bluuebunny/crossref_metadata_2025 huggingface dataset:
A Biochemical Analysis of the Membrane-Associated Gene Products of Major Histocompatibility Complex of the Rat
Authors: Sporer Ruth , Manson Lionel A. , Götze Dietrich
Published: December 2022

Thus far, the rat major histocompatibility complex (MHC), AgB or RTI, have been divided by recombinational analysis into two regions, AgB.A (RTI.A) coding for transplantation antigens (class I gene products) and AgB.B (RTI.B) controlling the humoral immune response (Ir genes) and the proliferative response to allogeneic cells (MLR, class II genes). In this study, an attempt was made by biochemical analysis to determine whether there exists more than one AgB.A (class I) gene product, as has been found in the mouse (H-2K and H-2D) and in the human (HLA.A and HLA.B antigens). NP40-solubilized 3H-fucose-labeled rat MHC membrane products were immunoprecipitated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Firstly, with a congenic anti-erythrocyte serum, only membrane structures with an approximate m.w. of 45,000 (AgB.A) were precipitated. A congenic anti-lymphocyte serum absorbed with erythrocytes precipitated antigens with an apparent m.w. of 33,000 and 28,000 (AgB.B) whereas the unabsorbed serum precipitated in addition a 45,000-dalton component (AgB.A). Secondly, sequential treatment of a DA (AgB4, RTIa) extract with a strongly cross-reactive serum followed by an (Lewis × BN.B2)F1 anti-AgB4-specific serum demonstrated that the AgB4 haplotype codes for at least two AgB.A and at least two AgB.B gene products.
Regulation of IgE Antibody Production by Serum Molecules
Authors: Katz David H. , Bargatze Robert F. , Bogowitz Cheryl A. , Katz Lee R.
Published: December 2022

Experiments presented in this paper were designed to test a new concept concerning the possible pathogenesis of the allergic phenotype. This concept, termed “allergic breakthrough” considers that one of the avenues toward the allergic phenotype involves coincidental sensitization combined with an imbalance in the normal damping mechanism that serves to limit IgE antibody production. The three predictions of this concept that can be tested experimentally are: 1) manipulations that are effective in heightening or re-establishing the damping mechanism should manifest persistence insofar as IgE antibody synthesis to the relevant allergen is concerned; 2) once allergic breakthrough has occurred, the height of production of IgE antibodies specific for the sensitizing agent should remain elevated at levels characteristic of the allergic phenotype, even after the threshold of damping activity has returned to a normal level; and 3) allergic breakthrough should display specificity in that breakthrough would occur in response to subsequent exposure to the specific antigen to which coincidental sensitization initially occurred, but not for other unrelated antigens. The studies presented herein confirm each one of these predictions, thereby providing substantial support for the validity of this concept as one possible distinguishing feature between individuals manifesting the nonallergic and allergic phenotypes, respectively.
Characterization of the Target Cell Receptor for IgE
Authors: Conrad Daniel H. , Helm Ricki M. , Yiu Suk Hing , Froese Arnold , Ishizaka Teruko , Ishizaka Kimishige
Published: December 2022

Previous studies had shown that isolation of the receptor for IgE from rat basophilic leukemia (RBL) cells by means of IgE-Sepharose yields two surface components having apparent m.w. of 55,000 and 45,000 daltons. In the present study, an antiserum to such a receptor preparation was induced in a rabbit. This antiserum, even though it could not initiate histamine release in vivo or in vitro, reacted with the RBL cell surface components bound to and eluted from IgE-Sepharose, as well as with these two components present in detergent extracts of surface-labeled RBL cells. It was also capable of reacting with IgE-receptor complexes. With this antiserum, it was possible to demonstrate that, when occupied by IgE, the 45,000 dalton component cannot be surface iodinated, whereas the 55,000 dalton component can. Moreover, it could be shown that both components are present in detergent lysates of surface-iodinated peritoneal rat mast cells (RMC). The results have also indicated that the 55,000 and 45,000 dalton components are present on both intact RMC and RBL cells. Moreover, the antibodies to the 45,000 dalton receptor component appeared to react with those parts of the receptor that do not make contact with IgE.
Tryptic-Peptide Identity of the GPLA-B.1 Alloantigens Derived from Noncongenic Inbred Strain 2 and Strain 13 Guinea Pigs
Authors: Rose Stephen M. , Hahn Yaffa , Schwartz Benjamin D.
Published: December 2022

The guinea pig MHC, the GPLA complex, codes for the B and S classic histocompatibility antigens. Four alleles have been postulated at the B locus and two alleles at the S locus. Strain 2 and strain 13 guinea pigs have serologically identical B.1 and S antigens but differ by the I region. However, since these two strains do not have common parentage, it was unknown whether their B.1 molecules were structurally identical. This is an important consideration since the large body of immunogenetic data attributed solely to I region differences is based on this premise and therefore would be subject to reinterpretation if it should prove untrue. We have demonstrated that the primary structures of these two B.1 molecules are identical by performing isoelectric focusing and comparative tryptic-peptide mapping analysis.

