bluuebunny/crossref_metadata_2025 huggingface dataset:
Preparation of an Alloantiserum to Murine C3 and Demonstration of Multiple Alleles
Authors: Natsuume-Sakai Shunnosuke , Amano Shigetoyo , Hayakawa Jun-Ichiro , Takahashi Morinobu
Published: December 2022

An antiserum was produced in C3H/He mice that can recognize the antigenic variations of murine C3. The alloantiserum is directed to the gene product of one of the alleles (C3-1a allele) of the C3-1 locus that controls genetic variations of murine C3 and is linked to H-2. The antiserum formed a single precipitin line in the Ouchterlony test with sera obtained from mice that carry the C3-1a/a or C3-1a/b genotype, but it did not react with sera from mice that do not have C3 coded for by the C3-1a allele. By the use of the alloantiserum, we detected a new C3 allotype (C3-1 CC) in SWR/J mice. The C3-1 CC is distinct from the other two allotypes on the basis of antigenicity and isoelectric point (pI). Therefore, at least three alleles have been identified for the C3-1 locus.
Secretion Rate Independent Evaluation of IgM Antibody Avidity at the Level of Single Immunocytes
Authors: Doria Gino , Mancini Camillo , DeLisi Charles
Published: December 2022

The hemolytic plaque inhibition assay has been performed on spleen cells from mice immunized with TNP-HRBC to evaluate avidity of anti-TNP IgM antibodies. At different times after immunization direct plaques were inhibited by soluble TNP-EACA, TNP61-BGG, or anti-µ antiserum. Analysis of the inhibition data provided independent estimates of antibody avidity and secretion rate. Avidity was found to increase with time, to reach a maximum when the antibody response attained the peak value, and then to decline as the response was waning. There was a decrease followed by increase of the secretion rate concomitant with the rise and fall of the antibody response and avidity.
Macrophage Activation for Tumor Cytotoxicity: Development of Macrophage Cytotoxic Activity Requires Completion of A Sequence of Short-Lived Intermediary Reactions
Authors: Ruco Luigi P. , Meltzer Monte S.
Published: December 2022

Macrophages from endotoxin (LPS)-unresponsive C3H/HeJ mice fail to develop tumoricidal activity after any of several in vivo or in vitro treatments that induce cytotoxicity with cells from LPS-responsive C3H/HeN mice. Under certain conditions, however, these macrophages could become tumoricidal: macrophages from in vivo immune reactions such as those induced by BCG infection, but not cells from irritant-induced peritoneal exudates, developed full cytotoxic capability after further exposure in vitro to microgram per milliliter concentrations of LPS or to supernatants from antigen-stimulated leukocyte cultures (lymphokines). Capacity of macrophages from BCG-infected C3H/HeJ mice to become tumoricidal after LPS exposure was gradually lost with time in culture and was absent by 24 hr. Requirements for at least two activation stimuli for cytotoxic activity with C3H/HeJ macrophages could also be fulfilled entirely in vitro: cells cultured in lymphokines plus LPS were fully tumoricidal; macrophages cultured in either agent alone were not cytotoxic. Similar interactions between lymphokines and LPS could be demonstrated with macrophages from LPS-responsive C3H/HeN mice. Tumoricidal activity by C3H/HeN macrophages cultured in lymphokines plus nanogram per milliliter concentrations of LPS was considerably greater than that by cells cultured in either agent alone. LPS and lymphokines were synergistic in their action on macrophage cytotoxicity. The synergistic effect of LPS on cytotoxicity by lymphokine-activated macrophages was evident after a 15-min pulse and was not dependent upon fluid-phase LPS. Synergistic action, however, was dependent upon 1.) treatment sequence: LPS was active only if given simultaneously with or after lymphokine treatment; LPS exposure before lymphokine treatment was ineffective, and 2.) treatment interval: capacity of lymphokine-treated macrophages to express cytotoxic activity after LPS exposure decayed with time in culture and was lost by 24 hr. Thus, macrophage activation for tumor cytotoxicity in both C3H/HeJ and C3H/HeN mice was the final result of a cascade of short-lived intermediary reactions in a defined sequence. Tumoricidal activity of fully activated cells and responsiveness of each noncytotoxic precursor cell to activation stimuli were all short lived macrophage reactions. Present evidence suggests that none of these reactions was reversible. Completion of each state of macrophage activation was completely dependent upon the simultaneous presence of localized activation signals and macrophage precursors able to respond.
Antigen-Initiated B Lymphocyte Differentiation
Authors: Shortman Ken , Howard Maureen C. , Baker Julie A.
Published: December 2022

The effect of specific and nonspecific stimuli on the cycle status of subsets of primary B lymphocytes was assessed by preinjecting donor CBA mice 1 to 2 days previously with various substances, and then incubating the isolated spleen cells with high specific activity 3H-TdR before assay. AFC-progenitor activity was assessed as a response to NIP-POL antigen, either by adoptive transfer to irradiated recipients or by cell culture. Previous studies showed these assays reflected the activity of different subsets of B cells, termed “pre-progenitors” (adoptive assay) and “direct progenitors” (culture assay).

