bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of castration on male-determined acceleration of autoimmune disease in BXSB mice.
Authors: Eisenberg R A , Dixon F J
Published: December 2022

The autoimmune disease of BXSB mice is markedly accelerated by a Y chromosome-associated male-specific factor. In order to investigate the mechanism of this effect, we castrated male and female BXSB mice at 10 days of age and compared the course of development of their autoimmune syndrome with that of sham-operated, sex-matched, littermate controls. Orchiectomy did not delay the early onset of fatal immunopathology in males, and oophorectomy did not exacerbate the indolent progression of morbidity in females. We conclude, therefore, that the male-specific effect that accelerates BXSB autoimmune disease is not hormonally mediated.
Mechanism of neonatal idiotype suppression. I. State of the suppressed B cells.
Authors: Fung J , Köhler H
Published: December 2022

Neonatal BALB/c mice, given small amounts of an anti-idiotype serum directed against the TEPC-15 idiotype, are chronically unresponsive to immunization with PC antigens. These mice recover from suppression slowly over a period of 10 mo, and their response is not of TEPC-15 idiotype. However, the PC-specific precursors, as analyzed by the splenic fragment culture technique, in 8-mo-old suppressed mice are normal with respect to their frequency and idiotype dominance. Furthermore, the PC-precursors in neonatally suppressed BALB/c are sensitive to tolerance induction, as are precursors from neonatal liver and spleen cells. When chronic suppressed mice are immunized with a novel TI-1 antigen, PC-LPS, shown to stimulate immature PC-specific precursors, their response to PC is 80% of TEPC-15 idiotype, whereas their response to a TI-2 PC antigen and to a TD PC antigen is not TEPC-15 dominant. These results indicate that the TEPC-15 positive B cells in neonatally idiotype suppressed mice are in a state of immaturity that resembles the developmental characteristics of normal neonatal BALB/c mice.
PGBX, a prostagandin derivative, mimics the action of the calcium ionophore A23187 on human neutrophils.
Authors: Serhan C N , Korchak H M , Weissmann G
Published: December 2022

The interactions have been studied of a water-soluble, polymeric derivative of prostaglandin B1, PGBX, with human polymorphonuclear leukocytes (PMN). PGBX, which is a potent ionophore of divalent cations, provoked superoxide anion (O2.-) generation and lysosomal enzyme release in cytochalasin B-treated PMN in the presence of extracellular divalent cations (Ca2+, Sr2+, Mg2+, Mn2+, Ba2+). Kinetic and dose-response studies showed that PGBX mimicked te action of ionophore A23187 in PMN. Both ionophores induced superoxide generation and release of enzymes from specific and azurophil granules (lysozyme > beta-glucuronidase) without provoking release of the cytoplasmic marker enzyme lactic dehydrogenase. In contrast, the precursor of PGBX, prostaglandin B1 (PGB1), and arachidonate did not mimic ionophore-induced stimulation of PMN. PGBX induced enzyme release both in the presence of extracellular Ca2+ and Ba2+ (both of which it translocates in model liposomes), whereas A23187 showed specificity for Ca2+ (which it translocates preferentially over Ba2+). These studies indicate that the actions of a water-soluble polymer (PGBX) derived from a naturally occurring prostaglandin (PGB1) on human neutrophils resemble those of a classical ionophore (A23187). Moreover, they provide additional evidence that increments in the intracellular levels of divalent cations may signal stimulus-secretion coupling in human neutrophils.
Human lymphocyte-activating properties of a purified polysaccharide from Candida albicans: B and T cell cooperation in the mitogenic response.
Authors: Piccolella E , Lombardi G , Morelli R
Published: December 2022

Purified human T and B lymphocytes were tested for their ability to show a mitogenic response to a purified polysaccharide (MPPS) extracted from C. albicans. It was found that neither T nor B cells alone could respond to MPPS. When a mixture of these two types of cells was tested, a normal response (comparable to the one of unfractionated peripheral blood lymphocytes) was observed. This response was not affected by prior depletion of macrophages. Results obtained with normal B or T cells mixed with mitomycin C-treated T or B cells indicated that a reciprocal helper effect exists between these two types of cells in their response to MPPS. This response could be inhibited by antisera against HL-A and Ia-like antigens.
Features of cells controlling H-2-restricted presentation of antigen to T helper cells in vivo.
Authors: Sprent J
Published: December 2022

Information was sought on the nature of cells presenting antigen (sheep red blood cells) in an H-2-restricted fashion to T helper cells in vivo. The approach used was to examine the capacity of F1 hybrid T cells to collaborate with parental strain B cells after prior positive selection of the T cells to antigen in irradiated parental strain mice in the presence of various sources of Thy 1-negative cells taken from F1 or parental strain mice. The results showed that H-2-restricted presentation of antigen to T cells is controlled by radioresistant, nylon-wool adherent. Thy 1-negative cells, which are enriched in spleen and peritoneal exudates, rare in lymph node cell suspensions, and undetectable in thoracic duct lymph; pretreatment in vitro with anti-Ia reagents directed either to I-A or I-E subregion determinants abolished antigen-presenting activity. These findings correlate closely with the known characteristics of cells that present antigen in vitro. Studies in which re-irradiated parent leads to F1 chimeras were used as hosts for positive selection suggested that nonhematopoietic cells do not play a discernible role in antigen presentation.
Identification of target antigen for antibody-dependent cellular cytotoxicity on cells carrying Epstein-Barr virus genome.
Authors: Takaki K , Harada M , Sairenji T , Hinuma Y
Published: December 2022

