bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Concanavalin A on the Classical Complement Pathway
Authors: Langone John J. , Boyle Michael D. P. , Borsos Tibor
Published: December 2022

Lysis of sheep erythrocytes (E) sensitized with anti-Forssman antiserum (EA) is inhibited by the action of concanavalin A (Con A) on whole guinea pig complement (GPC). The degree of inhibition observed for a given quantity of GPC was dependent on the Con A concentration. Specifically, Con A inhibits the activity of the early acting complement components C1 and C2 in the fluid phase, but has no significant effect on lysis once these components are bound to EA. Results of tmax experiments performed in the presence or absence of Con A showed that inhibition of C2 activity results from a direct interaction between Con A and C2 and not from a decreased number of effective EAC14 sites. Furthermore, since Con A pretreated or untreated EAC14 cells had the same tmax value, Con A and C2 apparently do not compete for the same binding site on the indicator cells. The lectin has no observable effect on either fluid phase or cell-bound C4 activity. Under similar conditions, wheat germ or soy bean agglutinin, leucoagglutinin or pokeweed mitogen did not inhibit hemolysis.
Inability of Choleragenoid-Sensitized Cells to Induce T Cell-Mediated Cytolysis
Authors: Forman James , Finkelstein Richard A.
Published: December 2022

Murine splenocytes and tumor cells bind cholera enterotoxoid (choleragenoid). Four hours after sensitization, choleragenoid-coated cells were lysed in the presence of anti-choleragenoid serum and complement, indicating that the binding was stable.

Choleragenoid-coated cells were unable to sensitize spleen cells from normal or choleragenoid primed syngeneic mice into displaying a cytotoxic effect against choleragenoid-coated target cells in the T cell-mediated cytotoxicity assay. Cells coated with both choleragenoid and trinitrophenyl (TNP) groups did sensitize syngeneic spleen cells to display a cytotoxic effect against target cells bearing choleragenoid and TNP or TNP alone, but not choleragenoid alone. These data demonstrate that the mere binding of a foreign component to lymphoid cells is not sufficient to allow sensitization of cytotoxic T cells.
Studies of Viral Antibody Responses among Amish Families
Authors: Hsia Shyuan , Howell David N. , Amos D. Bernard , Woodbury Max A.
Published: December 2022

Serum antibodies to adenovirus (ADN), cytomegalovirus (CMV), herpes simplex virus (HSV), influenza (INF), parainfluenza (PAR), mumps (MUM), coxsackie B4 (Cox B4) and B5 (Cox B5) viruses were measured from 584 individuals belonging to 21 Indiana Amish families. Sex and age effects on antibody responses to cytomegalovirus were observed. Age effect on CMV, HSV, INF, PAR, MUM responses were also found. The percentage of responders to some of the viruses was shown to be age dependent, but the levels of antibody response were not affected by the difference in age. A familial basis for the antibody response was demonstrated. Attempts at demonstrating association between HLA haplotypes and responses were not successful. The unlikelihood of a predominantly HLA-associated control of viral antibody response was discussed.
The Role of the Macrophage As the Stimulator Cell in Contact Sensitivity
Authors: Thomas David W. , Forni Guido , Shevach Ethan M. , Green Ira
Published: December 2022

In this study we examined the requirement for the type of stimulator cell for thymus-derived (T) lymphocyte activation to simple chemical haptens. T cells from picryl chloride-immune guinea pigs were challenged in vitro with various trinitrophenyl (TNP)-conjugated syngeneic stimulator cells and the extent of activation was determined by an increase in DNA synthesis. Hapten-specific T cell activation occurred with TNP-conjugated peritoneal exudate cells (PEC) and purified macrophages but not with TNP-conjugated erythrocytes, thymocytes, or nonadherent lymph node cells or PEC. In addition, T cell activation also occurred with TNP-conjugated guinea pig leukemia cells, but only in the presence of macrophages. Furthermore, it was shown that macrophages were required to process and/or present TNP-conjugated leukemia cell antigens rather than simply providing a growth-promoting function. These results suggest that a macrophage-like stimulator cell is required for hapten-specific T cell activation and that this particular stimulator cell may be important in contact sensitivity.
Murine Plasma Cells Secreting More Than One Class of Immunoglobulin Heavy Chain
Authors: Morse Herbert C. , Neiders Mirdza E. , Lieberman Rose , Lawton Alexander R. , Asofsky Richard
Published: December 2022

SAMM 368 is a BALB/c plasmacytoma which secretes IgG2b-κ and IgA-κ paraproteins. Immunofluorescence studies of ascites cells from the tumor with purified, heavy chain class-specific antiglobulins demonstrate that single cells contain both IgG2 and IgA heavy chains. Idiotypic antisera prepared in mice and rabbits indicate that the two paraproteins produced by the tumor do not share idiotypic determinants. Analysis of the purified paraproteins for allotype markers showed that SAMM 368 IgA bears the BALB/c A12, 13, 14 determinants. SAMM 368 IgG2b does not carry any detectable allotypic determinants in spite of the fact that heterologous antisera identify the paraprotein as IgG2b.
Immunologic Reactivity of the Lung
Authors: Hunninghake Gary W. , Fauci Anthony S.
Published: December 2022

