bluuebunny/crossref_metadata_2025 huggingface dataset:
Primary in Vitro Antibody Response of Human Peripheral Blood Lymphocytes: Role of Phagocytic Mononuclear Cells
Authors: Delfraissy Jean-François , Galanaud Pierre , Dormont Jean , Wallon Christine
Published: December 2022

We have studied the role of monocytes in the primary in vitro antibody response induced by the insolubilized conjugate of the hapten trinitrophenyl to polyacrylamide beads (TNP-PAA) in cultures of human peripheral blood lymphomonocytic cells (PBL). The depletion of adherent cells by passage on Sephadex G-10 columns does not modify the response. The functional elimination of phagocytic cells by the addition of silica is able, at the higher concentration used (1 mg/culture), to suppress the response profoundly. This response can be restored by the addition of 2-mercaptoethanol (2-ME). The lower concentration of silica (100 µg/ml) has an inconstantly suppressive effect, which, when present, is as marked as that of 1 mg/culture. One hundred micrograms of silica alone are able to stimulate the response in some cases, and when associated with 2-ME, has a reproducible enhancing effect. The addition of silica to the PBL of one of the few nonresponders to TNP-PAA allows a response from these cells. Silica does not allow the response of T-depleted cells to TNP-PAA. Thus, monocytes play a role in the response of human B cells to TNP-PAA, with two contrasting effects: Their presence is necessary in order to obtain a response, and in this respect they can apparently be replaced by 2-ME. They have an inhibitory effect which may, in rare instances lead to the absence of an in vitro response.
Human Leukocyte Inhibitory Factor (LIF): Use of Benzoyl L-Arginine Ethyl Ester to Detect LIF Activity
Authors: Rocklin Ross E. , Urbano Anthony M.
Published: December 2022

Human leukocyte inhibitory factor (LIF), a mediator elaborated by activated lymphocytes, has recently been shown to have the properties of an esterase. Further more, the LIF preparation contains an esteratic principle that hydrolyzes the amino acid ester, benzoyl arginine ethyl ester (3H-BAEE). In the present experiments, we sought to determine whether the esterase and biologic activities in the LIF preparation were due to the same or different molecules. This was accomplished by means of two series of experiments. With one approach, LIF was initially subjected to gel filtration and subsequently to polyacrylamide gel electrophoresis, sucrose density gradient electrophoresis, and affinity chromatography. The fractions obtained from each procedure were analyzed for biologic and esterase activities. The results indicate that both activities co-chromatographed with each technique. In addition, there are contaminating esterases present in the LIF preparation that can hydrolyze 3H-BAEE, some activity(ies) of which are eliminated by purification. The other approach directly demonstrated whether BAEE could serve as a substrate for LIF by substrate-competition experiments. BAEE and other structurally related amino acid esters were co-incubated with LIF and the ability of the former to protect the latter against inactivation by esterase inhibitors, or block biologic LIF activity on polymorphonuclear leukocytes was ascertained. It was shown that BAEE specifically prevented the destruction of LIF by diisopropylphosphofluoridate (DFP) and phenyl methyl sulfonyl fluoride (PMSF). Moreover, BAEE in a dose-dependent fashion, blocked LIF activity on PMN leukocytes in vitro when added to the culture supernatants. Taken together, these results strongly suggest that BAEE may serve as a substrate for LIF and point to the eventual development of a biochemical assay for this lymphocyte mediator.
Alloantigens Determined by A Second D Region Locus Elicit A Strong In Vitro Cytotoxic Response
Authors: Hansen Ted H. , Levy Robert B.
Published: December 2022

The newly defined L antigen (previously designated D') has been studied with the CML assay. BALB/c-H-2db mice, which carry a loss mutation in the D region, were found to generate anti-L cytotoxic effector cells when stimulated with wild-type (BALB/c) cells. When the target cells were treated with an antiserum against L, cytolysis by BALB/c-H-2db anti-BALB/c effectors was completely blocked. However, an antiserum against D, the antigen that bears the D region private specificity, had no effect on the anti-L cytotoxic response. In a control CML experiment that was specific for the D antigen, the antiserum to L had no effect, whereas the antiserum to D blocked completely. These results indicate that the L and D antigens are recognized in a CML response as separate entities on the cell surface. In an allogeneic response to the products of the D region, cytotoxic effectors were generated that were specific for L in addition to those specific for D. This conclusion was supported by blocking experiments with either antisera or unlabeled targets. These functional studies thus extend previous co-capping and immunochemical studies, and further support the hypothesis that L is a third transplantation antigen similar to the K and D antigens.
Specific Binding of T Lymphocytes to Macrophages
Authors: Ben-Sasson Shlomo Z. , Lipscomb Mary F. , Tucker Thomas F. , Uhr Jonathan W.
Published: December 2022

Peritoneal exudate lymphocytes (PEL) from immune guinea pigs that adhere to antigen-pulsed macrophages (MØ) were cultured for 1 week to yield a population enriched in antigen-specific (selected) T cells. These cells bind specifically within hours to fresh autologous antigen-pulsed MØ. The dissociation of these selected PEL from antigen-pulsed MØ was studied. No evidence was obtained that factors in the culture medium play a role in dissociation. Lymphocytes that have dissociated from antigen-pulsed MØ are usually fully capable of rebinding to MØ freshly pulsed with antigen, suggesting that there is no deficiency in the lymphocytes ability to bind. In contrast, readding antigen to cultures during incubation prevents the predicted dissociation. Moreover, repulsing MØ cultured without selected PEL restores their capacity to bind fresh selected PEL. These findings indicate that decay of antigen associated with MØ is the major mechanism underlying the observed dissociation.
Anti-Immunoglobulin Stimulation of Murine Lymphocytes
Authors: Weiner Howard L. , Scribner Duncan J. , Moorhead John W.
Published: December 2022

Anti-immunoglobulin (anti-Ig) stimulation of spieen cells from aged mice (7 to 9 months old) results in both blast cell transformation and increased incorporation of 3H-TdR. Younger animals rarely respond to anti-Ig. Using the above parameters to measure cell activation we investigated 1) the time requirements for anti-Ig-lymphocyte interaction which are necessary to induce stimulation, 2) the identity of the responding cells, and 3) the fate of surface receptors on the stimulated cells.

