamin D. C.
Published: December 2022

Spleen cells from mice made tolerant with high doses of human γ-globulin (HGG) specifically suppress the immune response of normal, syngeneic, spleen cells. These suppressor cells were found to be cross-reactive in that they would suppress the immune response of normal spleen cells to bovine γ-globulin (BGG) as well as to HGG. In contrast, suppressor cells could not be demonstrated in spleens of mice made tolerant with low doses of HGG (i.e., T-cell tolerance), nor could they be found in high dose tolerant mice following a second injection of DHGG at a time when the initial suppressor activity had waned. The role of suppressor cells in the induction, maintenance, and loss of tolerance is discussed.
Kinetic Studies on the Fragmentation of the Third Component of Complement (C3) by Trypsin
Authors: Minta Joe O. , Man Doreen , Movat Henry Z.
Published: December 2022

The kinetics of cleavage of C3 by trypsin was analyzed by electrophoresis in agarose and in polyacrylamide gels containing sodium dodecyl sulfate and the data obtained were used to construct an anatomical model for C3 showing the sites of tryptic attack, the fragments generated, and their composition.

Trypsin was shown to cleave C3 in a stepwise fashion. The attack was initially directed at the α-polypeptide chain and resulted in the generation of C3a and C3b. Further cleavage of the α-chain of C3b, converted it into C3bi and then into C3d and C3c. Cleavage of the β-chain by trypsin occurred only at the C3c stage with the release of a small peptide (m.w. 12,000) from C3c and the formation of C3c′. On immunoelectrophoresis, C3c′ had a less anodal mobility compared to the β1A mobility of C3c.

C3a, once formed could be further cleaved to give residual fragments with decreasing net positive charge. Exposure of C3 to acid conditions, pH 5.0 or below, rendered the molecule exceedingly susceptible to tryptic degradation.
Sequential Quantitation of Circulating Immune Complexes in Syngeneic and Allogeneic Rats Bearing Moloney Sarcomas
Authors: Jennette J. Charles , Feldman Joseph D.
Published: December 2022

A Raji cell radioimmunoassay was employed to quantitate serially circulating immune complexes (CIC) in the sera of syngeneic BN rats and allogeneic Lewis rats bearing BN Moloney sarcomas. In syngeneic BN hosts the levels of CIC attained and the time-course of detection were related to the tumor dose, tumor mass, and regressive or progressive course of the tumor. In general, syngeneic rats that received larger tumor doses developed larger tumors and greater maximum levels of CIC. However, the amount of CIC was not always directly proportional to the tumor size, although this was nearly the case with regressor BN and Lewis rats. In rats with regressing tumors, CIC decreased to insignificant levels as the tumors disappeared. Progressor BN rats that received 20 and 10 × 106 tumor cells had higher and more sustained levels of CIC, but, shortly before the hosts died, despite an increase of tumor size, there was a decline of CIC. Progressor BN rats that received an initial inoculum of 0.5 × 106 tumor cells that grew to 44 mm maximum mean diameter had levels of CIC which were only slightly above levels of control rats.

All allogeneic Lewis hosts rejected BN Moloney sarcomas, but had transient low levels of CIC coincident with tumor growth. Lewis rats had lower levels of CIC than BN rats bearing comparable masses of sarcoma.
Antigenic Studies of a VH Fragment: Demonstration of Three Sets of Antigens, Idiotypic, VH Subgroup, and VH Framework-Specific Antigens
Authors: Michaelsen Terje E. , Førre Øystein , Natvig Jacob B.
Published: December 2022