The response of the guinea pig respiratory tract to locally administered antigen as well as the regulatory effect of antibody on this response was studied. The immune response of the respiratory tract to locally administered antigen was assessed by enumerating the absolute numbers of direct and indirect plaque-forming cells in the lung, hilar node, and spleen after intratracheal immunization with various doses of sheep red blood cells. This immune response of the lung could be localized primarily to the lung and hilar nodes by maintaining the dose of immunizing antigen below a critical level. The modulation of the pulmonary immune response by antibody was assessed by its effect on the numbers of plaque-forming cells in the lung and hilar nodes after immunization. IgG or IgM anti-SRBC antibody or anti-SRBC guinea pig immune serum were administered either locally (i.e., complexed to the SRBC at the time of immunization) or were given intraperitoneally 24 hr before immunization. High doses of each type of antibody, administered either locally or systemically, resulted in a delay in the peak plaque-forming response and a marked decrease in the absolute numbers of plaque-forming cells in the lung and hilar node after immunization. In contrast, progressively smaller amounts of each type of antibody resulted in a marked enhancement in the absolute numbers of plaque-forming cells in the lung and hilar node after immunization. Thus, although the lung can mount an immune response distinct from the systemic immune response, specific antibody present either locally or systemically may modulate this pulmonary immune response in a manner similar to the regulation of the systemic immune response.
Sequential Proliferation Induced in Human Peripheral Blood Lymphocytes by Mitogen
Authors: Winger Larry A. , Nowell Peter C. , Daniele Ronald P.
Published: December 2022

A culture system is described in which 1000 human peripheral blood lymphocytes diluted in 2.5 × 105 mitomycintreated autologous cells respond to phytohemagglutinin (PHA). Proliferation data, including 3HTdR uptake, cell survival counts, and mitotic indices, indicate that this inoculum expands from 1000 to 40,000 cells by day 7, suggesting five or six sequential cell divisions. Chromosome markers in allogeneic cultures demonstrate that the dividing cells are derived from the original 1000 cells and not from the “feeder layer” of mitomycin-treated lymphocytes.

The time course of proliferation in this system is similar to that in other human lymphocyte culture systems with a low percentage of responding cells, as in the response to PHA of cells from patients with chronic lymphocytic leukemia or the response of normal lymphocytes to antigens. The conditions provided by the feeder layer which permit proliferation of this small number of lymphocytes are not precisely known, but erythrocytes, heat-killed lymphocytes, or inert particles do not provide a satisfactory substitute.
Thymic Maturation in Vitro by a Secretory Product from Macrophages
Authors: Beller David I. , Unanue Emil R.
Published: December 2022

Our results indicate that immature thymocytes can be induced by macrophage culture fluid (MCF) to differentiate in vitro. This maturation occurs during several days of culture but does not require DNA synthesis. It is accompanied by an augmentation of surface H-2 which precedes a loss of susceptibility to cytolysis with anti-TL and complement. Our evidence indicates that there is no physical loss of TL within the thymus. Thymocytes cultured with MCF also acquire the capacity to respond in an MLC.

Differentiation was shown not to be due to interferon nor could it be reproduced by 2-ME. The stimulating molecule is distinct from the principal mitogenic protein found in MCF.
The Class of Surface Immunoglobulin on Virgin and Memory B Lymphocytes
Authors: Coffman Robert L. , Cohn Melvin
Published: December 2022

The class of surface immunoglobulin receptors for antigen on B cell precursors of different classes of antibody-forming cells was determined by utilizing a technique of class-specific antigen suicide. Spleen cells are first treated with a class-specific antiserum under conditions that result in the stripping of that class from the cell surface. The cells are then permitted to bind a highly radioactive trinitrophenyl (TNP)-conjugated protein, which leads to lethal irradiation of all TNP-specific B cells except those whose TNP receptors had been removed by the class-specific stripping of surface immunoglobulin. In this way, the class of antibody-forming cells resulting from TNP stimulation of B cells with different classes of surface immunoglobulin can be examined. It was found that the virgin B cell precursors of IgM-producing cells are of two types: cells bearing IgM receptors only and those bearing both IgM and IgD receptors. All virgin B cells that gave rise to IgG1 antibody-forming cells had both IgM and IgD on their surfaces, demonstrating that an antigen-dependent switch from IgM and IgD to IgG1 production is a common feature of B cell maturation. In contrast, memory B cell precursors of IgG1 antibody-forming cells had predominantly IgG1 as their surface antigen receptor. The implications of these findings on current models of B cell maturation are analyzed.
Characterization of Cytolytic Effector Cells in Peripheral Blood of Healthy Individuals and Cancer Patients
Authors: Pape Gerd R. , Troye Marita , Perlmann Peter
Published: December 2022

To characterize the effector cells involved in ADCC in a model system (lysis of 51Cr-labeled chicken erythrocytes induced by rabbit IgG antibodies), in “spontaneous”, and in tumor-associated cytotoxicity, we prepared subpopulations of peripheral blood lymphocytes from healthy individuals and from cancer patients by fractionation on columns according to three different principles. Columns with anti-immunoglobulin activity but lacking Fc-structures (F(ab′)2/anti-F(ab′)2 columns) removed ∼99% of the SIg+ cells (B cells), but only ∼20% of the Fc-receptor-bearing cells, as characterized by rosette formation with IgG-coated bovine erythrocytes (EA+ cells), and ∼25% of the complement receptor-bearing cells (EAC+ cells). Cells with receptor for sheep erythrocytes (E+ cells) were enriched. The K cell activity of this fraction was unchanged. Columns charged with immune complexes (OA/anti-OA columns) which display no anti-immunoglobulin activity retained ∼70% of the EA+ cells but only ∼20% of the SIg+ cells. About 50% of the EAC+ cells and ∼10% of the E+ cells were also removed. The K cell activity of this fraction was strongly reduced. IgG/anti-IgG columns very efficiently remove SIg+ cells, EA+ cells, and EAC+ cells whereas E+ cells are enriched. Again K cell activity is strongly reduced. The results indicate that B cells with high density SIg are inactive in the K cell assay and that the majority of mature B cells either have Fc-receptors of low avidity or have receptors for immunoglobulin which are qualitatively different from the Fc-receptors of SIg- cells. Furthermore, a significant fraction of the lymphocytes with complement receptors is different from these B cells. It can also be inferred that a significant subpopulation of the cells with receptors for sheep erythrocytes also have Fc-receptors for IgG. The cytotoxicity of these different lymphocyte fractions against a variety of human tumor cells will be reported in the following paper.
Characterization of Cytolytic Effector Cells in Peripheral Blood of Healthy Individuals and Cancer Patients
Authors: Pape Gerd R. , Troye Marita , Perlmann Peter
Published: December 2022