When spleen cells were cultured with anti-Ig for various periods of time, it was found that the anti-Ig antibodies had to remain in culture for at least 48 hr of the 3-day culture period to induce maximum stimulation. Removing anti-Ig from culture at 24 or 4 hr resulted in a significantly lower response or an inhibition of proliferation. The cells responding to anti-Ig were B cells; 85 to 90% of anti-Ig-induced blast cells, cultured to allow re-expression of surface markers, were positive for surface Ig which was IgM.

The prolonged requirement for anti-Ig in culture indicated that surface Ig must be continuously re-expressed by the responding cells. However, free surface Ig was not detected on the cells during continuous culture with anti-Ig. Rather, the cells were found to have relatively large amounts of rabbit antibody bound to their surface. Similar cultures stimulated with F(ab′)2 fragments or cultures of spleen cells from young animals stimulated with intact anti-Ig did not have surface-bound rabbit antibody. The rabbit antibody is most likely in the form of antigen-antibody complexes, i.e., surface Ig-anti-Ig, bound to Fc receptors on the stimulated cells. These complexes are apparently produced by the continuous re-expression and redistribution of surface Ig by anti-Ig, a process that is required for anti-Ig stimulation of B cells.
Analysis of the Cellular Basis of Idiotype-Specific Suppression
Authors: Ward Kathleen , Cantor Harvey , Nisonoff Alfred
Published: December 2022

We have investigated the ability of B and T lymphocyte subclasses from donor mice that produce high levels of anti-Ar antibody but have been suppressed for one idiotypic component (CRI) to induce and maintain idiotypespecific suppression. Our studies indicate: 1) Memory B cells from such mice can preempt virgin CRI+ B cells present in the host from contributing to the anti-Ar response. 2) T cells can also adoptively transfer idiotypespecific suppression. 3) B and T cells do not act synergistically in this transfer of idiotype-specific suppression. 4) Extremely small numbers of Ly23 cells transfer suppression of idiotype and most probably represent true Ts cells. 5) Ly1 cells from hyperimmune idiotypically suppressed donors can induce idiotype-specific suppression. This latter result most likely reflects the induction of idiotype-specific suppressor cells in the host.
Regulation of IgE Antibody Production by Serum Molecules
Authors: Tung Amar S. , Chiorazzi Nicholas , Katz David H.
Published: December 2022

Exposure of mice to low doses of ionizing x-irradiation at or near the time of primary immunization with 2,4-dinitrophenyl (DNP)-Ascaris suum extract (ASC) results in substantial enhancement of IgE anti-DNP antibody responses; the IgG antibody responses of such mice do not increase after such manipulations. This selective enhancement of IgE antibody production occurs in mice of both high and low IgE responder phenotype, although the extent of enhancement compared to unmanipulated control animals is more striking in low IgE responder mice. The studies presented here demonstrate that the irradiation-enhanced IgE antibody responses of low responder SJL and C57BL/6 mice as well as of intermediate responder AKR mice can be effectively suppressed by passive transfer of CFA-immune serum obtained from isologous donor mice. Moreover, adoptive secondary IgE antibody responses in SJL recipients of primed syngeneic spleen cells can be totally abolished by passive transfer of CFA-immune serum or ascitic fluid from CFA-immune mice. The suppressive activity of CFA-immune serum can be diminished or eliminated by exposure of CFA-primed donor mice to low dose x-irradiation at an appropriate point during the priming regimen, after a single inoculation of CFA, and before collection of serum. Low dose x-irradiation was not effective in eliminating suppressive activity of CFA-induced ascites fluid obtained from donor mice inoculated repeatedly with CFA.

In contrast to the capacity of CFA-immune serum from isologous donors to suppress irradiation-enhanced IgE responses of low responder mice, similar sera or ascites fluids were ineffective in suppressing irradiation-enhanced responses of high responder BALB/c or (SJL × BALB/c)F1 hybrid mice. This was true irrespective of whether such sera or ascites were obtained from isologous donors or from donor mice of the low IgE responder phenotype. The implications of these findings with respect to clarifying some of the previously confusing observations concerning 1) the effects of adjuvant administration on IgE antibody production, and 2) differences in the capacities of different mouse strains to produce IgE antibodies in high vs low quantities are discussed.
Studies on the Question of Conventional Immunoglobulin on Thymocytes from Primitive Vertebrates
Authors: Yamaga Karen M. , Kubo Ralph T. , Etlinger Howard M.
Published: December 2022

The presence of antibodies directed against carbohydrate determinants in anti-trout immunoglobulin (Ig) and anti-keyhole limpet hemocyanin (KLH) antisera raised the possibility that these reagents might react with Ig and non-Ig membrane determinants. In order to discriminate between these two specificities, trout splenocytes and thymocytes were radioiodinated by the lactoperoxidase procedure and immunoprecipitated with anti-trout Ig and anti-KLH. With splenocyte membrane proteins, reduced anti-trout Ig immunoprecipitates, when analyzed on SDS-gels, resolved into a 100,000 m.w. component as well as the heavy and light chain constituents from membrane Ig. The anti-KLH, on the other hand, reacted preferentially with a 100,000 m.w. component. The binding to the 100,000 m.w. component could be removed from the anti-Ig sera by absorption of the reagent with KLH such that only the membrane Ig was precipitated. The lower reactivity of anti-KLH to membrane Ig, when compared with the 100,000 m.w. component, was probably due to a difference in carbohydrate content because specifically purified antibodies eluted with l-fucose were capable of reacting with both glycoproteins from splenocytes.