Most functional primary B cells, whether assayed in culture or by adoptive transfer, were not initially in rapid cell cycle in normal adult mice. However, nonspecific stimulation for 1 day caused NIP-specific adoptive transfer IgM AFC-progenitors to enter rapid cell cycle. This effect was independent of T cells and not related to the antigenicity of the stimulus: particulate peritoneal irritants were the most effective stimulants. In contrast to adoptive transfer results, AFC-progenitors assayed in cell culture were unaffected by nonspecific stimuli, but were activated into cell cycle by specific antigen.
Kinetic Studies of Cell Migration in a Modified Boyden Chamber: Dependence on Cell Concentration and Effects of the Chymotrypsin-Like Cationic Protein of Human Granulocytes
Authors: Venge Per
Published: December 2022

Migration of PMN cells was measured kinetically by the “leading front” method in a modified Boyden chamber. In some experiments, release of lactoferrin and of chymotrypsin-like cationic protein were measured simultaneously. Migration, both directed and random, was increased by increased cell concentration. Kinetically, directed migration consisted of two phases. The second phase was more influenced by the cell concentration. Preincubation of the cells with low concentrations of chymotrypsin-like cationic protein stimulated cell migration in the same way as did high cell concentrations, i.e., by primarily affecting the second phase. The effect of chymotrypsin-like cationic protein was time and dose dependent and dependent on the chymotrypsin-like activity of the protein. At high concentrations of protein, the cells were immobilized. Release of granular proteins did not take place during the first phase of migration but was parallel to the second phase. It is concluded that chymotrypsin-like cationic protein might be one of the substances responsible for the effect of cell concentration seen on migration in vitro.
Interactions of C-Reactive Protein and Complement with Liposomes
Authors: Richards Roberta L. , Gewurz Henry , Siegel Joan , Alving Carl R.
Published: December 2022

We found previously that interaction of C-reactive protein (CRP) with liposomal model membranes resulted in complement(C)-dependent membrane damage. In the present study, we investigated the influence of membrane composition on the interactions of CRP and C with liposomes. Adsorption experiments showed that binding of CRP was greatest to strongly positive liposomes. A lesser, but still substantial, extent of CRP binding also was observed with negative liposomes, but negligible amounts of CRP bound to neutral or weakly positive liposomes. CRP-mediated consumption of hemolytic C, and C-dependent glucose release from liposomes both were strongly influenced by liposomal charge, positive being superior to negative. Glucose release and, to a lesser extent, consumption of hemolytic C were inversely related to phospholipid fatty acyl chain length. Phospholipid fatty acyl unsaturation and liposomal cholesterol concentration both had strong influences on C consumption and glucose release. The data suggest that CRP-mediated C consumption and membrane damage require an optimum membrane fluidity.

Complement damage in the presence of CRP was enhanced by certain sphingolipids and also by digalactosyl diglyceride, but not by sphingomyelin. Our results thus demonstrate that CRP-mediated C consumption and C-dependent membrane damage both are influenced by the liposomal membrane composition.
Cell Differentiation in the Presence of Cytochalasin B: Studies on the “Switch” to IgG Secretion after Polyclonal B Cell Activation
Authors: Van Der Loo Wessel , Severinson Gronowicz Eva , Strober Samuel , Herzenberg Leonard A.
Published: December 2022

Mouse spleen cells were cultured with lipopolysaccharide in conditions that activate both IgM and IgG secretion. Addition of cytochalasin B (CB), an inhibitor of cytokinesis, lead to a high degree of polynucleation, with little effect on Ig secretion. Using cytoplasmic staining with fluorochrome conjugated antisera, we determined the numbers of IgG-containing cells that also contained IgM in their cytoplasm. Such double staining cells were relatively more frequent at early times of the cultures, but at all times single producing cells were in the majority. Addition of CB over the period when the IgG producing cells first appear, lead to a marked increased frequency of double staining, polynucleated cells. This characteristic was stable over a period of at least 42 hr, suggesting that each double staining cell actively synthesized both isotypes. When CB was added after IgG production had started, little increase in the numbers of double staining cells were observed, although polynucleation remained extensive. These data confirm previous findings that the lineage of one cell can produce both IgM and IgG. Furthermore, the results suggest that cells in the process of switching from IgM to IgG go through an asymmetric division leading to one IgM-producing and one IgG-producing daughter cell.
Isolation of a Sperm Membrane Sialoglycoprotein Autoantigen from Rabbit Testes
Authors: O'Rand Michael G. , Porter James P.
Published: December 2022

A single rabbit sperm membrane autoantigen has been isolated from rabbit testes by immunoadsorbent chromatography, agarose chromatography, and electrophoresis. This rabbit sperm autoantigen (RSA-1) is located in the sperm plasma membrane and blocks the sperm cytotoxic (immobilizing) activity of autoantisera. RSA-1 is a sialoglycoprotein, approximately 40% carbohydrate and containing 0.2 µg sialic acid/µg protein. Its m.w. is estimated at 13,000 ± 1200 by SDS-PAGE. RSA-1 can self-aggregate into higher m.w. forms. Approximately 0.6 mg of RSA-1 can be isolated from 100 rabbit testes. The isolated RSA-1 shows a 300-fold increase in specific cytotoxic (immobilizing) inhibitory activity over the starting testis pellet material.
Long Term Growth in Vitro of Human T Cell Blasts with Maintenance of Specificity and Function
Authors: Kurnick James T. , Grönvik Kjell-Olof , Kimura Arthur K. , Bertil Lindblom J. , Skoog Valdemar T. , Sjöberg Olof , Wigzell Hans
Published: December 2022

