bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship of Cellular Proliferation and the Generation of Cytotoxic Cells in an in Vitro Secondary Immune Response to Syngeneic Rat Lymphoma Cells
Authors: Bernstein Irwin D. , Cohen Edith F. , Wright Peter W.
Published: December 2022

The role of cellular proliferation for the generation of cytotoxic activity in an in vitro secondary immune response to syngeneic lymphoma cells was investigated. Spleen cells from W/Fu rats immunized with the syngeneic (C58NT)D tumor proliferate and generate cytotoxic potential for tumor targets after exposure to mitomycin C-treated (C58NT)D cells in vitro. Elimination of proliferating cells by exposure to high specific activity 3H thymidine at appropriate intervals impaired the generation of cytotoxic activity. Elimination of cells proliferating to either syngeneic lymphoma or BN rat alloantigens allowed the remaining cells to generate cytotoxic potential to the second set of antigens. Elimination of proliferating cells also abrogated the ability of the in vitro generated cells to adoptively confer anti-tumor protection on nonimmune recipients. These results demonstrate that cellular division is required for the generation of cells which are cytotoxic in vitro and can adoptively confer anti-tumor protection in vivo.
Cell-Mediated Immune Responses in Patients with Recurrent Herpes Simplex Infections
Authors: O'Reilly Richard J. , Chibbaro Anthony , Anger Edward , Lopez Carlos
Published: December 2022

Herpes simplex virus antigen-induced lymphocyte proliferation and production of leukocyte migration inhibitory factor (LMIF) and lymphocyte-derived interferon were studied in normal individuals and patients with recurrent Herpes labialis and Herpes progenitalis. Virus-specific lymphoproliferative responses were regularly detected in patients with recurrent infection irrespective of the clinical stage of infection. In contrast, transient deficiencies in herpes-specific lymphoid production of both LMIF and interferon were regularly documented at the time of and immediately before herpes simplex-induced vesicular eruptions. During convalescence, pronounced production of these mediators in response to antigenic stimulation with inactivated virus antigen preparations was regularly detected. The biology of these fluctuations in lymphokine production is evaluated and discussed.
A Neutrophil Chemotactic Factor and Its Inhibitor Found in DNCB-Induced Skin Inflammatory Lesions
Authors: Baba Toru , Tazaki Kayoko , Sonozaki Hidekichi , Torisu Motomichi
Published: December 2022

Neutrophil chemotactic factor(s) and their inhibitors were explored in the acute inflammatory skin lesions induced by the application of 4% DNCB solution with acetone in guinea pigs. Skin biopsies were taken periodically and tissue extracts were made from the biopsy specimens. Neutrophil chemotactic activity found in such extracts reached a peak at 12 to 24 hr after the induction of the inflammation, when the lesions were found to be infiltrated predominantly with neutrophils. After 24 hr, the activity gradually diminished. Physicochemical and antigenic characterization studies on the chemotactic substance indicated that the material was most likely the cleavage product of C3.

On the other hand, inhibitors against the neutrophil chemotactic factor were found in the tissue extracts which were obtained from the lesions at a later stage (48 to 96 hr after the induction of the inflammatory reaction). These inhibitors blocked not only the complement-derived chemotactic activity but also that obtained from bacterial culture filtrates. They were heat labile and showed striking heterogeneity in size on Sephadex gel filtration.
Characterization of Molecular Heterogeneity and Multispecificity in Homologous Idiotypic Antisera
Authors: Perlmutter Roger M. , Davie Joseph M.
Published: December 2022

The molecular heterogeneity of homologous anti-idiotypic reagents was characterized by a novel isoelectric focusing procedure. Idiotypic antisera directed against the PC-binding plasmacytoma protein T15 were raised in CE and A/J mice. These antisera were shown to be highly specific by hemagglutination with myeloma protein-derivatized sheep erythrocytes and by radioimmunoassay. Competition experiments performed with affinity-labeled T15 revealed that about 40% of the pooled CE antibody activity was directed against binding site-associated determinants. Further analysis of anti-idiotypic sera from individual animals with the use of isoelectric focusing disclosed heterogeneous populations of antibody molecules distinguishable by isoelectric point and by subspecificity. Each animal expressed a unique spectrotypic profile. In addition, clones reactive with binding site and non-binding-site determinants as well as some clones with specificity for other PC-binding mouse myeloma proteins were detected. These results emphasize the importance of careful selection and thorough absorption of idiotypic antisera.
Lipopolysaccharide-Induced Suppression of the Primary Immune Response to a Thymus-Dependent Antigen
Authors: Persson Ulla
Published: December 2022

The immune response to a thymus-dependent antigen was depressed in vivo and in vitro in spleen cells from mice injected with LPS i.p. a few days before challenge with the antigen. Spleen cells from LPS-injected mice could, however, respond with increased DNA synthesis after activation with polyclonal B and T cell activators in vitro. The LPS-activated spleen cells could actively suppress normal cells in their response to the antigen sheep red blood cells. The suppressor cells contained in the LPS-activated spleens were most likely B lymphocytes, and the possible mechanism for their inhibitory function is discussed.
Stimulation of Human Peripheral Blood Lymphocytes by Periodate, Galactose Oxidase, Soybean Agglutinin, and Peanut Agglutinin: Differential Effects of Adherent Cells
Authors: Novogrodsky Abraham , Stenzel Kurt H. , Rubin Albert L.
Published: December 2022