The antigenic properties of the VH region of immunoglobulin heavy chains were studied by means of a fragment corresponding to the variable part of the heavy chain of an IgG3 myeloma protein (KUP) and an antiserum made against this fragment. By hemagglutination, hemagglutination inhibition, and immunofluorescence techniques, it was shown that the anti-VH antiserum detected three sets of antigens in the VH region, namely idiotypic antigens, VH subgroup-specific antigens, and VH domain-(framework) specific antigens. The VH fragment inhibited in a VHII subgroup-specific hemagglutination inhibition test system. The VH fragment was thus antigenically similar to the VH region found in the intact molecules and the light chains were not needed to express the VH subgroup antigens or the VH framework antigens.
Splenic Influence of T Cell Function: The Immunobiology of the Inbred Hereditarily Asplenic Mouse
Authors: Fletcher Mark P. , Ikeda Richard M. , Gershwin M. Eric
Published: December 2022

The influence of the spleen on the maturation of T cell function was investigated by using hereditarily asplenic mice inbred on a B6.CBA background. Asplenic (Dh/+), in contrast to +/+ littermate control mice, manifest a major qualitative deficiency of T cell function as well as a quantitative reduction in serum immunoglobulins. In particular, Dh/+ thymocytes are not as efficient as +/+ thymocytes in synergizing with either Dh/+ or +/+ bone marrow cells in the primary response to SRBC in irradiated littermate +/+ controls. This latter defect in T cell-bone arrow synergism was markedly age dependent and was evident only in young mice. Similarly, the DNA synthetic response of Dh/+ thymocytes injected into lethally irradiated BALB/c mice was quantitatively diminished in comparison to +/+ thymocytes when spleen and lymph nodes were quantitated. Spleen cell-seeking patterns of Dh/+ thymocytes were reduced compared to +/+ control cells when injected into +/+ littermates. Surface marker and mitogen responsiveness of lymph node cells revealed an increase in ϑ-bearing and decrease in Ig-bearing cells in Dh/+ mice compared to +/+ controls. The mortality and morbidity of Dh/+ mice, when compared to +/+ mice, after the perinatal period were identical. However, Dh/+ mice were more susceptible than +/+ controls. This latter defect in T cell-bone marrow synergism cogenesis. Cell reconstitution studies and serial age studies in mice receiving either MSV or polyoma virus confirmed the above observations regarding an age-dependent qualitative defect in T cell function. It is concluded that the spleen significantly effects the rate of murine T cell maturation.
Human Monocyte-Lymphocyte Interaction: A New Technique
Authors: Fan Peng Thim , Yu David T. Y. , Pearson Carl M. , Bluestone Rodney
Published: December 2022

A rosette-type assay of the physical interaction between lymphocytes and monocytes after treatment with neuraminidase-galactose oxidase (NGAO) is reported. Monocyte-lymphocyte (ML) rosette formation and subsequent lymphocyte proliferation occurred when either lymphocytes or homologous monocytes were treated with NGAO and cultured together. Maximal ML rosette formation took place at 37°C 4 hr after culture in media containing 10% serum at lymphocyte to monocyte ratios of 10:1 to 20:1. The percentage of rosette formation correlated with the extent of thymidine incorporation when increasing concentrations of NGAO were used. When NGAO-treated monocytes were added to untreated T and non-T lymphocytes, they bound preferentially to T lymphocytes and induced proliferation only in the T subpopulation. These results indicate that the ML rosette assay measures a highly specific monocyte-lymphocyte physical interaction after a mitogenic stimulus which is an early event in lymphocyte activation since it reflects the degree of subsequent lymphocyte proliferation.
Erythrocyte Membrane-Associated Immunoglobulins during Malaria Infection of Mice
Authors: Lustig Hannah J. , Nussenzweig Victor , Nussenzweig Ruth S.
Published: December 2022

Immunoglobulin (Ig) is associated with erythrocyte membranes during infection of A/J mice with Plasmodium berghei. It was detected by agglutination of the cells with rabbit antibodies to mouse IgG and by a radioimmunoassay with 125I-labeled rabbit anbtibodies to mouse IgG. As shown by the degree of agglutination and of binding of radiolabeled antibodies to the erythrocyte membranes, the amount of Ig increased with time after infection and paralleled parasitemia and reticulocytosis.