The DHCB (PCA) guinea pig expresses the same I region as that of the strain 13 guinea pig, but has a different B alloantigen (i.e., B.3). Isoelectric focusing and comparative tryptic-peptide mapping analysis of this B.3 alloantigen have substantiated the serologic finding of nonidentity of the B.1 and B.3 molecules by showing that the two molecules do not share identical primary structure. However, the mapping data showed that there was 40 to 50% homology between the B.1 and B.3 molecules. This result is consistent with the findings in the mouse system for alleles of the same locus. Therefore, our study has demonstrated that the B.1 molecules from the two noncongenic strain 2 and strain 13 guinea pigs are structurally identical and that the B.3 molecule of the DHCBA guinea pig is most likely an allelic product of the B.1 molecule.
Neoantigen of the Complement Membrane Attack Complex on Cytotoxic Human Peripheral Blood Lymphocytes
Authors: Sundsmo John S. , Müller-Eberhard Hans J.
Published: December 2022

Neoantigenic determinants (neoAg) specific for the assembling membrane attack complex (MAC) of complement were detected by immunofluorescence microscopy on the surface of cytotoxic lymphocytes during the antibody-dependent cellular cytotoxicity (ADCC) reaction. This study employed antibody-sensitized chicken erythrocytes as target cells, human peripheral blood lymphocytes as effector cells, and RITC-conjugated rabbit F(ab′)2-anti-neoAg. NeoAg was present on 60% of ADCC plaque-forming lymphocytes (PFL). Eight out of 182 neoAg-positive PFL were observed in direct contact with their target cells. In these cases MAC-specific neoAg was visualized at the zone of contact between the cells. Anti-neoAg Ig was found to inhibit ADCC plaque assays up to 62%; and 51Cr-release assays up to 79%. Stimulation of lymphocytes by PHA or mixed lymphocyte culture increased the expression of neoAg. In the case of PHA, increased neoAg expression was correlated with an increased incorporation of 14C-leucine into C5, C6, C7, and C8 antigens, which was detected by immunodiffusion and autoradiography.
Generation on Infected Fibroblasts of Human T and Non-T Lymphocytes with Specific Cytotoxicity, Influenced by Histocompatibility, against Measles Virus-Infected Cells
Authors: Wright Larry L. , Levy Nelson L.
Published: December 2022

Human peripheral blood lymphocytes (PBL) were incubated (sensitized) for 6 days on autologous or allogeneic fibroblast monolayers either uninfected or infected with measles virus. The sensitized PBL were then tested, in a 6-hr 51Cr-release assay, against autologous and allogeneic measles virus-infected and uninfected fibroblast target cells. When the PBL had been sensitized on autologous infected monolayers, they were cytotoxic to both autologous and allogeneic measles-infected targets but not to uninfected targets or to targets infected with paramyxovirus I (Sendai virus). Likewise, PBL sensitized on uninfected autologous (or allogeneic) monolayers had no cytotoxic activity against any of the infected or uninfected target cells. When the PBL had been sensitized on allogeneic infected monolayers, they were cytotoxic to measles-infected targets that were autologous either to the PBL themselves or to the sensitizing monolayer. These sensitized PBL were not, however, cytotoxic against third-party infected targets that were totally allogeneic to both the PBL and the sensitizing monolayer.

Subpopulation depletion studies revealed at least two different effector lymphocytes in the sensitized PBL. One such effector was a T cell whose activity would be generated on autologous measles-infected monolayers but not on allogeneic infected monolayers. The cytotoxic activity of this cell was measles virus specific, and it required that the effector recognize as self antigens of the MHC on the target cell. The second effector cell also required in vitro generation but could be so generated on both allogeneic and autologous measles-infected, but not uninfected, monolayers. It was a non-T cell that had a receptor for the Fc fragment of IgG but lacked nonspecific esterase activity. This effector was measles virus specific, in that it failed to lyse either uninfected targets or targets infected with Sendai virus (the latter of which could be lysed by fresh PBL). When this non-T effector was generated on allogeneic infected monolayers, its cytotoxic activity showed an unusual form of MHC-related specificity, in that it lysed infected target cells autologous to either the PBL or the sensitizing monolayer but not infected third-party target cells allogeneic to both. Evidence was discussed which argued that our non-T effector was neither a conventional K nor NK cell.
Amplification of the Proliferative Response to Alloantigen by a Factor Present in an Extract of Syngeneic Thymic Lymphoid Cells: Demonstration of Optimal Conditions and Target Cell for Factor Activity
Authors: Wright Peter W. , Loop Stephen M.
Published: December 2022

A factor extracted from syngeneic thymic lymphoid cells (thymocytes) is shown to amplify the proliferative (MLC) response of syngeneic lymphoid cells to alloantigen in vitro. The optimal conditions for an effect of the thymus factor are quantitatively defined by kinetic and dose-response studies. Other variables that could potentially influence the activity of the thymus factor, such as the presence of 2-mercaptoethanol and the source of alloantigen, are identified. Factor activity can be recovered from semi-allogeneic thymocytes, as well as syngeneic thymocytes. The factor appears to predominantly effect the proliferative response of T cells localized in peripheral lymphoid organs. As such, this factor appears to be distinct from the variety of previously described factors derived from thymic reticuloepithelial elements that are thought to primarily induce the differentiation of T cell precursors found predominantly in bone marrow. Several possible mechanisms of action of this thymocyte-derived factor are considered.
Enhanced Immune Response Late in Primary Cytomegalovirus Infection of Mice
Authors: Tinghitella T. J. , Booss John
Published: December 2022