Human T lymphoblasts obtained via antigenic stimulation and gradient centrifugation purification can be maintained as perpetual dividing cells in vitro, retaining specificity and function. By repeated addition of supernatants obtained from in vitro lymphocyte cultures activated with T cell mitogens, one can avoid the need of having antigen continually present, while maintaining proliferation. We have utilized PHA to stimulate tonsil lymphocytes and harvested supernatants after 48-hr. Such supernatants contain growth-supporting factors for T blasts, and these factors are clearly distinguishable from the lectin used to induce the growth factors. Thus, the PHA can be removed from the supernatants via an anti-lectin immunosorbent coupled to Sepharose beads, without having any detectable negative impact on the T blast supporting ability of such supernatants. Conversely, although the lectin is capable of stimulating freshly isolated populations of lymphocytes, the supernatant after lectin removal will not activate detectable numbers of small lymphocytes. In the absence of supernatant factors, antigen-specific cells can also be maintained either by reexposure of the MLR blasts to the original stimulator cell population, or by the addition of the soluble antigen (PPD) to blasts in the company of irradiated syngeneic cells. Alternatively, syngeneic or allogeneic irradiated cells together with mitogen can generate growth-supporting factors and thus sustain blast proliferation.
IgE-Selective and Antigen-Specific Unresponsiveness in Mice
Authors: Usui Mitsuko , Matuhasi Tyoku
Published: December 2022

An experimental model was demonstrated in mice for the induction of IgE-selective unresponsiveness to ovalbumin, a protein antigen. An administration of ovalbumin conjugated with pullulan, a linear polymer of glucose, (OA-pullulan) into mice resulted in the induction of a long lasting, IgE-selective unresponsiveness to the subsequent immunization with native OA in the form optimal to elicit IgE antibody response. The IgE-selective unresponsiveness is antigen specific and is infectious to normal mice by transferring the spleen cells from mice receiving OA-pullulan conjugate at least 2 weeks before. In contrast to other modified antigens, OA-pullulan was found to elicit good IgM and IgG antibody responses, but not an IgE response, without the aid of an adjuvant.
Further Studies on the Activation of Lymphocytes by Endotoxin Protein
Authors: Goodman Gail W. , Sultzer Barnet M.
Published: December 2022

Endotoxin protein, an outer membrane protein of Gram-negative bacteria associated with lipopolysaccharide endotoxin, has been found to be a potent activator of lymphocytes. In the absence of T lymphocytes and macrophages, endotoxin protein can stimulate murine B lymphocytes to synthesize DNA and produce antibodies of diverse specificities. This stimulation is greater than that obtained with two well known murine B cell activators, lipopolysaccharide endotoxin or PPD-tuberculin. Splenic lymphocytes from other species, such as rats, rabbits, and guinea pigs also proliferate when cultured with endotoxin protein. Of particular significance is the finding that endotoxin protein is an activator of human peripheral blood lymphocytes.
The Expression of Ia Antigenic Determinants on Macrophages Required for the in Vitro Antibody Response
Authors: Niederhuber John E. , Allen Paul , Mayo Laura
Published: December 2022

A subpopulation of antigen-presenting macrophages required for an in vitro antibody response to burro erythrocytes was deleted by pretreating the splenic macrophages with anti-Ia serum and complement (C). The in vitro response of the macrophage depleted T-B cell population could not be restored by the addition of macrophages resistant to anti-Ia antibodies and C (Ia-). The response of Ia- macrophages and the macrophage-depleted T-B cells was only reconstituted by the addition of Ia+ macrophages. Macrophages pretreated with anti-Ia antibodies restricted to react with determinants of one I subregion could not support the in vitro antibody response when added to cultures whose macrophages were pretreated with anti-Ia serum and C specific for the I-J subregion. These results confirmed that Ia determinants of the I-A, the I-E, and the I-C subregions were all expressed on the I-J+ macrophage required for an in vitro antibody response.
Interaction of Human Cells with Prostaglandins and Cyclic AMP Modulators
Authors: Trofatter Kenneth F. , Daniels Charles A.
Published: December 2022

The release of prostaglandins (PG) has been incriminated in exacerbations of latent herpes simplex virus (HSV) infections in humans, but the mechanism(s) by which PG could precipitate a recurrence has not been adequately explored. In this study we determined the effect of PG-A1, B1, E1, E2, F2α, and other cAMP elevating agents had on complement (C)mediated lysis and antibody-dependent cell-mediated cytolysis (ADCC) of HSV-infected human fibroblasts (HuF). C-lysis was not inhibited by PG, phosphodiesterase inhibitors, or dibutyryl-cAMP regardless of the concentrations of reagent, human antiHSV, or C that was incubated with the HuF. As determined by 51Cr release, human mononuclear leukocytes (MNL) were inhibited from mediating ADCC by PGE1 or E2 (≥0.1 µg/ml), PGF2α or A1 (≥1.0 µg/ml), PGB1 (10 µg/ml), isoproterenol (≥10-6 M), theophylline (≥10-3 M), methyl-isobutyl-xanthine (≥10-4 M), and db-cAMP (≥10-4 M). The inhibitory effect these reagents had on ADCC was removed by washing the cells and no adverse effect on viability was noted. Another quantitative ADCC assay, antibody-dependent suppression of plaque formation, also indicated that PGE2 inhibited MNL-mediated ADCC, and showed the reagent had no effect on virus growth in HuF. Except for the 1.0 µg/ml concentrations of PGF2α and A1, the inhibition of ADCC produced by PG was correlated with significant elevations of cAMP in the human MNL. Finally, experiments showed that as the concentration of PGE2 added to the cultures increased, there was a decrease in both the number of MNL attached to the HuF and the amount of ADCC obtained. Thus, the data indicated that PGE2 inhibited cytolysis by interfering with attachment of the effector MNL to the antibody-coated target cells. In summary the studies support the proposition PG could cause exacerbation of HSV disease in patients by suppression of ADCC.
3H-5-Uridine Incorporation by Small Lymphocytes from Transplantation-Tolerant and Tolerance-Reversed Rats
Authors: Dennison David K. , Clancy John
Published: December 2022