The target antigen for antibody-dependent-cellular cytotoxicity (ADCC) on Epstein-Barr virus-(EBV) carrying lymphoblastoid cells expressing EBV-specific membrane antigen (MA) were examined with human serum antibody and adult human peripheral lymphocytes as effector cells. These studies confirmed that anti-MA-positive but not MA-negative sera were reactive in the ADCC. The ADCC reaction was positive with cells in which the MA consisted of late (LMA) and early (EMA) components. These included 1) MA-positive cells prepared by EBV antigen-adsorption, 2) cells carrying de novo-synthesized MA without adsorbed MA, and 3) EBV-producer cells expressing MA spontaneously. In all these preparations, the target cells were lysed roughly in parallel with the frequency of MA-positive cells. Inhibition of LMA synthesis in EBV-superinfected cells by phosphonoacetate (PA) reduced ADCC sensitivity significantly and to a far greater extent than MA synthesis as measured by immunofluorescence. This suggests that a target for ADCC is the PA-sensitive LMA. No ADCC reaction occurred with the cell preparation comprised of a high percentage of MA-positive cells induced by 5-iodo-2'-deoxyuridine, which is believed to be EMA only. These results strongly suggest that the target antigen for ADCC in EBV-positive cells is a late MA but not early MA.
Fc receptors on human T lymphocytes. V: Effects of colchicine and cytochalasin B on Fc receptor expression.
Authors: Reaman G H , Poplack D G , Broder S , Pichler W J
Published: December 2022

Previous studies have documented that the interaction of IgG immune complexes with the FcIgG receptor (Fc-IgGR) of human T lymphocytes induces alterations both in the expression of Fc receptors and in the functional capacities of these cells. After IgG immune complex exposure, Fc-IgGR-bearing T lymphocytes lose their Fc-IgGR and generate Fc-IgMR in subsequent culture. This transition phenomenon is mediated via patching and capping of the Fc-IgGR. Since Colchicine and Cytochalasin B have been demonstrated to interfere with capping of other membrane elements by their known effects on cytoskeletal structures, we investigated the effects of these agents on the expression of Fc receptors of human T cells. Neither Colchicine nor Cytochalasin B directly affects Fc-IgGR or Fc-IgMR expression. However, both agents inhibit the loss of Fc-IgGR subsequent to IgG immune complex exposure as well as the development of Fc-IgMR in culture. Immunofluorescent studies illustrate that this inhibition is mediated in part by interference with capping of the Fc-IgGR-immune complex ligand. This study emphasizes the inverse relationship between Fc-IgGR and Fc-IgMR expression on human T lymphocytes.
Development of functional complement receptors during in vitro maturation of human monocytes into macrophages.
Authors: Newman S L , Musson R A , Henson P M
Published: December 2022

The development of Fc and C3 receptor function was evaluated during differentiation of human peripheral blood monocytes into macrophages in an in vitro culture system. At the time of isolation, monocytes formed rosettes with C3-coated erythrocytes (EIgMC) but did nt internalize them, whereas IgG-coated erythrocytes (EIgG) were both bound and ingested. During the first 24 hr in culture, IgG-mediated phagocytosis and C3-mediated rosette formation declined. After 7 days of in vitro differentiation, monocyte-derived macrophages were capable of ingesting 2 to 4 times as many EIgG as uncultured monocytes. In addition, macrophages demonstrated a new function--the ability to ingest C3-coated erythrocytes, as well as to efficiently bind them. The time course of the reappearance of receptor functions varied between different individuals, but by day 7, the activity of Fc and C3 receptors was vigorous for all individuals tested. Monocytes underwent differentiation whether they were cultured on glass coverslips or on a plastic surface. However, macrophages demonstrated C3-mediated ingestion earlier when cultured on plastic. Culturing monocytes on the different surfaces did not affect the time course of the reappearance of Fc receptor function, but did have a small effect on the magnitude of the response. These experiments demonstrate for the first time, that a defined population of human phagocytic cells can acquire the ability to mediate ingestion through their C3 receptors.
Phenotypic expression of I-A and I-E/C subregion determinants on murine thymocytes.
Authors: Sharrow S O , Ozato K , Sachs D H
Published: December 2022

The expression of Ia antigens on murine thymocytes has been assessed by indirect immunofluorescence and flow microfluorometry analysis. Antisera specific for I-region determinants were found to react with at least 50% of thymus cells in single-cell suspensions. When these antisera were tested on thymus cells of an appropriate recombinant strain, it was found that both I-A and I-E/C subregion determinants could be detected. Monoclonal antibodies to I-A and to I-E determinants also reacted with thymus cells in a similar fashion, eliminating the possibility that contaminating specificities accounted for the reactivity of alloantisera. The majority of Ia-positive thymus cells appeared to express determinants encoded by genes of both I-A and I-E/C subregions. Ia expression on individual thymocytes was also correlated with cell size as measured by forward light scatter intensity. However, unlike antigens detected by rabbit anti-mouse brain antisera, Ia antigens did not appear to be associated with known subpopulations of thymocytes distinguishable by light scatter intensity. The presence of I-region encoded antigens on thymocytes may play a role in maturation of T cells in the thymus microenvironment.
Circulating immune complexes associated with naturally occurring lymphosarcoma in pet cats.
Authors: Day N K , O'Reilly-Felice C , Hardy W D , Good R A , Witkin S S
Published: December 2022