Blastogenic responses of normal human peripheral lymphocytes to three distinct groups of mitogens were studied: Group I—phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM); Group II—soybean agglutinin (SBA) and peanut agglutinin (PNA); and Group III—galactose oxidase (GO) and sodium periodate (IO4-). SBA was mitogenic for human cells, and this effect was enhanced by treating the cells with neuraminidase (NA). PNA was mitogenic only after cells had been treated with NA. GO was effective before and activity was increased after lymphocytes were treated with NA. Responses to Group II and III mitogens were more variable than were those to Group I mitogens. Studies with purified T and B cells indicated that SBA and PNA were T cell mitogens, whereas IO4- and GO failed to stimulate either T or B cells. Adding macrophages back to this system indicated that they were both T cell mitogens with strict macrophage requirements. T cell responses to SBA and PNA were enhanced over responses to unfractionated cells to a degree that could not be explained simply by enrichment of the cultures with T cells. Removal of adherent cells from unfractionated cell suspensions again revealed a marked enhancement of responses to SBA and PNA, a consistent decrease in responses to IO4-, and a variable decrease in responses to GO. Similar results were found with 14C-leucine and 3H-uridine incorporation, as well as 3H-thymidine for the assessment of blastogenic response. Mechanisms responsible for these differential effects of macrophage depletion on lymphocyte responses to different groups of mitogens are yet to be determined. Either different mitogens require different lymphocyte to macrophage ratios for optimal stimulation, or some mitogens (i.e., SBA and PNA) form inhibitory complexees in the lymphocyte-macrophage mixture. In any case, variability in response to mitogenic agents in normal as well as pathologic states may be dependent on adherent cell populations, rather than on the lymphocytes themselves.
Cross-Linking of T Cell Mitogens: Effects on B and T Cell Proliferation and Immunoglobulin Synthesis
Authors: Basham T. Y. , Waxdal M. J.
Published: December 2022

The T cell mitogens Pa-2, concanavalin A (con A) and its dimeric derivative succinyl-con A, were each cross-linked with the bifunctional reagent dimethyl suberimidate. Although the dose-response curves of these insoluble aggregated products were markedly changed from those of the soluble mitogens, each aggregate continued to stimulate DNA synthesis by murine thymus and T cells. Both aggregated Pa-2 and aggregated succinyl con A stimulated DNA synthesis by B cells from athymic (Nu/Nu) mice. Aggregated con A did not stimulate these cells and, like soluble con A, depressed the background incorporation of 3H-thymidine. Unlike soluble Pa-2, aggregated Pa-2 also greatly increased Ig production by both the B cell cultures and B + T cell cultures from normal (BALB/c) mice.
Localization of Receptors for Activated Complement on Visceral Epithelial Cells of the Human Renal Glomerulus
Authors: Shin M. L. , Gelfand M. C. , Nagle R. B. , Carlo J. R. , Green I. , Frank M. M.
Published: December 2022

Receptors for activated C3 have recently been demonstrated to be present in glomeruli of normal human kidneys. In the present communication, the precise location of these receptors within the renal corpuscle was studied with scanning electron microscopy. By this technique, the glomerular complement receptor (GCR) was found to be located on the visceral epithelial cell of the renal corpusle. This epithelial cell location was confirmed by comparison of in vitro GCR activity with the location of immunoglobulin and C3 deposited in vivo. Renal tissues in which Ig and C3 had been deposited in vivo diffusely in subepithelial loci had no in vitro GCR activity. Renal biopsies not showing Ig or C3 deposition or biopsies with Ig and C3 in the mesangium retained GCR activity. These results further confirm an important role for GCR in the trapping and deposition of C3 in some forms of immunologically mediated renal disease.
Mitogen Induced Cellular Cytotoxicity (MICC) in Multiple Myeloma
Authors: Yonkosky Donna M. , Silverman Stuart L. , Cathcart Edgar S.
Published: December 2022

Mitogen induced cellular cytotoxicity (MICC) was noted to be markedly increased in patients with multiple myeloma as compared to normal controls and to patients with chronic lymphocytic leukemia (CLL). Enhanced MICC was present at various effector-to-target cell ratios and at several mitogen concentrations. Removal of adherent, phagocytic cells by carbonyl iron, glass wool, or rayon columns abolished the MICC response from the peripheral blood of both multiple myeloma patients and normal controls. Thus, the effector cell mediating MICC may be monocytic in origin and closely resembles the suppressor cell for immunoglobulin synthesis described in patients with multiple myeloma. Our data suggest that the MICC assay with chicken red blood cells as targets may provide a convenient method for identifying pathologic conditions where this cytotoxic effector cell population plays an active role.
The Xenogeneic Effect
Authors: Simon Philip L. , Farrar John J. , Kind Phyllis D.
Published: December 2022

A supernatant from human mixed leukocyte cultures (MLC), Xenogeneic Reconstitution Factor (XRF), was added to one-way murine MLC. This supernatant greatly enhances the generation of cytotoxic cells from C57BL/6 or DBA/2 responder thymocytes, as assayed by a standard 51Cr-release assay. This enhancement is shown 1) to be dependent upon the presence of BDF1 semi-allogeneic stimulator cells and 2) to result in the generation of cytotoxic cells that are specific for the H-2 type of the stimulating alloantigen. In some experiments, alloantigen-stimulated murine spleen cells, when cultured with XRF showed increased induction of cell-mediated cytotoxicity. These experiments show that XRF contains a cytotoxic-cell activating factor(s) that is functionally similar to those found in murine-derived supernatants, and that this factor can participate in the generation of cytotoxic cells from precursors present in the murine thymus.
Multiple Occurrence of Spontaneous AKR/J Lymphomas with T and B Cell Characteristics
Authors: Greenberg Robert S. , Mathieson Bonnie J. , Campbell Pamela S. , Zatz Marion M.
Published: December 2022