The level of 3H-5-uridine (3H-Urd) incorporation by rat small lymphocytes (SL) taken from donors tolerant (T) or previously T to an allogeneic skin graft was monitored. DA rats were made T to Lewis (L) alloantigens by the injection at birth of semiallogeneic DA/L F1 bone marrow cells. Eight to 12 weeks after birth, allogeneic L skin was transplanted to the T rats. Animals accepting their skin graft for >100 days were either killed and 3H-Urd incorporation by SL from mesenteric lymph nodes (MLN) and spleen (SPL) was monitored, or the rats were injected with syngeneic DA MLN and SPL cells to abolish tolerance. Tolerance-abolished (TA) animals were killed and 3H-Urd incorporation was monitored autoradiographically at intervals before and after skin graft rejection. The level of graft-vs-host (GVH) reactivity of MLN cells from all T and TA rats was evaluated in a local popliteal lymph node GVH assay. T animals demonstrated a markedly depressed GVH reactivity when compared with control animals. Also, MLN SL from T rats autoradiographically exhibited 14.23 grains/SL, 226% above the average of 6.32 grains/SL observed in MLN SL from control animals. 3H-Urd incorporation by splenic SL from T rats was likewise elevated above control animals, representing a 221% increase. There was no increase in IgM+ cells in T vs. control animals. After i.v. injection of normal DA LN and SPL cells, the allogeneic grafts were rejected at 20 ± 2 days. However, from 15 to 21 days after the injection of syngeneic cells, the TA rat LN and SPL SL incorporated 3H-Urd at levels intermediate between those of T and control animals, but exhibited a level of incorporation not significantly different from controls 24 days after syngeneic cell injection. In addition, GVH reactivity of MLN cells taken from a T animal injected with syngeneic cells was restored gradually over a time period of >24 days. The fact that 3H-Urd levels are elevated in T animals indicates that an active phenomenon is related to the T state. The decrease in uridine incorporation during tolerance reversal provides further support for a metabolic change during tolerance that is different from that found in control animals.
The Activation of Guinea Pig T Lymphocytes by Anti-β2-Microglobulin Serum
Authors: Yamashita Uki , Lögdberg Lennart , Berggård Ingemar , Shevach Ethan M.
Published: December 2022

In order to study further the role of β2-m in the regulation of the immune response, we have examined the effects of a goat anti-guinea pig β2-m serum on a number of T lymphocyte functions in vitro. Anti-β2-m serum produced a marked inhibition of the response of peritoneal exudate T cells to antigen and mitogen stimulation. Surprisingly, a marked activation of lymph node T lymphocyte proliferation was observed in the absence of antigen or mitogen stimulation. This stimulatory effect of anti-β2-m serum was shown to be specific for β2-m and required the presence of macrophages. The T cell proliferative response induced by anti-β2-m could not be blocked by antisera to the antigens of the guinea pig MHC. These studies suggest that β2-m may play some critical role in the immune response at the level of T cell activation.
Nonspecific Activation of Murine Lymphocytes
Authors: Goodman Michael G. , Weigle William O.
Published: December 2022

The role of cellular interaction in the mitogenic activation of B and T lymphocytes by 2-mercaptoethanol was studied in cultures of C3H/St spleen cells. It has previously been shown that both B cell-enriched and T cell-enriched populations are capable of mitogenic response to 2-ME. In the present communication it is demonstrated that reactivity to 2-ME cannot occur when cells are cultured below a minimal permissive cell concentration. This permissive range exhibits an optimum beyond which responsiveness per cell declines. Analysis of the kinetic profile of cells cultured at different densities indicated that this observation was not the result of differential kinetics. When B and T cell-enriched populations were mixed in culture, 2- 3-fold enhancement of the mitogenic response to 2-ME resulted. Optimal responsiveness was shown to occur in the region of moderate T cell excess. Because the B cell-enriched population contained substantially more macrophages than did the T cell-enriched population, synergy was evaluated after the B cells had been depleted of macrophages. The collaborative interaction between B and T cells nevertheless remained intact. Furthermore, when macrophage-poor T cell populations were supplemented with additional macrophages, no synergistic response was observed. Treatment of the B cell-enriched population with anti-immunoglobulin antiserum plus complement completely abrogated the synergistic interaction between the two cell types. Similarly, treatment of the T cell-enriched populations with anti-thymocyte serum plus complement abolished synergy. Thus, these experiments confirm the identity of the collaborating lymphocyte types. T cells were furthermore shown to be capable of providing help to B cells in their response to poly-clonal B cell activation. The implications of these observations are discussed both with regard to the mechanism of activation of lymphocytes by 2-ME and with regard to the role of T cells in the process of polyclonal B cell activation.
Genetic Studies in NZB Mice
Authors: Raveche Elizabeth S. , Tjio Joe H. , Steinberg Alfred D.
Published: December 2022

These studies were performed to test the previously advanced hypothesis that the antibody response to ssDNA is controlled by an X-linked immune response gene. The antibody response to single-stranded DNA (ssDNA) complexed to MBSA was studied in NZB, DBA/2, F1 reciprocal hybrids, and backcross mice. We found that the anti-ssDNA response was not linked to genes located on the X chromosome, but rather that anti-ssDNA production was regulated by sex hormones. Reciprocal F1 hybrid males with either an NZB or a DBA/2 X chromosome were low responders; however, either castration or castration plus estrogen implants significantly increased the amount of antibody produced. Testosterone implants were immunosuppressive in both females and males. Backcross studies suggested that the ability to respond to ssDNA is inherited as an autosomal dominant trait that is further regulated by sex hormones.