Cats were classified into 4 categories by immunofluorescence antibody assay for the presence of feline leukemia virus (FeLV) and histologically as a) normal, FeLV-, b) normal, FeLV+, c) lymphosarcoma (LSA), FeLV+, and d) LSA, FeLV-. Determinations by Raji cell radioimmunoassay modified for cat serum revealed circulating immune complex (CIC) levels of healthy cats to be less than or equal to 50 micrograms equivalent aggregated cat immunoglobulin/ml (microgram/ml). In contrast, sera of cats in groups b, c, and d all contained significantly higher CIC levels (up to 12,000 micrograms/ml) associated with marked hypocomplementemia and C activation occurring via the classical pathway. Sera from FeLV+, LSA+ cats with high levels of CIC and sera of healthy cats were fractionated according to size and bouyant density by centrifugation through 10 to 40% sucrose gradients. Analysis of fractions from LSA+, FeLV+ sera revealed that both immune complexes (ICs), FeLV reverse transcriptase (RT), and IgG were present in fractions corresponding to a bouyant density of 1.15 to 1.18 g/ml. The CIC containing fractions activated C by the classical pathway. Sera from normal cats did not have CIC or RT and none of the fractions activated the classical pathway. These data suggest that vital antigen-antibody complexes are present in sera of viremic cats with LSA and these complexes activate the C system.
Augmented induction of tumor-specific resistance by priming with Mycobacterium tuberculosis (TBC) and subsequent immunization with PPD-coupled syngeneic tumor cells.
Authors: Takatsu K , Hamaoka T , Tominaga A , Kanamasa Y
Published: December 2022

The present study investigates the augmenting effect of tuberculin- (PPD) reactive amplifier T cells on the induction of syngeneic tumor immunity. PPD-reactive helper (amplifier) T cell activity was generated in C3H/HeJ mice by appropriate immunization with heat-killed Mycobacterium (Tbc). Immunization of these Tbc-primed mice with PPD-coupled syngeneic X5563 tumor cells led to augmented generation of in vivo tumor-neutralizing activity contingent on the presence of PPD-reactive amplifier T cell activity. Splenic T cells from these mice exhibited potent tumor-neutralizing activity using Winn's assay, whereas spleen cells from mice not primed with Tbc before PPD-X5563 immunization failed to neutralize viable X5563 tumor cells. After establishing that the neutralizing activity was tumor specific and mediated by T cells, the applicability of this augmentation of tumor-specific immunity to an immunotherapy model was explored. Immunization with PPD-X5563 in the early stages of the tumor-bearing state induced potent anti-tumor activity sufficient to reject the growing tumor. Pretreatment of mice with cyclophosphamide or light x-irradiation (250 R), procedures that eliminate suppressor cell activity nonspecifically, before priming with Tbc further potentiated the anti-tumor activity under these conditions. Thus, the present study elucidates the augmenting effect of PPD-reactive amplifier T cells in the induction of tumor-specific immunity and provides an effective method of immunotherapy in tumor-bearing animals.
Antigen and receptor-driven regulatory mechanisms. VI. Demonstration of cross-reactive idiotypic determinants on azobenzenearsonate-specific antigen-binding suppressor T cells producing soluble suppressor factor(s).
Authors: Dietz M H , Sy M S , Greene M I , Nisonoff A , Benacerraf B , Germain R N
Published: December 2022

The first detectable suppressor T cell (Ts) arising after i.v. administration of azobenzenearsonate- (ABA) conjugated syngeneic spleen cells to A/J mice has been studied for its receptor specificity and ability to produce soluble suppressor factor(s). This cell, termed Ts1, has a specific receptor for the eliciting antigen ABA, as demonstrated by selective binding to ABA protein- but not TNP protein-coated plastic dishes. The activity of ABA-Ts1 can be abrogated by treatment with anti-idiotypic antibodies made against anti-ABA antibodies of A/J mice (anti-CRI), indicating that these ABA-binding cells possess a surface receptor structure sharing idiotypic determinants with antibodies of the same specificity. Finally, soluble extracts from, antigen-adherent ABA-Ts1, but not nonadherent cells from the same spleen cell population, possess suppressive activity when assayed directly for afferent suppression or tested for their ability to trigger a second population of Ts (Ts2) in naive recipients. These findings demonstrate a close concordance between a T cell surface receptor, soluble T suppressor factors, and B cell derived antibody, all capable of direct recognition of the eliciting ABA antigen.
Specific antibody-dependent phagocytosis of lipid vesicles by RAW264 macrophages results in the loss of cell surface Fc but not C3b receptor activity.
Authors: Petty H R , Hafeman D G , McConnell H M
Published: December 2022

We have observed that the specific antibody-dependent phagocytosis of lipid vesicles (containing 1 to 2% phospholipid hapten) results in a loss of Fc surface receptor activity from RAW264 macrophages. The Fc-receptor activity was measured by the ability of the macrophages to form rosettes with antibody-coated sheep erythrocytes (E). C3b-receptor activity was monitored similarly by measuring rosetting of IgM and complement-coated sheep erythrocytes (EAC). Under the same conditions, there was no loss of C3b-receptor activity. This suggests that Fc but not C3b cell surface receptors are depleted during specific IgG-mediated phagocytosis. The rosette assay for receptors, in accord with a previous kinetic analysis of vesicle uptake, showed that the depletion of Fc receptors after phagocytosis of specific antibody-coated vesicles was highly dependent on the number of vesicles taken up but relatively independent of the density of IgG bound to the vesicle surface. In contrast to the above results the nonspecific phagocytosis of latex beads results in the loss of both Fc- and C3b-receptor activity as measured by rosetting experiments.
Functional analysis of human T cell subsets defined by monoclonal antibodies. I. Collaborative T-T interactions in the immunoregulation of B cell differentiation.
Authors: Thomas Y , Sosman J , Irigoyen O , Friedman S M , Kung P C , Goldstein G , Chess L
Published: December 2022

