bluuebunny/crossref_metadata_2025 huggingface dataset:
Change of Cell-Mediated Cytotoxicity with Aging
Authors: Shigemoto Shozo , Kishimoto Susumu , Yamamura Yuichi
Published: December 2022

Evidence has been accumulating that both cellular and humoral immunocompetences progressively decline with aging in man (1, 2) as well as in experimental animals (3, 4).

In a previous paper (5), we showed that the capacity of lymphoid cells from old C57BL/6J mice to induce graft-vs-host (GVH) reaction was reduced as compared to that from young mice. However, the proportions of θ.C 3H-bearing lymphocytes in the cell population of spleens and lymph nodes, and the indices of the response of lymph node cells to phytohemagglutinin showed little change with aging. In an extension of this observation, an attempt was made to elucidate the changes in cell-mediated cytotoxicity against allogeneic tumor cells with aging.

Materials and Methods. Female inbred mouse strains C57BL/6J(H-2b) and DBA/2-(H-2d) were used throughout the experiment. C57BL/6J mice were either 1.5 to 2 or 15 to 20 months old, and DBA/2 mice 2 to four months old.
Leukocyte Inhibitory Factor (LIF): Production by Purified Human T and B Lymphocytes
Authors: Chess Leonard , Rocklin Ross E. , MacDermott Richard P. , David John R. , Schlossman Stuart F.
Published: December 2022

Using column immunoabsorbent chromatography to isolate highly purified subpopulations of human lymphocytes, we have previously shown that T cells, but not B cells, are specifically triggered by antigens to proliferate in vitro (1, 2). Although B cells were not triggered to proliferate, they, like T cells, were specifically activated by soluble antigens to secrete migration inhibitory factor (MIF)3 (3). These initial studies prompted further investigations which showed that: 1) B cells produced quantitatively more MIF than did T cells, 2) B cell MIF is indistinguishable from T cell MIF by Sephadex G-100 chromatography, and 3) B cell MIF production, unlike T cell MIF production, was unaffected by BUdR and light, agents known to inhibit the proliferative response (3).

Recently, another mediator of cellular immunity, leukocyte inhibitory factor (LIF), has been described (4). This material, which is distinct from MIF, inhibits the migration of human polymorphonuclear leukocytes (PMN), but not human monocytes or guinea pig macrophages.
The Immune Response to a Synthetic Amino Acid Terpolymer in Man: Relationship to HL-A Type
Authors: Scher Irwin , Berning Alice K. , Strong Douglas M. , Green Ira
Published: December 2022

The immune response to the synthetic amino acid terpolymer (L-glutamic acid55 L-lysine33 L-tyrosine15)n (GLT) was studied in normal human volunteers. Delayed skin test reactivity to this antigen was seen in 34 of 61 subjects immunized with 150 µg of GLT. No antibody to GLT was detected in these responding individuals. There was a close correlation between the in vivo skin reactivity of volunteers to GLT and the ability of their lymphocytes to produce migration inhibitory factor (MIF) in response to GLT in vitro. However, a similar correlation was not seen when the in vitro proliferative response of lymphocytes to GLT, as measured by [methyl-3H] thymidine ([3H]TdR) incorporation, was assayed.

HL-A typing of volunteers was studied to determine if responsiveness to GLT was correlated to HL-A type. No statistical association was seen after correction was made for the number of individual HL-A antigens.
Characteristics of Mesenteric Lymph Node Cells Homing to Gut-Associated Lymphoid Tissue in Syngeneic Mice
Authors: McWilliams Michael , Phillips-Quagliata Julia M. , Lamm Michael E.
Published: December 2022

A subpopulation of cells in murine mesenteric lymph nodes, about 15% of those synthesizing DNA at any given time, homes specifically to the gut and mesenteric nodes of syngeneic recipients within 1 day of i.v. transfer. In contrast, cells from Peyer's patches or peripheral lymph nodes do not. A large proportion of the B blasts which home to the small intestine has surface Ig, but lacks complement receptors. Thy-1-positive T blasts home to the gut to a lesser extent than B blasts. However, it is probable that equal fractions of B and T blasts home to mesenteric nodes. Homing is not affected by measures calculated to interfere with the combination of cell surface IgA and secretory component.
The Modifications of the Final Stages of the Complement Reaction by Alkali Metal Cations
Authors: Dalmasso Agustin P. , Lelchuk Rosalia , Giavedoni Elda B. , de Isola Elvira D.
Published: December 2022

We studied the effects of alkali metal cations on the terminal stages of complement lysis of human and sheep HK erythocytes. Sensitized (EA) were reacted with limited amounts of complement for 1 hr at 37°C in buffer containing 147 mM NaCl (Na buffer), which resulted in 10-40% lysis. The unlysed cells were washed with Na buffer at 0-2°C and incubated for 1 hr at 37°C in buffers containing 147 mM of the various alkali metal cations. Although additional lysis (25 to 65%) occured with K, Rb, or Cs buffer, only minor degrees developed with Na or Li buffer. Intermediate levels occured with 100 mM of the divalent alkali cations. Halogen ions and SCN- (147 mM), Ca++ (0.15 mM), and Mg++ (0.5 mM) did not alter the effect of the alkali metal cations. Lysis occuring in K+, Rb+ or Cs+ proceeded without lag, was temperature dependent with an optimum of 43°C, and had a pH optimum of 6.5. Lysis in K and Na buffers was unaffected by 10-3 to 10-5 M ouabain. Experiments with mixtures of cations indicated that Na+ had a mild inhibitory effect that could be totally overcome by K+, partially by Rb+, and not at all by Cs+. Li+ had a strong inhibitory effect, 6 × 10-5 M causing 50% inhibition in buffers containing 147 mM K+, Rb+, or Cs+.