Three Thy 1.1-positive and surface IgM-positive (Thy 1+, SIg+) AKR/J lymphoma lines are described. These doubly marked tumors arose spontaneously in the peripheral lymphoid organs of 14- to 16-month-old AKR/J mice that either had spontaneous thymus atrophy or had been thymectomized at 1 month of age. All lines bore surface Thy 1.1, Ly, Ig (μ-chain) and Fc receptor (FcR), detectable by immunofluorescence. Immune response region (Iak) antigen was present on the two lines tested. Persistence of Thy 1.1 antigen and SIg after long-term tissue culture provided evidence that these markers were not passively acquired. One of these tumor lines, AkTB-1 always grows in lymph nodes as Thy 1.1-positive, SIg-negative tumor cells, whereas tumor cells growing in the spleen are initially Thy 1.1 positive, SIg negative, but they rapidly acquire SIg, FcR, and Ia between 18 and 21 days of passage.
In Vitro Secondary MLR
Authors: Fathman C. G. , Collavo D. , Davies S. , Nabholz M.
Published: December 2022

We have examined the kinetics and specificity of secondary in vitro mixed lymphocyte reactions (MLR). With limited numbers of primed responder cells (PRC) in the presence of “excess antigen” it was possible to obtain proliferative responses that were proportional to the number of PRC initially placed in culture. The responding cells, after an initial lag period, seem to grow exponentially until day 3 of culture. The responses of PRC (with the strain combinations and culture conditions described in this report) seemed to be directed toward stimulator cell determinants whose expression was determined by genes in the I region of the MHC. In one case, the relevant incompatibilities could be further restricted to the I-A region. Although PRC responded best to stimulator cells sharing the I region with the priming stimulator cell, apparent cross-reactivity could be observed by restimulating PRC with stimulator cells that did not carry the MHC haplotype of the priming stimulator cell. The rate of proliferation (measured as 3H-thymidine incorporation) in these apparent cross-reactions was reproducible and comparable to the rate observed in response to the priming stimulator cell. It was possible, therefore, to estimate the proportion of PRC that reacted in the presence of third party stimulator cells compared to the response of these PRC to the priming stimulator cells. We have estimated that the response of A (B6) PRC against H-2d and H-28 haplotype stimulator cells is about half of the response of these PRC to H-2b, the priming stimulator cell.
Influence of H-2 Complex on Susceptibility to Infection by Murine Leukemia Virus
Authors: Tucker Henry St. G. , Weens Joan , Tsichlis Philip , Schwartz Robert S. , Khiroya Raman , Donnelly Judith
Published: December 2022

Titers of infectious ecotropic MuLV in mouse spleen were examined after deliberate infection. In congenic mice differing only in H-2 haplotype, a gene (or genes) within the H-2 complex determined either a high virus titer (H-2k, H-2d, H-2a) or a low titer (H-2b, H-2q). Susceptibility to high virus titers was inherited as a dominant trait. Kinetic studies revealed similar initial patterns of infection in both groups, with a fall in titer in the “resistant” strain occurring from week 6 through 10 after infection. Anti-VEA antibody titers differed significantly between the groups, but no mechanistic role for antibody in eliminating virus was apparent. Genes outside the H-2 complex were shown to influence MuLV titers after infection as well.
Fractionation of Lymphocyte Subpopulations Which Regulate Mixed Lymphocyte Reactions
Authors: Rich Robert R. , Chu Lily , Solliday Rich Susan
Published: December 2022

Four days after injection of allogeneic lymphocytes BALB/c splenic T cells suppress proliferation of syngeneic cells in mixed lymphocyte reactions (MLR). Conversely, lymph node cells from the same mice amplify MLR responses. To further characterize these functional subpopulations, alloantigen-primed lymphocyte suspensions from both organs were fractionated by velocity sedimentation at unit-gravity. After fractionation MLR suppressor cells from spleens localized exclusively in rapidlly sedimenting fractions of large cells. MLR suppressor activity of cells from these fractions, as well as that of unfractionated spleen cell suspensions, was abolished by treatment with anti-Thy-1.2 serum and complement. Spleen cell fractions of similar sedimentation velocity also secreted a soluble MLR suppressor into culture supernatants. Although inhibitory of MLR, spleen cells of rapid sedimentation velocity did not suppress responses to T cell mitogens. In marked contrast with the effects of spleen cells, large 4-day-alloantigen-primed lymph node cells had no suppressive activity in MLR. MLR amplifier cells of uncertain derivation were found in fractions of medium sedimentation velocity from both spleens and lymph nodes. Fractionation of alloantigen-primed lymph node cell suspensions did reveal, however, a subpopulation of small cells with MLR suppressor activity which was unaffected by treatment with anti-Thy-1 serum and complement. The data thus indicate that large alloantigen-activated lymphocytes are not intrinsically suppressive nor are cells which suppress MLR necessarily large. We consequently conclude that regulation of MLR responses by alloantigen-primed lymphocytes involves a complex interaction between distinct functional subpopulations of cells which are separable both by physical and biologic properties.
Binding of Affinity Cross-Linked Oligomers of IgG to Cells Bearing Fc Receptors
Authors: Segal David M. , Hurwitz Esther
Published: December 2022

The binding of covalently cross-linked oligomers of rabbit IgG antibodies to tumor cells resembling macrophages and lymphocytes and to normal spleen cells has been measured. With all cells trimeric IgG binds with greater affinity than does the dimer, which in turn binds more tightly than does the monomer. However, as the oligomers increase in size, the individual monomeric subunits bind with decreasing energy. The macrophage tumor line, P388D1, binds oligomers with greater affinity than does the lymphocyte line, AKTB-1, but the differences in affinities are not great, differing by at most a factor of 5 in equilibrium constant. Normal spleen cells bind oligomers in the same concentration range as the tumor cells.