T-B and T-T interactions involved in the regulation of PWM-triggered human B cell differentiation were studied in vitro. Functionally distinct human T cell subsets were isolated by C-mediated lysis by using the monoclonal antibodies OKT4 and OKT8. Graded numbers of either untreated or irradiated T cell subsets were added to autologous B cells, and total antibody synthesis was measured after 5 to 6 days of culture by using a highly sensitive reverse hemolytic plaque assay. The data indicate that a) the helper activity that is exclusively contained within the OKT4+ population is radiosensitive. Only at high T/B ratios can this radiosensitivity be overcome; b) the OKT8+ population contains radiosensitive cells important in suppressing B cell differentiation, and c) the suppression induced with OKT8+ cells requires the presence of radiosensitive OKT4+ cells. Thus, OKT8+ cells added to cultures containing B cells and irradiated OKT4+ cells do not suppress the PFC response. Addition of unirradiated OKT4+ cells to these cultures permits reexpression of suppression by OKT8+ cells. It is concluded that two radiosensitive cells, one within the OKT4+ population and the other within the OKT8+ population, collaborate to induce suppression. Possible mechanisms for this suppressive interaction including induction of suppressor precursor cells within the OKT4+ population or inhibition of OKT4+ helper cells by OKT8+ cells are discussed.
Structural comparisons of serologically indistinguishable H-2K-encoded antigens from inbred and wild mice.
Authors: Arden B , Wakeland E K , Klein J
Published: December 2022

H-2K glycoproteins from 2 wild-derived strains, B10.KPB128 and B10.GAA37, and from standard congenic lines B10.S(9R) and B10.A were compared by ion-exchange chromatography of tryptic peptides. The H-2K products of B10.S(9R) and B10.A were found to differ in about 33% of their peptides. The H-2K molecule of B10.S(9R) was indistinguishable from the H-2K molecules of B10.GAA37 and B10.KPB128. These results indicate that the H-2K-encoded products of the B10.S(9R), B10.KPB128, and B10.GAA37 lines are structurally very similar or, perhaps, identical, suggesting that some alleles of the H-2K locus may be maintained in a stable form in allopatric populations of wild mice.
Polyclonal activation of immunoglobulin secretion in B lymphocytes induced by staphylococcal peptidoglycan.
Authors: Dziarski R
Published: December 2022

Staphylococcal peptidoglycan (PG) is a B cell mitogen and immunomodulator in mice. The ability of PG to induce the secretion of polyclonal antibodies in murine lymphocyte cultures was studied using the protein A hemolytic plaque assay. PG was as effective as or more effective than lipopolysaccharide as an inducer of polyclonal antibodies in spleen, lymph node, and bone marrow lymphocytes. The highest numbers of immunoglobulin (Ig) secreting cells were induced in the spleen, and the lowest in the bone marrow cell cultures. All major classes of Ig (IgM, IgG, and IgA) were induced, and the maximal numbers of Ig-secreting cells were detected between days 4 and 5 of culture. The induction of Ig secretion by Pg was T cell, 2-mercaptoethanol, and to a large extent macrophage independent.
Alteration of T and null lymphocyte frequencies in the peripheral blood of human opiate addicts: in vivo evidence for opiate receptor sites on T lymphocytes.
Authors: McDonough R J , Madden J J , Falek A , Shafer D A , Pline M , Gordon D , Bokos P , Kuehnle J C , Mendelson J
Published: December 2022

Street opiate addiction produces a significant depression in the absolute number of total T lymphocytes in peripheral blood as measured by the ability of the lymphocytes to rosette sheep red blood cells (SRBC). Associated with the decrease in T cells, there is an increase in the absolute number of null lymphocytes but no significant changes in B lymphocytes or total white blood cell count. The T cell values for 2 different populations of addicts (n = 12 and 32) are 31.8% and 23.1%, whereas the null cell values are 51.1% and 57.6%, respectively. The values for comparable control populations (n = 18 and 10) are: T% = 70.7% and 67.4%, and null % = 9.2% and 14.5%. Self-reported use of marihuana does not significantly alter the distribution of cell populations. A 1- to 3-hr incubation of addicted-derived lymphocytes with 10(-6) to 10(-7) M Naloxone reverses both T cell depression and null cell increase by allowing the null cells to express SRBC receptors. Cyclic AMP and dibutyryl cyclic AMP can also convert the null cells to T cells. The conversion of null to T lymphocytes has additionally been measured by monitoring the increase in PHA-stimulated growth in 72-hr cultures as determined by tritiated thymidine incorporation into DNA. These results support the hypothesis that opiates can alter T lymphocyte number and function in vivo, and that this alteration may produce a significant degeneration in the immune competence of street opiate addicts.
Gut-associated lymphoid tissue in the chicken. I. Morphology, ontogeny, and some functional characteristics of Peyer's patches.
Authors: Befus A D , Johnston N , Leslie G A , Bienenstock J
Published: December 2022