The erythrocyte-associated immunoglobulins are mainly IgG but IgM was also present on the cells of some mice. A large proportion of the Ig could be eluted at 37°C and was analyzed by immunoprecipitation and acrylamide gel electrophoresis.

A radioautographic study with 125I-labeled anti-mouse IgG revealed that both parasitized and nonparasitized reticulocytes of infected mice had much larger amounts of membrane-bound immunoglobulin than did mature, nonparasitized erythrocytes.

The nature of the bonds between the Ig and the surface membrane of the reticulocytes is not known. The Ig could be part of immune complexes nonspecifically bound to the cell surface. However, since we have not detected Fc or C3d receptors on reticulocytes, it is possible that the Ig constitutes autoantibodies against reticulocytes or antibodies against parasitic antigens present on the cell membrane.
Enhancement of Weak Mixed Lymphocyte-Type Reactions by Hydrophilic Polymers
Authors: Ben-Sasson Shmuel A. , Henkart Pierre A.
Published: December 2022

Two hydrophilic polymers, dextran and polyethylene glycol, were found to enhance in vitro MLR-type reactions which are weak or nonexistent in normal culture media. BALB/c and C57BL/6 thymocytes were cultured for 4 days in Eagle's MEM + 10% FCS, with or without the x-irradiated lymphomas LSTRA and EL-4, respectively. By thymidine incorporation no stimulation was observed with EL-4 in plain culture whereas occasionally a very small stimulation was observed with LSTRA. Four percent dextran (w/v, m.w. ≥ 40,000) improves stimulation (E/C) by about 2-fold. The effect of polyethyleneglycol m.w. 6,000 (PEG-6) is much more pronounced. At the optimal concentration of 4 to 5%, PEG-6 causes an increase of 5- to 20-fold in the stimulation index. The PEG-6 effect can also be observed microscopically by the appearance of numerous blasts clustered around the tumor cells. PEG-6 by itself has little effect on thymocytes in the absence of tumor cells. In the case of one-way allogeneic MLR under suboptimal ratios, dextrans (m.w. ≥ 40,000) enhance the reaction dramatically. We speculate that these effects are occurring via a change in the solvent (such as solvent exclusion) in a way which enhances immunologic interactions under conditions which are tolerable to cells in vitro.
Neutrophil Aggregation and Swelling Induced by Chemotactic Agents
Authors: O'Flaherty J. T. , Kreutzer D. L. , Ward P. A.
Published: December 2022

An assay is described for measuring neutrophil aggregation and cell size with a Coulter Counter equipped with a mean cell volume computer and volume channalyzer. With this assay, it was found that several chemotactically active agents rapidly induce rabbit peritoneal polymorphonuclear neutrophils in suspension to aggregate reversibly. The aggregating activities of these chemotactic agents paralleled their in vitro chemotactic- and in vivo neutropenia-inducing potencies. Neutrophil aggregation may underlie the neutropenia which follows the infusion of these agents into rabbits.

Chemotactic agents may also induce polymorphonuclear neutrophils to increase in volume; this effect appears to be independent of aggregation. The volume change develops more slowly and does not appear reversible under the conditions employed. The volume increase may be an expression of pseudopodia formation and membrane ruffling known to develop in neutrophils exposed to chemotactic agents.
Immunologic Tolerance to Heterologous Immunoglobulin: Its Relation to In Vitro Filtration by Macrophages
Authors: Cowing Carol , Harris Dorothy , Garabedian Charles , Lukic Miodrag , Leskowitz Sidney
Published: December 2022

The cellular and molecular basis for the difference in ability of BGG to induce tolerance in BALB/c and DBA/2 mice has been examined by in vitro biofiltration. It was found that incubation with the adherent cells from BALB/c but not DBA/2 spleens could remove the material from BGG which inhibited tolerance induction in BALB/c mice. This material was shown to represent only a trace component in BGG, was present in only certain commercial batches of BGG, and was apparently unrelated to the presence of aggregates or endotoxin.
Studies of Tolerance to DNA in NZB/NZW F1 Mice
Authors: Tanphaichitr Kongsak , Hahn Bevra H.
Published: December 2022