By using intermediate complexes of EA and purified complement components we demonstrated that K+ enhances the lytic action of C8 on EAC1-7 as well as that of C9 on EAC1-8. It was known that Li+ facilitates lysis when acting on the entire complement reaction. We found that Li+ enhanced the lytic action of C8 on EAC1-7, with a kinetic that differed from that of the K+ effect. In addition, Li+ inhibited the enhancing effect of K+ upon lysis of EAC1-8 by C9. This occured at concentration of Li+ similar to those which inhibited the additional lysis by K+, Rb+, Cs+ of cells that were pretreated in Na buffer with the entire complement sequence. We propose that the major effects of alkali metal cations on complement lysis are due to their interaction with C8 and/or membrane constituents.
Macrophage-Lymphocyte Interaction: Antigen-Independent Binding of Guinea Pig Lymph Node Lymphocytes by Macrophages
Authors: Lipsky Peter E. , Rosenthal Alan S.
Published: December 2022

The antigen independent binding of guinea pig lymph node lymphocytes by glass-adherent macrophages was investigated. Binding was found to be mediated by a trypsing digestible, divalent cation-dependent, temperature-sensitive macrophage receptor mechanism that was not competitively inhibited by excess immunoglobulin. Data are presented to indicate that in the absence of antigen, macrophages were capable of binding both thymus-derived and bone marrow-derived lymphocytes without apparent selectivity, and further, that the binding of neither cell was mediated by surface membrane-associated immunoglobulin.
T Cell Activation and Cellular Cooperation in Autoimmune NZB/NZW F1 Hybrid Mice
Authors: Roder John C. , Bell David A. , Singhal Sharwan K.
Published: December 2022

Old (6 months) overtly autoimmune female NZB × NZW F1 (B/W) mice were markedly hyporesponsive to sheep erythrocytes (SRBC). The response to SRBC was restored by a) simultaneously injecting lipopolysaccharide (LPS) with SRBC or b) transferring bone marrow and thymocytes with SRBC into lethally x-irradiated (1000 R) syngeneic old recipients. The in vitro PFC response of old B/W spleen cells to SRBC was restored by adding in culture a) theta-positive and radioresistant spleen cells from old B/W mice primed with homologous antigen or b) activated T cells from the spleens of lethally x-irradiated (1000 R) old B/W mice injected with old syngeneic thymocytes and SRBC but not horse erythrocytes. Various populations of unprimed lymphoid cells from young (4 to 6 weeks) female B/W mice, which respond normally to SRBC, were not capable of restoring the response of old syngeneic mice in vitro or in vivo. These results suggest the existence of a suppressor of T cell activation and/or B and T cell interaction in old autoimmune B/W mice.
Cell-Mediated Cytotoxicity in Vitro of Human Lymphocytes against a Tissue Culture Melanoma Cell Line (IGR3)
Authors: Peter Hans H. , Pavie-Fischer Jeannine , Fridman Wolf H. , Aubert Christian , Cesarini Jean-Pierre , Roubin Régine , Kourilsky François M.
Published: December 2022

Purified peripheral blood lymphocytes from 13 healthy donors, 6 melanoma patients and 1 halo nevus patient were tested for cytotoxic activity against an allogeneic melanoma cell line (IGR3) in, at least, one of the following assays: cell-mediated cytotoxicity (CMC), antibody-dependent cellular cytotoxicity (ADCC) and microcytotoxicity assays (MA). The lymphocytes were isolated by Ficoll-Triosil gradient centrifugation (fraction F) followed by removal of iron-phagocytosing and adherent cells (fraction FFF) and by subsequent passage through anti-IgG columns (fraction FFF-C). Leukocytes of each fraction were identified by different methods including morphology, rosetteformation, phagocytic activity, and membrane fluorescence.

CMC activity paralled ADCC activity at a log lower level of sensitivity. In both assays lymphocytes of fractions F and FFF had the highest activity, whereas in fraction FFF-C cytotoxicity was strongly reduced. In all three lymphocyte fractions CMC and ADCC activity could be blocked by preincubation of the effector cells in aggregated IgG. Furthermore, depletion of E rosette-forming lymphocytes slightly increased ADCC and CMC activity, whereas depletion of EA and EAC rosette-forming lymphocytes strongly decreased it. Our results therefore indicate that in both CMC and ADCC assays, non-adherent, non-phagocytic Fc receptor-bearing lymphocytes (“K” cells) were the active cytotoxic cells. In MA, on the other hand, mononuclear phagocytes seemed to be the most active cell population. So far no significant difference was observed in CMC, ADCC, and MA between control persons and melanoma patients.
The Antigens and Autoantigens of the Seminal Vesicle
Authors: Shulman Sidney , Orsini Frank R.
Published: December 2022

Rabbit vesicular fluid (RVF) was shown to contain only one major electrophoretic component. This component, isolated by starch block electrophoresis, was found to have a molecular weight, by determination of sedimentation and diffusion coefficients, of approximately 17,300. This major component was also effective in isoimmunization. Another component, possibly a minor one, had similar antigenic properties. The major component, and the “minor” one also, were found to be highly tissue-specific (that is, accessory glands-specific) and also species-specific. Isoimmunization procedures have led to the production of autoantibodies. No delayed hypersensitivity response or lesions of the accessory glands of reproduction resulted. It was concluded that the major reason no lesions were found was that the rabbit was not able to develop a delayed hypersensitivity type of response to the components of RVF.