Lymphoid aggregates with many of the characteristics of mammalian Peyer's patches (PP) were identified in the chicken intestine. These chicken PP could not be seen with the naked eye at the time of hatching, but by 10 days of age, 1 or 2 PP were identified in 50% of the birds. Up to 16 wk of age, the numbers of PP increased to a maximum of 5 per animal and they were widely scattered in the intestine, apart from one that was regularly found about 5 to 10 cm anterior to the ileocecal junction. As the birds aged, the number of PP declined so that in older birds (58 wk) only a single PP was evident near the ileocecal junction. Chicken PP possess: a distinct lymphoepithelium with M cells and strong pinocytotic activity; follicles and a subepithelium zone and central zone similar to those of the cecal tonsil (CT). Studies with in ovo hormonal bursectomy and age-associated involution of PP suggested that the subepithelial zone is a B-dependent area and the central zone a T, or thymus-dependent region. These observations extend our knowledge of gut-associated lymphoid tissue (GALT) in the bird to include the bursa of Fabricius, CT, PP, and aggregates in the urodeum and proctodeum. They raise issues about the roles of chicken PP that may aid our understanding of the functions of mammalian GALT.
Interaction of reovirus with cell surface receptors. II. Generation of suppressor T cells by the hemagglutinin of reovirus type 3.
Authors: Fontana A , Weiner H L
Published: December 2022

In studying reovirus interactions with lymphocytes, we have found that reovirus type 3, but not type 1, inhibits the in vitro proliferative response of murine splenic lymphocytes to concanavalin A (Con A). By analyzing recombinant clones containing genes from both reovirus types 1 and 3, we found that the S1 gene, the gene that encodes the viral hemagglutinin, is responsible for the inhibitory effect. In addition we found that type 3, but not type 1, generates suppressor T cells in vitro capable of suppressing Con A proliferation. By analyzing recombinant clones, we also found that the viral hemagglutinin is responsible for the generation of suppressor T cells by reovirus type 3. These effects were observed whether UV-inactivated or live virus was used. Reovirus type 3 inhibition of the proliferative response of murine splenic lymphocytes to Con A was blocked by anti-reovirus type 3 antibody but not by anti-reovirus type 1 antibody. Antiviral antibody had no effect on the ability of reovirus type 3 induced suppressor cells to inhibit Con A proliferation. We have previously demonstrated a receptor on murine lymphocytes for the hemagglutinin of reovirus type 3, and our results suggest that the in vitro suppression of Con A proliferation of murine lymphocytes by reovirus type 3 is secondary to the interaction of the viral hemagglutinin with a receptor on the surface of murine lymphocytes, which results in the generation of functionally active suppressor T cells.
Macrophage dependency of in vitro B cell response to influenza virus antigens.
Authors: McLaren C , Pope B
Published: December 2022

Specific B cell responses to influenza viruses were induced by in vitro stimulation of spleen cells from normal or influenza-immune mice with inactivated influenza virus. Both primary and secondary responses to A/Pt Chalmers/73 (A/Pt Ch/73) and A/New Jersey/76 (A/NJ/76) viruses were detected in a hemolytic plaque assay, with peak numbers of specific plaque-forming cells (PFC) present after 4 days of incubation. Depletion of macrophages from BALB/c spleen cell suspensions by treatment with either carrageenan or carbonyl iron markedly reduced the primary and secondary B cell responses to influenza virus. The responses could be restored by reconstitution with either BALB/c or CBA/J macrophages. Specificity of the antibody response was demonstrated in cross-reaction experiments with A/Pt Ch/73 and A/NJ/76 viruses. Evidence of recall to A/NJ/76 virus was, however, found in A/NJ/76-immune spleen cells stimulated in vitro with A/Pt Ch/73 virus. Greatest in vitro stimulation of A/Pt Ch/73-immune spleen cells was produced with inactivated whole A/Pt Ch/73 virus preparations; specific responses were low or absent after stimulation with preparations of partially disrupted virus vaccine, a mixture of purified hemagglutinin (HA) and neuraminidase antigens or soluble hemagglutinin containing similar amounts of viral HA antigen. This lack of response to A/Pt Ch/73 HA could not be overcome with A/NJ/76 whole virus together with purified A/Pt Ch/73 HA.
IgD allotypic determinants. II. Determinants expressed on IgD of mice of the b, e, and o Igh-C haplotypes.
Authors: Woods V L , Scher I , Lieberman R , Paul W E
Published: December 2022

Allotypic determinants expressed on membrane IgD of 27 strains of mice representing each of the major Igh-C haplotypes of inbred mice were examined with reagents specific for the IgD of mice of the b, e, and o Igh-C haplotypes. The distribution of two specificities, IgD.44 (Ig5.2) and IgD.37 (Ig5.3), recognized by two monoclonal anti-delta reagents was determined. In addition, a new specificity, IgD.46, expressed on mice of the o Igh-C type, but not of the b or a types, was detected by a BALB/c anti-AL/N spleen cell antiserum. The results also confirmed the description of the new Igh-C haplotype o, by demonstrating that the mice of this group express the IgD.37 determinant. This determinant is also expressed by mice of the b and e types, but not by mice of the d haplotype, from which the o haplotype was subdivided.
Human B lymphocyte colony responses. I. General characteristics and modulation by monocytes.
Authors: Bobak D , Whisler R
Published: December 2022

