bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunologic Properties of Bacterial Lipopolysaccharide (LPS)
Authors: Skidmore Barry J. , Chiller Jacques M. , Weigle William O.
Published: December 2022

Lymphoid cells obtained from the C3H/HeJ mouse strain respond abnormally to LPS in vitro, as shown by the fact that they are unable to make a mitogenic response to some LPS preparations and make only a low mitogenic response to other LPS preparations. In contrast, cells from a closely related C3H substrain, the C3H/St, are highly responsive to both types of LPS preparations. Experiments were carried out to determine the cellular basis of these genetically determined LPS response differences. This question was approached by studying the mitogenic response to LPS in cultures containing mixtures of various combinations of B cells, T cells, and macrophages from C3H/HeJ and C3H/St mice. Experiments utilizing an LPS preparation to which the C3H/HeJ is totally unresponsive (negative LPS) revealed, first, that either spleen cells, or partially purified T cells and/or macrophages obtained from C3H/St, could not restore the ability of C3H/HeJ spleen cells to respond to LPS, indicating that the C3H/HeJ is not deficient in an LPS-specific helper cell population which may be required for mitogenesis. Secondly, the addition of either spleen cells or partially purified T cells or macrophages from the C3H/HeJ to spleen cells from the C3H/St did not inhibit the mitogenic response to LPS, suggesting that the presence of suppressor cell activity is also not involved. Experiments analogous to those described, except utilizing another LPS preparation to which the C3H/HeJ is partially responsive (positive LPS), also failed to demonstrate reconstitutive or suppressive effects when C3H/HeJ and C3H/St spleen cells were admixed.

The results obtained indicate that the defect in the C3H/HeJ mouse strain that limits its responsiveness to positive LPS and which renders it totally unresponsive to negative LPS appears to be an intrinsic defect in the capacity of B cells to react to the mitogenic stimulus of LPS.
Effect of Selective Complement Deficiency on the Rate of Neutralization of Enveloped Viruses by Human Sera
Authors: Leddy John P. , Simons Ruth L. , Douglas R. Gordon
Published: December 2022

The capacity of human sera genetically deficient in selective complement (C) components to enhance neutralization of enveloped viruses was examined by kinetic plaque reduction assays. Vaccinia virus, a DNA virus, and vesicular stomatitis virus (VSV), an RNA virus, were studied. Exogenous rabbit or human antibody to vaccinia virus, and guinea pig or human antibody to VSV were provided in limiting, C-dependent concentrations. IgG antibodies predominated in most of the antisera employed.

C5-deficient and C6-deficient human sera consistently supported normal rates of neutralization of either virus; this effect was heat-labile. C4-deficient human serum did not exceed heat-inactivated serum in any neutralization assay. C1r-deficient serum displayed slight heat-labile neutralizing capacity against vaccinia but none against VSV. C2- and C3-deficient sera consistently exhibited measurable but clearly subnormal rates of neutralization. Two fresh agammaglobulinemic sera failed to inactivate either virus in the absence of added antibody.

These results confirm and extend earlier evidence, based on neutralization of herpes simplex and Newcastle disease viruses in the presence of early (IgM) antibody and functionally pure guinea pig C components or C-deficient animal sera, that the late-acting components C5–C9 are not required for C-dependent neutralization. Data on four enveloped viruses now agree that this function is mediated by C1–C3, although C1 plus C4 appear to have some neutralizing capacity. This requirement for C1–C3 is overcome, however, in the presence of higher antibody concentrations, suggesting that the contribution of the C system to viral neutralization in vivo may be chiefly in the early phase of infection when antibody is limited.
Cell-Mediated Cytotoxicity Against Measles Virus in SSPE
Authors: Kreth Hans W. , ter Meulen Volker
Published: December 2022

In patients with subacute sclerosing panencephalitis (SSPE) virus persistency has been explained by a blocking factor of specific cell-mediated immunity in SSPE serum and cerebrospinal fluid (CSF). In the present study the effect of SSPE serum and CSF on cell-mediated cytotoxicity against 51Cr-labeled allogenic measles virus-infected target cells was tested.

Pretreatment of SSPE peripheral lymphoid cells by serum or CSF neither blocked nor increased measles-specific cytotoxicity when cells were subsequently tested in FCS containing tissue culture medium. However, killing activity was always enhanced after pretreatment of target cells or direct addition of serum or CSF to the cytotoxic assay. Enhancement was also observed with non-SSPE-derived measles antibodies. The effect was dependent on measles antibody concentrations. These results suggest that antibody-dependent killing by K cells is intact in patients with SSPE. It is possible that virus-infected brain cells are protected from an immune attack in vivo by antibody-induced antigenic modulation.
Generation of Human Plaque-Forming Cells in Culture: Tissue Distribution, Antigenic and Cellular Requirements
Authors: Dosch Hans-Michael , Gelfand Erwin W.
Published: December 2022

A system for the induction of specific, hemolytic plaque-forming cells from normal human lymphocytes in vitro (HcPFC) has been established and cells from various normal lymphoid tissues have been investigated. Normal values for anti-SRBC HcPFC responses in cultures of 107 Ficoll-Hypaque separated lymphocytes range from 2000 (bone marrow) to 7000 (spleen) and 15,000 (tonsillar and peripheral blood lymphocytes). HcPFC responses to ovalbumin were lower by a factor of 2 to 4. Anti-SRBC as well as anti-ovalbumin responses required the cooperation of T lymphocytes and IgM-bearing B lymphocytes and the magnitude of the response was antigen dose dependent. Addition of adherent cells as well as of 2-mercaptoethanol enhanced the response. On the basis of the data obtained in experiments examining the role of B and T lymphocytes, a tentative model of cellular interaction has been postulated, suggesting a major role for antigen concentration in the modulation of the response via reactive T lymphocytes.
Affinity in Radioimmunoassay of Antibody Cross-Reactive with Carcinoembryonic Antigen (CEA) and Colon Carcinoma Antigen-III (CCA-III)
Authors: Primus F. James , Newman Edward S. , Hansen Hans J.
Published: December 2022