Administration of sDNA-poly-D-lysine (DNA-PDL) to newborn NZB/NZW F1 mice (B/W) was previously shown to prolong survival and to decrease nephritis and DNA antibodies. In this study, B/W mice treated from birth with DNA-PDL were found to be tolerant to immunization with sDNA on PDL or mBSA carriers in adjuvants. Tolerance to sDNA was present and was hapten-specific carrier-dependent. IgG and IgM anti-nDNA circulating antibodies were suppressed. Continuous tolerization was necessary to maintain tolerance. Tolerance to sDNA could be transferred by spleen cells, by tolerized thymus cells, and by tolerized bone marrow cells, suggesting that both T and B cells participated in this phenomenon.
Enhancement of the DNA Synthetic Response of Antigen-Primed Lymph Node Cells by Splenic Promoter Cells: Characterization of the Splenic Promoter Cells
Authors: Shek Pang N.
Published: December 2022

Rabbits were immunized in the footpad with diphtheria toxoid in complete Freund's adjuvant. At various times after immunization, cells harvested from the spleen, the draining (immune) and the opposing (control) lymph nodes (LN) were assayed for their proliferative response to incorporate tritiated thymidine upon exposure to the priming antigen. Although the immune LN cells responded by a substantial incorporation of thymidine, cells from the control LN and the spleen either did not respond or responded feebly. An enhancement in the response of the immune LN cells was observed when they were cultured in the presence of nonresponsive spleen cells. Pretreatment of spleen cells with mitomycin C did not abolish the enhancement. This suggests that the target cells which respond to the antigen are derived from the immune LN whereas the promoter cells which enhance the response are present in the spleen. The removal of adhering cells by glass wool columns and of Ig-bearing cells by anti-Ig immunoabsorbent columns from the spleen did not reduce the enhancing capacity of the nonadhering cells. Conversely, the killing of splenic T cells by specific heterologous antiserum directed against rabbit thymus lymphocyte antigen abolished the enahncement. Thus, the promoter cell which facilitates the enhancement has been characterized as a nonadherent, splenic T cell.
Variation in DNA and RNA Synthetic Responses during Activation of Lymphocytes from Inbred Strains of Mice
Authors: Buckley Patrick J. , Wedner H. James
Published: December 2022

Previous studies have suggested that interstrain variations in the mitogenic response of murine lymphocytes to concanavalin A (Con A), phytohemagglutinin (PHA) and bacterial lipopolysaccharide (LPS) are genetically controlled. DNA and RNA synthesis in splenic lymphocytes from six inbred strains of mice were examined in response to PHA, Con A, and LPS along with other measures of lymphocyte activation. Marked variations in nucleic acid synthesis in mitogen-stimulated lymphocytes from these strains were noted when assessed by the incorporation of radiolabeled precursors into DNA or RNA. Cells from PL/J incorporated two to four times more radiolabel into DNA and RNA in response to PHA and Con A than cells from the other strains whereas cells from CBA/J, AKR/J and C57BL/6J exhibited the highest response to LPS. Variation in optimum mitogen concentration, time of peak response in culture, proportion of B and T lymphocytes, extent of the S-phase of the cell cycle, or differences in mitogen receptor affinity and number could not account for these experimental observations. There was, however, no correlation between the increases in nucleic acid synthesis and the actual number of activated lymphocytes when mitotic indices, lymphoblast numbers or number of S-phase cells were measured. Little strain variation in the number of lymphocytes activated in response to each mitogen was observed with the exception of the low response of C3H/HeJ to LPS. These studies suggest that some of the genetic variation in the incorporation of labeled precursors into nucleic acid does not reflect the number of lymphocytes activated. Measurement of DNA and RNA synthetic responses alone may not be an accurate assessment of the activation process.
Effects of α-Fetoprotein on Murine Immune Responses
Authors: Sheppard H. W. , Sell S. , Trefts P. , Bahu R.
Published: December 2022