The present study investigated the ability of human B cell-enriched subpopulations to focally proliferate and form colonies in semisolid cultures after stimulation with staph protein A (SpA). After 6 days of incubation, cultures of B-enriched populations exhibited distinct colonies, the number being dependent on the concentration of SpA and the cell density. Optimal colony responses were 1.6 x 10(3) per 1 x 10(6) B lymphocytes, and greater than 83% of the colony-forming cells expressed surface immunoglobulin (sIg). The depletion of adherent monocytes from the B cell-enriched preparations decreased the colony responses approximately 3-fold compared with the nondepleted B cell populations. Adding optimal numbers of adherent monocytes to the monocyte-depleted B cells restored the colony responses; however, less augmentation was observed in single-layer co-cultures containing greater than optimal numbers of monocytes. Identical experiments in double-layer semisolid cultures revealed that relatively greater numbers of monocytes were required to enhance B cell colony responses. Thus, progressively higher ratios of monocytes to B cells resulted in increasing numbers of colonies and failed to demonstrate the diminished colony responses observed in the single-layer system. These studies demonstrate that human B cells form distinct colonies when activated by SpA and that normal adherent monocytes modulate the magnitude of colony responses. Although monocytes predominately enhance B cell clonal differentiation, the evidence presented also suggests that, to a lesser extent, soluble inhibitory materials are elaborated.
HLA-DR epitope region definition by use of monoclonal antibody probes.
Authors: Grumet F C , Charron D J , Fendly B M , Levy R , Ness D B
Published: December 2022

Definition of HLA-DR epitopes has been attempted by utilizing monoclonal antibody probes. Hybridoma antibodies L203 and L227, known to bind different epitopes on human Ia-like molecules, were tested for their ability to block cytotoxicity of monoclonal and allogeneic anti-DR antibodies. Monoclonal cytotoxic antibodies segregated into two groups: those more effectively blocked by L203, and those more effectively blocked by L227. Alloantisera also segregated into two groups, but according to their DR specificity. Anti-DR1, -2, and -3 alloantisera were effectively blocked by both L203 and L227, whereas anti-DR7, -w9, -w10, and MT1 alloantisera were not blocked by either. Blocking was not correlated with immunoglobulin class of the alloantibody and further definition of the mechanism of cytotoxicity blocking remains to be elucidated. Based on these data and prior binding and immunochemical studies with L203 and L227, a model is proposed in which the tertiary structure of each DR molecule, or complex of associated molecules on the cell surface, has two reference epitopes, one defined by L203, and another defined by L227. HLA-DR epitopes defined by the cytotoxic monoclonal or alloantibodies to the L203 or L227 epitope in order to begin epitope mapping or grouping.
Studies on stimulation of cell-mediated cytotoxicity by skin test antigens. I. Candida antigen stimulation of cell-mediated cytotoxicity in vitro correlated with the skin test response to candida antigen in vivo.
Authors: Tartof D , Check I J , Matutis A , Hunter R L , Fitch F W
Published: December 2022

The magnitude of the skin test response in a group of normal volunteers to intradermal Candida antigen correlated closely with the level of cytolytic activity stimulated by Candida antigen in vitro in the peripheral blood lymphocytes from those same individuals. The cytolytic activity stimulated by Candida antigen was cell mediated, and cold target inhibition studies demonstrated that Candida antigen-stimulated effector cells were capable of cross-species (i.e., nonspecific) killing. Analysis of the Candida antigen-induced effector cell population for various cell markers did not enable absolute identification of the cell responsible for the killing. The findings in this study indicate that the stimulation of cell-mediated cytolysis by Candida antigen may be related to the delayed-type hypersensitivity response produced by the same antigen.
Establishment and characterization of a murine hybridoma secreting monoclonal anti-DNA autoantibody.
Authors: Tron F , Charron D , Bach J F , Talal N
Published: December 2022

Twenty-five clones of hybridoma cells secreting anti-DNA autoantibodies were isolated after fusions between plasmacytoma cells (P3-x 63 Ag 8.653) and spleen cells from nonimmunized autoimmune NZB/NZW F1 (B/W) mice. A monoclonal anti-DNA autoantibody was immunochemically analyzed.
Defective Fc-mediated phagocytosis in C3H/HeJ macrophages. II. Correction by cAMP agonists.
Authors: Vogel S N , Weedon L L , Oppenheim J J , Rosenstreich D L
Published: December 2022

Peritoneal macrophages from LPS hyporesponsive C3H/HeJ mice lose the capacity to bind and phagocytose opsonized sheep erythrocytes (EA) over a 48-hr culture period. This loss in Fc receptor capacity is markedly different from the progressive increase in phagocytic ability exhibited by cultured macrophages derived from LPS-responsive C3H/HeN mice. Since dibutyryl-cyclic adenosine monophosphate (DBcAMP) has previously been reported to modulate membrane receptor expression in lymphocytes and certain macrophage-like cell lines, we examined its effects on EA binding and phagocytosis by C3H/HeJ macrophages. DBcAMP not only reverses the binding defect in C3H/HeJ macrophages but also restores EA phagocytosis to the level of control C3H/HeN cultures. 8-Bromo-cAMP, as well as other agents known to elevate intracellular cAMP (i.e., isoproterenol plus isobutylmethylxanthine or prostaglandin E2) also corrected the phagocytic defect. Since the C3H/HeJ macrophage phagocytic defect can also be reversed by in vitro stimulation with a lymphokine-rich culture supernatant, we examined the effect of this treatment on intracellular cAMP levels. Lymphokine treatment produced a 60% increase in the levels of macrophage intracellular cAMP. These findings suggest that the C3H/HeJ differentiation defect may be secondary to some abnormality in a cAMP dependent pathway.
Antigen-specific human T cell factors. I. T cell helper factor: biololgic properties.
Authors: Heijnen C J , Uytdehaag F , Pot K H , Ballieux R E
Published: December 2022