Peripheral blood lymphocytes from patients with transitional cell carcinoma of the urinary bladder, from control tumor patients, and from healthy individuals were fractionated on columns retaining 1) cells with Fc-receptors for IgG, as defined by rosette formation with IgG-coated erythrocytes, 2) cells with surface-bound immunoglobulin, or 3) cells with both types of surface markers. The spontaneous cytotoxicity of these lymphocytes against allogeneic or autochthonous tumor cells of bladder carcinoma or unrelated origin was studied in a 51Cr-release assay. The cytotoxicity to allogeneic target cells of lymphocyte fractions depleted of B cells (SIg+ cells) was only slightly reduced, unchanged, or, occasionally, augmented when compared to that of unfractionated lymphocytes. Fractions containing the majority of the B cells present initially but depleted of most of the Fc-receptor-bearing lymphocytes (EA+ cells) displayed a strongly reduced cytotoxicity against the same tumor cells. These effector cell fractions were also depleted of a majority of the complement receptor-bearing lymphocytes (EAC+ cells) and of a minor but significant fraction of T cells (E+ cells) and of a minor but significant fraction of T cells (E+ cells). Removal of both SIg+- and Fc-receptor-bearing cells gave the same results. Thus, in most instances, the spontaneous cytotoxicity against allogeneic tumor cells of fractionated lymphocytes was similar to their K cell activity against chicken erythrocytes (Ec) in the presence of rabbit anti-Ec antibodies (preceding paper). In conjunction with other data on the immunoglobulin dependence of this spontaneous cytotoxicity these results suggest that a significant fraction of this reactivity, displayed by both tumor patients' and normal individuals' lymphocytes, may be antibody dependent. The antibodies may either be carried over cytophilically during lymphocyte isolation or may be formed and released by some antibody-producing cells during the in vitro incubation. In two cases, in which bladder cancer patients' lymphocytes were tested with both autochthonous and allogeneic target cells, the effector cells killing the autochthonous target cells were neither B cells nor Fc-receptor-bearing cells. These results suggest that different effector mechanisms were involved in these cases.
Target Cells for Interferon Production in Human Leukocytes Stimulated by Sendai Virus
Authors: Yamaguchi Tai , Handa Kazuko , Shimizu Yoshinobu , Abo Toru , Kumagai Katsuo
Published: December 2022

Whole leukocytes, mononuclear cells, polymorphonuclear cells (PMN), monocytes, purified lymphocytes, and T (rosette-forming cells, RFC) and non-T (nonrosette-forming cells, nonRFC) lymphocytes isolated from the human peripheral blood were stimulated by Sendai virus, respectively, and examined for interferon production in their culture fluids. High levels of interferon were produced by mononuclear cells, but not by PMN. Removal of monocytes from the mononuclear cell population did not affect at all the levels of interferon produced, although it strongly suppressed interferon induction by polyinosinic-polycytidylic acid (poly IC) and mitogenic response to phytohemagglutinin (PHA) of the lymphocytes. Purified monocytes and T lymphocytes were unresponsive to the virus. In contrast, a population of purified non-T lymphocytes produced high levels of interferon. Addition of monocytes to the interferon-producing non-T lymphocytes did not affect the levels of interferon produced. No detectable levels of interferon were produced in the mixture of T lymphocytes and monocytes. It is concluded that non-T lymphocytes may be a major target for interferon induction of human leukocytes by Sendai virus.
Somatic Cell Hybrids Between Mouse Peritoneal Macrophages and SV40-Transformed Human Cells
Authors: Cicurel Lucienne , Croce Carlo M.
Published: December 2022

Somatic cell hybrids between SV40-transformed human cell lines and mouse peritoneal macrophages (MPM) containing either human chromosome 7 or 17 carrying the SV40 genome were injected into mice syngeneic to the mouse parental cells. Since either chromosome 7 or 17 was the only human chromosome present in the hybrids used as immunogens, the humoral immune response to gene products coded for by either chromosome was assayed. Using a sensitive radioimmunoassay, we were able to identify noncross-reactive cell-surface antigen(s) specifically coded for by either human chromosome 7 or 17, and present in normal, tumorderived and virus-transformed human cells. However, no reactivity against SV40 tumor-specific surface antigen (TSSA) could be detected in the antisera.
IgE Formation in the Rat After Infection with Nippostrongylus Brasiliensis
Authors: Urban Joseph F. , Ishizaka Teruko , Ishizaka Kimishige
Published: December 2022