Because of the current interest in the possible “immunosuppressive” effects of α-fetoprotein a systematic study of the effect of normal sera, amniotic fluid (AmF), sera from hepatoma-bearing animals (Tu sera), and AFP isolated by immunoabsorption from AmF or Tu sera on various models of the immune response of the mouse was carried out. Whereas some in vitro reactions were inhibited by AFP, other in vitro systems were only weakly inhibited, unaffected, or even enhanced by the addition of AFP. AFP by itself was mitogenic for murine lymphoid cells, inhibited the LPS mitogenic response 3-fold, but did not affect the response to Con A. Very high concentrations of AFP partially inhibited primary in vitro plaque-forming cell (PFC) responses to sheep red blood cells (SRBC). The secondary in vitro PFC response to keyhole limpet hemocyanin (KLH) conjugated to trinitrophenol (TNP) was inhibited 2- to 4-fold. The induction of cytotoxic effector cells directed against P815 mastocytoma cells was increased 9-fold in the presence of AFP. There was no significant difference between the effect of AmFAFP and TuAFP on any of the in vitro reactions studied. Normal and tumor sera produced a similar reduction in secondary PFC responses in spite of a 6000-fold difference in AFP concentration. The in vivo PFC response to SRBC in mice bearing a transplantable hepatoma was greater than that of normal mice even though the serum AFP concentration of the tumor-bearing mice was 1000 times normal. In addition, no AFP synthesis or coating of lymphocytes was observed during the course of a graft-vs-host or mixed lymphocyte reactions. Therefore, although some effects on immune phenomena occur upon addition of serum proteins and AFP, these data do not support a general immunosuppressive role for AFP in the mouse.
Antibodies from Healthy Cats Exposed to Feline Leukemia Virus Lyse Feline Lymphoma Cells Slowly with Cat Complement
Authors: Grant Chris K. , DeBoer D. J. , Essex M. , Worley M. B. , Higgins Joanne
Published: December 2022

Antisera from cats naturally exposed and resistant to feline leukemia virus lysed feline lymphoma cells in vitro with cat complement. Lymphoma cell lysis was detected by 51Cr release and by viable cell counts with phase-contrast microscopy. The rate of target cell lysis varied according to the nature of the antiserum and the origin of the complement used. Cat alloantisera, raised by immunization with target cells, lysed lymphoma cells rapidly with cat complement, but immune sera from cats naturally infected with feline leukemia virus caused slow lysis requiring up to 20 hr to reach maximum effect. Slow lysis of lymphoma cells in homologous serum mixtures was complement dependent, specific for leukemia virus-infected cells, and was found to occur in cat-immune whole serum. When 143 cat sera were examined for presence or absence of lytic antibody by 51Cr release and for antibody to feline oncornavirus-associated cell membrane antigen by immunofluorescence, the qualitative correlation obtained varied according to the complement used in the 51Cr release assay. With heterologous absorbed complement sources the correlation was 79% with guinea pig and 65% with rabbit; with homologous complement the correlation was 92%. The close association seen between high antibody levels and tumor resistance in this outbred species under natural conditions serves to emphasize the potential significance of these observations.
Studies on Receptors for IgG on Epithelial Cells of the Rat Intestine
Authors: Borthistle Bruce K. , Kubo Ralph T. , Brown William R. , Grey Howard M.
Published: December 2022