The in vitro induction and assay of an ovalbumin-specific human T cell helper factor are described. Peripheral blood T cells, cultured with ovalbumin in a Marbrook-Diener system, produce an antigen-specific factor(ThF120-OA), which can be purified by affinity chromatography. The in vitro studies with ThF120-OA pointed out that in the production of the factor as well as in the factor-B cell interaction the adherent cell determines the genetic restriction. The results of kinetic studies on T helper activities demonstrated that Thf120-OA provides an auxiliary activity at various moments during the differentiation of the human peripheral B cell into an antibody-secreting cell. The observed differences in the mode of action of Th cells and Th factor are discussed.
Identification and characterization of protein antigens of Leishmania tropica isolates.
Authors: Handman E , Mitchell G F , Goding J W
Published: December 2022

Promastigotes of 4 Leishmania tropica isolates were biosynthetically labeled with 35S-methionine or surface radioiodinated, and the detergent lysates were analyzed by 2 dimensional gel electrophoresis. The protein patterns of cytoplasmic and membrane proteins detected in two independent isolates from simple cutaneous leishmaniasis cases were similar to each other, but were different from 2 isolates taken over a 20-yr interval from a case of leishmaniasis recidiva. The analysis of radioiodinated membrane proteins of L. tropica promastigotes revealed a simple pattern of 6 to 7 labeled major proteins and some minor ones. Unlike cytoplasmic proteins, membrane proteins were highly resistant to digestion by proteases. Partial cleavage of radioiodinated surface proteins by staphylococcal V-8 protease in the presence of 4.5 M urea and peptide mapping confirmed the presence of shared and isolate-specific proteins. The significance of these findings for the classification of Leishmania spp. and the pathogenesis of the different disease states that they cause are discussed.
Serologic and immunochemical characterization of the specificity of four monoclonal antibodies to distinct antigenic determinants expressed on subpopulations of human Ia-like antigens.
Authors: Quaranta V , Pellegrino M A , Ferrone S
Published: December 2022

Serologic and immunochemical assays showed that the monoclonal antibodies Q2/70, Q2/80, Q5/6, and Q5/13 react with human Ia-like antigens. Each monoclonal antibody recognizes distinct antigenic determinants that are different from those defining the serologic polymorphism of Ia-like antigens defined by conventional alloantisera and are expressed on subpopulations of Ia-like antigens. The determinants recognized by the MoAb Q2/70 and Q5/13 are expressed on all HLA-DR allospecificities tested, whereas those reacting with the MoAb Q2/80 and Q5/6 are not detectable on HLA-DR5 and HLA-DR7 allospecificities, respectively.
Measurement of IgG blocking antibodies by interference in the radioallergosorbent test.
Authors: Gleich G J , Zimmermann E M , Gleich M C , Yunginger J W
Published: December 2022

We previously found that sera of patients immunized with ragweed pollen extract contained a factor that interfered with the binding of IgE antibodies to solid-phase allergens in the radioallergosorbent test (RAST). We now describe an assay, RAST interference, to measure this factor, and we present evidence that the factor is IgG blocking antibody. Sera from immunized allergic patients were heated at 56 degrees C for 4 hr to destroy heat-labile Fc determinants on IgE and were tested for their ability to prevent binding of additional IgE antibody to solid-phase allergens in the RAST. Eight of 10 sera from allergic immunized patients gave RAST interference dose-response curves that did not differ from the arbitrary standard. The factor causing interference showed specificity for the immunizing antigen, was heat-stable, eluted from Sephadex G-200 in the 7S peak, was present only in sera of immunized patients, and rose after initiation of immunization. These results indicated that RAST interference can be used to measure IgG blocking antibodies with the same reagents employed for the measurement of IgE antibodies, provided the antiserum to IgE is specific for the heat-labile FC determinants on IgE.
Ir gene control of the immune response to insulins. I. Pork insulin stimulates T cell activity in nonresponder mice.
Authors: Bucy R P , Kapp J A
Published: December 2022

Immune responses by mice to heterologous insulins are controlled by H-2-linked Ir genes. In studies to determine the mechanism(s) of nonresponsiveness, we found that although pork insulin fails to stimulate antibody or proliferative responses in H-2b mice, it does stimulate enhanced responses to subsequent challenge with an immunogenic species of insulin, such as beef insulin. Experiments described in this communication analyze the cell type primed in H-2b mice by pork insulin using an adoptive transfer protocol. The results demonstrate that pork insulin primes T cells that can express helper activity when recipient mice are challenged with beef but not pork insulin. This helper T cell activity is insulin specific in both elicitation and effect but is dependent upon stimulation by beef insulin for expression. Our interpretation of these results is that 2 antigen-specific T cell subpopulations are required for the generation of insulin-specific antibody responses and that the Ir gene defect in this case is expressed as a failure of specific interaction of these 2 T cell populations.
Capping of DNA on phytohemagglutinin-stimulated human lymphoblasts.
Authors: Rogers J C , Kerstiens J W
Published: December 2022