Sprague-Dawley rats were infected with the helminth, Nippostrongylus brasiliensis, and the presence of IgE-bearing cells in various lymphoid tissues was examined by indirect immunofluorescence. An increase in IgE-bearing cells was detected in all lymphoid tissues except the thymus. In order to examine the importance of T cells in the proliferation of IgE-bearing cells and in the enhancement of IgE synthesis, neonatally thymectomized rats, which were unable to produce antibodies against a hapten-protein conjugate, were infected with N. brasiliensis. After infection, IgE-bearing cells appeared in all lymphoid tissues including the bone marrow. The kinetics of IgE-bearing cells and IgE synthesis were comparable in both thymectomized and nonthymectomized controls, however, the proportion of IgE-bearing cells in the mesenteric lymph nodes and spleen of thymectomized animals was about twice as high as that of nonthymectomized rats. At 14 days after infection of thymectomized rats, about 17% of total nucleated cells in the mesenteric lymph nodes bore IgE on their surface. These results as well as the appearance of IgE-bearing cells in the bone marrow of thymectomized animals suggested that T cells were not required for the development of IgE-bearing cells. By contrast, the serum IgE concentration and the number of IgE-forming cells detected in the mesenteric lymph nodes of thymectomized rats were significantly lower than that of nonthymectomized rats, indicating that the enhancement of IgE synthesis after infection was impaired by the absence of T cells.
Functional, Structural, and Antigenic Similarities of Guinea Pig Anti-Phosphorylcholine Antibodies
Authors: Schroer Joyce , Davie Joseph M.
Published: December 2022

The humoral immune response to PC was measured in guinea pigs. PC-vaccine stimulated IgM and IgG2, but little IgG1, anti-PC antibodies. No memory was induced and immunization in CFA produced tolerance. PC-KLH, on the other hand, stimulated IgM, IgG2, and IgG1 anti-PC antibodies with carrier-specific memory. Hapten inhibition of plaque formation showed uniform binding patterns with minor, but significant, differences between anti-PC-vaccine and anti-PC-KLH antibodies. The antibodies were characterized by IEF and idiotypic analyses. Early after immunization with PC-vaccine, guinea pigs had restricted IEF patterns which in inbred, but not outbred animals were indistinguishable between individuals. These patterns remained restricted but more individualized with time after immunization. Anti-PC-KLH antibodies showed more heterogeneity and individuality. However, these structurally heterogeneous antibodies reacted equivalently with rabbit anti-idiotype antisera and therefore must share common structural features, regardless of isotype or the genetic background of the guinea pig.
The Necessity for T Cell Help for Human Tonsil B Cell Responses to Pokeweed Mitogen: Induction of DNA Synthesis, Immunoglobulin, and Specific Antibody Production with a T Cell Helper Factor Produced with Pokeweed Mitogen
Authors: Insel Richard A. , Merler Ezio
Published: December 2022

Human B lymphocytes obtained from tonsils do not proliferate when stimulated with pokeweed mitogen. A soluble factor produced from T cells cultured with pokeweed mitogen stimulates B cells to synthesize DNA and differentiate into immunoglobulin producing cells. This PWM produced supernatant induced a PFC response to SRBC. The T cell supernatant activity is produced within 12 hr of stimulation in the presence of serum and without a requirement for T cell division. Optimal stimulation of B cells occurred at 7 to 9 days of culture. This helper factor activity eluted postalbumin from a column of Sephadex G-200. Insolubilized pokeweed mitogen was not mitogenic for B cells. The continuous presence of the lectin in culture was not required for B cell proliferation or for immunoglobulin synthesis.
Cellular Components of Allograft Rejection: Identity, Specificity, and Cytotoxic Function of Cells Infiltrating Acutely Rejecting Allografts
Authors: Strom Terry B. , Tilney Nicholas L. , Paradysz Jean M. , Bancewicz John , Carpenter Charles B.
Published: December 2022

Functioning Mononuclear cells have been harvested from heterotopic rat cardiac allografts during maximal transplant cellular infiltration. T cells, identified by a T cell-specific absorbed rabbit anti-rat brain serum, constituted two-thirds of the total cells recovered. Approximately 20% of the infiltrating cells bear and synthesize surface immunoglobulin. Macrophages, identified by latex ingestion and morphologic and cytochemical techniques, comprise 9% of the graft infiltrate. Donor-specific cytotoxic T lymphocytes are concentrated within the graft. A separate population of Fc receptor-positive recovered cells mediate antibody-dependent LMC (Ab-LMC). Neither effector cell was adherent or phagocytic. These studies have conclusively established that cytotoxic T lymphocytes accumulate within rejecting allografts; however, the enriched presence of cytotoxic T cells within the grafts is not fully dependent upon antigen recognition per se, since Lew animals grafted with both BN and BUF hearts have Lew anti-BN and Lew anti-BUF killer cells in each graft.
Heterologous Antiserum to Rat Lymphohemopoietic Precursor Cells
Authors: Goldschneider Irving
Published: December 2022

Lymphohemopoietic precursor cells in rat bone marrow are members of a subset of lymphocyte-like cells that bears the bone marrow lymphocyte antigen (BMLA) and that lacks antigens present on peripheral T and B cells. This was demonstrated by two experimental approaches. In the first, bone marrow cells with the potential to form hemopoietic colonies in spleen (CFU-S), to repopulate lymphoid tissues and blood, and to rescue lethally irradiated recipients were enriched approximately 10-fold by a fractionation procedure designed to isolate a “null” population of bone marrow lymphocytes. In the second approach, the lymphohemopoietic precursor cell activity in bone marrow was completely abrogated by opsonization with rabbit antiserum (ALSBM) raised against this “null” population of bone marrow cells. Precursor cell activity was not affected by treatment with antiserum to T and B cells. Quantitative crossabsorption studies showed that the antigen detected by ALSBM on lymphohemopoietic precursor cells had the same cellular distribution as did the previously described bone marrow lymphocyte antigen. It is likely that this antigen is present both on pluripotent stem cells and on committed progenitors of the myelocytic, erythrocytic and lymphocytic series.
Antigen Handling by Guinea Pig Macrophages: Further Evidence for the Sequestration of Antigen Relevant for Activation of Primed T Lymphocytes
Authors: Ellner Jerrold J. , Lipsky Peter E. , Rosenthal Alan S.
Published: December 2022