When rabbit vesicular fluid was ejaculated as part of the seminal plasma, its molecules were still able to induce an autoantibody response by isoimmunization techniques. This was the same as the response induced by rabbit vesicular fluid, itself.
Immunochemical Analysis of a Smith-Like Antigen Isolated from Two Human Strains of Staphylococcus Aureus
Authors: Karakawa Walter W. , Kane Judith A.
Published: December 2022

A surface antigen consisting of aminoglucuronic acid and N-acetyl-L-alanine was isolated from the culture filtrates of two human strains of Staphylococcus aureus. Double diffusion analysis in agar suggested that the antigen is immunologically similar to the alanyl-aminoglucuronic acid capsule of the Smith strain of S. aureus. Quantitative precipitain inhibition studies indicated that N-acetyl-L-alanine is the immunodominant determinant of the acidic antigen. In addition, conjugates consisting of N-acetyl-L-alanine coupled to bovine serum albumin gave a significant precipitin reaction with anti-staphylococcal serum which is rich in alanyl-aminoglucuronic acid polymer antibodies. Antibodies with N-acetyl-L-alanine specificity were isolated from N-acetyl-L-alanine-Sepharose immunoabsorbent columns. Double diffusion analysis in agar indicated that the eluted antibodies were serologically reactive and belonged to the IgG class of immunoglobulins.
Analysis of Specificity of Poliovirus Inhibitors with Inhibitor-Resistant Mutants of a Strain of Type 1 Poliovirus
Authors: Hashimoto Nobuo
Published: December 2022

Attempts were made to analyze the specificity of inhibitory activities of normal bovine and equine sera to the Mahoney strain of type 1 poliovirus. A total of five inhibitory factors were postulated to explain the complicated results. Two of the three bovine inhibitors were identical in specificity to certain equine inhibitors despite differences in their mode of virus inactivation and their molecular size. In addition to this, inhibitors that could inactivate certain resistant mutants, but not the parent virus, were newly detected in a number of normal bovine and equine sera.

Antigenic variation of the resistant mutants against equine sera containing an inhibitory factor h-11 was determined by means of the kinetic neutralization test by using both anti-Mahoney and anti-M-H11 sera. These results offer evidence that some inhibitors, at least in part, are indistinguishable from specific antibody.
Cellular Distribution of Rabbit IgA Allotypes
Authors: Hanly W. Carey , Knight Katherine L.
Published: December 2022

Intestinal tissue from rabbits heterozygous at one or both of the α-chain subclass loci, i.e., f or g, was examined with mixtures of rhodamine-labeled and fluorescein-labeled antibodies each directed against one of the allelic products of the f or g locus. Individual plasma cells were stained by only one fluorescent reagent and thus exhibited both allelic exclusion and subclass exclusion. In contrast, the epithelial cells of the Lieberkühn's glands did not exhibit allelic exclusion. Studies with rabbits heterozygous at the f and g loci revealed that the ratio of the number of cells expressing maternal-type f cells to paternal-type f is equal to the ratio of the number of maternal-type g cells to paternal-type g cells. These data are used to suggest that the expression of each of the genes in the heavy chain chromosomal region of a particular allogroup is under the same control.
3H-Uridine Incorporation by Small Lymphocytes of Tolerant Rats: Relationship to T and B Lymphocytes
Authors: Bottomly H. Kim , Perkins William D. , Schwarz M. Roy
Published: December 2022

Tissue tolerance was induced in neonatal rats by the intravenous injection of bone marrow cells from adult allogeneic rat donors. After 6 to 8 weeks, lymphoid cells from rats in which tolerance had been induced were tested for mixed lymphocyte reactivity (MLR), 3H-uridine uptake, and the relationship of uridine incorporation to B and T lymphocytes.

Lymph node (LN) and spleen (SPL) cells from the adult inoculated rats showed no reactivity in the MLR or normal lymphocyte transfer reaction (NLTRx), indicating that the animals were tolerant. After in vitro exposure to 3H-uridine, an abundance of small lymphocytes (SL) from these same tolerant rats were heavily labeled, in contrast to nontolerant controls, where relatively few SL were heavily labeled.

In order to determine whether the heavily uridine-labeled cells were T cells or B cells, lymphoid cells from the LN and SPL of tolerant animals were exposed to either rabbit anti-AKR brain serum or rabbit anti-rat Ig conjugated with ferritin. The results showed that the heavily uridine-labeled SL of the tolerant rats were mainly Ig-positive cells.
B Lymphocyte Differentiation Induced by Lipopolysaccharide
Authors: Kearney John F. , Lawton Alexander R.
Published: December 2022

Cultured cells from mouse thoracic duct, spleen, lymph node, Peyer's patches, and bone marrow gave rise to cells containing large amounts of cytoplasmic IgM, IgG1, and IgG2 when stimulated by bacterial lipopolysaccharide (LPS). Differentiation of IgA producers occurred in bone marrow only, and to a lesser extent than other classes. Significant IgM responses preceded development of cells containing other classes. The differentiation of IgG and IgA producers did not appear to depend on T cells, since cultures from nu/nu or thymectomized-irradiated, bone marrow-protected mice responded as well as normals. Cultures from mice rendered deficient in B cells by anti-µ treatment responded normally to T cell mitogens, but did not proliferate or give rise to immunoglobulin-secreting cells when stimulated with LPS. Bone marrow cultures gave relatively meager proliferative responses to LPS, but generated as many or more immunoglobulin-secreting cells as did other tissues.
Lymphocyte Plasma Membranes
Authors: Zimmerman Barry
Published: December 2022