On thymocytes, anti-Ig and anti-KLH reacted with membrane proteins that gave qualitatively similar SDS-gel patterns. The anti-Ig sera absorbed with KLH failed to react with any thymocyte membrane proteins indicating an absence of Ig-specific determinants on these cells. By immunofluorescence, 100% of the thymocytes were stained with anti-Ig and anti-KLH reagents. Some faint residual fluorescence was found when anti-Ig was absorbed with erythrocytes, KLH, and trout Ig glycopeptides. Thus, although some minor reactivity of anti-Ig on thymocytes cannot be accounted for by cross-reactive determinants by immunofluorescence, most of the reactivity was due to carbohydrate moieties. It does not appear that conventional Ig is found on thymocytes, at least in amounts that are detectable by the lactoperoxidase labeling method.
Genetic Analysis of Ia Determinants Expressed on Con A-Reactive Cells
Authors: Ahmann Gerald B. , Sachs David H. , Hodes Richard J.
Published: December 2022

Pretreatment of mouse lymphoid cells with anti-Ia sera and C abrogated the proliferative responses of these cells to Con A. Studies were carried out with several anti-Ia reagents and intra-H-2 recombinant mouse strains to map the I subregion(s) whose products are expressed on Con A-reactive cells. Treatment with a (B10.A × A)F1 anti-B10 reagent and C abrogated the ability of BALB/c cells to respond to Con A. Absorption studies on this reagent demonstrated that Con A-reactive cells express Ia determinants coded by the I-A subregion. The results with two additional reagents, B10.A(4R) anti-B10.A(2R) tested on B10.BR cells and (B10 × D2.GD)F1 anti-B10.D2 absorbed with B10.A cells and tested on BALB/c cells, demonstrated that Con A-reactive cells also express Ia determinants encoded to the right of I-A. Several antisera and strain combinations were evaluated in which the antisera could contain antibodies specific for products of genes encoded by the I-J subregion, but the results were inconclusive. These data demonstrate that there are at least two different I subregions, one in I-A and one to the right of I-A, that code for antigens expressed on Con A-reactive cells.
Studies on the Mechanism of Lymphocyte-Mediated Cytolysis
Authors: Siliciano Robert F. , Henney Christopher S.
Published: December 2022

These studies were addressed to the hypothesis that interactions between a target cell's plasma membrane and its nucleus might be of significance in determining the susceptibility of the cell to lysis by cytotoxic lymphocytes. To this end, P815 mastocytoma cells were enucleated by centrifugation through a discontinuous Ficoll gradient containing 20 µg/ml cytochalasin B. Two types of vesicles were isolated, both enclosed by plasma membrane: one fraction, termed cytoplasts, contained significant quantities of the cytoplasmic marker lactate dehydrogenase, but lacked nuclei by both biochemical and phase-contrast microscopic criteria. The other type, karyoplasts, contained nuclei. Cytoplasts displayed surface H-2 antigens but “capped” these molecules poorly; karyoplasts showed a “capping” capacity similar to that of intact cells. Despite a deficient capacity to “cap” surface H-2, cytoplasts were susceptible to the action of cytotoxic T cells. There was no indication that cytoplasts were more resistant to lysis than either karyoplasts or intact cells. Moreover, sources of NK, K, and lectin-dependent cytotoxic cells were all capable of inducing specific 51Cr release from both cytoplast and karyoplast preparations.

Thus, in keeping with our earlier suggestion that the target cell plays a passive role in the lytic process, these studies show that possession of a nucleus is not a prerequisite for susceptibility to cell-mediated cytoxicity.
Trinitrophenylated Red Cells (E-TNP) as a Model for Antibody-Independent Activation of the Complement System via the Classical Pathway
Authors: Loos Michael , Thesen Rolf
Published: December 2022

Recently, it was shown that DNP-substituted human serum albumin (DNP-HSA) bound directly C1q, a subcomponent of C1. Evidence is presented that TNP-conjugated erythrocytes (E-TNP) were lysed by autologous sera as source of complement (C). The lysis was strongly dependent on the TNP substitution rate so that the TNP-carrying erythrocytes can be used directly as an indicator for the TNP-C interaction. The TNP-induced lysis of erythrocytes required the presence of C1q, C4, and Ca++, indicating a dependency on the classical pathway of C. Purified C1̄ is directly bound to E-TNP; C1̄ bound to E-TNP could be transferred to EAC4. From E-TNP-1̄ the intermediates E-TNP-C1̄4 and E-TNP-C1̄42̅ can be formed by successive addition of C4 and C2 similarly to EAC1̄4 and EAC1̄42̅. Native C1 can be activated to C1̄ by E-TNP in the same fashion as by EA. The E-TNP model provides evidence that the “activated” Fc part of the antibody molecule is not unique in its ability to trigger the internal activation of C1. Molecules with negatively charged groups, like polyanions, or molecules with high electron dense groups, like DNP or TNP, can activate the C system via the classical pathway by binding directly to C1q.
Theory of Equilibrium Binding of Symmetric Bivalent Haptens to Cell Surface Antibody: Application to Histamine Release from Basophils
Authors: Dembo Micah , Goldstein Byron
Published: December 2022

We present a theory of equilibrium binding of symmetric bivalent haptens to cell surface antibody in the presence or absence of monovalent hapten. Bivalent haptens can link together antibodies to form linear chains or rings on cell surfaces. We show how to calculate the amount of any complex of bound bivalent hapten, monovalent hapten, and antibody.