Control studies were performed in hybrids of the CBA/N strain, which has a well established X-linked defect in the ability to respond to a variety of thymic independent antigens including polyriboinosinic·polyribocytidylic acid (rI·rC). F1 hybrid males that received the CBA/N X chromosome were low-responders to rI·rC despite the hormonal status of the animal. On the other hand, females were high responders to rI·rC but testosterone implants in females decreased the amount of antibody produced.

These studies indicate that it is possible to separate sex hormone effects from X-linked genetic effects on immune responses. We found that the response to ssDNA is controlled by sex hormones but not an X-linked immune response gene.
Many Bacterial Species Bind Human IgD
Authors: Forsgren Arne , Grubb Anders O.
Published: December 2022

Forty-four bacterial strains belonging to 19 species were tested for their IgD-binding capacity by incubation with radiolabeled human IgD. A high binding of IgD to Neisseria catarrhalis and Hemophilus influenzae and a moderate binding of IgD to streptococci of the groups A, C, and G were found. Two strains of N. catarrhalis were tested for their ability to bind selectively the IgD in normal pooled serum and in three serum samples with IgD M components and were found to possess this property. Binding studies with radiolabeled IgD Fab and Fc fragments indicated that the binding mainly but not exclusively involves the CH1 region of the IgD molecule.
Cytotoxicity by Cultured Human Lymphocytes: Characteristics of Effector Cells and Specificity of Cytotoxicity
Authors: Ortaldo J. R. , Bonnard G. D. , Kind P. D. , Herberman R. B.
Published: December 2022

The specificity and the cells involved in natural cytotoxicity against tumor cells by fresh normal human peripheral blood mononuclear leukocytes have been extensively examined. Recently we have observed that removal of Fc receptor-positive cells from peripheral blood mononuclear leukocytes on day 0 removed all natural cytotoxic activity against K-562 (a myeloid leukemia cell line that is particularly susceptible to natural cytotoxicity) but after in vitro culture for 7 days, cytotoxic activity against K-562 had regenerated to near its original level.

In the present study, we examined the specificity of the cytotoxicity of both fresh and cultured peripheral blood mononuclear leukocytes against a panel of target cells susceptible to natural killing and the characteristics of the cultured effector cells. By testing various donors against a checkerboard panel of labeled target and cold target inhibitor cells, it was found that the reactivity after culture was similar to that seen on day 0. Small variation in rank order of the level of inhibition was observed in most experiments, but none of the reactivity patterns of the donor lymphocytes demonstrated major changes after culture. Furthermore, the surface characteristics of the effector cells before and after culture were similar with regard to surface immunoglobulin, Fc receptors, complement receptors, sensitivity to trypsin and to specific anti-T antiserum. However, the cultured effector cells did have some distinctive characteristics, rosetting less with sheep red blood cells, adhering to some extent to rayon wool, and being partially sensitive to an anti-Ia-like antiserum. These features may indicate that the cultured cells are at a different stage of differentiation than in vivo generated natural killer (NK) cells. These results indicate that the natural cytotoxicity of human lymphocytes and the reactivity of cultured cells are mediated by similar effector cells, i.e., NK cells, and are directed against similar specificities.
Studies of the Mechanisms for the Induction of in Vivo Tumor Immunity
Authors: Igarashi Tsuyoshi , Rodrigues Dennis , Ting Chou-Chik
Published: December 2022

The effect of macrophages on the induction of the cell-mediated cytotoxicity against a leukemia in a syngeneic system was investigated. The addition of exogenous peritoneal cells from normal C57BL/6 mice enhanced the in vitro secondary cell-mediated cytotoxic response of both spleen and lymph node cells as responding cells against syngeneic FBL-3 leukemia. Peritoneal phagocytic macrophages seemed to be responsible for the enhancement. No inhibitory effect was demonstrated by the addition of peritoneal macrophages at a concentration as high as 20%, whereas the primary cytotoxic allograft response was significantly suppressed. In the present studies, there was no absolute restriction of macrophage-T cell interaction by an H-2 barrier. Supernatants of peritoneal macrophage cultures also enhanced this cell-mediated cytotoxic response. There was no difference between the effects of syngeneic or allogeneic peritoneal macrophage culture supernatants.
Formation of Thromboxane B2 and Hydroxyarachidonic Acids in Purified Human Lymphocytes in the Presence and Absence of PHA
Authors: Parker Charles W. , Stenson William F. , Huber Mary G. , Kelly James P.
Published: December 2022

The metabolism of exogenous and endogenous [14C] arachidonc acid was studied in purified human peripheral blood lymphocytes carefully freed of contaminating platelets. Formation of products co-migrating in a number of different solvent systems with 5-hydroxyarachidonic acid (5-HETE), thromboxane B2 (TB2), prostaglandins and probably 12-hydroxyarachidonic acid (12-HETE) was demonstrated. In cells prelabeled with [14C] arachidonic acid, phytohemagglutinin (PHA) produced substantial (3.5- to 12-fold) increases in 5-HETE, 12-HETE, and TB2 radiolabeling. The metabolism of exogenous [14C] arachidonic acid was much less affected by PHA. Since PHA releases cell-bound arachidonic acid, it appears that the response involving endogenous label is due to increased availability of free arachidonic acid rather than induction of arachidonic acid-metabolizing enzymes. Various inhibitors of arachidonic acid metabolism exerted similar effects in lymphocytes to those described previously in other tissues providing a possible basis for interpreting their inhibitory effects on mitogenesis, described in the preceding paper.
In Vivo Generation of Hapten-Specific Killer T Cells without Elimination of Suppressor Cells
Authors: Butler Larry D. , Battisto Jack R.
Published: December 2022