Anti-lymphocyte sera against human thymus [ALS-(THY)] were absorbed serially with cultured human lymphoblasts (CHL) or thymus and residual antigen-binding activity was tested. The absorbed ALS were used to bind 125I-labeled antigens from lymphocytes labeled by the lactoperoxidase catalyzed iodination technique. Absorption of ALS(THY) with CHL led to the absorbed serum having less than 5 to 10% of its original antigen-binding activity against labeled CHL antigens while maintaining from 20 to 40% of its original activity against labeled THY. Serial absorption of ALS(THY) with THY led to an equal decrease in activity against both THY and CHI. When the immunoprecipitates from these experiments were examined on polyacrylamide gels containing SDS it was found that serial absorption of ALS(THY) with THY first removed activity against a component of m.w. ∼ 48,000 leaving relatively greater activity against material of apparent high molecular weight. In contrast, absorption of ALS(THY) with CHL removed the antibodies against the high molecular weight material while leaving activity against the component of m.w. 48,000. When these absorbed ALS were used to induce in vitro lymphocyte proliferation, it was found that ALS(THY) absorbed with human THY lost its mitogenicity whereas ALS(THY) absorbed with CHL, did not. The retention or loss of mitogenicity seemed to correlate with retention or loss of binding activity against the component(s) of m.w. ∼ 48,000.
Immunologic Effects of Neonatal Infection with Mouse Thymic Virus
Authors: Cohen Philip L. , Cross Sue S. , Mosier Donald E.
Published: December 2022

Mouse thymic virus is a herpesvirus that causes extensive thymic necrosis when given to newborn mice. During the time of acute infection spleen cells have markedly diminished reactivity to T cell phytomitogens and to allogeneic cells and are incapable of effecting a primary in vitro response to a “T-dependent” antigen; responses to B cell mitogens and to a T-independent antigen are unimpaired. Spleens from acutely infected mice have low ϑ antigen content and depletion of T-dependent areas, but have normal numbers of immunoglobulin-bearing cells. Surprisingly, despite widespread necrosis and cellular depletion, thymic cell reactivity to mitogens is unimpaired. However, the ability of thymocytes to proliferate and to generate cytotoxic killer cells in response to allogeneic cells is diminished.
The Circulating Life Span, Immunocompetence and 3H-Uridine Uptake of Small Lymphocytes from Thymus-Grafted, Neonatally Thymectomized Rats
Authors: Willis Judith I. , Schwarz M. Roy
Published: December 2022

By 7 weeks post-grafting, the number of small lymphocytes in the thoracic duct lymph (TDL) and blood of the thymus-grafted neonatally thymectomized adult rats had increased to 60% of the number of cells in sham controls, or 2-1/2 times thymectomized control values. This increase consisted almost exclusively of long-lived, recirculating small lymphocytes and corresponded to a 60% recovery of cellular immunocompetence as measured by the mixed lymphocyte reaction (MLR). Associated with the return of cellular immunocompetence was an increased incorporation of 3H-uridine by the small lymphocytes. Cells from thymectomized animals grafted with lymph node fragments demonstrated no significant increase in lymphocyte numbers nor was there a return of immunocompetence as compared to thymectomized controls.
Studies on the Human T Lymphocyte Population
Authors: Owen Frances L. , Fanger Michael W.
Published: December 2022

The IgG fraction of a goat antibody specific for human T lymphocytes (ATG) and that of a rabbit antibody directed toward the purified T lymphocyte receptor for SRBC (anti-Ts) were used to study the synthesis, display, and release of the receptor for SRBC (Ts) by normal human peripheral lymphocytes. A large percentage of lymphoblasts obtained by stimulation of lymphocytes with PHA or PWM formed multilayered rosettes with SRBC. The vast majority of these rosetting lymphoblasts were T lymphocyte derived, whereas few B lymphoblasts formed rosettes. Significant increases were found in the quantity of Ts on the lymphoblast membrane and in the supernatants from PHA-stimulated cultures, especially late in transformation. Furthermore, lymphocytes cultured with PHA and 3H-leucine synthesized and released radiolabeled Ts.
Determinants of Differential Migration of Mouse Spleen Cells Treated with Concanavalin A
Authors: Rodríguez Beverly A. , Rich Robert R. , Rossen Roger D.
Published: December 2022

The effects of Con A on homing patterns of mouse spleen cells were investigated. After 1 to 18 hr incubation with or without the mitogen, cells were labeled with 51Cr and injected i.v. into syngeneic hosts. Treatment of spleen cells with Con A decreased localization in lymph nodes 75 to 95% and inhibited homing to recipient spleens 17 to 50%. Conversely, localization in liver and lungs was increased. Migration of cells labeled with both 125I-Con A and 51Cr revealed that cells binding the largest amount of Con A were trapped in lungs whereas cells migrating to lymph nodes and spleen had a much lower Con A content. Elution of Con A from lymphocyte surfaces with methyl-α-D-mannoside (MAM) reduced the percentage of cells localizing in lungs, with a concomitant increase in the percentage of cells migrating to lymph nodes. However, even after MAM treatment, a cell population with a relatively large Con A content localized in the lungs. These data indicate that both surface-bound Con A and Con A-induced changes in lymphocyte membranes inhibit migration of Con A-activated cells to recipient lymph nodes. They suggest that after Con A treatment lymph node-seeking cells do not migrate normally because of preferential sequestration in lungs of lymphocytes with either Con A-binding sites of increased density or avidity or with an increased propensity for Con A internalization.
Modulation of the Immune Response by Antigen-Reactive Lymphocytes after Cultivation with Gangliosides
Authors: Miller Harold C. , Esselman Walter J.
Published: December 2022