We are particularly interested in the mole fraction of antibody involved in complexes made up of two or more antibodies, i.e., the fraction of antibody that is cross-linked (xpoly). We treat the case when the antibody on the cell surface, which is specific for the hapten, is homogeneous. For this case we prove a number of general properties about xpoly: 1) xpoly approaches zero at both high and low bivalent hapten concentration. 2) xpoly becomes a maximum when the bivalent hapten concentration equals Amax, where Amax = 1/H + B/2. H is twice the equilibrium constant for the binding of a single hapten site to a single antibody site and B is the monovalent hapten concentration. 3) a plot of xpoly vs the log of the bivalent hapten concentration is symmetric about the maximum value of xpoly. We use these and other properties of xpoly in this paper to clarify the relationship between cross-link formation and histamine release.
Intra- and Interspecies Reactivity of Human and Mouse Natural Killer (NK) Cells
Authors: Hansson Mona , Bakacs Klas Karre Tibor , Kiessling Rolf , Klein George
Published: December 2022

Natural killer (NK) cells can kill certain syngeneic, allogeneic and xenogeneic tumor targets in short-term 51Cr release assays. In the present study, intra- and interspecies NK activity was analyzed. Ten mouse and five human tissue culture lines were used. In direct cytolytic assays with mouse spleen cells or human PBL effectors, intraspecies was much stronger than interspecies reactivity, as a rule. A certain interspecies activity was obtained, stronger in mouse anti-human (M α H) than in human anti-mouse (H α M) combinations. In the H α M system, activity was associated with the same type of PBL-derived non-B-non-T cell fraction as in the intraspecies H α H system. The non-B-non-T cell nature of the M α H killer cell has been demonstrated previously.

Nonlabeled tumor cells were allowed to compete with isotope-labeled targets in intra- and interspecies cytolytic NK tests. NK-sensitive tumor lines of the same species were superior to xenogeneic competitors in both M α M and H α H tests. In the M α M assay, the competing ability of the same human tumors varied, depending on the genotype of the mouse effector cells. None of the human lines tested competed effectively with strain CBA effectors but some showed a certain competition with C57BL effectors. In the H α H assay, strong competition was seen with two of the 10 xenogeneic mouse tumors tested.
Suppression of Humoral and Delayed Hypersensitivity Repsonses by Distinct T Cell Subpopulations
Authors: Whisler R. L. , Stobo J. D.
Published: December 2022

Immunization of mice with a supraoptimal dose of sheep red blood cells (SRBC) results in splenic T cell populations capable of specifically suppressing recipients' plaque forming cell (PFC) and delayed-type hypersensitivity (DTH) responses to SRBC when tested in an adoptive transfer system. By localization on discontinuous bovine serum albumin (BSA) gradients and relative sensitivity to Cytoxan, two distinct T cell subpopulations suppressing DTH reactivity were identified. One population could not be distinguished from T cells capable of inhibiting direct and indirect PFC responses. However, another population appeared quite distinct and capable of inhibiting DTH, but not PFC responses.
Macrophage Activation for Tumor Cytotoxicity: Control of Macrophage Tumoricidal Capacity by the LPS Gene
Authors: Ruco Luigi P. , Meltzer Monte S. , Rosenstreich David L.
Published: December 2022

In contrast to cells from C3H/HeN mice, macrophages from LPS-unresponsive C3H/HeJ mice fail to develop tumoricidal capacity after a variety of in vivo of in vitro activation stimuli. The macrophage-tumoricidal defect of C3H/HeJ mice was evident with treatments not dependent upon exogenously added LPS: BCG (see Abbreviations) infection, T cell mitogens, or in vitro exposure to lymphokines. This study was designed to determine if macrophage-tumoricidal capacity was controlled by the LPS gene. Tumoricidal responses of macrophages from BCG-infected F1 hybrids (C3H/HeJ × C3H/HeN) were intermediate to those of the parental strains. There were no differences in cytotoxic responses between macrophages from BCG-infected male or female F1 hybrids. These responses suggest an autosomal codominant pattern of inheritance for control of macrophages cytotoxicity similar to that previously reported for B cell responsiveness to LPS. In a backcross linkage analysis (C3H/HeJ × F1), there was complete concordance in the expression of macrophage-tumoricidal capacity and spleen cell capacity to proliferate in response to LPS. Furthermore, macrophage-tumoricidal capacity correlated with LPS responsiveness in other inbred mouse strains. Macrophages from BCG-infected, LPS-unresponsive C57BL/10ScCR mice were not tumoricidal in vitro, whereas cells from BCG-infected, LPS-responsive C57BL/10Sn mice were. Moreover, analysis of tumoricidal responses in eight different BXH recombinant inbred strains of mice revealed that the development of cytotoxic macrophages during BCG infection was associated with the capacity of each strain to respond to LPS. These findings strongly suggest that the gene for control of macrophage-tumoricidal capacity is either closely linked or identical to the LPS gene.
Inductive Requirements for the Generation of Virus-Specific T Lymphocytes
Authors: Hapel Andrew J. , Bablanian Rostom , Cole Gerald A.
Published: December 2022