Several antisera raised against purified carcinoembryonic antigen (CEA) were evaluated for their content of antibody cross-reactive with colon carcinoma antigen-III (CCA-III). All antisera gave a reaction of partial identity between CEA and CCA-III and demonstrated a high titer in CEA radioimmunoassay (RIA). Between 50 and 70% of the CEA RIA activity was removed, however, by absorption with soluble CCA-III or adsorption onto CCA-III-containing immunoadsorbents. Immunoadsorbent retained antibody gave a line of complete identity between CEA and CCA-III. Purified CCA-III (2 µg) only partially depressed CEA binding by this common site antibody, whereas nanogram quantities of CCA-III inhibited the reaction between specific CCA-III antibody and radioiodinated CCA-III. In addition, low levels of CEA were equally effective in depressing CEA binding by the common site or CEA-specific antibody. The higher affinity in RIA of the common site antibody for CEA over CCA-III suggests that the common determinant expressed on CEA is stereochemically different from that on CCA-III. The results further demonstrate that interference by plasma CCA-III is not a significant factor in the measurement of CEA by RIA.
Studies on the Mechanism of Lymphocyte-Mediated Cytolysis
Authors: Kuppers Rudolf C. , Henney Christopher S.
Published: December 2022

The relationship between antigen recognition and lytic expression by killer T cells was studied by co-culturing two effector cell populations. When antigen recognition was bidirectional (e.g., b anti-d cells cultured with d anti-b) there was a loss of lytic activity in both populations. In contrast, when antigen recognition was unidirectional (e.g., a anti-d co-cultured with d anti-b) then the loss of lytic activity only occurred in that direction; i.e., there was a marked decline in the d anti-b activity but no change in the a anti-d population.

These studies suggest: i) that mere proximity to a killer cell does not lead to target cell death; ii) that accommodation of the T cell's antigen receptor is necessary for the cell to express its lytic potential; and iii) there is direct linkage between the T cell's antigen receptor site and its killing mechanism.
Cutaneous Basophil Responses in Neonatal Guinea Pigs: Active Immunization, Hapten Specific Transfer with Small Amounts of Serum, and Preferential Elicitation with Phytohemagglutinin Skin Testing
Authors: Haynes J. David , Askenase Philip W.
Published: December 2022

Previous studies have demonstrated a failure to elicit delayed hypersensitivity skin responses in neonatal guinea pigs despite apparent immunologic competence of their lymphocytes and monocytes. The ability of neonatal guinea pigs to manifest cutaneous basophil hypersensitivity (CBH) responses was examined. Neonates were competent to express macroscopic and microscopic aspects of these delayed reactions. Strong cutaneous basophil responses were elicited in newborns (0 to 2 days old) in a hapten-specific manner after intravenous transfer of small amounts of immune serum (0.5 ml) obtained from adult donors. Also, neonatal guinea pigs actively immunized at birth with hapten-carrier conjugates emulsified with incomplete Freund's adjuvant (IFA) and skin tested when 1 week old had 24-hr hapten-specific cutaneous basophil reactions and CBH reactions to the carrier protein as well. Compared with adult CBH reactions, neonatal responses had equal concentrations of basophils, fewer mononuclear cells, less macroscopic erythema, and almost no induration. Hapten-specific CBH reactions also contained significant infiltrates of eosinophils which were more prominent in the ear skin vs flank skin and in neonates vs adults.

Immunization with complete Freund's adjuvant (CFA) resulted in a marked difference between adults and neonates; adult PPD reactions were quite indurated and contained many more mononuclear cells and few basophils, whereas neonatal PPD reactions were flat, erythematous, nonindurated, and contained relatively few mononuclear cells and more basophils. Thus, neonatal tuberculin reactions elicited by PPD in animals immunized with CFA were examples of CBH. Skin testing nonimmune guinea pigs with phytohemagglutinin (PHA) also revealed marked differences in 24-hr cutaneous reactions between adults and newborns. Adults had indurated and erythematous reactions which contained approximately 20% basophils and 80% mononuclear cells, whereas similar PHA skin tests in newborns elicited small macroscopic reactions, which microscopically showed large infiltrates containing approximately 80% basophils and 20% mononuclear cells.