It has been previously shown that immunoglobulins of the IgG class are absorbed and transported across the proximal small intestine of neonatal rats during the first 18 to 21 days of life. In the present work, we examined the binding of mouse immunoglobulins to the intestinal epithelium of neonatal rats. Radioiodinated mouse immunoglobulins of various classes and IgG subclasses, Fab and Fc fragments of IgG, and monomeric and heat-aggregated forms of IgG were instilled into loops of intestine prepared from rats of various ages. The specificity of the radiolabeled immunoglobulin binding was evaluated by competitive inhibition with unlabeled proteins. Monomeric IgG bound specifically to the apical regions of villi in the proximal jejunum. Specific binding progressively decreased from the 12th day of life and could not be demonstrated after about the 21st day of life. Mouse IgA and IgM did not bind specifically. Radioiodinated Fc fragments of IgG bound specifically, whereas radioiodinated Fab fragments did not. Treatment of 14-day-old rats with cortisone acetate caused binding of monomeric IgG to cease prematurely by day 16. Intestinal loops treated with trypsin no longer bound radioiodinated IgG.

Heat-aggregated IgG also bound specifically to jejunal epithelium. In contrast to the binding of monomeric immunoglobulin, this binding continued into adulthood and increased in response to cortisone treatment. Aggregated immunoglobulin did not inhibit the binding of monomeric immunoglobulin, but did inhibit the binding of aggregates.

These studies: 1) confirm the presence of receptors for the Fc region of monomeric IgG on absorptive epithelial cells of the rat jejunum; 2) demonstrate that the receptors disappear at the time when immunoglobulin transport is known to cease, and therefore appear to be critically involved in transport of monomeric IgG; 3) demonstrate that the receptors for monomeric IgG are sensitive to trypsin and disappear prematurely after cortisone treatment; 4) demonstrate specific binding of aggregated IgG, perhaps by receptor mechanisms different from those involved in the binding of monomeric IgG.
Similarity of the Carbohydrate Structures of H-2 and Ia Glycoproteins
Authors: Freed John H. , Nathenson Stanley G.
Published: December 2022

The glycopeptides produced by pronase digestion of two H-2K, two H-2D, and three Ia glycoprotein antigens were examined for size and charge. The glycopeptides derived from all of the antigens examined were found to have m.w. of 3250 ± 200 daltons with a similar and variable composition of sialic acid residues. These data, when combined with the similarity in monosaccharide incorporation, suggest that the general parameters of the carbohydrate structure of the Ia glycoproteins from different I subregions and H-2 glycoproteins are highly similar if not identical.
Induction of Unresponsiveness to Bone Marrow Grafts
Authors: Lotzová Eva
Published: December 2022

Unresponsiveness to Hh incompatible bone marrow grafts was induced in mice by single or multiple injections of various tissues from a prospective donor before irradiation and bone marrow grafting. The results show that lymph node cells and splenocytes (both adherent and nonadherent) were the most effective in inducing unresponsiveness; thymocytes showed only a marginal effect in female and no effect in male mice, and hepatocytes had no effect. There was a direct relationship between the number of cells required for unresponsiveness induction and the strength of incompatibility between donor and recipient, i.e., the stronger the donor-recipient incompatibility, the more cells were required to induce unresponsiveness. The rapidity of unresponsiveness induction and its duration were also dependent on the number of cells in the “immunizing” inoculum. In general, unresponsiveness was induced sooner and persisted longer when larger cell doses were used. The unresponsiveness was highly specific with regard to donor antigens.
Hereditary C5 Deficiency in Man: Genetic Linkage Studies
Authors: Rosenfeld Stephen I. , Weitkamp Lowell R. , Ward Frances
Published: December 2022

Genetic linkage studies were performed on the only reported kindred with genetic deficiency of the fifth component of complement (C5). Thirty family members in four generations were studied for C5 deficiency and 32 genetic marker systems. Of these marker loci, 13 were informative in this pedigree. Most importantly, C5 deficiency was excluded (lod score < -2.0) from linkage with the major histocompatibility locus (HLA) for a recombination frequency of <15% (in females). Other marker systems excluded from linkage with C5 deficiency included the ceruloplasmin and Duffy loci at a recombination frequency of <15%, and the erythrocyte glyoxalase, MN, and Lewis loci at a recombination frequency of <5%. The most positive lod score (1.07, ϑ=0.05) was for linkage between C5 and haptoglobin, but this score does not reach statistical significance.