GM1 ganglioside isolated from mouse brain was found to be capable of inhibiting the thymocytotoxicity of AKR anti-Thy 1.2 antiserum and to a lesser extent anti-BAϑ antiserum. GD1b ganglioside of mouse brain and thymocytes inhibited the thymocytotoxicity of rabbit anti-BAϑ antiserum for mouse thymocytes. When GM1 ganglioside, in the form of cholesterol-lecithin liposomes, was incubated with spleen cell cultures in the presence of SRBC, depressed anti-SRBC hemolytic plaque responses were observed. This effect could be neutralized by first absorbing the GM1 liposomes with anti-Thy-1.2 antibodies. The kinetics of GM1 suppression of spleen cell cultures reveal reduced PFC numbers when the ganglioside was added even as late as the 4th day of culture. Results of experiments in which GM1 ganglioside was incubated with either bone marrow cells or with thymocytes suggest that the target cell for GM1 liposomes is the B lymphocyte.
Two Novel Stimuli of Cyclic Adenosine 3′,5′-Monophosphate (cAMP) in Human Lymphocytes
Authors: Atkinson John P. , Wedner H. James , Parker Charles W.
Published: December 2022

Polystyrene latex particles (PLP) and zymosan particles (ZP), two commonly employed phagocytic stimuli, were noted to bind to purified human peripheral blood lymphocytes. This interaction was not accompanied by ingestion but did lead to a marked increase in intracellular cyclic AMP. The cAMP response to PLP was proportional to the particle cell ratio which, in turn, correlated with the number of membrane-associated particles. After the addition of PLP to lymphocytes, the cAMP response occurred within 2 min, peaked between 4 and 15 min, and returned to baseline by 30 to 60 min. The cAMP response to ZP was similar in onset and duration to that seen with PLP but was less marked (2- to 4-fold vs 25- to 50-fold) and more variable in magnitude. This is probably a reflection of the smaller number of cells interacting with ZP. At high PLP to cell ratios almost all of the lymphocytes bound PLP but only 10 to 28% of the mixed lymphocyte population bound ZP.

Two lines of evidence established conclusively that the cAMP response was taking place in the lymphocytes themselves rather than in contaminating cells. 1) When lymphocytes were purified additionally by filtration through a nylon wool column (99 to 100% lymphocytes), they were found to undergo a similar cAMP response to PLP. Since the nylon filtration procedure also removes almost all of the B cells, this further indicates that T cells are capable of undergoing the response. 2) Immuno-fluorescence studies with anti-cAMP antibody revealed an increase in intralymphocytic cAMP which was primarily adjacent to the site of PLP or ZP attachment.

The likely explanation of this data is that PLP and ZP perturb the lymphocyte surface leading to regional activation of membrane-bound adenylate cyclase and subsequent cAMP accumulation. Although the physiologic significance of these observations remains to be determined, the results: 1) provide histologic confirmation for the concept of cAMP compartmentalization, 2) clarify conflicting results regarding the localization of cAMP accumulation during the phagocytosis of PLP by mixed leukocyte populations, and 3) suggest that this experimental system may allow an analysis of the mechanism by which perturbations of the lymphocyte surface modulate cAMP.
Mechanism of Complement-Induced Cell Lysis Demonstration of a Three-Step Mechanism of EAC1-8 Cell Lysis by C9 and of a Non-Osmotic Swelling of Erythrocytes
Authors: Valet Günter , Opferkuch Wolfgang
Published: December 2022

Sheep erythrocytes were sized by an electrical method during lysis by antibody and complement. No volume changes were observed up to the EAC1–8 cell intermediate. Three steps could be distinguished in the reaction of C9 with EAC1–8 cells: fixation of C9 to EAC1–8 cells, a process which proceeds also at 0°C; temperature-dependent swelling of the EAC1–9 cells; and lysis of the cells.

We could show that swelling of the erythrocytes induced by the complement reaction is independent of protein osmotic forces and is caused by an alteration of the erythrocyte membrane.
Antibody-Induced Redistribution of CEA on the Cell Surface: Utilization in Separation of CEA and Isoantigen A
Authors: Rosenthal K. L. , Palmer J. L. , Harris J. A. , Rawls W. E. , Tompkins W. A. F.
Published: December 2022

Antibodies specific for carcinoembryonic antigen (CEA) induced polar redistribution of CEA which was expressed on the surface of human intestinal carcinoma cells grown in tissue culture. Polar redistribution occurred at 23°C and 37°C but not at 4°C and was inhibited by sodium azide and cytochalasin B. Antibodies to isoantigen A did not react with CEA at the cell surface but did react with isoantigen A which was not redistributed by the antibody. Cells which expressed both CEA and isoantigen A were used to demonstrate that polar redistribution of CEA could be induced without altering the distribution of isoantigen A. The data indicate that CEA and isoantigen A can be expressed on the same cell as separate molecules.
Evidence for an Abnormal Microenvironment in the Thymus of New Zealand Black Mice
Authors: Dauphinée Michael J. , Palmer Donald W. , Talal Norman
Published: December 2022

The proliferative response of NZB and DBA/2 thymocytes and spleen cells to allogeneic cells and mitogens was studied in intact and lethally irradiated bone marrow-repopulated syngeneic recipients. The major findings were complete recovery of function in NZB spleen cells and premature recovery in thymocytes compared to the control DBA/2 strain. This recovery was not influenced by the administration of thymosin to the recipients, and was not due to functional thymocytes present in the donor marrow inoculum.