It was concluded that neonatal guinea pigs were not only competent to manifest basophil-containing delayed-type reactions, but that cutaneous basophil responses were preferentially elicited in these animals under a variety of circumstances. These results underline the fact that basophil accumulations are one aspect of delayed skin test responses and that the regulation of the arrival of these cells in neonates is different from that in adult guinea pigs.
Role of the Immune Response in Recovery from Sindbis Virus Encephalitis in Mice
Authors: Griffin Diane E. , Johnson Richard T.
Published: December 2022

The role of the immune response in recovery from acute encephalitis was studied by using a neuroadapted strain of Sindbis virus. Intracerebral inoculation of this virus caused fatal disease in weanling mice. Passive transfer of homologous immune serum protected mice, but transfer of immune spleen cells and lymph node cells failed to alter mortality. Transfers were made 24 hr after intracerebral infection of the recipient, when central nervous system infection was established. The protective factor in serum was present 7 days after donor infection, was specific for Sindbis virus, and appeared to be IgG; however, the protection did not correlate with virus-neutralizing activity.
Isolation of Mouse C-Reactive Protein from Liver and Serum
Authors: Bodmer Barbara , Siboo Russell
Published: December 2022

Purified proteins have been isolated from the sera and livers of mice. These proteins are antigenically identical but share antigenic determinants in common with human C-reactive protein and do not share antigenic determinants in common with mouse or human immunoglobulins. The proteins interact with C-polysaccharide, are precipitated by calcium ions, migrate electrophoretically with gamma mobilities, and have isoelectric points of 4.8 and 5.62. Since these properties are characteristic of C-reactive protein of man, monkey, rabbit and dog, the pure proteins isolated from mice are designated mouse C-reactive protein.
Human Monocyte-Macrophage-Mediated Antibody-Dependent Cytotoxicity to Herpes Simplex Virus-Infected Cells
Authors: Kohl Steve , Starr Stuart E. , Oleske James M. , Shore Steven L. , Ashman Robert B. , Nahmias André J.
Published: December 2022

Human peripheral blood mononuclear cells which mediate antibody-dependent cellular cytotoxicity (ADCC) against herpes simplex virus- (HSV) infected target cells consist of both adherent (MA) and nonadherent (MNA) effector cell populations. These two cell populations can be distinguished by their different phagocytic properties and morphologic appearance, their requirement for antibody in the ADCC reaction, and the rapidity with which they lyse target cells in the presence of immune serum. The MA cells are predominantly phagocytic and have the morphologic characteristics of monocyte-macrophages, whereas the MNA cells are nonphagocytic and appear to be small to medium-sized lymphocytes. Optimal expression of ADCC by MA cells requires higher concentrations of immune serum than does MNA cell-mediated ADCC. MA-mediated cell killing is first detectable by 8 hr and reaches completion after 24 hr of incubation. In contrast, MNA-mediated ADCC produces target cell damage by 2 hr and reaches completion at 8 hr of incubation. Unlike MNA effector cells, the MA effector cells are profoundly inhibited after preincubation with either latex or silica particles. The HSV immune status of the donor had no effect on the ability of either cell population to mediate ADCC.

These data demonstrate the participation of both nonadherent mononuclear cells, presumably K cells, and monocyte-macrophages, in ADCC directed against HSV-infected target cells.
Mitogen Activation of Human Chronic Lymphatic Leukemia Cells
Authors: Maino Vernon C. , Kurnick James T. , Kubo Ralph T. , Grey Howard M.
Published: December 2022

The response of CLL (chronic lymphatic leukemia) lymphocytes to PHA, PWM, and Con A with respect to changes in surface markers and synthesis and secretion of immunoglobulin were examined. After PHA stimulation the percentage of cells bearing readily detectable surface immunoglobulin (SmIg) diminished rapidly whereas cells forming rosettes with sheep erythrocytes (E-rosettes) increased from <1% to 30 to 50%. The great majority of blast-transformed cells were E-rosette-positive cells with a small population of SmIg-positive blast cells also observed. Ig production in four of seven CLL lymphocyte populations was increased 2.5 to greater than 40-fold after 4 to 6 days of culture in the presence of PHA. In contrast, pokeweed mitogen did not affect Ig synthesis. Furthermore, the Ig secreted into the culture supernatant fluids from seven of eight CLL cases examined consisted almost entirely of free light chain molecules. In contrast, the cell lysates contained a significant proportion of intact Ig molecules. These results indicate that CLL cells can, under certain circumstances, be stimulated to synthesize and secrete increased amounts of Ig, but that there is a basic defect in the biosynthetic mechanism of these cells which result in the secretion of free light chains rather than intact immunoglobulin molecules.
Characterization of IgG Receptors of the Fetal Rabbit Yolk SAC Membrane: Localization to Subcellular Fraction and Effects of Chemical Agents and Enzymes on Binding
Authors: Hillman Kathleen , Schlamowitz Max , Shaw Anita R.
Published: December 2022

Previous studies have established the existence of IgG receptors on the endodermal cells of the fetal rabbit yolk sac membrane (YSM). The present study partially characterizes these cell-associated receptors. The specific binding of rabbit IgG (IgGR) to freshly prepared cell homogenates, nuclei-free brush border preparations, and plasma membrane-rich fractions confirms that receptor function is associated with the endodermal cell, and indicates that this function is localized on its apical brush border, specifically on its plasma membrane. The protein nature of the receptor is demonstrated by the loss of specific binding capacity after treatment of formalin-fixed YSM with papain and trypsin. Evidence has also been adduced which indicates that membrane carbohydrate is not involved in receptor function. Thus, treatment of YSM with neuraminidase, β-galactosidase, mixed glycosidases, as well as oxidation of YSM with periodate all are without effect on its capacity to bind IgGR. The interaction of IgGR with the receptor elements of formalin-fixed YSM is partially ionic in character. NaCl reversibly inhibits binding of IgGR by 60%. Divalent ions such as Ca++ are not involved in this receptor-ligand interaction since EDTA-treated YSM binds IgGR to the same extent as do controls. Receptor material on fixed YSM resists extraction by non-ionic detergents, deoxycholate, and chaotropic agents.
Age-Related Decrease in Mouse T Cell Progenitors
Authors: Tyan Marvin L.
Published: December 2022