Murine cytomegalovirus (MCMV) infection has been previously shown to depress humoral and cell-mediated functions to non-MCMV antigens. In this report we show that in C3D2 mice undergoing nonlethal primary infection the depressed anti-sheep RBC plaque-forming cell (PFC) response is followed by an enhanced PFC response. Infected mice often generated twice the number of PFC per spleen than that of control mice. Total numbers of spleen cells as well as the recovery of virus from spleens of infected mice did not distinguish the depressed from the enhanced phase of the response. Investigation of the kinetics of the response revealed a defect in shutdown regulation. This enhanced PFC response during primary MCMV infection was not reflected in measurements of serum hemagglutinin. These findings suggest that MCMV induces an impairment of immunoregulation.
Phenotypic and Physical Characteristics of the Lymphoid Cells Involved in the Immunity to Syngeneic UV-Induced Tumors
Authors: Daynes Raymond A. , Schmitt Mary K. , Roberts Lee K. , Spellman Craig W.
Published: December 2022

Recent studies have indicated that the inability of UV-irradiated mice to reject UV-induced fibrosarcomas is, at least in part, due to the presence of suppressor T lymphocytes. These cells appear to exert their influence by inhibiting the functional development of effector cells with specificity for cross-reactive tumor-associated antigens. In these experiments, we present evidence as to the nature of the cell types involved in the immunologic responses elicited against UV-induced tumors. The effector cells capable of establishing a state of tumor immunity are T lymphocytes that appear to lack demonstrable Ia determinants. The regulatory suppressor cells, previously shown to be T lymphocytes, have been determined to be Ia+ and functionally radiosensitive. The in vivo administration of anti-Ia antibody or the exposure of UV-irradiated tumor-bearing animals to low dose gamma radiation was capable of influencing the tumor susceptible state. This type of therapy, theoretically designed to selectively depress suppressor T cell function in vivo, appears to enhance the degree of tumor immunity by removing regulatory influences. We believe that these studies establish the importance of immunoregulatory mechanisms in host-UV tumor interactions.
Antigen-Initiated B Lymphocyte Differentiation
Authors: Shortman Ken , Howard Maureen , Teale Judy , Baker Julie
Published: December 2022

Both primary IgM and secondary IgM and IgG adoptive immune responses to the thymus-independent antigen NIP-POL were found to proceed in two stages. The first 3 to 4 days of these responses required a stimulus, but this could be provided by an unrelated antigen such as HRC. Only the second 3- to 4-day stage of the response, leading to an AFC peak at day 8, required the specific antigen NIP-POL. This novel priming schedule of nonspecific followed by specific antigenic stimulation in fact produced a significantly greater AFC response than the standard specific immunization procedure. With the schedule of nonspecific, then specific antigen injection, the primary IgM response was T cell independent. Two sequential injections of specific antigen also gave an elevated adoptive response from the same B cell subset, but this time the increased component of the response was T cell dependent.

The results corroborate earlier work, and support a model of two sequentially related and “antigen”-responsive subsets of both primary and secondary B cells, termed “pre-progenitors” and “direct AFC progenitors”. Responses to antigens are thus two-tiered. In the first stage, all “pre-progenitor” subsets are nonspecifically activated to proliferate and differentiate into “direct-progenitor” B cells. This stage is different from “pre-B” to B cell development. Although it may occur continuously at a lower level in normal animals, it requires an environmental or experimentally introduced stimulus, and is probably triggered via macrophage activation. In the second stage the conventional rules apply, and antigens specifically stimulate antigen-restricted clones of direct-progenitor B cells to produce AFC.
Production of Human T Cell Colony-Inducing Activity (TCI) by Malignant Cells from Patients with Lymphocytic and Myeloid Leukemia
Authors: Douer Dan , Sachs Leo
Published: December 2022

Normal peripheral blood lymphocytes can be induced to form T cell colonies in culture by a T cell colony-inducing activity (TCI) found in conditioned medium (CM) induced by phytohemagglutinin (PHA) in normal T lymphocytes. TCI was also found in CM induced by PHA in peripheral blood cells from patients with untreated B cell chronic lymphocyte leukemia (CLL) and untreated acute myeloid leukemia (AML). The CM from these leukemic patients showed a much higher TCI activity for T lymphocytes both from normal persons and from CLL patients, than the CM from normal lymphocytes. This more active TCI was produced in the CLL patients by the peripheral blood B lymphocytes and in the AML patients by the peripheral blood myeloblasts. The CM from normal persons and CLL patients also contained an activity that induced the formation of granulocyte colonies from normal bone marrow cells. Although the CM from CLL showed a higher TCI activity than the CM from normal persons also with normal bone marrow cells, CM from both sources induced a similar number of granulocyte colonies.