Guinea pig macrophages can take up sufficient 2,4 dinitrophenyl guinea pig albumin during a brief in vitro exposure at 37°C to trigger proliferation and lymphokine production with primed T lymphocytes on subsequent co-culture. Treatment of such antigen-bearing macrophages with trypsin, a procedure which removes surface antigen, does not alter the ability of such macrophage to initiate the release of migration inhibition factor from sensitized T lymphocytes. In addition, formation of antigen-specific rosettes between primed T cells and antigen-bearing macrophages is not blocked by high concentrations of antibody directed against the antigen mediating this interaction. Similarly, primed T lymphocyte DNA synthesis induced by antigen-bearing macrophages is not inhibited by specific antibody to that antigen. These data support the conclusion that the fraction of macrophage-associated antigen which is relevant to T lymphocyte activation does not reside on the macrophage surface but rather remains in a restricted compartment from which it is accessible to the T cell but unavailable to either blockade by specific antibody or removal by proteolytic enzymes.
Increased Responsiveness to Macrophage-Activating Factor (MAF) after Alteration of Macrophage Membranes
Authors: Piessens Willy F. , Remold Heinz G. , David John R.
Published: December 2022

Guinea pig macrophages pretreated with the esterase inhibitor, antithrombin III (AT III) show increased responsiveness to macrophage-activating factor (MAF) as demonstrated by their enhanced cytotoxicity for tumor cells. Other proteins that are not esterase inhibitors did not enhance the effect of MAF on the macrophage.

Enhancement of MAF activity was also obtained when macrophages were preincubated with the cell surface reactant, diazotized sulfanilic acid (DSA).

These studies indicate that the effect of MAF can be enhanced by chemical modifications of the macrophage membrane. They also provide further evidence to support the hypothesis that an esterase on the macrophage membrane modulates this cell's responsiveness to lymphocyte mediators.
Ag-B-Linked Analogue of Ia Antigens in the Rat
Authors: Shinohara Nobukata , Cullen Susan E. , Sachs David H.
Published: December 2022

The reactions of Lewis rat lymphocyte membrane antigens with two alloantisera, BN anti-Lewis and BN anti-Fischer have been studied. Three lines of evidence indicated that these antisera reacted with cell surface antigens homologous to Ia antigens of the mouse. 1) After absorption with Lewis platelets, the antisera killed only 40 to 50% of Lewis spleen cells. The majority of such cells were shown to be Ig-positive B cells by the examination of reaction patterns on lymphocytes after separation on nylon wool into T cell- and B cell-enriched subpopulations. 2) SDS-PAGE analysis of solubilized Lewis spleen cell antigens precipitated with these antisera revealed that the platelet-absorbed antisera reacted with molecules comparable in size to mouse Ia antigens (mw ≡ 35,000 and 28,000). The unabsorbed sera reacted with these molecules and with additional molecules corresponding in size to mouse K and D antigens (m.w. ≡ 45,000). 3) Neither of these antisera killed significant numbers of spleen cells from the partially congenic strain F.BN (seventh backcross homozygotes), a Fischer rat to which the Ag-B.3 allele is being transferred by repetitive backcrossing, indicating that the genes coding for these Ia-like antigens in the rat are linked to the Ag-B locus.
Sites of “Spontaneous” Degradation of IgD
Authors: Goyert Sanna M. , Hugli Tony E. , Spiegelberg Hans L.
Published: December 2022

When 10 IgD myeloma proteins were examined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), the δ-chains of each protein showed multiple banding; the average extrapolated m.w. for the two major bands was 63,900 ± 735 and 60,550 ± 675 daltons. When compared to δ-chains, only ε-chains and not σ-, α- or µ-chains showed a multiple banding pattern. Double reciprocal plots of the observed m.w. versus varying acrylamide gel concentrations demonstrated that of all the Ig heavy chains δ-chain mobility was most variable. Digestion of δ-chains with yeast carboxypeptidase caused an initial release of methionine and proline, followed by leucine, tyrosine, valine, and serine. Carboxypeptidase A did not release any amino acids, whereas carboxypeptidase B released fractional molar quantities of both lysine and arginine. Therefore, the native δ-chain probably has a Pro-Met-COOH carboxy-terminal sequence analogous to the Pro-Gly-COOH sequence of δ-chains. A portion of the δ-chains may have undergone degradation near the COOH-terminus by an enzyme with tryptic like specificity, thus explaining the nominal release of basic residues by CPB as well as the multiple forms of the δ-chain seen by SDS-PAGE analysis. Tryptic Fc but not Fab or Fd fragments of IgD showed multiple banding by SDS-PAGE analysis which provides further evidence that the COOH-terminal region of the δ-chain is responsible for the apparent heterogeneity.