Mice were given a lethal dose of whole-body γ-radiation and injected with 105 or 106 marrow cells from 10- to 143-week-old syngeneic donors. Nine days later, colony-forming units (CFU) were counted in the spleens of mice given 105 cells, and 15 to 21 days after irradiation thymus weights and in some experiments 3H-thymidine uptake or total thymic cellularity were determined in the recipients of 106 cells. It was found that in the majority of mouse strains studied there were no significant changes with age in marrow CFU. In contrast, thymic regeneration was significantly impaired when the recipients received marrow cells from donors 100 weeks of age or older. These observations and results obtained in dose-response and time-course studies are best explained by an age-related decrease in marrow T cell progenitors; however, certain findings suggest that in addition the proliferative capacity of these stem cells may at times be moderately impaired.
Heterologous Antigenic Stimulation in Induction of Delayed Hypersensitivity
Authors: Brunda Michael J. , Raffel Sidney
Published: December 2022

An influence of a delayed hypersensitive reaction to a primary antigen on the induction of delayed hypersensitivity to a second unrelated antigen was observed in guinea pigs immunized with azobenzenearsonate-N-acetyl-L-tyrosine (ABAT), and injected intradermally 3 weeks later with a mixture of ABAT and secondary antigen. Animals so treated developed delayed hypersensitivity to sheep red blood cells (SRBC) or Type II pneumococcal polysaccharide as secondary antigens, as measured by skin test reactivity and inhibition of macrophage migration, whereas ABAT unsensitized control groups did not. However, attempts to induce delayed reactivity to proteins as secondary antigens were unsuccessful. The injection of secondary antigen into a mineral oil-induced inflammatory lesion did not induce delayed hypersensitivity, suggesting that specific reactivity to ABAT is a prerequisite for heterologous induction. Possible mechanisms for the observed phenomenon, including a role for macrophages, are discussed.
Immunoregulatory Properties of Fractions from Human Pregnancy Urine: Evidence That Human Chorionic Gonadotropin is Not Responsible
Authors: Muchmore A. V. , Blaese R. M.
Published: December 2022

The immunologically privileged position of the histoincompatible fetus and placenta is a striking example of a physiologic immunoregulatory mechanism. This study was designed to examine the effects of human chorionic gonadotropin (HCG) on the recognitive proliferative phase and the cytotoxic effector phase of in vitro cell-mediated immune responsiveness, since HCG has previously been reported to be immunosuppressive in vitro and in vivo. Commercial preparations of HCG were found to be potent inhibitors of lymphocyte proliferative responses to nonspecific mitogens like phytohemagglutinin (PHA), specific antigens such as streptolysin-O (SLO), and allogeneic cells as measured in the one-way mixed leukocyte response. Cytotoxic effector function of lymphocytes as measured by antibody-dependent cellular cytotoxicity (ADCC) and mitogen-induced cellular cytotoxicity were also markedly inhibited by these preparations. However, the 50% inhibitory concentration varied widely from lot to lot of these commercial materials. After dialysis, a portion of the inhibitory activity was lost from some but not all HCG lots. The dialysate from those lots with diminished activity was found to be immunosuppressive in vitro but contained no HCG detectable by radioimmunoassay. Following dialysis, the immunosuppressive activity of the various HCG lots remained variable and correlated poorly with values for HCG obtained by a double antibody radioimmunoassay. HCG preparations purified to a homogeneity sufficient for amino acid sequence were found to be only minimally immunosuppressive to the in vitro PHA response and had almost no effect on proliferative responses to antigens and allogeneic cells.

These data do not support the concept of a primary immunoregulatory role for HCG, but they suggest that other uncharacterized compounds partially co-purified from pregnant urine along with HCG may have such immunoregulatory activity. Further characterization and identification of this immunoregulatory material(s) is essential, since it appears to have many of the properties of an ideal immunosuppressive compound: a) nontoxicity, b) ready reversibility, c) activity at very low concentration, and d) activity on a broad range of cellular immune functions.
Distribution of Alloantigens on Human Fc Receptor-Bearing Lymphocytes: The Presence of B Cell Alloantigens on sIg-Positive but Not sIg-Negative Lymphocytes
Authors: Nelson David L. , Strober Warren , Abelson Leslie D. , Bundy Bonita M. , Mann Dean L.
Published: December 2022

Human peripheral blood lymphocyte subpopulations were analyzed for the presence of B cell alloantigens with a microcytotoxicity assay. B cell alloantigens were found exclusively on sIg-positive lymphocytes and were not present on sIg-negative, Fc receptor-bearing lymphocytes or sIg-negative, Fc receptor-negative T lymphocytes.
Effect of Concanavalin A on Lymphocyte Interactions Involved in the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Markham Richard B. , Stashak Philip W. , Prescott Benjamin , Amsbaugh Diana F. , Baker Phillip J.
Published: December 2022

Concanavalin A (Con A) administered at the time of immunization induces suppression of the in vivo splenic plaque-forming cell (PFC) response to type III pneumococcal polysaccharide (SSS-III). As with low dose paralysis of the PFC response to SSS-III, Con A-induced suppression could not be demonstrated in congenitally athymic (nu/nu) mice and could be eliminated partially by treatment with anti-lymphocyte serum (ALS). The kinetics for Con A-induced suppression paralleled those for low dose paralysis of the antibody response to SSS-III. These findings support the view that Con A-induced suppression is produced in vivo by suppressor T cells and that this form of suppression shares with low dose paralysis a common pathway through which suppression is mediated.
Inhibition of Antibody-Dependent Cellular Cytotoxicity by Protein A from Staphylococcus Aureus
Authors: Rosenblatt Joe , Zeltzer Paul M. , Portaro Joe , Seeger Robert C.
Published: December 2022