The kinetics of binding do not occur as single first or second order reactions and suggest a multistage mechanism for oligomer binding. The presence of large concentrations of monomeric IgG tends to weaken oligomer binding and increases the exchange rate of bound oligomer from the cells. Since plasma and extracellular fluid also contain large concentrations of monomeric IgG, it is suggested that many immune complexes will bind weakly to the types of cells examined here and will rapidly exchange with IgG from the external medium.
The Role of an Activatable Esterase in Immune-Dependent Phagocytosis by Human Neutrophils
Authors: Musson Robert A. , Becker Elmer L.
Published: December 2022

Diisopropylphosphofluoridate, cyclohexyl alkylphosphonofluoridates, and cyclohexyl phenylalkylphosphonofluoridates, which are potent, irreversible inactivators of serine esterases, inhibit the phagocytosis of opsonized sheep erythrocytes by human neutrophils. Two types of inhibition were observed: a) “cell-dependent” inhibition, which is determined by measuring the ingestion of neutrophils pretreated with the esterase inhibitors and washed; and b) “phagocytosis-dependent” inhibition, which is due to the presence of the inhibitors during phagocytosis.

With the cyclohexyl alkylphosphonofluoridates, phagocytosis-dependent inhibition was always greater than cell-dependent inhibition. Cell-dependent inhibition was irreversible and dependent on the duration of the incubation of neutrophils with inhibitor. Both types of inhibition were dependent on the concentration of the inhibitor. Poorly or non-phosphorylating analogues of the cyclohexyl alkylphosphonofluoridates of DFP were not inhibitory; nor did fluoride, the hydrolysis product of these inhibitors, inhibit ingestion under either condition. In addition, neither method of treating the neutrophils resulted in a decrease in neutrophil viability. Furthermore, pretreating the EAC1423 with the inhibitors did not decrease ingestion.

We conclude that cell-dependent inhibition is due to the inactivation of an esterase required for phagocytosis which is in or on the neutrophil in an active form, and thus is susceptible to inhibition by the esterase inactivators before contact of the neutrophil with the phagocytic stimulus. Phagocytosis-dependent inhibition is interpreted as being due to inactivation of an esterase required for phagocytosis which is normally in an inactive precursor proesterase form that is activated by the interaction of the neutrophil with the phagocytic stimulus. The distinctly different inhibition profiles of the active and activatable esterases indicate that they are two different activities.
The Reactions of Antibodies to Paragloboside (Lacto-N-Neotetraosyl Ceramide) with Human Erythrocytes and Lymphocytes
Authors: Schwarting Gerald A. , Marcus Donald M.
Published: December 2022

Paragloboside (lacto-N-neotetraosyl ceramide) is a biosynthetic precursor of the ABH and P1 blood group glycosphingolipids and of one class of gangliosides. This study concerns the specificity of rabbit anti-paragloboside antibodies, and their reactions with human erythrocytes and lymphocytes. Lacto-N-neotetraose was approximately 10 times as effective as lacto-N-tetraose in inhibiting complement fixation by purified IgG antibodies to paragloboside. The purified antibodies cross-reacted weakly with lactosyl ceramide, asialo GM1, and lacto-N-neotetraosyl polylysine. The purified IgG and IgM antibodies exhibited weak hemagglutinating activity which was enhanced by treatment of erythrocytes with papain or neuraminidase. In contrast to a macroglobulin described by Tsai et al. (J. Immunol. 117:717, 1976), which also binds paragloboside and lacto-N-neotetraose, anti-paragloboside antibodies reacted equally well with normal adult, Oi, and umbilical cord erythrocytes. The IgG antibodies to paragloboside reacted with about 60% of B lymphocytes of peripheral blood and 5 to 10% of T cells. The relationship between antibodies to paragloboside and other lectins and antibodies directed against β-galactosyl groups, including anti-T (Thomsen-Friedenreich) antibodies, is discussed.
Effect of Cytochalasin B on Human Lymphocyte Responses to Mitogens: Time and Concentration Dependence
Authors: Hoffman Rosemary , Ferguson Ronald , Simmons Richard L.
Published: December 2022