We have provided evidence to demonstrate that hapten-specific killer cells can be generated in vivo toward hapten-conjugated syngeneic splenic cells. The critical aspect is to provide an auxiliary cellular antigenic stimulus in addition to the hapten-conjugated syngeneic cells. In our experiments this stimulus was CBA/J splenic cells that possess Mls disparate but H-2 compatible antigens with C3H/HeN hosts. The Mls antigen has been shown by others to activate helper T cells in vitro to synthesize KAF, a signal that prekiller cells require besides target antigen. Killer cells (shown to be T cells) as well as helper T cells were found to be derived from the C3H host. Induction in the host animal of partial tolerance to the auxiliary cells possessing Mls antigen abrogated the response.

This system, besides providing a better understanding of the control mechanisms involved in the development of T killer cells in vivo, points to a way in which the latter may be generated toward haptenic moieties on syngeneic cells without the necessity for eliminating suppressor cells. The latter principle may prove highly useful in certain clinical situations.
Defective Tumoricidal Capacity of Macrophages from A/J Mice
Authors: Boraschi Diana , Meltzer Monte S.
Published: December 2022

Macrophages from A/J mice fail to develop tumoricidal activity after any of several in vivo or in vitro treatments that activate cells from C3H/HeN mice. Peritoneal macrophages from A/J mice treated i.p. with viable Mycobacterium bovis, strain BCG, killed Corynebacterium parvum, or pyran copolymer fail to develop in vitro tumoricidal activity; varying the numbers of macrophages from treated mice added to target cells, or the dose and time of treatment, or the treatment schedule of these in vivo activation stimuli did not evoke cytotoxic activity. Moreover, cytotoxic activity by macrophages from A/J mice was not observed with any of four target cell lines derived from three different mouse strains. In vitro treatment of peritoneal exudate macrophages from A/J mice with lymphokine-rich supernatants, bacterial endotoxins, or T cell mitogens was also ineffective; varying the numbers of treated macrophages added to target cells, the dose of in vitro activation stimuli, or the time of treatment did not evoke cytotoxic activity. Thus, A/J mice exhibit a profound defect in macrophage tumoricidal capacity to both in vivo and in vitro activation stimuli over a wide range of experimental conditions.
Macrophage-Lymphocyte Clusters in the Immune Response to Soluble Protein Antigen In Vitro
Authors: Braendstrup Otto , Werdelin Ole , Shevach Ethan M. , Rosenthal Alan S.
Published: December 2022

We have assessed the genetic restrictions on physical interactions between macrophages and central lymphocytes and between central and peripheral lymphocytes in antigen-specific macrophage-lymphocyte clusters with respect to I-region differences of inbred strains 2 and 13 guinea pigs. When using lymphocytes from guinea pigs immunized with DNP-OVA or DNP-GL in CFA, the antigen-specific interaction between central lymphocyte and macrophage requires that both cells be derived from animals syngeneic at the I-region of the major histocompatibility complex. In studies using antigens, the response to which is under the control of MHC-linked Ir genes, macrophages from the responder, but not from the nonresponder parental strain support cluster formation with responder × nonresponder F1(2 × 13) T cells. In contrast, the physical interactions between central and peripheral T lymphocytes are not restricted by the I-region of the MHC and the peripheral lymphocyte need not be from an animal immune to the antigen used to drive macrophage central lymphocyte interactions.
Hypocomplementemia Associated with Naturally Occurring Lymphosarcoma in Pet Cats
Authors: Kobilinsky Lawrence , Hardy William D. , Day Noorbibi K.
Published: December 2022

Eighty cats were classified by indirect immunofluorescence and histologic diagnosis into four categories: normal, feline leukemia virus (FeLV) infected; normal noninfected; lymphosarcoma-FeLV infected; lymphosarcoma, no FeLV present. All viremic cats with lymphosarcoma were found to be hypocomplementemic and activation of the complement system had occurred via the classical pathway. Sera of cats with lymphosarcoma in the absence of FeLV had varying levels of total hemolytic complement (TCH50) ranging from normal to hypocomplementemic. Approximately 50% of the cats that were viremic but histologically and clinically free of disease had TCH50 levels within normal range, and the remainder exhibited varying degrees of hypocomplementemia.
PPD-Induced Immunoglobulin Production in Human Peripheral Blood Lymphocytes
Authors: Nishikawa Shin-Ichi , Hirata Takeo , Nagai Tomoko , Mayumi Mitsufumi , Izumi Takateru
Published: December 2022

The ability of PPD to induce Ig production in human PBL was investigated. PPD proved to be a good B cell activator for inducing polyclonal Ig production in PBL from healthy Japanese. Comparative studies of this response with PWM-induced Ig production showed that the cellular mechanisms involved in the two responses were different. First, PBL from an atypical individual with a deficient IgM production to PWM responded normally to PPD with IgM production as well as IgG production. Secondly, in IgG production, the effects of the two mitogens (PPD and PWM) were additive.