Protein A, a cell wall constituent of several strains of Staphylococcus aureus, binds strongly to the Fc portion of immunoglobulins. This investigation demonstrated that such binding can inhibit antibody-dependent cellular cytotoxicity (ADCC). The degree to which ADCC was inhibited depended upon the relative concentrations of protein A and anti-target cell antiserum. Protein A also inhibited the formation of rosettes between antibody-coated sheep red blood cells and lymphoid cells with Fc receptors. We, therefore, conclude that protein A inhibits ADCC by preventing the binding of antibody-coated target cells to Fc receptors on cytotoxic effector cells.
Ia Antigens on Murine Thymocytes: Detection by Cellular Radioimmunoassay
Authors: Elkins William L. , Klinman Norman R. , Mayol Renato
Published: December 2022

An indirect radioimmunoassay was used to detect Ia antigenic determinants at the cell surface of thymic and splenic cells. An A.TH anti-A.TL serum was used as a source of anti-Ia antibodies which could bind to H-2a or H-2k target cells. Bound alloantibody was then detected with 125I anti-mouse IgG1 antibody. This assay system revealed Ia antigens on thymic T cells which are not usually detectable by cytotoxicity assays, and has the potential for quantitative measurement of such determinants on cells. Critical adsorption studies were performed to confirm that Ia antigens were in fact the targets of the alloantiserum tested.
Interactions between Human Lymphocytes and Paramyxovirus-Infected Cells: Adsorption and Cytotoxicity
Authors: Härfast Bengt , Andersson Torbjörn , Stejskal Vera , Perlmann Peter
Published: December 2022

The capacity of human lymphocytes to adhere to paramyxovirus-infected monolayers and their capacity to kill paramyxovirus-infected cells was investigated. A large fraction of human lymphocytes was found to adhere firmly to the paramyxovirus-infected monolayers. Preadsorption of lymphocytes on mumps virus-infected cells impaired their adsorption to a second cell monolayer of the same type. The cytotoxic activity of lymphocytes against mumps virus-infected cells was also reduced after preadsorption on mumps virus- or Newcastle disease virus-infected (NDV) cell monolayers. Exposure of lymphocytes to trypsin did not significantly decrease either adsorption or cytotoxicity. Pretreatment of lymphocytes with neuraminidase (NANase) partly inhibited adsorption whereas cytotoxicity was not decreased. Cell fractionation experiments after rosetting of the lymphocytes with sheep erythrocytes (E) indicated that T cells were equally or better adsorbed than “non-T” cells. Taken together with previous experiments which showed that the majority of T lymphocytes are not cytotoxic against mumps virus-infected cells these results suggest that adherence of lymphocytes to infected cells and cytotoxicity may be unrelated phenomena.
Stem Cell Lines from Old Immunodeficient Donors Give Normal Responses in Young Recipients
Authors: Harrison David E. , Astle Clinton M. , Doubleday John W.
Published: December 2022

Two different immune responses were compared in spleen cells obtained from old and young CBA/HT6J mice. Spleen cells from old mice (23 to 33 months) responded about half as well as did spleen cells from young mice (4 to 10 months) in the adoptive transfer anti-sheep red blood cell (SRBC) plaque-forming assay, and caused slightly less than half the uptake of tritiated thymidine in response to phytohemagglutinin (PHA) in vitro.

Marrow stem cells from some of the old and young mice whose splenic immune responses were tested were transplanted into irradiated young CBA/CaJ recipients. Seven to 17 weeks later these same immune responses were tested in the spleen cells of these young recipients, and the T6 chromosome marker was used to identify donor cells. Old animals' responses varied greatly, perhaps due to suppressing cells or factors in some individuals. Therefore, cells were never pooled and the responses of recipients were compared to the responses of the donor whose marrow had populated them. The response for a particular old donor, or for the recipients of its stem cells, was divided by the response for the young control used with that donor, or for its stem cell recipients. This was called the old/young ratio.

With original donors with an old/young ratio for the SRBC response of (mean ± S.D.) 0.35 ± 0.14, the old/young ratio for that same response in the recipients was significantly improved to 1.26 ± 0.71. In original donors with an old/young ratio for the PHA response of 0.44 ± 0.17, the old/young ratio in the recipients improved significantly to 0.86 ± 0.27. Thus, little or none of the decline with age in these immune responses was intrinsic to the old lymphoid stem cells.
Immunosuppressive Activity or IRA on the Plaque-Forming Response to SRBC in Vitro: Reversal with Educated T Cells
Authors: Badger Alison M. , Merluzzi Vincent J. , Mannick John A. , Cooperband Sidney R.
Published: December 2022

The α-globulin-rich fraction of Cohn Fraction IV, designated IRA, suppresses the in vitro antibody response to sheep red blood cells (SRBC) without cytotoxicity. IRA was effective if added before or up to 24 hr after antigen exposure. The suppression could be reversed after 24-hr treatment by washing the cells two to three times; after 48 hr of IRA treatment, however, suppression could only be partially reversed. The addition of a population of thymus-derived cells educated to the antigen SRBC could effectively reverse the IRA-induced inhibition of antibody production, whereas BSA-educated T cells could not.
A Human Alloantiserum Precipitates Ia-Like Molecules from Chronic Lymphocytic Leukemia Cells
Authors: Nilsson Sten , Schwartz Benjamin D. , Waxdal Myron , Green Ira , Cullen Susan E. , Mann Dean L.
Published: December 2022