The results indicate that malignant B lymphocytes from CLL patients and malignant myeloblasts from AML patients can produce a higher TCI activity than that produced by normal lymphocytes. It is suggested that induction of high TCI activity in certain malignant cells may selectively increase the number of normal T lymphocytes, and that the high TCI activity that can be produced by malignant B cells from CLL patients may explain the higher number of T cells that have been reported in these patients.
Induction of Autoreactive Cells by the Preculture of Human Peripheral Blood Mononuclear Cells with the Autologous Fresh Plasma
Authors: Tomonari Kyuhei , Aizawa Miki
Published: December 2022

An AMLR in which precultured cells proliferated in response to fresh non-T cells is described. In our system, the responder is human peripheral blood mononuclear cells precultured in the autologous fresh plasma for up to 16 days, and the stimulator is fresh autologous non-T cells. Results suggested that there were two subpopulations of autoreactive cells obtained from the preculture; the high and low density small lymphocytes, both having ERF activity. The autoreactivity of low density cells was augmented when either macrophages or N-ERF-cells were depleted from PBM and thereafter precultures were performed. A survey of the functional characteristics of the responding cells showed that the responding cells had NK activity against Molt-4 cells but had no significant ADCC activity against target CRBC. Mechanisms for the induction of autoreactive cells by the preculture in the presence of autologous fresh plasma are discussed.
Characterization of a Circulating Subpopulation of Spontaneous Antitetanus Toxoid Antibody Producing B Cells Following in Vivo Booster Immunization
Authors: Stevens Ronald H. , Macy Eric , Morrow Casey , Saxon Andrew
Published: December 2022

The appearance of circulating B lymphocyte subpopulations responsible for the in vitro synthesis of IgG or IgM antitetanus toxoid antibody after in vivo booster immunization has been analyzed. Before booster immunization, only background levels of IgG antitetanus toxoid antibody were synthesized by B lymphocytes alone, or in combination with T cells and pokeweed mitogen, as measured by quantitative radioimmunoassay. Within 5 days after booster immunization, a population of B lymphocytes appeared within the circulation that had the capacity to synthesize IgG antitetanus toxoid antibody in the absence of T lymphocytes or pokeweed mitogen. These cells could be detected in the circulation until 10 to 12 days after immunization, and the antibody synthesized was inhibited by the inclusion of 50 µg/ml cyclohexamide in the culture medium. At the peak of this response, between 10 and 60% of the total IgG synthesized was IgG directed against tetanus toxoid. The cells responsible had a precursor frequency of up to 10-3 in the non-E-rosetting cell fraction obtained from peripheral blood and sedimented at 1 × G with velocities ranging from 4 to 11 mm/hr. The synthesis of IgM antitetanus toxoid antibody by this cell fraction was not detected.

In contrast to the above B cells, lymphocytes capable of responding to pokeweed mitogen and T lymphocytes with the synthesis of IgG antitetanus toxoid antibody did not appear in the circulation until 10 days after immunization. Limiting dilution analysis of the pokeweed mitogen responsive B lymphocytes 2 to 4 weeks after booster immunization gave a precursor frequency of 1 × 10-4. In the presence of T cells and PWM, the synthesis of IgM anti-Tet remained at constant levels before and for 3 weeks post-booster immunization. These results indicate that multiple subsets of circulating B cells can be functionally defined in humans.
Expression of MHC Antigens by Mouse Thymic Dendritic Cells
Authors: Rouse Robert V. , van Ewijk Willem , Jones Patricia P. , Weissman Irving L.
Published: December 2022

Thymic epithelial cells express MHC antigens in several different patterns. I-A is present throughout the thymic cortex on dendritic cells. The remainder of the I region and H-2K/D are expressed on dendritic cells apparently only variably in the cortex (at least in some haplotypes). All MHC antigens tested are present in the medulla on epithelial cells; expression on medullary lymphocytes cannot be evaluated. Monoclonal anti-MHC antibodies confirm these results. The significance of these findings to T cell maturation is discussed.
Natural Killer Cells, Bone, and the Bone Marrow: Studies in Estrogen-Treated Mice and in Congenitally Osteopetrotic (mi/mi) Mice
Authors: Seaman William E. , Gindhart Thomas D. , Greenspan John S. , Blackman Marcia A. , Talal Norman
Published: December 2022

Mice lose natural killer cells after 6 weeks of treatment with 17β-estradiol. We here demonstrate that the same protocol leads to loss of genetic resistance to bone marrow transplantation and to significant osteoproliferation with loss of bone marrow. We also show that mice with reduced marrow because of congenital osteopetrosis are deficient in natural killing. These findings are consistent with previous evidence that natural killing and genetic resistance to bone marrow transplantation are dependent upon the marrow.

Temporal studies of bone histology and radiology during and after treatment with estrogen reveal that alterations in natural killing proceed more rapidly than changes in bone marrow volume. These studies also demonstrate that estrogens induce osteoproliferation only at endosteal surfaces that are adjacent to hematopoietic marrow. From these observations, we conclude that estrogens do not reduce natural killer cells simply by reducing the volume of bone marrow. Estrogens may instead have an effect on bone marrow cells that leads both to osteoproliferation and to a deficiency of marrow-dependent cells.
T Cell Regulation of Polyclonal B Cell Responsiveness
Authors: Goodman Michael G. , Weigle William O.
Published: December 2022