Priming CBA mice (H-2k) with infectious ectromelia virus (EV) and C3H mice (H-2k) with noninfectious vaccinia virus (VAC) resulted in a transient virus-specific splenic cytotoxic T lymphocyte (CTL) response followed by persisting CTL memory. When unfractionated spleen cells from these mice were cultured with mitomycin-C-treated stimulator L fibroblasts (H-2k) exposed previously to the homologous noninfectious virus, potent secondary (2°) virus-specific CTL were induced. CTL induction was not affected by depleting the responders of phagocytic and/or adherent cells (presumed to be macrophages) if either virus was presented on L cells. However, only unfractionated responders yielded 2° CTL when cultured with free EV or VAC, indicating that fibroblasts could substitute for responder macrophages as antigen-presenting cells. The failure of virus to induce 2° CTL when presented on either allogeneic fibroblasts or Sepharose provided clear evidence that triggering of CTL precursors required the recognition of antigen in some closely linked association with a “self” marker. This requirement also held for stimulating the responders to undergo virus-specific proliferative responses although proliferation was not a prerequisite for 2° CTL induction.

By exposing L cells to defined numbers of purified noninfectious VAC for only 10 min and then after increasing intervals fixing them with glutaraldehyde, it was shown that viral adsorption alone was necessary for the cells to become capable of both stimulating the induction of 2° CTL and inhibiting competitively their lytic activity. After adsorption, this dual capability of the cells increased at a rate that correlated inversely with the concentration of virions to which they were initially exposed. Thus, the stimulator or competitor efficacy of cells exposed to a poxvirion multiplicity of 5000 to 10,000 for 10 min was reached 90 min later by cells exposed to a multiplicity of 50 to 100.

Collectively, these experiments demonstrate that only a brief exposure of host cell membranes to intact poxvirions is required to form the antigenic configuration recognized by poxvirus-specific CTL or their precursors. Since this event can occur under conditions that do not favor viral modification of native host cell membrane antigens, it supports the concept that antigen recognition occurs through T lymphocyte receptors with separate specificities for “self” and viral determinants.
In Situ Cytotoxic T Cells in a Methylcholanthrene-Induced Tumor
Authors: DeLustro Frank , Haskill J. Stephen
Published: December 2022

The in situ localization of cytotoxic T-cells was examined in a methylcholanthrene-induced fibrosarcoma. The MCA 2 tumor was initiated in this laboratory and utilized before extensive in vivo passage. Tumor cell suspensions were separated by unit velocity sedimentation. The T cell-enriched tumor-derived fractions and spleen cells from MCA 2-bearing mice were cytotoxic for in vivo isolated MCA 2 and in vitro cultured MCA 2 tumor cells, but not for SAD2 tumor cells. The cytotoxic activity of effector cells isolated from MCA 2 tumors was abrogated by treatment with anti-ϑ serum plus complement. The significance of cytotoxic T cells with in a progressing tumor is discussed.
Quantitative Analysis of Spleen Cell and Immunoglobulin Allotype Composition of (CBAxCBA/H-T6) → C57BL/6 Allophenic Mice
Authors: Warner Carol M. , McIvor Judith L. , Berntson Tom J.
Published: December 2022

Fifteen (CBAxCBA/H-T6) → C57BL/6 allophenic mice were analyzed for the parental composition of their spleen white blood cells and serum immunoglobulin allotypes. The spleen compositions, assessed by a microcytotoxicity test, showed that the animals covered a range of parental cell mixtures. The allotype compositions of the IgG2a and IgG2b subclasses were determined by a solid phase radioimmunoassay. Although the absolute amount of IgG2a and IgG2b varied markedly from animal to animal, the percentage of the two allotypes in each subclass was remarkably constant. This suggests that the synthesis of IgG2a and IgG2b may be under a coordinate control system. Also, the spleen cell compositions were in excellent agreement with both the IgG2a and IgG2b subclass compositions. The quantitative analysis of the immune systems of allophenic mice may be a first step toward the understanding of the mechanisms of tolerance of histoincompatible cells in allophenic mice.
Effect of in Vitro Virus Infection on Response of Human Monocytes and Lymphocytes to Mitogen Stimulation
Authors: Roberts Norbert J. , Steigbigel Roy T.
Published: December 2022

Viral infection and immunization can depress certain immune functions, including in vitro transformation response of lymphocytes to mitogens and antigens. Since optimal lymphocyte response occurs only in the presence of macrophages, we sought to determine whether the depression of transformation response could result from macrophage dysfunction induced by the viral infection.

Highly purified human macrophages and lymphocytes were infected in vitro with influenza A virus. There was no significant difference in transformation response between uninfected and infected lymphocytes either alone or when combined with uninfected macrophages, suggesting, by exclusion, a macrophage defect. Direct evidence for macrophage dysfunction was provided by the observation that virus-infected macrophages were significantly less effective than uninfected macrophages in enhancing the transformation response of either uninfected (p < 0.025) or infected (p < 0.01) lymphocytes. Adverse effects of virus infection could not be attributed to an effect of the virus cultivation medium, to cell separation procedures, or to changes in cell viability. Adverse effects on macrophage function were evident as early as 24 hr after virus infection and were directly related to the virus inoculum. Depressed enhancement of response by macrophages was demonstrated to a greater extent with suboptimal than optimal dose phytohemagglutinin stimulation, and was shown by using allogenic as well as autologous macrophages. Macrophage phagocytosis of staphylococci was similarly depressed but not eliminated by virus infection. Depressed transformation of unseparated mononuclear leukocytes was only manifested when virus infection occurred before or at the time of phytohemagglutinin stimulation, again suggesting a primary effect on macrophages.