Alloantigens specific for human B lymphocytes can be identified with selected antisera. These antigens have similarities to murine Ia antigens in that they are found on human B lymphocytes and are controlled by genes linked to genes controlling HLA. Chronic lymphocytic leukemia cells bearing B cell antigens were labeled with 3H leucine and the membrane components reacting with the B cell antisera isolated by immunoprecipitation. These membrane components had m.w. of 33,000 and 24,000 daltons similar to the murine Ia antigens. The results complete the homology of murine Ia and human B cell alloantigens.
Genetics of Cell-Mediated Lympholysis in Man
Authors: Geha Raif S. , Malakian Artin , Geha Orietta , Yunis Edmond
Published: December 2022

Cell-mediated lympholysis (CML) was studied in a family containing two siblings in whom genetic recombination had occurred in the HLA complex. In one sibling, recombination occurred between the HLA-A locus and the HLA-B locus. In the second sibling recombination occurred between the HLA-B locus and the HLA-D locus.

Strong CML activity was generated in mixed lymphocyte cultures (MLC) when stimulator and responder cells differed in HLA-A, B, and D antigens. MLC involving HLA-D differences alone did not generate CML. Weak, but definite CML activity was generated during MLC with cells differing at HLA-A and HLA-B but sharing HLA-D.

HLA-B antigens were good targets for lysis in all combinations studied. HLA-A antigens were poor targets in some but not in all combinations. However, combinations where HLA-A antigens seemed to be good targets could have involved HLA-B differences due to polymorphism of HLA-B7 antigens each inherited from a different parent. HLA-D antigens did not serve as targets for lysis.

In three cell experiments, excellent CML activity was generated when responder cells were stimulated by HLA-D antigens and by HLA-A and B antigens present on separate stimulator cells.
Measurement of IgE on Human Basophils: Relation to Serum IgE and Anti-IgE-Induced Histamine Release
Authors: Conroy Mary Carol , Adkinson N. Franklin , Lichtenstein Lawrence M.
Published: December 2022

The number of IgE molecules bound to human basophils was calculated from direct measurements of the IgE dissociated after exposing leukocytes to pH 3.7 acetate buffer in the cold. In 18 donors studied, cell-bound IgE ranged from 4000 to 500,000 molecules/basophil and correlated with the serum IgE concentration (r = 0.89, p < 0.001) which ranged from 5 to 3,000 ng/ml. Sensitivity of these cells to anti-IgE was tested to explore the relationship between cell-bound IgE and the concentration of anti-IgE required for histamine release. Cells from some nonatopic donors (4000 to 100,000 IgE molecules/basophil) were as sensitive as cells from allergic donors (100,000 to 500,000 IgE molecules/basophil). Moreover, cells from donors having approximately the same cell-bound IgE concentration varied widely in their sensitivity to anti-IgE. We conclude that an intrinsic property of human basophils (“releasability”) is an important parameter in determining mediator release.
Hapten-Specific Leukocyte Migration Inhibition
Authors: McMaster Philip R. B. , Owens James D. , Asofsky Richard
Published: December 2022

Guinea pigs were immunized with dinitrophenyl-keyhole limpet hemocyanin (DNP-KLH) or with hemocyanin in complete Freund's adjuvant. The migration of oil-induced peritoneal exudate cells was measured in the presence and absence of ε-dinitrophenyl-lysine-tyrosinyl Ficoll (DNPL-F). When the cells came from animals immunized with DNP-KLH their migration was inhibited by DNPL-F. Control cells from animals immunized with KLH or not immunized migrated normally in the presence of DNPL-F.
The Biologic Activity of Affinity Cross-Linked Oligomers of Rabbit Immunoglobulin G as Tested by Antibody-Dependent Cell-Mediated Cytolysis (ADCC)
Authors: Hurwitz Esther , Zatz Marion M. , Segal David M.
Published: December 2022

The abilities of normal spleen cells and macrophage-like and lymphocyte-like tumor cells to serve as effectors in antibody-dependent cell-mediated cytolytic (ADCC) reactions were examined. It was observed that those cells which contained Fc receptors on their surfaces were effective mediators of ADCC whereas those not bearing Fc receptors were inactive. Oligomers of IgG were able to inhibit the ADCC reaction and, with the macrophage line, inhibition and binding closely paralleled each other. With the lymphocyte lines, a large degree of inhibition of ADCC occurred when only small amounts of oligomers were bound; with normal spleen cells inhibition occurred in the same range of oligomer concentrations required to inhibit the tumor-mediated ADCC reactions. The data suggest that the interaction between antibody-coated erythrocytes and effector cells is unexpectedly weak.
The Quantitative Production of Interferon by Mitogen-Stimulated Mouse Lymphocytes as a Function of Age and Its Effect on the Lymphocytes Proliferative Response
Authors: Heine Jochen W. , Adler William H.
Published: December 2022

Interferon production by mitogen-stimulated mouse spleen cells increases as the age of the cell donor increases and also varies with the strain of the cell donor.