Human peripheral blood mononuclear cells were exposed to various concentrations of concanavalin A (Con A) and cytochalasin B (CB). Low concentrations of CB (0.5 to 2.0 μg/ml) augmented, whereas high concentrations of CB (τ;2.0 μg/ml) inhibited the mitogenic response to Con A when Con A and CB were added at the initiation of culture. Augmented responses could also be found by adding normally inhibitory high concentrations of CB after 72 to 96 hr incubation with Con A. Preincubation of the cells with CB 5 min to 4 hr before adding Con A also resulted in potentiation of the mitogenic response. CB did not increase spontaneous transformation and worked as well in various solvents (dimethyl sulfoxide, acetone, and ethanol). The effect was totally reversible by removing CB before adding Con A. CB had no effect on 3H-Con A binding to cells. The potentiating effect of CB was still evident after removal of CB, Con A, or both from cultures that had been simultaneously exposed to Con A and CB for at least 2 days. CB augmented the response of human lymphocytes to Con A-adsorbed, x-rayed autologous lymphocytes. In the presence of optimal concentrations of CB, x-ray-damaged cells respond better to Con A; the response to suboptimal and supraoptimal concentrations of Con A is better, and the response with delayed addition of α-methyl-D-mannoside is better. CB also potentiates the response of human lymphocytes to pokeweed mitogen and phytohemagglutinin. Direct observation of cells in culture demonstrated the development of huge clusters of cells during mitogenic responses to Con A in the absence of CB. Circumstances which inhibited the mitogenic response to Con A totally prevented the development of cellular aggregates (prolonged incubation with high concentrations of CB). Each of the conditions which augmented the mitogenic response to Con A was accompanied by the formation of multiple small, relatively uniform cellular aggregates. These observations suggest that CB affects cell-cell interaction possibly by affecting distribution of Con A on the cell surface, by affecting cell surface net negative charge, or by increasing the response to chemotactic stimuli. CB may be acting in some manner to enable certain nonresponsive cells to respond to a mitogenic stimulus, although an increase in the response of individual cells cannot be ruled out.
Detection of Plaque-Forming Cells in the Peripheral Blood of Actively Immunized Humans
Authors: Thomson Philip D. , Harris Nick S.
Published: December 2022

Ten adult human volunteers were immunized with Salmonella typhi and their peripheral blood leukocytes were collected for 14 days after immunization. These peripheral blood leukocytes, rich in lymphocytes, were plaqued in a modified Jerne assay against sheep erythrocytes coated with either Salmonella or Escherichia lipopolysaccharide. A specific direct and indirect PFC response developed in immunized individuals by day 7 and peaked at day 10. This vigorous PFC response rapidly declined to normal levels by day 14. This marked and specific PFC response of human peripheral blood leukocytes may be developed as a useful tool for monitoring the humoral immune response of patients with Gram-negative bacterial infections.
Evidence that Human α-Lymphotoxin Is Not an RNase
Authors: Moy Jeffrey , Rosenau Werner
Published: December 2022

Conclusion

In view of the results presented above, we conclude that human a-LT has no demonstrable RNase activity, and that the enzTmatic activity described by others was a contaminant. Ribonucleases are widely distributed throughout biologic systems. This notion is supported by our observation of RNase activity in crude supernatants of human lymphoid cells, whether stimulated with PHA or not. At this time, we have no knowledge of the precise mechanism of this lymphokine, although there is evidence that it binds to target cells (6), alters the cellular membrane (7), and increases target-cell synthesis of RNA (8, 9). This last phenomenon may be involved in a reparative process, since inhibitors of RNA synthesis greatly enhance LT-induced cytotoxicity (5).
A Study of the Variable Heavy Chain (VH) Region of Membrane-Bound Ig on Human Chronic Leukemic Lymphocytes
Authors: Førre Øystein , Frøland Stig S. , Natvig Jacob B. , Michaelsen Terje E. , Johnson Peter M. , Ly Bernt , Laake Knut
Published: December 2022

Lymphocytes from 20 patients with chronic lymphocytic leukemia (CLL) were studied for membrane staining by direct immunofluorescence by employing anti-F(ab′)2, anti-VHI, anti-VHII, anti-VHIII subgroup-specific antisera, as well as light chain-specific antisera. Some lymphocyte preparations were also studied in indirect immunofluorescence with an antiserum raised against a fragment (VH) corresponding to the variable region of the heavy chain of a human IgG3 myeloma protein (Kup). Lymphocytes from each CLL patient demonstrated a restriction of VH subgroups expressed on the cell membrane; six were restricted to the VHI subgroup, seven to VHII, and seven to the VHIII subgroup. This restriction gave further evidence for monoclonality of the membrane-bound Ig and the leukemic cell proliferation. Antiserum to the VH fragment stained closely similar percentages of CLL lymphocytes to that obtained with anti-F(ab′)2 antiserum. Furthermore, double staining revealed that the same cells were stained with anti-VH antiserum as were stained with anti-F(ab′)2 antiserum, i.e., only the B lymphocytes.
Characteristics of the Immature Cells Involved in T Cell-Mediated Enhancement of Syngeneic Tumor Growth
Authors: Small Myra
Published: December 2022

Enhancement of tumor growth was observed when nonsensitized thymocytes were injected together with tumor cells into syngeneic mice, although this tumor enhancement was less pronounced than that caused by tumor-sensitized T lymphocytes. The cells within the thymus which are responsible for this tumor enhancement were found to be rapidly dividing and to be absent from the thymus 1 day after cortisone administration. At a longer time interval the cortisone-depleted thymus was repopulated by dividing cells which exhibited tumor-enhancing reactivity. The characteristics of these cells suggest that they are in the early stages of thymic processing. The enhancing thymocytes were sensitive to treatment with the thymic humoral factor which functions in T cell maturation, and their enhancing activity was cancelled by such treatment. These results are compatible with our hypothesis that exposure of immature T cells to a tumor stimulus may lead to tumor enhancement whereas interaction between mature T lymphocytes and tumor cells may be required for tumor inhibition.
The Cutaneous Basophil Response to Phytohemagglutinin in Chickens
Authors: Stadecker Miguel J. , Lukic Miodrag , Dvorak Ann , Leskowitz Sidney
Published: December 2022