Polyclonal activation of murine splenic B lymphocytes to secrete immunoglobulin was shown to be subject to regulation by splenic T cells. By admixture of separated B and T cell populations it was demonstrated that normal fresh splenic T cells were able to augment polyclonal B cell responsiveness to LPS up to several-fold. Optimal collaboration between these two cell types ensued when they were co-cultured in equal numbers. T cell-mediated enhancement of polyclonal B cell responses was dependent upon the ability of T cells to divide and was manifested upon T cell interaction with B cells soon after culture initiation. Originally expounded as a one-signal phenomenon, polyclonal activation of lymphocytes by LPS is, under the circumstances described, attributable instead to two distinct, nonspecific signals acting in concert. The observation that T cells from LPS-nonresponder (C3H/HeJ) mice were deficient in the capacity to enhance polyclonal B cell responsiveness of B cells derived from responder (C3H/HeN) mice implied a direct action of LPS on the involved T cells as well as an active role for the T cell signal in this immunoregulatory event. The novel observation of a functional T cell defect in LPS responsiveness in the C3H/HeJ mouse is discussed in terms of its other cellular defects.
Maturation of the Lymphoid System
Authors: Etlinger Howard M. , Chiller Jacques M.
Published: December 2022

A/J mice displayed a striking ontogenetic difference in the capacity to respond to DNP-Ficoll, a T-independent antigen, and to aggregated human γ-globulin (AHGG), a T-dependent antigen. Thus, whereas responses to DNP-Ficoll of 4-day-old mice were similar in magnitude to those of adult animals, responses to AHGG did not become pronounced until mice were some 30 to 40 days of age. The inability of young animals to respond to AHGG was reflective of a negative consequence of lymphocyte/antigen interaction, since such mice became specifically unresponsive to subsequent challenges with AHGG. Unresponsiveness induced by neonatal injection of AHGG lasted 50 to 60 days, in contrast to that induced by deaggregated HGG, which persisted some 100 days longer. The unresponsive state induced by injection of neonates with AHGG maintained itself upon adoptive transfer and did not appear to be linked to suppressive factors associated with either serum or lymphoid cells for its maintenance. Finally, AHGG was also shown to be capable of inducing unresponsiveness in neonatal, athymic mice. These results demonstrate that AHGG, the normally immunogenic form of HGG in adult mice, can serve as an effective tolerogen when administered into a neonatal environment.
Maturation of the Lymphoid System
Authors: Etlinger Howard M. , Chiller Jacques M.
Published: December 2022

It has been previously shown that AHGG, a form of HGG that is highly immunogenic in euthymic adult mice, is capable of inducing specific unresponsiveness when injected into neonatal animals. This report extends this finding and indicates that such a neonatal treatment results in the induction of tolerance in T as well as B cells. Furthermore, a similar conclusion was reached regarding specific T lymphocyte function in animals treated as neonates with OVA. The ability of LPS to modulate responses of neonatal animals to AHGG or DHGG was also examined. It appeared that such mice were not susceptible to the adjuvant effects of LPS until the 4th week of life. Furthermore, LPS was incapable of inhibiting the unresponsiveness induced in mice by either DHGG or AHGG until the 3rd or 4th week of life.
Requirement for a Bacterial Flora before Mice Generate Cells Capable of Mediating the Delayed Hypersensitivity Reaction to Sheep Red Blood Cells
Authors: MacDonald Thomas T. , Carter Philip B.
Published: December 2022

A delayed-type hypersensitivity (DTH) reaction can be elicited by an injection of 108 sheep red blood cells (SRBC) into a rear footpad of conventional (CV) mice previously immunized with small doses of SRBC. In contrast, immunization of germ-free (GF) mice with the same doses of SRBC produced no DTH when immunization was by the intravenous (i.v.) route, and only weak reactions when immunization was by the subcutaneous (footpad) route. Varying the immunizing dose of SRBC, or the time at which DTH was elicited, did not produce a state of DTH responsiveness in i.v. immunized GF mice. However, the transfer of lymphocytes from CV mice, immunized 4 to 5 days previously with SRBC, into GF mice, conferred on GF mice the capacity to express DTH. Although DTH was not readily demonstrable in GF mice immunized with SRBC, they nevertheless produced normal levels of hemagglutinating antibody to SRBC. Finally, it was shown that GF mice could generate a normal DTH response to SRBC if they were first monoassociated with a Gram-negative bacterial flora.
Eosinophil-Mediated Killing of Schistosomula of Schistosoma Mansoni in Vitro: Synergistic Effect of Antibody and Complement
Authors: McLaren Diane J. , Ramalho-Pinto F. J.
Published: December 2022

The capacity of rat peritoneal eosinophils to kill schistosomula of Schistosoma mansoni coated with antibody plus complement (C) (fresh immune rat serum) has been compared with that recorded when the parasites are coated with C alone (fresh normal rat serum) or antibody alone (heat-inactivated immune rat serum). Time course studies of adherence and killing, together with serum and cell dilution experiments, have revealed the superior efficiency of the system involving eosinophils, antibody, and C. Cell migration assays have shown that the combined factors generated when both the classical and alternative C pathways are activated at the parasite surface are more potent eosinophil chemoattractants than those generated from the alternative pathway alone. Moreover, faster and higher levels of eosinophil-mediated killing accompany the generation of such factors; 70 to 80% mortality being recorded within 3 hr as compared to 25% or less when the factors are absent. Experiments with fIRS chelated with EGTA indicate that more C3 molecules are probably bound to the parasite surface when C is activated via the classical rather than the alternative pathway, and that factors stimulating eosinophil migration generated from the classical pathway promote faster adherence and, as a consequence, higher levels of eosinophil-mediated killing. IgE does not seem to play an important role in eosinophil-mediated cytotoxicity in vitro, in the presence of fIRS, but the possible participation of the anaphylatoxins and ECF-A is discussed. It is suggested that the rapid and enhanced parasite killing effected by the combination of eosinophils, antibody, and C may be of paramount importance in the destruction of the schistosomula of a secondary infection in the immune host.
Some Invariant Properties of IgE-Mediated Basophil Activation and Desensitization
Authors: Goldstein Byron , Dembo Micah , Sobotka Anne K. , Lichtenstein Lawrence M.
Published: December 2022