Exogenous interferon added to mouse spleen cell cultures at dose levels known to be produced by the cells causes a reduction in the proliferative response of T cells to mitogen stimulation. Since the spleen cells from old mice respond poorly to mitogen stimulation, it may be possible that the interferon elaborated by these cells is adversely affecting the mitogen assay.
Regulatory Substances Produced by Lymphocytes
Authors: Namba Yuziro , Jegasothy Brian V. , Waksman Byron H.
Published: December 2022

The conditions necessary for production of inhibitor of DNA synthesis (IDS) by rat lymphocytes were investigated. In concanavalin A (Con A)-stimulated lymph node cell (LNC) cultures, IDS production was not detected in the culture supernatant during the first 24 hr, and it increased gradually after that to reach a maximum at 3 to 4 days. When the cells were pretreated with mitomycin C, IDS was not produced, suggesting that DNA synthesis of LNC or a LNC subpopulation is necessary for IDS production. In contrast, Con A-stimulated spleen cells produced a high level of IDS within 24 hr, and its production fell off sharply thereafter. Con A-stimulated rat thymocytes also produced IDS reaching a maximum at 2 to 3 days. However, thymus cells from rats treated with hydrocortisone 48 hr previously did not produce IDS. This finding implies that cortisol-sensitive (cortical) thymocytes are capable of producing IDS and cortisol-resistant (medullary) thymocytes are not. IDS production by lymphoblasts was proportional to cell number and unaffected either by cell density (1 to 10 × 106/ml) or by the concomitant presence of normal cells from spleen, lymph node, or thymus. Thus Con A-stimulated cells, after becoming blasts, appear to produce IDS automatically without affecting or being affected by other cells.

Both spleen and thymus cells from rats injected with a large dose of antigen (ovalbumin, 100 mg, i.p.) 24 hr in advance produced substantial amounts of IDS in culture within 24 hr in the absence of mitogen or additional antigen, but not the cells from rats injected with an immunizing dose (1 mg) of the same antigen. The cells producing IDS in the spleen were shown to be adherent to glass wool, and those in the thymus were partially so. IDS production by antigen-stimulated spleen cells was abrogated by injecting rats with bromodeoxyuridine (BUdR) at 0 and 12 hr after the ovalbumin. These findings suggest that a subpopulation of adherent spleen cells (possibly resembling cortical thymocytes), which begins to proliferate within a few hours after a large dose of systemic antigen, produces IDS. This may account for increased nonspecific suppressor activity observed at the same time.
Hypogammaglobulinemia in Experimental Myeloma: The Role of Suppressor Factors from Mononuclear Phagocytes
Authors: Krakauer Randall S. , Strober Warren , Waldmann Thomas A.
Published: December 2022

We have investigated the pathogenesis of the polyclonal hypogammaglobulinemia associated with BALB/c plasmacytomas TEPC-183 and SPQC-11 to gain insight into the hypogammaglobulinemia observed in human myeloma. With pokeweed mitogen-driven IgM biosynthesis by mouse splenocytes as the indicator system for suppression, we found that a protein extract of ascites cells obtained from these tumor-bearing animals could suppress immunoglobulin production, whereas like extracts from a non-suppressing plasmacytoma, modified RPC-5, caused no suppression in vitro. Extracts of tumor ascites depleted of mononuclear phagocytes by iron carbonyl treatment showed little suppressor activity. The active extract was not cytotoxic and contained no mycoplasma or common murine viruses. Furthermore, the active suppressor factor appears to be a low m.w. protein that is not affected by treatment with ribonuclease. These results and others are consistent with the idea that the hypogammaglobulinemia of myeloma is due to the formation of immunoregulatory macrophage-like cells which synthesize a suppressor substance.
Isolation and Enumeration of Peripheral Blood Monocytes
Authors: Sanderson Richard J. , Shepperdson Frances T. , Vatter Albert E. , Talmage David W.
Published: December 2022

Monocytes were isolated by counterflow centrifugation from human peripheral mononuclear cells obtained from whole blood by the Ficoll-Hypaque technique. Examination of volume spectra of the monocytes showed a bimodal distribution with peaks at 330 and 370 μ3. The upper peak contained 61.4 ± 3.3% of the total monocyte population, whereas the remainder were in the lower peak which overlaps the lymphocyte spectrum. The purity of the monocyte preparations from six normal adult donors, identified by peroxisomes observed by electron microscopy, was 90.4 ± 1.6%.
The Allogeneic Effect on Tumor Growth
Authors: Osborne David P. , Katz David H.
Published: December 2022

The allogeneic effect has been employed as a potent immunopotentiator in preventing the growth of a murine plasmacytoma and prolonging host survival. Parental BALB/c spleen cells were passively transferred to (BALB/c × A/J)F1 hybrid mice, who were then given a highly lethal dose of MOPC 315 plasmacytoma, a tumor of BALB/c origin. The resultant graft-vs-host reaction protected the recipient mice against growth of the tumor and significantly prolonged survival. This phenomenon was dependent upon the dose of BALB/c lymphoid cells employed, the route of administration, and the time interval between lymphoid cell transfer and tumor inoculation. A wide range of lymphoid cell doses and time intervals were effective, and repeated doses of allogeneic cells provided better protection than a single dose.
The Effect of Cyclophosphamide on the Specific Unresponsiveness to Skin Allografts Induced in Alstreated Mice Infused with Donor Bone Marrow
Authors: Wood Mary L. , Monaco Anthony P.
Published: December 2022

Specific unresponsiveness to skin allografts can be induced in ALS-treated mice by the injection of bone marrow from the graft-donor strain. Mice bearing long-term grafts in perfect condition have evidence of cell-mediated immunity against donor antigens and also serum-blocking factors. The effect of cyclophosphamide on graft prolongation was investigated in this model. Cyclophosphamide was given either before or after marrow. Cyclophosphamide given before marrow abrogated the enhancing effect of marrow possibly due to a depletion of antibody-forming cells. Cyclophosphamide given after marrow potentiated the effect of marrow probably due to the destruction of immunocompetent cells responding to the challenge of marrow.
Mitogenic Activity of Actinomyces Viscosus
Authors: Engel David , Clagett James , Page Roy , Williams Betsy
Published: December 2022