The NH2-terminal sequence of Fc fragments isolated from IgD that had undergone “spontaneous” fragmentation during isolation was analyzed. The fragments formed by serum enzymes showed multiple sequences in contrast to Fc fragments generated by trypsin which gave a singular NH2-terminal sequence. Three cleavage sites were deduced for lysyl and arginyl residues 6, 8 and 12 residues NH2-terminal to the common half-cystine residue which participates in the inter-heavy chain disulfide bond. These data demonstrate that the δ-chain is highly susceptible to degradation both near the COOH-terminus and in the inter-Fd-Fc region by plasma enzymes of trypsin-like specificity.
Niridazole Suppression of Experimental Allergic Encephalomyelitis in Lewis Rats
Authors: Paterson Philip Y. , Harvey Johanne M. , Webster Leslie T.
Published: December 2022

Administration of niridazole to Lewis rats beginning 2 days before sensitization with guinea pig spinal cord in combination with immunologic adjuvants exerted a dose related suppressive influence on development of experimental allergic encephalomyelitis (EAE). A daily dose of 75 mg/kg completely prevented clinical neurologic signs as well as markedly suppressed occurrence of immunohistopathologic manifestations of this autoimmune disease. A higher daily dose level of niridazole, i.e., 100 mg/k, also inhibited EAE but was associated with neurotoxic manifestations.
The Elimination of Herpes Simplex Plaques by Antibody and the Emergence of Resistant Strains
Authors: Pavan Peter Reed , Ennis Francis A.
Published: December 2022

Established type 1 HSV plaques were eliminated by antibody. Antibody had to remain in contact with infected cells for several days to have a maximum effect. It did not prevent the production of viral induced antigens in a cell when applied after the cell was infected but did prevent the transmission of infection to contiguous cells. Strains which were resistant to elimination by antibody formed syncytia, did not grow to significantly higher titers than nonresistant strains, and were as easily neutralized by antiserum as nonresistant strains.
Mitogenic Receptors on Human Peripheral Blood Lymphocytes: The Interaction of Phaseolus Vulgaris Erythroagglutinating Phytohemagglutinin and Anti-Thymocyte Globulin on the Human Peripheral Blood Lymphocyte Membrane
Authors: Kelly James P. , Wedner H. James , Parker Charles W.
Published: December 2022

A horse anti-human thymocyte antibody (ATG) obtained from the Upjohn Company was shown to stimulate DNA synthesis in human lymphocytes with a time course and magnitude of radioactive thymidine uptake comparable to that seen with phytohemagglutinin (E-PHA) and concanavalin A (Con A). Low mitogenic or nonmitogenic concentrations of intact ATG or its Fab fragments inhibited E-PHA-induced mitogenesis, whereas the response to Con A was unaffected. Competitive binding studies with ATG and E-PHA revealed mutual inhibition of binding to lymphocytes suggesting that E-PHA and the ATG share a common receptor site on the cell surface. ATG binding was unaffected by Con A. From the analysis of the binding data and the inhibition of mitogenesis, it appears that at least part of the E-PHA response in human lymphocytes involves receptors that are not acted on by Con A.
Antibody-Dependent Eosinophil-Mediated Damage to 51Cr-Labeled Schistosomula of Schistosoma Mansoni: Mediation by IgG, and Inhibition by Antigen-Antibody Complexes
Authors: Butterworth Anthony E. , Remold Heinz G. , Houba Vaclav , David John R. , Franks David , David Peter H. , Sturrock Robert F.
Published: December 2022

The properties of the antibodies responsible for complement-independent, eosinophil-mediated damage to schistosomula have been characterized in serum samples from Schistosoma mansoni-infected baboons and human patients. The antibody is opsonic for the target larvae, but is not detectably cytophilic. It is associated with IgG-rich fractions after DEAE-cellulose chromatography, and is removed by anti-IgG immunoadsorbents. In studies on the specificity of the antibody, it was found that activity was also removed by absorption with whole schistosomula, but not with an insoluble adult worm membrane preparation. In this preparation, however, membrane antigens may have been masked; and immunofluorescence studies showed that absorption with schistosomula was associated with loss of activity against the adult worm membrane, indicating a cross-reactivity between schistosomular and adult worm antigens.

A finding that activity was also lost after incubation of sera with soluble adult worm antigens was possibly attributable to the formation of immune complexes which blocked effector cell activity. In further studies, it was shown that preincubation of effector cells with exogenous immune complexes resulted in a loss of cytotoxic activity, supporting the hypothesis that Fc receptors are involved in the initial interaction between eosinophils and antibody-coated larvae. The blockade of eosinophil-enriched effector cells by immune complexes was not reversed by washing and subsequent overnight culture before assay. A possible relationship between circulating immune complexes and the relative inactivity of cells from patients with schistosomiasis and eosinophilia is discussed.
Characterization and Specificity of Murine Anti-Tumor Cytotoxic Effector Cells within an Ascitic Tumor
Authors: Biddison William E. , Palmer Jon C. , Alexander Michael A. , Cowan Elliot P. , Manson Lionel A.
Published: December 2022

The anti-tumor immune reactivity of cells which are contained within a progressively growing ascitic tumor has been analyzed by studying the cell-mediated response of DBA/2 mice to the syngeneic mastocytoma P815Y. Peritoneal cells (P.C.) which were harvested from these P815Y tumor-bearing hosts were fractionated by velocity sedimentation at unit gravity. Cell-mediated cytotoxicity (CMC) of fractionated and unfractionated cells was measured by a 4-hr51Cr-release assay. The cell separation procedure revealed significant levels of CMC to P815Y in P.C. populations at 8 to 16 days following i.p. inoculation of 1 × 103 viable P815Y cells. The tumor-bearer P.C. which mediated CMC against P815Y were generally confined to P.C. populations that sedimented between 2 and 6 mm/hr. The cytolytic activity against P815Y mediated by tumor-bearer P.C. was sensitive to treatment with an anti-Thy-1.2 preparation and complement, but not to treatment with anti-immunoglobulin serum and complement. CMC against P815Y by tumorbearer P.C. was not inhibited by the presence of heat-aggregated human γ-globulin. In addition, it was shown that the cytolytic activity of tumor-bearer P.C. against P815Y was not mediated by “natural” killer cells, since the cytolytic activity was not labile at 37°C in culture. These results indicate that T lymphocytes which are cytotoxic for tumor cells in vitro can be found within a growing ascitic tumor.