These results suggest normal or increased support of T cell differentiation by the radioresistant thymus in 4-month-old NZB mice. Thymic epithelial cell products regulating T cell maturation may be disordered or unbalanced but do not appear to be broadly deficient at this age.
The Role of Cyclic AMP in the Chemotactic Responsiveness and Spontaneous Motility of Rabbit Peritoneal Neutrophils
Authors: Rivkin Israel , Rosenblatt Joseph , Becker Elmer L.
Published: December 2022

Agents known to affect intracellular levels of cyclic AMP in many diverse systems have been tested for their effect on the chemotaxis induced by Escherichia coli culture filtrates, spontaneous motility and cyclic AMP levels of rabbit peritoneal neutrophils. Prostaglandin E1 and A1 but not prostaglandin F2α increased neutrophil cyclic AMP levels and, correspondingly, only the former two prostaglandins inhibited chemotaxis. Nevertheless, a quantitative relationship between prostaglandin stimulation of cyclic AMP and inhibition of chemotaxis could not be found. Epinephrine, isoproterenol, and, to a much lesser extent, norepinephrine increased neutrophil cyclic AMP through beta adrenergic stimulation. Only epinephrine and isoproterenol inhibited chemotaxis, but the inhibition was variable and not related to the ability of these catecholamines to increase intracellular cyclic AMP. Cholera toxin increased neutrophil cyclic AMP after a 30-min lag period which paralleled its inhibitory effect on chemotaxis and spontaneous motility. However, the effect on chemotaxis required 50 ng/ml of toxin whereas the effect on cyclic AMP was manifested at 2 ng/ml of toxin. Prior to 30-min preincubation there was no effect of even 1250 ng/ml of toxin on either cyclic AMP or chemotaxis. Choleragenoid prevented the effects of toxin on both cyclic AMP and chemotaxis.

The bacterial chemotactic factor obtained from E. coli culture filtrates did not effect a measurable change in levels of neutrophil cyclic AMP.

The data indicate that even though cyclic AMP is not, in the main sequence of events, triggering the chemotactic response, increases in neutrophil cyclic AMP may modulate the movement and thus the chemotactic responsiveness of the neutrophil.
Genetically Controlled Autologous Immune Complex Glomerulonephritis in Rats
Authors: Stenglein Bodo , Thoenes Gunther H. , Günther Eberhard
Published: December 2022

A study was conducted on the disease susceptibility of inbred strains of rats to experimental autologous immune complex glomerulonephritis (AIC). Three strains representing two different H-1-haploytpes developed severe glomerulonephritis within 3 months in response to a single injection with equal doses of an autologous primary tubular epithelial fraction and complete Freund's adjuvant (Lewis, AS (H-1l)) and Lew.BDV (H-1d)). By contrast, two strains of the H-1 haplotype H-1n (BN and Lew.BN) showed no proteinuria and no immunohistologic findings during that time. Hybrids of Lew.BN and Lewis, subjected to the same immunizing procedure, showed a later onset of the disease as compared to the responder parent. The possible relationship between responder status and the major histocompatibility complex is discussed.
Immunosuppression by Spleen Cells from Moloney Leukemia. Comparison of the Suppressive Effect on Antibody Response and on Mitogen-Induced Response
Authors: Cerny Jan , Stiller Ronald A.
Published: December 2022

The inhibitory effect of cells from leukemic spleens on the immune functions of normal lymphocytes was studied. Suppressor cells were obtained as the nonadherent fraction (NA) from splenic tumors of mice infected with MuLV-Moloney. This fraction (NAMuL V-M) contained less than 10% membrane Ig-positive (Ig+) cells, 45 to 60% θ-positive cells (θ+) and 40 to 50% naught cells (θ-, Ig-). Similarly prepared fractions from normal control spleens (NAc) containing 75 to 90% θ+ cells and less than 10% Ig+ and naught cells were utilized in control cultures. Addition of the NAMuL V-M cells into cultures (Marbrook system) of normal spleen cells with sheep red blood cells suppressed the specific antibody response determined by the number of hemolytic plaque forming cells (PFC). The PFC response was significantly suppressed at a suppressor cell to responder cell ratio of 1:100, and was completely abolished at a ratio of 1:10 or higher. The control NAc fraction showed some inhibitory effect only at high suppressor to responder ratios (1:2 or 1:1). In contrast, the suppressive effect of NAMuLV-M on mitogen-induced 3H-thymidine incorporation in normal B and T cells was much weaker. Very little, if any, suppression occurred at the ratio of 1:100 or 1:10, however, about 50% decrease in DNA synthesis was observed at the ratio 1:2 or 1:1. On the basis of this differential suppressive effect, it is suggested that leukemic spleen cells can suppress the function of immunocompetent cells by more than one mechanism.
Antigenic Analysis of Chlamydiae by Two-Dimensional Immunoelectrophoresis
Authors: Caldwell Harlan D. , Kuo Cho-Chou , Kenny George E.
Published: December 2022

Two-dimensional immunoelectrophoresis was used for the antigenic analysis of Chlamydia. Antigens of chlamydiae were found to be successfully solubilized with the nonionic detergent Triton X-100. Nineteen and 16 distinct components were observed for Chlamydia trachomatis strain L2/434/Bu and C. psittaci strain Meningopneumonitis/Cal-10, respectively. The triton-solubilized antigens of chlamydiae differ strikingly between species since only one strong reciprocal cross-reacting antigen was observed between 434 and Cal-10. However, the antigens of Trachoma strain K/UW-31/Cx and LGV strain 434 were found to be very similar.
Triggering Mechanism of B Lymphocytes
Authors: Kishimoto Tadamitsu , Miyake Tadao , Nishizawa Yoshio , Watanabe Takeshi , Yamamura Yuichi
Published: December 2022