We investigate certain general properties of antigen induced degranulation of sensitized basophils by analyzing two types of experiments: Experiments in which we expose basophils to two antigens sequentially and then determine the fraction of histamine released; and experiments in which we obtain time-dependent release and desensitization curves. To analyze the latter type of experiments we introduce a new way to plot release and desensitization data that depends on the nature of the interactions of histamine-containing units (histamine quanta) with themselves or the cells degranulation apparatus, but not on any specific properties of the antigen.

From our analysis we conclude that: 1) A fraction of histamine within a population of basophils is nonreleasable by antigenic stimulation. 2) When a basophil degranulates the initial release of histamine appears to inhibit subsequent release. 3) The rate of histamine release is proportional to the amount of releasable histamine remaining in the cells when the amount remaining is small, as expected if released of histamine granules is a stochastic process. 4) There is no dependence of desensitization on the extracellular calcium concentration.
Young F1 Mice Spontaneously Generate Cytotoxic T Cells against Parental Targets
Authors: Ishikawa Hiromichi , Dutton Richard W.
Published: December 2022

Spleen cells from young (AKR/J♀ × BALB/c) or (BALB/c♀ × AKR/J)F1 mice can spontaneously generate effector cytotoxic T lymphocytes (CTL), in a 5-day primary in vitro culture, which lyse target cells from AKR/J and BALB/c but not allogeneic mice. These spontaneous CTL responses first appear when spleen cells are taken from F1 mice at 3 to 4 weeks of age, are maximum at about 5 weeks, and have declined by week 7. The fact that these spontaneous CTL responses are never detectable in the spleen cell cultures from any ages of parental AKR/J and BALB/c mice makes them unique properties of the F1 mice.
Surface Ig Isotypes on Cells Responding to Lipopolysaccharide by IgM and IgG Secretion
Authors: Gronowicz Eva Severinson , Doss Carol , Assisi Francis , Vitetta Ellen S. , Coffman Robert L. , Strober Samuel
Published: December 2022

Using the fluorescence-activated cell sorter (FACS), we have investigated the Ig isotypes present on murine B cells, which can be polyclonally activated by lipopolysaccharide (LPS) in low cell density cultures. The LPS response was partly inhibited as a result of staining with anti-IgD and anti-IgM reagents, but not with anti-IgG reagents. The IgM+, IgD+, or IgG- fractionated cell populations gave both an IgM and an IgG response comparable to controls, whereas the response of the IgM-, IgD- cells was 5- to 20-fold lower. IgG- cells separated 1 day after LPS stimulation could still mount an IgM and IgG response indistinguishable from controls at the peak of the response. It is concluded that IgM+, IgD+, IgG- cells constitute the major LPS-sensitive cell population in the low cell density culture system and that IgG is not a necessary cell surface isotype for precursors of IgG-secreting cells.
Human T Cell Differentiation Antigens and Correlation of Their Expression with Various Markers of T Cell Maturation
Authors: Boumsell Laurence , Bernard Alain , Coppin Hélène , Richard Yolande , Penit Claude , Rouget Pierre , Lemerle Jean , Dausset Jean
Published: December 2022

Two sets of differentiation antigens are demonstrated on human T cells by using 11 heterologous anti-human antisera raised against various normal and malignant T cells. The two antigenic determinants from the first set of differentiation antigens are expressed only on thymus cells and on T lymphoblasts, whereas the two antigenic determinants from the second set are expressed on blood T cells, Sezary cells, T.CLL cells, and thymus cells. Four T cells phenotypes are thus defined; two phenotypes are expressed only by T lymphoblasts, whereas the other two phenotypes are expressed both by normal and malignant T cells. Moreover, a clear-cut relationship exists between the four T cell antigenic phenotypes and two other markers of T cell differentiation: terminal deoxynucleotidyl transferase and peanut agglutinin. Two phenotypes are linked with the presence of TdT, one phenotype is linked with the affinity for PNA, and the fourth phenotype is correlated with the absence of both markers.
Increased Prostaglandin Synthesis by Macrophages from Tumor-Bearing Mice
Authors: Pelus Louis M. , Bockman Richard S.
Published: December 2022