Analysis of the specificity of DBA/2 anti-P815Y cytotoxic T lymphocytes (CTL) demonstrated that these effector cells could lyse three of four DBA/2 tumor lines and four of four C57BL/6 tumor lines which were tested. Cytolysis of two strain A tumors and Con A- and LPS-induced DBA/2 or C57BL/6 lymphoblasts was not observed. No correlation was found between the surface expression of the gp69/71 or p30 components of murine leukemia virus and the susceptibility of tumor cells to lysis by these CTL. These results indicate the T cell killing directed against shared antigenic determinants on P815Y and other tumor cells is not H-2 restricted and that gp69/71 and p30 are not the target specificities recognized by DBA/2 anti-P815Y CTL.
The Formation of Stable E-Rosettes by Human T Lymphocytes Activated by Phytohemagglutinin
Authors: Katoh Arthur K. , Charoensiri Surin
Published: December 2022

Sheep red blood cells (SRBC), when incubated with human peripheral blood lymphocytes (PBL),2 form rosettes (1). The lymphocytes that participate in rosette formation by adhering to SRBC are generally believed to be T lymphocytes (2). These rosettes have been shown to disintegrate rapidly if incubation of the reaction is carried out at 37°C (3). In striking contrast, however, is the behavior of rosettes formed by human thymus cells, which are highly stable and do not disintegrate even after prolonged incubation at 37°C (4).

Galili and Schlesinger (5) reported that human PBL, when treated by allogeneic stimuli in mixed lymphocyte cultures, were resistant to the degradative effects of 37°C incubation on the rosettes formed with SRBC. They suggested that the T lymphocytes were activated in mixed lymphocyte cultures to form stable E-rosettes.
The Role of Activated Accessory Cells in Preventing Immunosuppression by Hydrocortisone
Authors: Mishell Robert I. , Lucas Alexander , Mishell Barbara B.
Published: December 2022

The generation of humoral immunity in vitro by normal and antigen-primed mouse spleen cells was suppressed by in vitro treatment with hydrocortisone. Functions of normal and antigen-activated helper T lymphocytes and of accessory cells were inhibited by the corticosteroids. Spleen cells cultured overnight in medium containing fetal bovine serum became highly resistant to the effects of hydrocortisone. Similar resistance was found to occur when spleen cells were cultured with accessory cells that previously had been activated with bacterial lipopolysaccharide. These studies show that immunologically nonspecific processes significantly alter the effects of the steroids on specific immune responses and suggest that accessory cell products modulate T cells in ways which differ from antigen induction.
The Binding of Antibodies to the Solubilized and Membrane-Integrated Mouse and Rat Receptor for IgE
Authors: Isersky Chaviva , Mendoza Guillermo R. , Metzger Henry
Published: December 2022

Rabbit antiserum to rat basophilic leukemia cells (anti-RBL) inhibited the binding of 125I-IgE to and caused agglutination of RBL and mouse mastocytoma cells. Immunoglobulin G, Fab′, and Fab derived from this antiserum inhibited binding of 125I-IgE to whole rat and mouse tumor cells, normal rat peritoneal mast cells, and to detergent-solubilized receptors from either cell line. The inhibition was less efficient with receptors from the mouse mastocytoma and no inhibition of binding of 125I-IgE to normal mouse peritoneal mast cells was observed. The degree of inhibition of IgE binding to whole cells and solubilized rat and mouse receptors was inversely proportional to the time of incubation with and concentration of 125I-IgE. In a double immune precipitation system, anti-RBL reacted with detergent-solubilized receptors and IgE — receptor complexes derived from either mouse or rat cells. However, although nearly 100% of the free rat receptors could be precipitated no more than 70% of free mouse receptor were maximally precipitated. The efficiency of precipitation of detergent-solubilized free receptors, and the maximal precipitation of receptor — IgE complexes by anti-RBL were decreased when the detergent was above the critical micelle concentration. However, neither the precipitation of receptor — IgE complexes by anti-IgE, nor the binding of 125I-IgE to the free receptor was significantly affected under the same conditions.

The results suggest that the anti-RBL contains antibodies to at least two discrete determinants on the receptor. One determinant (or group of determinants) is sufficiently close to the combining site for IgE to interfere with IgE binding and in turn is blocked from binding by the presence of IgE. The second is distal from the first, is exposed at low detergent concentration even in the presence of IgE, but is blocked when the receptor interacts with larger amounts of detergent. Taken together with other data these findings suggest that the receptor is substantially embedded in the surface membrane.
Conditions Affecting the Immunosuppressive Properties of Human α-Fetoprotein
Authors: Goeken Nancy E. , Thompson John S.
Published: December 2022

The hypothesis that α-fetoprotein (AFP) is immunosuppressive in vitro was tested with immunoabsorbent purified human cord AFP in human lymphocyte cultures. Albumins were purified identically from cord plasma and pooled adult plasma. No preparations were suppressive in phytohemagglutinin-stimulated cultures. Both AFP and human cord albumin (HCA) produced 50% suppression of allogeneic lymphocyte cultures at 50 to 100 µg/ml final concentrations, whereas adult and commercial albumins were not suppressive. When AFP was eluted from the immunoabsorbent column in 0.15 M NaCl rather than the conventional 0.5 M NaCl, activity was greatly diminished or lost, but could be restored by dialysis against 0.5 M Kcl. Previously, inactive lots of HCA also demonstrated the same phenomenon. It appears, therefore, that the immunosuppressive activity of AFP (and albumin) may depend both on its source and the procedure by which it is isolated. Whether analogous conditions occur in vivo is unknown at this time.
Suppression of Lymphoproliferation by High Concentrations of Normal Human Mononuclear Leukocytes
Authors: Laughter Arline H. , Twomey Jeremiah J.
Published: December 2022