A cellular infiltrate rich in basophils was observed in chickens in skin sites injected locally with the mitogen PHA. These infiltrating metachromatically granulated cells exhibited features common to mast cells of some species (monolobed nucleus), but also resembled basophils of other species in that they circulated in the blood, and they responded to the PHA mitogen by infiltrating skin test sites. Previous literature suggests that the chicken has a single metachromatic granulated cell and our data support this notion. PHA, a known T cell mitogen, may act by causing the stimulated T lymphocyte to produce a lymphokine that is responsible for the attraction of basophils to the test site.
Age-Related Changes in T Cell Function
Authors: Krogsrud R. L. , Perkins E. H.
Published: December 2022

A comparison was made of the abilities of carrier (BGG)-primed T cell populations from young (4-month old), middleaged (14- and 19-month old) and old (31- and 34-month old) mice to collaborate with hapten (DNP)-primed B cells from young mice in a cell-transfer system. The plaque-forming cell responses to 2, 4-dinitrophenol (DNP) were measured by a modification of the Jerne plaque assay. The DNP-specific antibody-forming cell responses of old T cell/young B cell combinations were significantly lower than those of young T cell/young B cell combinations, both in the number of T cells needed for peak response and in the size of that response. These data indicate that the primed T cell populations of old mice are deficient by a factor of 6 in their ability to initiate B cell proliferation and differentiation into antibody-forming cells.
Studies on the Role of the Thymus and T Cells in the in Vivo Suppression of Delayed Hypersensitivity (Desensitization): Radiosensitivity of the Mechanism Inducing Nonspecific Anergy
Authors: Thestrup-Pedersen Kristian , Dwyer John M. , Askenase Philip W.
Published: December 2022

Guinea pigs immunized with two or more antigens can have their ability to manifest delayed hypersensitivity (DH) reactions to all these antigens temporarily blocked by the administration of large (mg) amounts of one or more of these antigens 9 days after immunization. This nonspecific anergy is called desensitization. This study presents evidence for the induction of the desensitized state by an active, radiosensitive, immunoregulatory mechanism involving the peripheralization of thymocytes. Desensitization was associated with 1) a marked reduction in thymic weight; 2) an increase in mature T cells in the peripheral blood; 3) decreased responsiveness of the lymphocytes in the spleen to concanavalin-A; and 4) markedly reduced numbers of mononuclear cells, basophils, and polymorphs at the skin sites of specific and nonspecific desensitization. Small doses of whole-body irradiation which left DH capacity intact prevented nonspecific desensitization, but did not prevent specific desensitization, suggesting that a radiosensitive cell was involved in the production of anergy.
In Vitro Augmentation of Lymphocyte Sheep Cell Rosette Formation by Leukocyte Dialysates
Authors: Holzman Robert S. , Lawrence H. Sherwood
Published: December 2022

We have studied in vitro the effects of leukocyte dialysates containing transfer factor on the formation of sheep cell rosettes by human lymphocytes. Dialysate had no effect on the total rosettes, but increased the number of rapidly forming (“active”) rosettes. This was due to an increased affinity of the lymphocytes for the sheep red cell. Trypsin-treated lymphocytes regained the ability to form rosettes more rapidly when cultured with leukocyte dialysates than with control media. These experiments suggest that leukocyte dialysate acts to increase the number or arrangement of sheep cell receptors on the lymphocyte surface.
Immunosuppressive Activity of Human Seminal Plasma
Authors: Lord Edith M. , Sensabaugh George F. , Stites Daniel P.
Published: December 2022

A high molecular weight fraction prepared from human seminal plasma by gel filtration chromatography suppresses human lymphocyte transformation and DNA synthesis induced by mitogens (PHA, Con A, PWM), antigens (Candida albicans, tetanus toxoid), and allogenic cells. This same fraction also suppresses the stimulated response of mouse lymphocytes to allogenic cells and to various mitogens, including T cell-dependent and T cell-independent mitogens. The induction, but not the expression, of cell-mediated cytotoxicity is also suppressed. Similar high molecular weight fractions suppress the in vitro humoral response of mouse spleen cells to both a T cell-dependent (SRBC) and a T cell-independent (DNP-F) antigen. The high m.w. fraction exhibited in vitro suppressive activity at concentrations of 0.1 to 1.0 mg/ml which corresponds to a 1/50 or greater dilution of human seminal plasma. These observations support the concept that a local immune response against sperm in the female reproductive tract is actively suppressed by a component in seminal plasma.
Histamine-Induced Suppressor Factor (HSF): Effect on Migration Inhibitory Factor (MIF) Production and Proliferation
Authors: Rocklin Ross E.
Published: December 2022

Histamine added in vitro to cultures of sensitized lymphocytes suppresses antigen-induced production of migration inhibitory factor (MIF) and proliferation by these cells. Recent studies have suggested that lymphocytes bearing histamine type-2 receptors play a regulatory role in these in vitro responses. The present studies were undertaken to determine if suppressor function by cells having histamine receptors was mediated through a soluble product. It was found that lymph node cells from nonimmune or immune strain 2 guinea pigs elaborate a nondialyzable factor into the culture supernatant when incubated with 10-3 to 10-5 M histamine (histamine-induced suppressor factor or HSF). HSF, when cocultured with sensitized lymphocytes, suppressed their MIF and proliferative responses to antigen. HSF was made by lymphocytes but not macrophages. Its production could be blocked by an H2 receptor antagonist (burimamide) but not an H1 receptor antagonist (chlorpheniramine). Furthermore, the inhibitory effect of HSF was reversible as lymphocytes washed free of the factor after 24 hr and recultured with fresh medium and antigen were able to produce MIF. Gel filtration by Sephadex G-100 chromatography indicated that HSF had an approximate m.w. of 23,000 to 40,000. These results suggest that the release of histamine at the sites of immediate hypersensitivity reactions, possibly by generating HSF activity, may play a regulatory role in the subsequent development of cellular-immune reactions at the same site.
Complement-Dependent and -Independent Pathways of T Cell-B Cell Cooperation
Authors: Lewis George K. , Ranken Raymond , Goodman Joel W.
Published: December 2022