Prostaglandin E2 (PGE2) synthesis by macrophages was measured in normal and tumor-bearing mice. Splenic macrophages from C57BL mice bearing a transplantable, nonmetastasizing methylcholanthrene-induced fibrosarcoma produced significantly more PGE than cells from normal nontumor-bearing littermates. Augmented PG synthesis was found basally and after nonspecific stimulation of macrophages with endotoxin (LPS) and concanavalin A (Con A) as measured by radioimmunoassay. Resident peritoneal macrophages showed similar differences in radioimmunoassayable PGE levels. The conversion to prostaglandins of radiolabeled arachidonic acid previously incorporated into membrane phospholipids was followed over time. Macrophages from both normal and tumor-bearing animals incorporated equivalent amounts of tritiated arachidonic acid, showed similar distribution of label in phospholipid pools, and released equivalent amounts of labeled Arachidonic Acid and phospholipid. However, at both 3 and 18 hr of incubation, unstimulated macrophages from tumor-bearing animals released significantly more PGE2 and 6-keto PGF1α (the metabolite of prostacyclin) compared to macrophages from normal animals. Nonspecific stimuli (Con A and zymosan) greatly augmented PG production in both animal groups, with tumor-bearers greater than with normals. LPS caused little augmentation in basal PG secretion at 3 hr, but it greatly increased synthesis measured 18 hr later. In similar experiments, Moloney sarcoma virus (MSV) could be shown to augment PG synthesis in BALB/c mice bearing a transplantable MSV-transformed tumor line, but not in normal animals. No significant difference was noted in adherence, morphology, phagocytosis, or enzymatic activity when the resident macrophages from normal and tumor-bearing animals were compared, suggesting that they were at the same relative state of activation but differed in their capacity to synthesize prostaglandins.
The Clearance, Tissue Distribution, and Cellular Localization of Intravenously Injected Lipopolysaccharide in Rabbits
Authors: Mathison John C. , Ulevitch Richard J.
Published: December 2022

The initial interaction of bacterial lipopolysaccharide (LPS) with the blood and tissues of rabbits was studied by using radioiodinated LPS purified from Escherichia coli O111:B4 and Salmonella minnesota R595. After i.v. injection of 250 µg of 125I-LPS, a rapid clearance phase (t1/2 < 30 min) was observed with substantial uptake of LPS by liver, spleen, and lung during this initial period. With autoradiography, LPS was found to be concentrated in phagocytic vacuoles of hepatic Kupffer cells, splenic macrophages, and leukocytes. LPS remaining in plasma beyond 30 min was converted to a form with a density of less than 1.2 g/cm3, which disappeared from the blood with a half-life of 12 hr. Except for a marked increase in the concentration of O111:B4 LPS in the adrenal glands, tissue-bound LPS concentrations did not change appreciably between 5 and 180 min. When the low density form (ρ < 1.2 g/cm3) of either LPS was injected i.v., the initial rapid clearance phase was absent and approximately one-half of the LPS was cleared from blood during the next 4 hr followed by a slower clearance rate (t1/2 = 15 hr). In contrast to findings with the parent LPS, LPS concentrations in the adrenals at 180 min were 3- to 4-fold increased over those in other tissues. Results from SDS-PAGE of LPS recovered from plasma and liver homogenates indicated that no major cleavage of the glycolipid structure of LPS occurred within 180 min. Thus, i.v. injected LPS is rapidly partitioned into tissuebound LPS and that remaining in plasma, neither of which shows evidence of major degradative change within 180 min. Accumulation of low density LPS in the adrenal may compromise the ability of the host to survive in LPS-induced shock.
Effect of Cyclophosphamide on Suppressor Cell Activity in Mice Unresponsive to EAE
Authors: Lando Zeev , Teitelbaum Dvora , Arnon Ruth
Published: December 2022

Protection against experimental allergic encephalomyelitis (EAE) was induced in susceptible mice of (SJL/J × BALB/c)F1 hybrid, by injection of either mouse spinal cord homogenate, the small mouse basic protein, or Cop 1 in incomplete Freund's adjuvant, before EAE induction. It was demonstrated that the unresponsiveness induced by the three antigens is mediated by suppressor T cells residing in the spleen cell population and can be adoptively transferred to normal syngeneic recipients. Low dose of cyclophosphamide (20 mg/kg) administered 2 days before the encephalitogenic challenge abrogated the unresponsiveness to EAE and reverted the protected mice sensitive to disease induction. Cyclophosphamide was also active on adoptively transferred unresponsiveness, thus donors that had been treated with cyclophosphamide were unable to further transfer unresponsiveness to EAE. These results indicate the elimination by cyclophosphamide of suppressor cells that interfere with the effector mechanisms leading to EAE.
B Lymphocyte Determinants in Immunoglobulin a Nephropathy
Authors: Friend Peter S. , Yunis Edmond J. , Noreen Harriet J. , Reinsmoen Nancy , Dubey Devendra , Michael Alfred F.
Published: December 2022

Because of our prior demonstration of a strong association between a specific B lymphocyte determinant and the occurrence of chronic membranoproliferative glomerulonephritis and the demonstration by others of the frequent concurrence of an IgA deposit disease, anaphylactoid purpura nephritis, with HLA-BW35, we conducted an investigation of the association among Minnesota Caucasians of the HLA markers BW35, DW1 and B lymphocyte determinants identified by three alloantisera with IgA nephropathy and the syndrome of recurrent macroscopic hematuria. The highly significant association between HLA-BW35, DW1, and B cell antigens identified by alloantisera L, B, and F, present in 50 normal Minnesota Caucasians, was not observed in 18 Minnesota patients with IgA nephropathy. The frequency of HLA-DW1 was the same among controls and patients, whereas the relative risk of developing IgA nephropathy was demonstrated to be increased 4- to 5- fold in the presence of the B cell antigens identified by alloantisera L and B. These sera appear to identify determinants on B lymphocytes associated with the disease state and possibly not intimately related to the HLA-D region. These findings emphasize the possible informativeness of defining B cell alloantisera on disease panels.
Production of Chemotactic Activity in Mixed Leukocyte Cultures: Maximum Effect Caused by H-2I Region Disparity
Authors: Cheung Hou Tak , Sundharadas G.
Published: December 2022