Mononuclear leukocytes (MNL) include cells that suppress lymphoproliferation in unstimulated and antigenstimulated cultures. Suppression is demonstrated by increasing the concentration of cells added to cultures and does not require preactivation of suppressor cells. Suppression of 3H-thymidine incorporation occurred if the high concentrations of MNL were added to cultures before the proliferative responses commenced. This suppressive effect of high cell concentration upon 3H-thymidine incorporation is removed by depleting MNL present in high numbers of cells that adhere to foreign surfaces or by preincubating these cells with cycloheximide, puromycin, or pactamycin. The suppressor cell, which only functions when present in a viable state, is radioresistant, adheres to foreign surfaces, remains active through 5 days in culture, and equates with the presence of a cell that is rich in cytoplasmic esterase. The suppressor cell may be of the monocyte series and did not appear to belong to either the T or the B lymphocyte series. This study provides additional evidence that normal immune reactivity in man is under regulatory control. The suppressor mechanism identified herein with normal human MNL is probably related to a similar type of suppression (but at a much lower cell concentration) that has been described with Hodgkin's disease and solid tumors.
Genetic Polymorphism of the Sixth Component of Complement (C6) in the Rhesus Monkey
Authors: Hall Joseph E. , Alper Chester A.
Published: December 2022

With isoelectric focusing, the complement protein C6 has been shown to be genetically polymorphic in the rhesus monkey. Three codominant alleles of a single autosomal locus, Rh C6, have been recognized: C6A, C6B, and C6R, with gene frequencies of 0.592, 0.354, and 0.053 in a random rhesus monkey population. Hardy-Weinberg analysis of the phenotypic frequencies in this population yielded observed values very close to those expected. Both natural mating between individuals carrying the various alleles and artificial combinations of sera of the different C6 types demonstrate patterns consistent with this model. Analysis of several families of monkeys confirmed the Mendelian autosomal codominant inheritance with numbers of offspring very close to expected values and no offspring types inconsistent with the mating pair types.
Identification of RNA as a Complement Inhibitory Component in an Extract of Ehrlich Ascites Tumor Cells
Authors: Renk Clifford M. , Hoffmann Edward M.
Published: December 2022

A factor capable of inhibiting complement was obtained from intact Ehrlich ascites tumor cells by mild extraction with phosphate-buffered saline (PBS). The inhibitor caused a decrease in extent of lysis of EAC with a concomitant extension of Tmax. EA, EAC1̄, EAC4 and were all less susceptible to complement-mediated lysis after treatment with the tumor cell extract. Partial purification of a complement inhibitor was accomplished. The inhibitor was rich in RNA and its activity was totally destroyed by RNAase but not DNAase. RNA from mouse tissues, yeast, and Escherichia coli also inhibited complement hemolytic activity. The partially purified material only inhibited lysis of EAC1̄ and EAC. Slow inhibition of fluid phase C1̄ was also demonstrated. In addition, RNA-rich partially purified tumor cell extract was capable of precipitating with purified human C1q.
Evaluation of Radioimmunoprecipitation for the Detection of Bovine Leukemia Virus Infection in Domestic Cattle
Authors: Devare Sushilkumar G. , Chander Satish , Samagh Bakhshish S. , Stephenson John R.
Published: December 2022

The potential application of a recently developed radioimmunoprecipitation test for antibody directed against the major structural protein of bovine leukemia virus was evaluated for use in detection of BLV-infection in domestic cattle. This technique was found to be considerably more sensitive than serologic procedures currently being utilized for this purpose. Radioimmunoprecipitation was also shown to have distinct advantages as compared to hematologic criteria, such as specified by Bendixen's index, for identification of BLV-infected animals. By the use of radioimmunoprecipitation, high levels of antibody to BLV were demonstrated in sera of animals with confirmed adult lymphosarcoma, but not in animals with a less common sporadic form of the disease which occurs in calves.
The Structural Basis of Rabbit VH Allotypes: Serologic Studies on a1 H Chains with Defined Amino Acid Sequence
Authors: Margolies Michael N. , Cannon L. Edward , Kindt Thomas J. , Fraser Blair
Published: December 2022

The amino acid sequences for the VH regions of three homogeneous antibodies elicited by type III pneumococcal vaccine were determined. All three antibodies had the group a allotype a1. Two of the antibody H chains (3374, 3381) had identical amino acid sequences in all framework positions that are considered correlates of the VH allotype, whereas the third H chain (3T72) differed from these at positions 15 and 16. The a1 allotypic specificities of the three homogeneous antibodies were compared by quantitative radiobinding and inhibition assays by using both insolubilized anti-a1 antisera and allotypic antiserum fractions rendered specific for the homogeneous antibody 3374. It was found that antibodies 3374 and 3381 are allotypically indistinguishable and have in common an a1 allotypic specificity that predominates in pooled a1 IgG. The allotypic specificity of the 3T72 antibody, on the other hand, was markedly deficient to those of 3374, 3381, and the a1 IgG pool. This correlation of allotypic difference with amino acid sequence variation at position 15 and 16 of the H chain indicates the involvement of these two residues in a major a1 allotypic determinant.