Thus, among the genes for complement components which can be mapped because of deficiency states or polymorphic gene products, C5 joins C1r, C3 and C6 in not being closely linked to HLA. In contrast, close HLA linkage has been demonstrated for C2, C4, properdin factor B and, in one of two families, C8.
In Vitro Lysis of Target Cells by Rat Polymorphonuclear Leukocytes Isolated from Acute Pyelonephritic Exudates
Authors: Williams Terry W. , Lyons Joseph M. , Braude Abraham I.
Published: December 2022

Polymorphonuclear (PMN) leukocytes from acute pyelonephritic exudates of rats were examined for cytoxicity against various target cells. Pyelonephritic PMN leukocytes caused lysis in vitro of syngeneic renal epithelial cells in 24 to 48 hr. They also caused lysis of allogeneic and xenogeneic target cells. The susceptibility to PMN-induced cytolysis was different among various target cell lines. Cell lysis proceeded at low ratios of effector to target cells and was not dependent on the addition of antibody or lectins. PMN isolated from peritoneal exudates (starch or endotoxin induced) were also cytoxic for the target cells. Cell-free supernatants from pyelonephritic PMN-target cell mixtures, or pyelonephritic PMN leukocytes cultured alone contained cytolytic substances which were nondialyzable and heat stable Finally, the urine of pyelonephritic rats was cytotoxic. These results suggest that PMN leukocytes and their cytotoxic mediators are directly involved in the pathogenesis of renal injury in pyelonephritis.
Hapten-Specific IgE Antibody Responses in Mice
Authors: Chiorazzi Nicholas , Tung Amar S. , Eshhar Nomi , Katz David H.
Published: December 2022

Administration of an appropriate dose of anti-lymphocyte serum (ALS) results in a selective enhancement of the primary, secondary, and adoptive secondary IgE anti-2,4-dinitrophenyl (DNP) antibody responses of mice to DNP-key-hole limpet hemocyanin (KLH); in contrast, responses of the IgG class were not similarly affected under the conditions employed. The enhancing activity of ALS could be completely absorbed with mouse thymocytes, but not with erythrocytes, plasma cells, or brain cells; in fact, preabsorption with brain cells consistently increased the enhancing activity of ALS.

Although the initial interpretation of such findings was the ALS enhanced IgE antibody production by preferential elimination of suppressor cells, further analysis revealed that such an interpretation is inadequate to explain the phenomenon. Thus, lymphoid cells from otherwise unprimed donors that had been pretreated with various forms of ALS were found to be consistently capable of exerting some type of enhancing activity on the IgE responses of syngeneic DNP-KLH-primed spleen cells when the two populations were adoptively transferred concomitantly into irradiated recipients. Such observations indicate that ALS may facilitate antibody production not only by possible elimination of suppressor T cell activity, as heretofore concluded, but also by induction of lymphoid cells capable of actively potentiating the IgE response. Such ALS-induced “potentiator” cells were not readily susceptible to depletion by anti-ϑ serum plus complement treatment and do not appear to act directly on DNP-specific IgE B lymphocytes in the primed cell population.
Expression and Function of I Region Products on Immunocompetent Cells
Authors: Henry Claudia , Chan Eva L. , Kodlin Dankward
Published: December 2022

The anti-Lac B precursor cells from BALB/c (H-2)d mice which survive cytotoxic treatment with anti-Iak and complement will respond to Lac-KLH in culture, but require more KLH helper T cells than unselected B cell populations or B cells surviving anti-Ig killing. These findings are not explainable by the classical Poisson assumption of a constant target of T-B ionteraction. We propose a T-B interaction theory with variable Ia target on the B cell surface. The theory quantitatively predicts the observed dose response relationships, and implies that Ia molecules on B cells are cell interaction structures.