An analysis of the cellular requirements in PPD-induced Ig production clearly demonstrated that T cells played a role in this response as well as in the PWM-induced response. However, the head-to-head comparative study on the titration curves of helper T cells in the two responses showed that PWM-induced helper activity was 2 to 5 times more effective than PPD-induced helper activity. Moreover, PPD-induced helper activity was shown to be more sensitive to ionizing radiation than was PWM-induced helper activity. Thus, this system of PPD-induced Ig production may provide a useful tool for understanding the human antibody production system as well as the PWM-induced response.
Regulation of IgE Antibody Production by Serum Molecules
Authors: Katz David H. , Bargatze Robert F. , Bogowitz Cheryl A. , Katz Lee R.
Published: December 2022

Two biologically active serum molecules manifesting precisely opposite biologic effects, both of which are selective for IgE antibody synthesis, can be detected in the serum and ascites fluids of CFA-immune mice. One activity, described previously, is suppressive and hence termed suppressive factor of allergy (SFA); the other, reported for the first time herein, is enhancing and has been termed enhancing factor of allergy (EFA). The ability to detect one vs the other activity requires certain special manipulations such as different doses of low dose x-irradiation. Conclusive evidence for the existence of two distinct factors mediating these two opposing biologic effects was obtained in studies demonstrating that affinity chromatography on concanavalin A-Sepharose segregated the two molecular entities. Thus, SFA binds poorly or not at all to Con A-Sepharose, whereas EFA binds to Con A and can be recovered in the eluate eluted with the competitive sugar α-methyl-D-glucopyranoside.
Local Production of IgG, IgA, and IgE Antibodies in Grass Pollen Hay Fever
Authors: Platts-Mills Thomas A. E.
Published: December 2022

An antigen-binding radioimmunoassay was used to measure binding activity for the group I protein of Rye grass pollen (BA). Most patients with grass pollen hay fever were found to have IgG, IgA, and IgE BA in both their serum and nasal secretions. When these results were related to the quantities of total immunoglobulin of each class, it was clear that the IgG and IgA in nasal secretions had a higher proportion of pollen-specific ab than the IgG and IgA in serum. This calculation showed that most of the IgG BA and IgA BA in nasal secretions must have been produced locally. In contrast, IgG ab to diphtheria toxin was found to represent a lower proportion of nasal IgG than serum IgG. These findings for IgG anti-toxin supported the view that the pollen-specific IgG BA must have been produced locally. Specific antipollen IgE ab in nasal secretions was compared with the other classes. The results suggested that the IgE ab in nasal secretions was also locally produced. Furthermore, saliva samples from the patients that contained similar total quantities of IgG and IgA to nasal secretions were found to have little or no pollen-specific IgG, IgA, or IgE BA. After injections of grass pollen extract, the major antibody response was an increase in serum IgG BA, which was not matched in nasal secretions. Calculations in relation to total IgG showed that the IgG BA response to injections was unlikely to have been produced in the same place as the IgG BA produced in response to natural exposure. The findings show that the ab response to pollen antigens is truly local. The possibility that serum IgG BA and IgE BA was produced predominantly in local lymph nodes whereas the BA found in nasal secretions was derived from plasma cells in the nasal mucosa is discussed.
Macrophage Activation by Mycobacterial Water Soluble Compounds and Synthetic Muramyl Dipeptide
Authors: Wahl S. M. , Wahl L. M. , McCarthy J. B. , Chedid L. , Mergenhagen S. E.
Published: December 2022

The adjuvant effects of mycobacteria can be replaced by more chemically defined isolates of the cell walls including a water soluble fraction (WSA) and by the synthetic analog N-acetyl-muramyl-L-alanyl-D-isoglutamine (MDP), which is the minimal structure required for adjuvanticity. These compounds can directly activate macrophages as determined by an increase in spreading and adherence and by an elevated synthesis of the enzyme collagenase. Moreover, this increase in collagenase production is modulated by enhanced production of prostaglandins that influences intracellular levels of cyclic AMP. In addition, both MDP and WSA induced macrophages to produce a biologically active mediator that triggers quiescent fibroblasts into active proliferation. It thus appears that a mechanism for mycobacterial adjuvant action as determined with MDP and WSA is via activation of macrophages, which may then precipitate a multiplicity of other reactions resulting in enhanced immune phenomena. Furthermore, the granulomatous and fibrotic reactions associated with mycobacterial infection may be a consequence of this direct activation of macrophages.
Lyb-7, a New B Cell Alloantigen Controlled by Genes Linked to the IgCH Locus
Authors: Subbarao Bondada , Ahmed Aftab , Paul William E. , Scher Irwin , Lieberman Rose , Mosier Donald E.
Published: December 2022

Anti-Lyb-5.1 serum contains antibodies against two different B cell surface antigenic determinants, Lyb-5.1 and Lyb-7.1, which are defined in cytotoxicity and functional tests, respectively. The antibody against Lyb-7.1 is identified by its ability to specifically inhibit in vitro primary antibody responses to TNP-Ficoll. Anti-Lyb-5.1 serum can be made monospecific for anti-Lyb-7.1 activity by absorption with spleen cells from AL/N mice which have been typed as Lyb-5.1+, Lyb-7.1-. Lyb-5.1 and Lyb-7.1 are each under control of one or one set of closely linked genes. The loci specifying Lyb-5.1 and Lyb-7.1 are not linked to each other nor to M1s and H-2 loci. However, the gene controlling the expression of Lyb-7.1 is linked to the genes coding for the constant region of immunoglobulin heavy chains.
Antigen-Induced Changes in the Proportion of Antigen-Binding Cells Expressing IgM, IgG, and IgD Receptors
Authors: Kanowith-Klein Susan , Vitetta Ellen S. , Korn Edward L. , Ashman Robert F.
Published: December 2022

Changes in the isotype of the antigen receptor after immunization were assessed by using antigen-binding cells (ABC) from mouse spleen specific for sheep erythrocytes (SRC). Rabbit antisera specific for mouse heavy chains µ, γ, and δ, as well as polyspecific anti-Ig were used to inhibit SRC binding to the receptors of nonimmune ABC and immune ABC 5 days after immunization with SRC.