Stimulation of rabbit mesenteric lymphocytes with antiimmunoglobulin antibody (anti-Ig) for 24 hr (first stage of culture) followed by the addition of enhancing soluble factor (second stage of culture) induced the formation of IgG. The presence of mitotic inhibitors, such as cytosine arabinoside (Ara-C), hydroxyurea (HU), with anti-Ig for the first 24 hr of incubation did not inhibit the production of IgG induced by anti-Ig and soluble factor. The addition of cytochalasin-B in the first stage of culture with anti-Ig did not inhibit, but rather enhanced, the formation of IgG. On the other hand, the presence of Ara-C, HU, or cytochalasin-B with soluble factor in the second stage of culture completely inhibited the production of IgG. Soluble factor preferentially induced the uptake of thymidine of anti-Ig-stimulated cells. Lipopolysaccharide (LPS) also induced the proliferation of rabbit lymphocytes, but did not induce the IgG production of anti-Ig-stimulated cells. However, LPS-stimulated cells formed a significant amount of IgG when they were cultured with soluble factor. Anti-Ig-stimulated cells absorbed the enhancing activity of soluble factor, but non-stimulated cells did not absorb the activity. These results showed that cross-linkage of Ig receptors or LPS receptors with anti-Ig or LPS induced the formation of acceptor sites for soluble factor without the requirement of cell division and these activated cells started to proliferate and maturate to antibody-forming cells under the influence of soluble factor.
Secondary IgG Responses to Type III Pneumococcal Polysaccharide
Authors: Braley-Mullen Helen
Published: December 2022

Mice primed with a thymus-(T)3 dependent form of Type III pneumococcal polysaccharide (S3), i.e., S3 coupled to sheep or horse erythrocytes (S3-RBC), produce S3-specific IgG antibody after secondary challenge with either the T-dependent (S3-RBC) or T-independent (S3) form of the antigen. The potential to produce IgG antibody after challenge with S3-RBC appears earlier after priming than the potential to produce IgG after challenge with S3, suggesting that different ‘memory’ cells may be involved in the two responses. The ‘memory’ cells were shown to be S3-specific since S3 had to be present on the carrier in order for priming to occur and carrier specificity was not required for elicitation fo the secondary response by S3-RBC.
Splenic Suppressor Macrophages Induced in Mice by Injection of Corynebacterium Parvum
Authors: Kirchner Holger , Holden Howard T. , Herberman Ronald B.
Published: December 2022

Spleen cells from C57BL/6N mice injected with killed Corynebacterium parvum (CP) had a marked growth inhibitory effect on the in vitro proliferation of RBL-5 murine lymphoma cells. It was most marked 12 to 14 days after injection and was usually no longer detectable later than 21 days. It could be demonstrated at effector cell to target ratios between 20:1 and 5:1 at which normal spleen cells had a growth-promoting effect. Addition of CP to an in vitro mixture of spleen cells and tumor cells augmented the inhibitory effect of spleen cells from CP-injected mice although it conferred no inhibitory potential on normal spleen cells. Growth inhibition by CP spleen cells was not medicated by T cells and various depletion experiments suggested that the effect or cells of the phenomenon were macrophages. Spleen cells of CP-injected mice also showed strongly depressed responses to the T cell mitogens PHA and Con A and suppressed the mitogen responses of syngeneic normal spleen cells. The characteristics of the suppressor cells mediating this effect appeared to be very similar to those inhibiting lymphoma cell growth. The responses to LPS were also strongly suppressed in mice injected with 2.1 mg of CP. However, after injection of one-tenth of the dose a relative sparing of the LPS response was noted, whereas the PHA response was still suppressed.
Anomalous Reactions of Mouse Alloantisera with Cultured Tumor Cells
Authors: Klein Paul A.
Published: December 2022

Mouse alloantisera produced against different specificities of the K, I, and D regions of the H-2 gene complex reacted as immunogenetically anticipated with normal lymphoid target cells of different haplotypes in cytotoxicity and indirect immunofluorescence tests.

These same alloantisera, however, produced anomalous positive reactions when tested on cultured MCA-induced sarcoma cells from B10 background H-2 congenic mice.

Absorption experiments demonstrated that the anomalous activity in these sera was directed against a tumor membrane antigen(s) which was distinct from H-2 region specificities against which the reference alloantisera were produced, and which was shared in common by multiple cultured sarcoma lines.

Similar anti-tumor antibody activity could be demonstrated in the serum of older (τ 12 weeks) but not younger normal unimmunized mice of the strains used as recipients for alloantiserum production.

It is suggested that the observed anomalous anti-tumor activity in these alloantisera may be due to the presence of antibodies reactive with envelope antigens of murine leukemia virus which are expressed on sarcoma cells maintained in culture.
Immune Response to a Syngeneic Mammary Adenocarcinoma
Authors: Kuperman Oded , Fortner G. William , Lucas Zoltan J.
Published: December 2022

Measurement of the development of cytolytic activity by mammary tumor primed or unprimed syngeneic spleen cells on in vitro monolayers of the 13762 rat mammary tumor operationally defined several subpopulations of lymphoid cells involved in the cytotoxic response. In vitro sensitization of cells from Fischer 344 animals injected 2 to 10 days earlier with 2 × 107 viable tumor cells always resulted in a higher and earlier lytic response than cells from non-inoculated animals. Adoptive transfer of the same in vivo primed cells for 5 days in irradiated syngeneic hosts removed any cytotoxic cells originally present but subsequent in vitro sensitization still resulted in a higher and earlier cytolytic response. We defined such cells as “memory” cells for cytotoxicity. Memory cells were radiosensitive and specific for the immunizing target cell. In contrast to cells from animals inoculated for 3 to 10 days, cells obtained 11 and 12 days after immunization had a lower response than unprimed cells on in vitro sensitization. The anamnestic response could be restored either by culturing 12-day primed cells in vitro for 2 days without antigen or by adoptive transfer for 5 days into irradiated syngeneic rats. This suggests that another population of cells is present in spleen and suppresses the conversion of memory to cytotoxic cells. A more direct measurement of suppressor cell function was obtained by coincubating tumor-primed and unprimed cells on monolayers during in vitro sensitization. Cells from animals bearing tumors for 5 to 10 days always caused an increase in the response of the mixed lymphocyte groups, whereas 11- to 13-day tumor primed cells always caused a marked decrease in the cytolytic response.