The inability of virus-infected human macrophages to enhance lymphocyte response to mitogen can account for the depression of human lymphocyte transformation seen during influenza virus infection. Viral infection had no adverse effect on lymphocyte response per se.
Cytotoxic Activities of Gut Mucosal Lymphoid Cells in Guinea Pigs
Authors: Arnaud-Battandier Franck , Bundy Bonita M. , O'Neill Michael , Bienenstock John , Nelson David L.
Published: December 2022

Populations of lymphoid cells were obtained from the gastrointestinal tract of guinea pigs and compared with splenic cells for effector cell activity in assays of spontaneous cell-mediated cytotoxicity (SCMC), mitogen-induced cellular cytotoxicity (MICC), and antibody-dependent cellular cytotoxicity (ADCC). Lymphoid cells were isolated from mesenteric lymph nodes (MLN), Peyer's patches (PP), and the small intestinal mucosa. Mucosal lymphoid cells were separated into a population of cells containing intraepithelial lymphocytes (IEL), and a second population of cells containing lamina propria (LP) lymphocytes by a combination of physical and enzymatic methods. All populations of lymphoid cells from the gastrointestinal tract were capable of mediating MICC. Relative to spleen cells, IEL possessed more MICC effector cell activity whereas cells from MLN, PP, and LP possessed less activity. In contrast, SCMC and ADCC effector cell activities were present only in the IEL population and these cytotoxic activities were comparable to that of splenic cells. These results demonstrate that mucosal lymphoid cells possess cytotoxic effector cell activities that may contribute to local mucosal immunologic reactivity.
Autoreactive Antibody-Forming Cells Directed against Thymocytes and Thymus-Derived Lymphocytes
Authors: McHugh Yvonne E. , Bonavida Benjamin
Published: December 2022

Antibody-forming cells with specificities against syngeneic and allogeneic thymocytes are detected in the spleens of normal mice after activation in vitro or in vivo with lipopolysaccharide (LPS). The activity of such cells was measured in a complement-dependent plaque assay employing trypan blue dye to assess zones of lysis. Plaques were rarely seen in the absence of LPS treatment. Anti-immunoglobulin added to the plaque assay abrogated the appearance of plaques, but the addition of LPS had no effect. Furthermore, plaque formation was 2-mercaptoethanol sensitive indicating that the antibody responsible was of the IgM class. Plaque forming cells (PFC) were also detected against syngeneic and allogeneic lymph node cells and to a much lesser extent against splenocytes. The numbers of PFC found against syngeneic, allogeneic, or a mixture of thymocytes was similar and ranged from 1000 to 3000 PFC/108 viable spleen cells tested. All murine strains tested, including congenitally athymic nude mice, exhibited anti-thymocyte PFC after LPS activation. C3H/HeJ mice, genetically unresponsive to LPS, did not respond mitogenically to LPS and no anti-thymocyte plaques were observed. These findings suggest that clones of autoreactive B cells are present in normal mice and can be activated by LPS.
Properties of the Cell Surface Fc-Receptor Induced by Herpes Simplex Virus
Authors: Nakamura Yoshiko , Costa Jose , Tralka Tommie Sue , Yee Carole L. , Rabson Alan S.
Published: December 2022

Detection of peroxidase-antiperoxidase soluble complexes (PAP) bound to the surface of herpes simplex virus-infected cells has been used to demonstrate virusinduced Fc receptors and to study their distribution. The PAP method is more sensitive than hemadsorption with immunoglobulin-coated sheep red blood cells, and can be used to study localization by light and electron microscopy. Our results indicate that capping takes place after the receptor is engaged by antigen-antibody complexes and that at least a portion of the bound ligand is internalized.
Monocyte-Mediated Suppression of Human B Lymphocyte Differentiation in Vitro
Authors: Knapp Walter , Baumgartner Gerhard
Published: December 2022

Adherent mononuclear cells (AC) from the peripheral blood of myeloma patients and also of healthy individuals in numbers usually present in mononuclear cell (MNC) preparations isolated by the Ficoll-Hypaque method markedly suppress the in vitro plasma cell response of human B cells to PWM. In myeloma patients the mean number of recovered cytoplasmic Ig-positive cells was 4.3 × 103/culture when unfractionated MNC were cultured with PWM and 24.6 × 103 when partially AC-depleted MNC were used. In healthy individuals the mean total number of recovered cytoplasmic Ig-positive cells increased from 19.1 × 103 to 46.2 × 103/culture. Readding partially AC-depleted MNC to plastic adherent cells and also co-cultivation of strongly adherent MNC with partially AC-depleted MNC led to a significant suppression of plasma cell differentiation. Partial depletion of AC not only led to an increased differentiation and proliferation of human B cells but also to a shift in the Ig class distribution of cytoplasmic Ig-positive cells from IgM to IgG.

A complete removal of AC, on the other hand, completely abrogated PWM-induced B cell differentiation in vitro. Contrary to the significantly increased B cell proliferation and differentiation after partial depletion of AC, no significant difference could be observed when tritiated thymidine incorporation of unfractionated and AC-depleted MNC after stimulation with PWM or PHA was measured.