Nonimmune ABC were more heterogeneous than immune ABC in their sensitivity to inhibition by polyspecific anti-Ig and anti-µ in that they included a greater proportion of cells inhibited by low concentrations of these antisera. The maximum percent of cells inhibitable by anti-µ (60 to 80%) did not change after immunization. Whereas IgG was not detectable on nonimmune ABC, after immunization 40 to 50% of the ABC expressed IgG. The proportion of ABC with IgD receptors fell from about 45% in nonimmune mice to around 25% in 5-day immune mice.

From the inhibition produced by two or three antiheavy chain sera present individually or simultaneously, the proportion of cells bearing various combinations of surface Ig isotypes were estimated. Thus, nonimmune and immune ABC-bearing IgD also appeared to bear IgM although a few immune ABC expressed only IgM. In immune ABC, IgG-bearing ABC overlapped extensively with the IgM-bearing population but not with the IgD-bearing population. ABC bearing IgD alone were not detected, but immune animals probably contained µ+δ+γ+ ABC.

These results suggest that antigen contact with specific ABC triggers changes of receptor isotypes during the expansion of the ABC population including a partial loss of IgD and the appearance of IgG.
Shared Idiotypes on Human Peripheral Blood B and T Lymphocytes
Authors: Lea Tor , Førre øystein T. , Michaelsen Terje E. , Natvig Jacob B.
Published: December 2022

In a patient with an IgG λ monoclonal serum component possessing anti-streptolysin O activity, we have demonstrated peripheral blood B and T lymphocytes with shared or similar idiotypes. The idiotypic T lymphocyte membrane structure was capable of binding the specific antigen (SLO). After radioiodination and subsequent detergent solubilization of the same T cell population, immunoprecipitation of the lysate by employing anti-idiotypic antibodies, resulted in the isolation of a polypeptide chain with a m.w. of 70,000 on SDS polyacrylamide gels under reducing conditions. The polypeptide expressed no isotypic immunoglobulin markers. Internal labeling experiments indicated that this membrane structure was actively synthesized by the T lymphocytes.
An Anticomplementary Agent, K-76 Monocarboxylic Acid: Its Site and Mechanism of Inhibition of the Complement Activation Cascade
Authors: Hong Kyongsu , Kinoshita Taroh , Miyazaki Wasei , Izawa Taketoshi , Inoue Kozo
Published: December 2022

A monocarboxylic acid derivative (K-76 COOH) of K-76, purified from the culture filtrate of Stachybotrys complementi nov. sp. K-76, inhibits complement (C) activity. Its inhibitory action is mainly on C5 step. It strongly inhibits the generation of EAC1̄,4b,2a,3b,5b from C5 and EAC1̄,4b,2a,3b, and accelerates the decay of EAC1̄,4b,2,3b,5b. It also causes some inhibition of the reactions of C2, C3, C6, C7, and C9 with their respective preceding intermediate cells. It has no effect on the generation of EAC1̄,4b from C4 and EAC1̄, or of EAC-8 from C8 and EAC-7, and apparently increases the generation of EAC1̄,4b from C1 and EAC4b probably by inhibiting transfer or turnover of C1. It does not affect the rate of decay of EAC1̄,4b,2a or the T max of generation of EAC1̄,4b,2a, and it inhibits immune adherence only at high concentration.

K-76 COOH also strongly inhibits hemolysis through the alternative pathway of C activation by cobra venom factor, but it does not seem to inhibit the early steps of the alternative pathway, because it has little affect on the consumption of C3 or the conversion of β1C to β1A on treatment of C serum with zymosan.

K-76 COOH probably combines with C5 molecules, forming the inactive complexes, or it causes the structural alteration of C5.
Manipulation of Cytotoxic T Lymphocyte Responses to Syngeneic Tumors
Authors: Altman Amnon , Bechtold Thomas E. , Katz David H.
Published: December 2022

This study was designed to extend a recent report about the ability of hapten-primed T lymphocytes to serve as helper cells, capable of augmenting the generation of tumor-specific CTL in vivo when the priming hapten was coupled to the immunizing tumor cells. Spleen cells of mice primed with 2,4,6-trinitrophenyl-mouse γ-globulin (TNP-MGG) conjugate under conditions known to elicit hapten-reactive T lymphocytes, and subsequently immunized with TNP-derivatized, mitomycin C-treated syngeneic tumor cells, generated, upon in vitro stimulation with the relevant tumor cells, substantially higher CTL responses than did spleen cells from mice that were not primed with TNP-MGG. Furthermore, we demonstrated directly that such help resulted from T-T cell cooperation by showing that the addition of TNP-MGG-primed spleen cells in culture augmented the in vitro induction of a secondary tumorspecific CTL response upon sensitization of tumor-primed spleen cells against TNP-derivatized tumor cells. These experiments, therefore, establish a convenient in vitro model system that allows study of the regulatory influences of hapten-reactive T lymphocytes on tumor-specific, cell-mediated immune responses, and open obvious new avenues for the potential immunotherapy of growing tumors.