BDF1 mice treated with CoV had markedly reduced levels (< 20%) of native serum C3 32 hr later, whereas the frequency of splenic CR+ cells was normal. CoV treatment before immunization reduced the IgM PFC response to a T-dependent antigen (TNP-SRBC) by more than 60%. Inclusion of highly specific anti-C3 antibody in spleen cell cultures decreased the IgM PFC response by approximately 80%. In contrast, anti-C3 antibody had no effect on the T-dependent IgM response of CR- B cells. The residual PFC responses in cultures of unfractionated spleen cells treated with anti-C3 could be largely or completely accounted for by CR- B cells in the cultures. The effect of anti-C3 antibody was not due to cytotoxicity. These data collectively indicate that the effect of CoV on T-dependent antibody responses is due to decreased C3 in serum rather than to interaction of C receptors directly with CoV or with C3 cleavage products. They suggest the existence of at least two distinct pathways of T-B cooperation, one in which C3 is an obligatory participant and another in which it may be uninvolved.
T Cell-Mediated Cytotoxicity against Trinitrophenyl-Modified Cells: Effect of Glutaraldehyde Treatment on the Immungenicity and Antigenicity of Trinitrophenyl-Modified Cells
Authors: Forman James
Published: December 2022

Cells treated with low concentrations of glutaraldehyde for a 10-sec interval were unable to incorporate 3H-leucine into TCA precipitable protein, respond to H-2 allogeneic cells in mixed lymphocyte reactions (MLR) and cell-mediated lympholysis (CML) assays, or display capping of cell surface immunoglobulin (Ig) with a fluoresceinated anti-Ig reagent.

Such cells could stimulate and specifically block H-2 allogeneic CML activity but could not stimulate an H-2 allogeneic MLR response.

Trinitrobenzenesulfonic acid (TNBS) treated spleen cells were used to sensitize syngeneic splenocytes into displaying a cytotoxic effect against trinitrophenyl (TNP)-modified target cells. Treatment of the stimulator cells with glutaraldehyde immediately after modification with TNBS did not impair their immunogenic activity. Similar treatment of TNP-modified concanavalin A-stimulated lymphoblasts that were used as inhibitors in a CML cold target competition assay allowed such cells to retain their antigenicity. Cells treated with glutaraldehyde before TNP-modification, however, were not antigenic in the cold target competition assay.

These data are compatible with TNBS acting on plasma membrane molecules directly to cause cells to be antigenic and immunogenic in the CML assay rather than affecting internal cellular components.
An Evaluation of Elution Techniques in the Study of Immune Complex Glomerulonephritis
Authors: Woodroffe Andrew J. , Wilson Curtis B.
Published: December 2022

Antigen and antibody from glomerular immune complex deposits in rabbits with experimental bovine serum albumin-(BSA) induced chronic serum sickness (CSS) were quantitated in eluates from kidneys in which a portion of the antigen and antibody had been radiolabeled. The largest quantities of 125I BSA eluted with 1 M proprionic acid at pH 2.7 (86%) and 0.1 M borate buffer at pH 11.25 (80%). However, these buffers yielded less functional anti-BSA antibody than 0.02 M citrate buffer at pH 3.2 (344 µg/g kidney). Citrate buffer-eluted anti-BSA antibody was reactive in immunodiffusion, immunofluorescence, and radiolabeled BSA binding test systems, but complement fixation was impaired relative to chaotropic ion-eluted antibody. It was found that up to 75% of the eluted antibody was lost to further study by recombination with eluted BSA. This could be prevented by fractionation of the dissociated eluate before neutralization. IgG fractionated eluates were successfully fluorescein conjugated or radiolabeled for use as reagents. Elution of cryostat sections of CSS kidney was also studied; BSA, IgG, and complement (C3) eluted in parallel, and sub-microgram quantities of anti-BSA antibody were recovered.
Specificity of Rat Xenogeneic Cell-Mediated Cytolysis for the Products of the K and D Loci of the Mouse H-2 Complex
Authors: Burakoff Steven J. , Dorf Martin E. , Benacerraf Baruj
Published: December 2022

Lewis rat lymph node (LN) cells, when cultured with irradiated mouse spleen cells, develop cytotoxic effectors that were shown to be specific for the sensitizing mouse strain. Alloantisera directed to the products of the K and D loci of the target inhibited cytolysis whereas an antiserum to the I region gene products did not inhibit cytolysis even though LPS-induced spleen blasts were used as targets. Thus, it appears that even when effector cells from another species are generated to mouse spleen cells, the gene products of the K and D loci continue to play a predominant role.
Studies on Cellular Inhibition and Serum-Blocking Factors in 28 Human Patients Given Marrow Grafts from HLA Identical Siblings
Authors: Tsoi Mang-So , Storb Rainer , Weiden Paul L. , Thomas E. Donnall
Published: December 2022

Fifteen patients with aplastic anemia and 13 with acute leukemia were studied 36 to 1547 days after treatment with high-dose cyclophosphamide and/or total-body irradiation and marrow transplantation from HLA identical siblings. Peripheral blood lymphocytes from patients and normals (marrow donors and healthy unrelated individuals) were tested for cell inhibition (CI) of cultured skin fibroblasts from both patients and donors by using the microcytotoxicity assay. In addition, blocking of CI by factors in patient serum was studied.