Actinomyces viscosus homogenate (AVIS) contains substance(s) which cause spleen cells from conventional and germfree mice to undergo increased DNA synthesis. This mitogenic effect is primarily on B cells since spleen cells from nude mice or T-depleted spleen cells from conventional mice respond as strongly as conventional (T + B) spleen cells. Mouse thymocytes do not respond mitogenically to AVIS. It is unlikely that the mitogenic activity is due to the presence of LPS, since A. viscosus is Gram-positive and is not known to have an LPS cell wall component. Also, AVIS is not inactivated by polymyxin B, as are some preparations of LPS, and C3H/HeJ mouse splenocytes respond strongly to AVIS but not to LPS. The activity is heat stable, is not lost upon dialysis, and is not affected by lysozyme. Mitogenic activity is partially lost when AVIS is digested with nonspecific bacterial protease or treated with metaperiodate. Sodium hydroxide treatment completely abolishes mitogenic activity. Actinomycotic lesions are characterized by a long-term inflammatory response involving a dense plasma cell infiltrate. We suggest that B cell mitogens from Actinomyces may play a role in the elicitation of the plasma cell component of these lesions.
In Vitro Reactivity of Splenic Lymphocytes from Normal and UV-Irradiated Mice Against Syngeneic UV-Induced Tumors
Authors: Fortner G. William , Kripke Margaret L.
Published: December 2022

Skin tumors induced in mice by chronic ultraviolet (UV) irradiation are highly antigenic and are frequently immunologically rejected upon transplantation to normal syngeneic recipients. In this study we characterized this immune response with an in vitro microcytotoxicity test. Cytotoxic activity was present in the spleen cells of mice given a single injection of syngeneic UV-induced fibrosarcoma cells. After removal of adherent spleen cells, the remaining splenic lymphocytes were specifically cytotoxic for the immunizing tumor and showed no cross-reactivity with other syngeneic UV-induced or methylcholanthrene-induced tumors of similar histologic type. The level of cell-mediated reactivity against UV-induced tumors was quite high compared to that obtained with syngeneic tumors induced by methylcholanthrene, and the cytotoxicity was attributable to a population of θ antigen-bearing lymphocytes.

With this in vitro test, we compared the response of normal mice, which reject a syngeneic tumor challenge, with that of UV-irradiated mice, in which the syngeneic UV-induced tumors grow progressively. After tumor cell inoculation, lymphocytes from the unirradiated (regressor) mice showed a high degree of cytotoxicity that reached a maximum level 8 days after injection. In contrast, no reactivity could be detected in the spleens of tumor-challenged UV-irradiated (progressor) mice.
Ala-1: A Murine Alloantigen of Activated Lymphocytes
Authors: Feeney Ann J. , Hämmerling Ulrich
Published: December 2022

Ala-1 (activated lymphocyte antigen-1) is a murine alloantigen expressed only on activated peripheral T and B lymphocytes. The presence or absence of Ala-1 on specific functional lymphocyte subsets was determined by treating the relevant cell population with anti-Ala-1 and complement, and assaying for residual functional activity. By this method, Ala-1 was shown to be on in vivo primed killer T cells cytotoxic for allogeneic tumor cells. It was also found on helper T cells generated in vivo to sheep red blood cells, and on IgM and IgG plaque-forming cells (PFC) to sheep red blood cells. In contrast, splenic precursors of helper cells and of IgM PFC to sheep red blood cells were completely resistant to treatment with anti-Ala-1 and complement. These findings indicate that effector cells can be distinguished from their nonactivated precursors by their expression of Ala-1.
Excretion of DNA by Purified Human Lymphocyte Subpopulations
Authors: Boldt David H. , MacDermott Richard P. , Speckart Stephen F. , Nash Geoffrey S.
Published: December 2022

A large proportion of DNA synthesized in vitro by human lymphocytes stimulated with plant mitogens or specific antigens is selectively excreted from the cells. To determine if DNA excretion differs among various types of lymphocytes, we examined purified human lymphocyte subpopulations for DNA synthesis and excretion in response to stimulation by L-PHA. The relative proportion of newly synthesized DNA that is excreted by unseparated mononuclear cells, macrophage-depleted cells, T, and B lymphocytes is identical despite great differences in the magnitude of their responses. Low levels of both DNA synthesis and excretion by macrophage-depleted cells and B cells can be increased by reconstitution with macrophages and T cells, respectively. These data indicate that DNA excretion is a general property of lymphocytes stimulated to undergo DNA synthesis by plant mitogens.
Microwaves Induce an Increase in the Frequency of Complement Receptor-Bearing Lymphoid Spleen Cells in Mice
Authors: Wiktor-Jedrzejczak Wieslaw , Ahmed Aftab , Sell Kenneth W. , Czerski Przemyslaw , Leach William M.
Published: December 2022

A single 30-min exposure of mice to 2450 MHz microwaves (12 to 15 mW/g body weight) in an environmentally controlled waveguide facility induced a significant increase in the proportion of complement-receptor positive lymphoid cells in the spleen. This effect was further enhanced by repeated (three times) exposures, which in addition produced a significant increase in the proportion of Ig+ cells. The proportion of θ-positive cells and the total number of spleen cells remained unchanged.