Adherent mononuclear cells from healthy individuals with suppressor activity for PWM-induced B cell differentiation were shown to be relatively radioresistant. They have to be viable to mediate the suppression; they cannot be removed by depletion of EN-RFC; they are sensitive to carrageenan; and their suppressive activity is not abrogated by treatment with anti-Ia-type serum plus complement under conditions that kill all B cells. These cells therefore were considered to belong to the monocyte/macrophage series.
Biochemical Criteria for Activated Macrophages
Authors: Karnovsky Manfred L. , Lazdins Janis K.
Published: December 2022

The designation “activated macrophage” was originally introduced to describe the state of macrophages that have an enhanced ability to phagocytize microorganisms and exert antimicrobial activity. This effect was found to be dependent upon particular infections and involves the participation of lymphocytes (1, 2). “Activation” has a specific immunologic basis, but its expression is nonspecific. Mackaness has written lucidly about activated macrophages and has referred readers back to the concepts of Lurie in tuberculosis, and further to the fons et origo of thinking in this context, i.e., to concepts of Metchnikoff (3). The latter's oft quoted phrase regarding a “perfecting of the phagocytic and digestive powers of the leukocytes” might well be, broadly speaking, synonymous with “activation.” Once it was determined that macrophages taken from animals at some appropriate time after infection, for example, with Listerria or Bacillus Calmette-Guérin2 (BCG), were superior to control cells with respect to their antimicrobial powers, it was natural for inquiry to be made into the differences between such cells and normal (control) cells, in terms of their morphology, cellular chemistry, and biochemical attributes.
Ig-Isotype Diversity Generated in Antibody-Forming Cells of the Mouse
Authors: Ventura Michel , Bleux Christian , Crepin Yvonne , Liacopoulos Panayotis
Published: December 2022

Generation of Ig-isotype diversity was studied in hemolytic plaque-forming cells (PFC) of mice immunized with sheep erythrocytes (SRBC). Spleen cells were taken at various days after immunization, cultured in vitro for 24 hr in the presence of bacterial lipopolysaccharide (LPS), and PFC occurring in these mass cultures were micromanipulated and individually cultured for 48 hr. Daughter PFC were revealed by direct or indirect procedures in order to study the Ig-isotype they produced.

Parental PFC of all three Ig isotypes studied, IgM, IgG, and IgA, generated daughter PFC, producing the same isotype but with maximum yields on different days: postimmunization day 3 for IgM PFC, day 4 for IgG PFC, and day 5 for IgA PFC. A proportion, but not all, of parental IgM PFC generated daughter IgG producers only during the first 3 days after immunization and IgA producers during the 4th to 6th post-immunization days. Parental, especially 4th day IgG PFC also generated daughter IgA producers, thus showing that IgA producers could stem from both IgM and IgG precursors in addition to IgA-producing precursors. It was also found that IgG PFC could present a reverse switching and generate IgM producers. This suggests that the overall orderly IgM, IgG, IgA conversion is possibly not strictly programmed by an internal control of B cells but submitted to some extent to microenvironmental influences.
Genetic Control of Immune Response to Sperm Whale Myoglobin in Mice
Authors: Okuda Kenji , Christadoss P. Remkumar , Twining Sally , Atassi M. Zouhair , David Chella S.
Published: December 2022

Studies on the genetic control of immune response to sperm whale myoglobin were initiated. As demonstrated in this paper, the T lymphocyte proliferative response to whale myoglobin is under H-2-linked Ir gene control. Mice of H-2d, H-2f, and H-2s haplotypes were high responders to the myoglobin, whereas haplotypes H-2b, H-2k, H-2p, H-2q, and H-2r were low responders. The Ir gene(s) was localized between H-2K and H2D regions, since the recombinant strain A.TL (KsIkSkDd) was a low responder and A.TH (KsIsSsDd) was a high responder. Further studies with recombinant strains revealed that the expression of the high-responder I-Ad or I-As alleles was sufficient to give a good response, since strains D2.GD (d d b b b b b b) and B10.HTT (s s s s k k k d) were high responders. The expression of the I-Cd allele in strains B10.A (k k k k k d d d) and B10.A(5R) (b b b k k d d d) also gave high response, and thus suggested a second Ir gene, derived from the H-2d haplotype. The finding that expression of the I-Cs allele in B10.S(8R) (k k ? ? s s s s) did not result in high response suggests the lack of the second Ir gene in the high-responder H-2s haplotype.
Regulatory Substances Produced by Lymphocytes
Authors: Wagshal Alan B. , Waksman Byron H.
Published: December 2022

The effects of IDS and of other agents that raise intracellular cyclic AMP levels on a variety of lymphocyte functions occurring during various phases of the cell cycle were compared. IDS, which inhibits DNA synthesis as a result of increasing cyclic AMP levels, has no effect on cyclic AMP until 12 hr after adding mitogen. Evidence that IDS does not affect unstimulated lymphocytes or lymphocytes exposed to mitogen for less than 12 hr was obtained by showing that elevated cyclic AMP, but not IDS, caused increased early calcium uptake. IDS failed to inhibit RNA synthesis until 8 to 12 hr after addition of mitogen. Although prostaglandin E1 and dibutyryl cyclic AMP inhibited release of lymphotoxin, a lymphokine produced by lymphocytes during the early G1 phase, IDS actually increased lymphotoxin release. IDS was unable to duplicate the effects of other cyclic AMP agents in enhancing mitogen responses of unstimulated spleen and thymus cells via a thymus hormone-like maturational effect. In a comparison of lymphocytes either 3 hr (G0 or early G1) or 20 hr (middle to late G1) after addition of mitogen, only 20-hr lymphocytes were able to absorb IDS. These findings suggest that the cell cycle specificity of IDS may be due to appearance of cell receptors for IDS only during a limited phase of the lymphocyte cell cycle in mature cells, and that this cell cycle specificity may be an important factor regulating IDS activity.
Suppression of Growth of Guinea Pig Line-10 Hepatocarcinoma
Authors: Minden Percy , Brunda Michael J. , Rodney Ferguson H.
Published: December 2022