Three groups of patients were studied. Patients in group I were stable long-term survivors without evidence of graft-vs-host diseases (GVHD) between 250 to 1547 days postgrafting. Patients in group II were short-term survivors with or without acute GVHD between 36 and 144 days postgrafting. Patients in group III had chronic GVHD either at the time of testing or developed chronic GVHD subsequent to CI testing between days 61 and 960 postgrafting. Eleven of 14 patients in group I showed absence of both CI and serum blocking and three showed CI and blocking. Patients in group II without acute GVHD showed absence of CI and serum blocking on three occasions, presence of CI and blocking on four occasions, and CI without blocking on three occasions. Patients in group II with acute GVHD showed absence of CI on one occasion and presence of CI and blocking on three occasions. Patients in group III showed absence of CI and blocking on seven occasions and CI without blocking on two occasions.

These results suggest that the maintenance of stable graft-host tolerance in long-term survivors after marrow grafting from HLA identical donors does not depend on the presence of serum-blocking factors. Short-term survivors with and without GVHD showed a spectrum of in vitro reactivity with 50% of the patients showing serum-blocking factors, and these results did not appear to be correlated with presence or absence of acute GVHD. Finally, results of the microcytotoxicity assays failed to provide insight into the mechanism of chronic GVHD.
Inherited Polymorphism of Guinea Pig Factor B and C4: Evidence for Genetic Linkage between the C4 and Bf Loci
Authors: Bitter-Suermann Dieter , Krönke Martin , Brade Volker , Hadding Ulrich
Published: December 2022

An inherited structural polymorphism of the guinea pig complement protein C4 and of factor B of the alternative pathway was demonstrated by use of high voltage agarose electrophoresis combined with a subsequent immunofixation technique. No polymorphism of guinea pig C3 could be shown. Three common allelic genes were proven to code for C4-S, C4-Sl, and C4-F, respectively. Two common allelic genes are responsible for the Bf-F and Bf-S phenotype expression. A strong linkage disequilibrium between both C4 and Bf was demonstrated.
Enhancement of Carrier-Mediated Transport after Immunologic Activation of Peritoneal Macrophages
Authors: Bonventre Peter F. , Straus David , Baughn Robert E. , Imhoff John
Published: December 2022

Immunologically activated peritoneal macrophages from inbred mice and Hartley strain guinea pigs demonstrate a markedly greater than normal transport of 2-deoxy-d-glucose and l-leucine. The degree of nutrilite transport enhancement was greatest when animals were injected with the appropriate eliciting antigens before harvesting and also, if antigen was included in the tissue culture medium during the initial hours of in vitro culture. Enhanced hexose and amino acid uptake could also be achieved by exposure of macrophages from nonimmunized animals for 48 hr to supernatants of sensitized splenic lymphocyte cultures incubated with specific antigens. The animal systems in which this pheomenon was observed included CBA/J and C57BL/6J mice immunized with Staphylococcus aureus or sub-lethal doses of Listeria monocytogens, and the Hartley strain, albino guinea pig immunized with S. aureus or BCG. In all cases, immunization resulted in a state of delayed hypersensitivity as measured by skin testing or footpad swelling. Splenic cell supernatants contained lymphokines as detected by the presence of macrophage inhibitory factor (MIF), and by the supernatants' capacity to stimulate incorporation of 14C-glucosamine by macrophages in vitro. No increase of glucose or leucine transport by macrophages was observed in the absence of appropriate antigen stimulation in vivo or in vitro.

We previously showed that a phagocytic stimulus results in a significant increase in hexose transport by normal macrophages; leucine transport by these same cells was unaltered after phagocytosis. In contrast, immunologically activated macrophages do not transport measurably more 2-deoxy-d-glucose after particle ingestion; activation or the phagocytic stimulus enhance 2-deoxy-D-glucose uptake to approximately the same extent. Analysis of nutrilite transport kinetics revealed that immunologic activation of macrophages increases the initial velocity (V1) and Vmax but does not change the Km values of hexose or amino acid transport. The kinetics of transport by the immunologically activated macrophages do not change measurably after phagocytosis. We conclude that either immunological activation or phagocytosis results in augmented 2-deoxy-D-glucose transport via identical or related mechanisms and that transport of the sugar can't be increased above that level induced by either event. The reasons why immunologic activation increases both glucose and leucine transport but phagocytosis increases only the former are not yet understood.
Mechanisms of Humoral Cytotoxicity Testing: The Role of Rabbit Serum, Xenoantibody, and Human IgM
Authors: Quindlen Eugene A. , Wood William C. , Kornblith Paul L.
Published: December 2022

The reproducibility and sensitivity of a humoral microcytotoxicity assay which detects anti-astrocytoma antibodies depends on a consistent, effective complement preparation. Rabbit serum mixed 1/1 with human cord serum proved to be an ideal preparation with full complementarity and low nonspecific cytotoxicity. Human cord serum provided consistent small amounts of human IgM and did not contribute additional complement activity. The inhibition of xenoantibody by human IgM is the critical mechanism which determines the efficacy of the complement. Human IgM in a final concentration of 10 mg/dl in the rabbit serum produced a preparation with 10 to 15% nonspecific cytotoxicity against astrocytoma target cells. This residual nonspecific cytotoxicity serves a catalytic function and is necessary for the detection of weak specific antibodies.