These results suggest the following interpretation of the kinetics of cell-mediated cytotoxicity to syngeneic tumor inoculation. Cytotoxic cells appear about 6 days after immunization, reach peak levels 2 days later, and then decrease rapidly. Memory cells are generated at a faster rate, reach peak levels before maximum cytolytic activity, but are then functionally inhibited from converting into differentiated cytotoxic cells by a new population of suppressor cells which reach peak activity about 12 days after immunization.
The Immune Adherence Receptor: Dissociation Between the Expression of Erythrocyte and Mononuclear Cell C3b Receptors
Authors: Rothman Ivan K. , Gelfand Jeffrey A. , Fauci Anthony S. , Frank Michael M.
Published: December 2022

Human erythrocytes have a surface receptor for the third component of complement (C3b). This receptor mediates the immune adherence (IA) reaction. In this study, erythrocytes from over 100 normal donors and from donors with rare blood group types were examined for IA reactivity. All donors had red cells with high titer IA activity, with the exception of one Rh null individual whose erythrocytes had barely detectable IA activity. Five other RH null donors had normal IA titers. Because of the apparent defect in the expression of the red cell C3b receptor in this one individual, the mononuclear cell C3b receptors of this donor were studied and found to be normal. These findings suggest that expression of the erythrocyte C3b receptor may be controlled by factors which differ from those determining expression of the mononuclear cell C3b receptor.
Immunosuppression Effected by Macrophage Surfaces
Authors: Ptak Wlodzimierz , Gershon Richard K.
Published: December 2022

Heat-killed macrophages (but not six other types of cells tested) suppress Mishell-Dutton cultures by a mechanism that does not adversely affect cell viability. The suppressive effect is abolished by treatments that inactivate receptors for cytophilic antibody. We suggest macrophages may absorb, and thus render inoperative, factors which are necessary for lymphocyte cooperation.
The Elaboration of Leukotactic Mediators during the Interaction between Parental-Type Lymphocytes and F1 Hybrid Cells
Authors: Ward Peter A. , Volkman Alvin
Published: December 2022

Mixed leukocyte cultures consisting of white blood cells from (Lewis × BN) F1 hybrids and Lewis parents produced monocyte chemotactic factor. Elaboration of this material preceded incorporation of 3H-thymidine. Local graft vs host (GVH) reactions were induced by subcapsular injection of parental thoracic duct cells into F1 hybrids. Homogenates from these kidneys, but not from kidneys injected with syngeneic thoracic duct cells, contained monocyte chemotactic factor. Little or no neutrophil chemotactic factor was present. Ultracentrifugal analysis of the monocyte chemotactic factor indicated a distribution similar to that found previously in culture fluids of lymphoid cells stimulated by soluble antigens. Differential counts of inflammatory cells extracted in suspension form from kidneys undergoing a GVH reaction indicated the majority of cells to be lymphocytic in type, but with a significant proportion of monocytes. Virtually no neutrophils were present. These findings indicate that a monocyte chemotactic factor is produced by cultures of parental leukocytes stimulated by semiallogeneic cells and that a similar factor appears in GVH reactions in the rat kidney. This chemotactic factor may be relevant to the character of the cellular exudates.
The Immune Response against an Encephalitogenic Fragment of Guinea Pig Basic Protein in the Lewis and Brown Norway Strains of Rat
Authors: McFarlin Dale E. , Hsu Sharon Chung-Ling , Slemenda Susan B. , Chou Selene C.-H. , Kibler Robert F.
Published: December 2022

Lewis rats which have the Ir-EAE gene develop EAE after challenge with either GPBP or GPEF, a 42-residue fragment. BN rats which lack the Ir gene do not develop EAE after challenge with these proteins. The immune response against GPEF was compared in these two strains of rats. It was found that Le rats produce small amounts of antibody and a cell-mediated immune response against GPEF; in contrast BN rats did neither. These findings support the concept that the Ir-EAE gene exerts its effect by controlling the immune response against GPEF.
Detection of Hepatitis a Viral Antigen by Radioimmunoassay
Authors: Hollinger F. Blaine , Bradley Daniel W. , Maynard James E. , Dreesman Gordon R. , Melnick Joseph L.
Published: December 2022

In 1973, Feinstone et al. (1), with the technique of immune electron microscopy (IEM)2 (2), detected virus particles in acute-phase stool suspensions obtained from patients with type A hepatitis. This technique was useful in further characterizing hepatitis A virus, but required the expertise of a skilled electron microscopist. The need for a rapid, more sensitive technique that could qualitatively evaluate large numbers of specimens became apparent. Recent success in our laboratory with a microtiter solid phase immunoradiometric assay (micro-SPIRA) for detecting hepatitis B core antigen in the sera of patients with post-transfusion hepatitis B suggested that a similar application to the hepatitis A problem might be appropriate (3). The method involves the coupling of unlabeled antibody to an insoluble matrix which can then extract immunologically reactive antigen from the test specimen. Detection of the specific antigen is measured after interaction with a second radiolabeled antibody.