Complete or partial protection against the line-10 hepatocarcinoma was conferred to a significant number of strain-2 guinea pigs by the passive transfer of antibodies reactive to antigens from line-10 cells. Immunoglobulins were administered intradermally and injected sites were subsequently challenged with line-10 cells. Immunoglobulins prepared from the sera of rabbits that had been immunized to line-10 cells and from the sera of guinea pigs that had been rendered immune to this tumor by previously described methods were more effective than immunoglobulins obtained from normal rabbit or guinea pig sera. Protection was abolished if immunoglobulins were absorbed with line-10 cells. Antitumor effects were dependent on the close contact of antibodies and tumor cells. Protection was greater if normal syngeneic peritoneal exudate cells were tranferred in addition to the immunoglobulins. A possible mechanism for the antitumor effects observed is that of antibody-dependent cellular cytotoxicity.
Comparison of the Developmental Kinetics of Antibody- and Immunoglobulin-Forming Cells in Normal and Tolerant Mice
Authors: Carelli Claude , Antoine Jean-Claude , Petit Christine , Rodrigot Maryvonne , Avrameas Stratis
Published: December 2022

The developmental kinetics of antibody-forming cells (AFC) and of cells synthesizing immunoglobulins without detectable antibody function against the antigen injected (IFC) were compared in lymph nodes of normal mice and tolerant mice receiving an immunogenic challenge. Tolerant states to bovine serum albumin (BSA), induced in adult mice, and to human γ-globulin (HGG), induced in newborn mice, were studied. Whatever the tolerant state induced, the following main results were obtained: 1) both AFC and IFC were suppressed in tolerant mice, and this to approximately the same extent for the two populations; 2) both the suppression of the AFC and of the antigen-induced IFC were specific to the tolerogen used.
Transplantable IgD Immunoglobulin-Secreting Tumors in Rat
Authors: Bazin H. , Beckers A. , Urbain-Vansanten G. , Pauwels R. , Bruyns C. , Tilkin A. F. , Platteau B. , Urbain J.
Published: December 2022

The LOU/C/Wsl rat inbred strain presents a high incidence of spontaneous malignant ileocecal immunocytomas or monoclonal immunoglobulin-secreting tumors. Some tumors have been transplanted in histocompatible animals over years without any change in their secretion products. Among approximately 600 different monoclonal proteins we have studied so far, we recognized six showing properties different from those of rat IgM, IgA, IgE, or IgG classes, and characteristic of the IgD class.
A New Method for the Purification of Human Eosinophils and Neutrophils, and a Comparison of the Ability of These Cells to Damage Schistosomula of Schistosoma Mansoni
Authors: Vadas Mathew A. , David John R. , Butterworth Anthony , Pisani Nancy T. , Siongok Timothy A.
Published: December 2022

Centrifugation of human white blood cells over either Ficoll-Hypaque or slightly hypertonic Metrizamide discontinuous gradients reliably produces separate fractions that are enriched for either neutrophilic or eosinophilic granulocytes. This single step purification routinely yields 90 to 100% pure neutrophils and 85 to 100% pure eosinophils. Metrizamide gradients, in particular, reproducibly provide high yields of 90 to 100% pure eosinophils from normal subjects with 2 to 3% eosinophils in their peripheral blood. The method does not damage cells as judged by morphologic or functional criteria.

The purified cell populations were tested for their ability to damage antibody-coated schistosomula either by the measurement of 51Cr release from labeled organisms, or by direct morphologic assessment. Neutrophils were superior in their ability to release 51Cr from labeled organisms, but eosinophils adhered to the organisms to a greater extent and induced microscopically detectable damage.
Plaque-Forming Cell Response in BALB/c Mice to Two Preparations of LPS Extracted from Salmonella Enteritidis
Authors: Hepper Kelton P. , Garman Richard D. , Lyons Michael F. , Teresa George W.
Published: December 2022

The pattern of development of antibody-forming cells in BALB/c mice after immunization with PW-LPS or TCA-LPS was shown to be different. On days 10 and 20, the primary response to PW-LPS was characterized by a low level of IgM synthesis. The plaque-forming cell (PFC) response to TCA-LPS, however, increased from day 10 to day 20. Initially, IgM was the only detectable antibody synthesized but by day 20 a significant number of IgG-producing spleen cells had developed. After a secondary immunization with the appropriate lipopolysaccharide (LPS) preparation, IgG-producing spleen cells were detected in mice immunized with either PW- or TCA-LPS. Partial removal of the LAP of TCA-LPS with phenol or trypsin and pronase significantly reduced the PFC response, suggesting that the protein moiety played an influential role in the immunogenicity of TCA-LPS. The TCA-LPS contained the same antigenic determinants as PW-LPS, so any difference observed between the PFC response was not due to any associated immunogenic moiety.