A portion of the DNA released by PHA-stimulated human lymphoblasts becomes bound to the plasma membranes of the cells and is actively capped. Capping is accompanied by shape change, and caps are localized to uropods. DNA can be exchanged between cells for capping.
In vivo generation of suppressor T cells by thymosin in congenitally athymic nude mice.
Authors: Marshall G D , Thurman G B , Rossio J L , Goldstein A L
Published: December 2022

Treatment of nude mice with thymic factors such as thymosin has been mostly ineffective in generating effector T cells. This study examined the effects of treating nude mice with thymosin fraction 5 on the induction of cells that could participate in and/or regulate cytotoxic T lymphocyte (CTL) generation by normal spleen cells in vitro. Splenic lymphocytes from BALB/c nude mice injected with thymosin fraction 5 every other day for 2 wk were tested for their ability to generate CTL in vitro. Two days after the last subcutaneous injection of thymosin, nude spleens were removed, mixed with normal BALB/c spleen cells, and placed into a mixed lymphocyte tumor culture (MLTC) against allogeneic RBL 5 tumor cells. After a 5-day incubation, cultures were tested for the presence of CTL in a 4-hr 51Cr-release assay. Spleen cells from thymosin-treated nude mice did not generate CTL but suppressed the ability of normal spleen cells to generate CTL in vitro. Characterization of the thymosin-induced nude mouse suppressor cells showed them to be Thy 1 positive, nonadherent, cyclophosphamide-sensitive T cells. These data demonstrate that some T cell maturation occurs in vivo under thymosin influence. However, the activity of these cells is initially limited to a regulatory function. These studies suggest that maturation of functional suppressor T cells occurs before CTL. Further immunologic manipulation appears to be necessary in order to induce CTL effector cells in nude mice.
Macrophage T cell interaction in man: binding of antigen-specific human proliferating and helper T cells to antigen-pulsed macrophages.
Authors: Geha R S , Jonsen M E , Ault B H , Yunis E , Broff M D
Published: December 2022

Adsorption of T cells over monolayers of autologous, but not allogeneic, macrophages (M phi) pulsed with tetanus toxoid (TT) antigen resulted in the specific depletion of the capacity of the T cells to proliferate and to release T cell helper factor in response to stimulation, with TT antigen. T cells that bound to the TT-pulsed M phi monolayers were specifically enriched in their capacity to proliferate in response to TT. Turkey antiserum to the human B cell glycoprotein p 29,34, but not turkey antiserum to human beta 2 microglobulin, inhibited the binding of TT-reactive T cells to TT-pulsed M phi. This inhibition resulted from the binding of the anti-p 29,34 antiserum to DR determinants expressed on the surface of the M phi. Immunosorbent purified IgG anti-TT, added in great excess, did not inhibit the binding of TT-reactive T cells to TT-pulsed M phi monolayers. Soluble TT antigen and rabbit antiidiotypic IgG, directed against IgG F(ab')2 anti-TT obtained from the cell donor, minimally inhibited the binding of TT-reactive T cells to TT pulsed M phi monolayers. M phi pulsed with urea-denatured TT, which contained less than 1% native TT, depleted T cell reactivity to TT to an extent almost equivalent to that achieved by M phi pulsed with native TT. These results indicate that human antigen-reactive T cells bind to the DR determinants of M phi pulsed with antigen and that the immunogenic moiety of antigen recognized by the majority of antigen reactive human T cells may differ from native antigen.
Malignant cell arrest in thymus and spleen of mice bearing transplanted tumors.
Authors: Haran-Ghera N , Krauthgamer R , Peled A
Published: December 2022

The relationship between the local outgrowth of different subcutaneous transplantable tumors (plasmacytoma MOPC-315, mastocytoma P-815; Lewis lung carcinoma-3LL; fibrosarcoma; lymphomas and mammary adenocarcinoma) and tumor cell spread in different organs was evaluated. The presence of tumor cells in thymus, spleen, lung, and liver was demonstrated by using both in vivo and in vitro methods. Sequestration of tumor cells derived from the transplanted solid tumor was indicated within 3 to 7 days after tumor graft, shortly before or after early palpable outgrowth of the primary tumor was observed. Tumor cells present in thymus and spleen might affect immune responses of tumor-bearing mice.
In vitro regeneration of resting lymphocytes from stimulated lymphocytes and its inhibition by insulin.
Authors: Kumagai J , Akiyama H , Iwashita S , Iida H , Yahara I
Published: December 2022

Concanavalin A- (Con A) stimulated mouse lymphocytes were separated from unstimulated cell populations by a bovine serum albumin (BSA) density gradient centrifugation 2 days after the stimulation. The stimulated cells divided, but did not initiate a new round of DNA replication when cultured further in the absence of Con A. The divided cells became similar in various properties to unstimulated lymphocytes during a prolonged incubation, suggesting that they entered the resting state. In the present study, the process toward the resting state of lymphocytes from the proliferating state was manifested by: a) a decrease in a cell volume, b) lowering of protein content, c) a loss of responsiveness to T cell growth factor (Interleukin 2) (TCGF(IL2)) probably as a result of disappearance of TCGF(IL2) receptors, d) disappearance of insulin receptors, and e) a duration of time lag between the addition of Con A and the onset of DNA synthesis. In addition, it was found that insulin was not mitogenic for recently divided lymphocytes, but prevented these cells from entering the resting state. This suggests that there may be an in vivo system that keeps stimulated lymphocytes highly responsive to antigens for at least several days without initiating DNA synthesis and cell division.