Primary mixed mouse leukocyte culture supernatants contain an activity chemotactic for mouse peritoneal exudate cells and it can be detected within 72 h after initiation of the culture. Disparity for H-2I region leads to maximum production of chemotactic activity whereas H-2K or H-2D region differences result in the production of significantly less activity. The rate of production of chemotactic activity follows closely the rate of incorporation of 3H-thymidine, and both attain the peak on day 4 after initiation of the culture. But whereas proliferation is sensitive to gamma-irradiation, chemotactic activity production is not. It is our hypothesis that proliferating cells are primarily responsible for the production of chemotactic activity. The possible relevance of chemotactic activity production to graft rejection is discussed.
In Vitro Killing of S. Mansoni Schistosomula by Eosinophils from Infected Rats: Role of Cytophilic Antibodies
Authors: Capron Monique , Torpier Gérard , Capron André
Published: December 2022

The cytotoxic effect of peritoneal cells from Schistosoma mansoni-infected rats against antibody-opsonized or nonopsonized schistosomula in vitro has been studied during the course of infection. Eosinophil-enriched cell preparations were shown to have a high cytotoxic effect on schistosomula in the absence of antibody. The killer cells were identified as eosinophils. As in the ADCC mechanism previously described, mast cell-eosinophil interaction was required for eosinophil cytotoxicity.

Rosette formation using S. mansoni antigen-coated erythrocytes was used to demonstrate the presence of anti-S. mansoni IgG2a antibody at the surface of infected eosinophils. Passive sensitization of normal eosinophils with ultracentrifugation pellets of immune rat serum resulted in a significant cytotoxicity of sensitized eosinophils. A close relationship was found between the cytotoxic activity of infected cells and the ability of the corresponding infected serum to arm normal eosinophils.

At certain periods after infection, eosinophils from infected rats were less effective than normal eosinophils on antibody-coated schistosomula. EA- (rat) rosetting assay and blockade experiments with homologous immune complexes have revealed in a kinetic study that the blocking of cytotoxic activity of infected eosinophils was related to heat-stable circulating immune complexes.

The possible role of immune complexes either in arming or inhibiting effector cells is suggested.
In Vitro Primary Immunization of Human Peripheral Blood Lymphocytes to KLH: Evidence for HLA-D Region Restriction
Authors: Rodey Glenn E. , Luehrman Lisa K. , Thomas David W.
Published: December 2022

Human T cells were primed in vitro by culture for 12 days with autologous adherent monocytes and KLH. KLH-primed cells recovered from primary cultures were restimulated in a second culture containing autologous or allogeneic monocytes in the presence or absence of KLH, and proliferation was determined by incorporation of tritiated methyl-thymidine. Secondary proliferation of primed T cells requires both autologous adherent cells and KLH in the cultures. Allogeneic adherent cells work as effectively as autologous adherent cells in supporting proliferation only if the allogeneic cell shares an HLA-D region determinant in common with the autologous adherent cells used during the priming culture. Proliferative responses in cultures containing allogeneic adherent cells with no shared D-region specificities were markedly diminished or absent. This D region restriction was further confirmed by demonstrating that the addition to the secondary cultures of an anti-DR antiserum directed against the shared DR determinant abrogates proliferation whereas anti-DR antisera against the nonshared DR determinants on either primed T cell or allogeneic adherent cell do not.

In conclusion, this study provides additional evidence that the genetic restrictions governing human cell-interactions of in vitro immune responsiveness are analogous to those observed in other animal models. By extension, it is likely that these in vitro techniques can also be utilized to define Ir genes, which control immune responsiveness in humans; and ultimately, to characterize better the H-linked genetic factors that contribute to the susceptibility to certain disease states that are associated with HLA-D region gene products.
Selective Effects of Anti-la Serum and Complement Treatment upon Polyclonal B Lymphocyte Responses to Dextran Sulfate and Lipopolysaccharide
Authors: Bick Peter H. , Shreffler Donald C.
Published: December 2022

Subpopulations of B lymphocytes have been shown to vary in their expression of Ia alloantigens and polyclonal responsiveness to thymic independent antigens. We have demonstrated that the polyclonal B cell antibody response to dextran sulfate is less sensitive to removal of Ia-positive cells than is the response to LPS. This is a consistent finding whether alloantibody and complement (C) pretreatment is directed toward cells bearing Ia antigens coded for by the entire I region or by the I-A or I-E subregions.

Heterogeneity appears to exist within the dextran sulfate-sensitive population in that using high antibody:cell ratios during antibody and C-mediated cell selection results in an inhibition of the proliferative but not the antibody response. This result may indicate a differential expression of Ia antigens on dextran sulfate-sensitive B cells that respond by proliferation versus those cells that produce antibody. Alternatively, proliferative responses to dextran sulfate may be more dependent upon Ia-positive accessory cells than is the polyclonal antibody response.
Postnatal Development of T Cells
Authors: Papiernik Martine , Bach Jean-François
Published: December 2022

We have explored the thymus dependency of T cell precursors in mouse neonatal spleen. Neonatal spleen was grafted under the kidney capsule of adult syngeneic recipients [normal, adult thymectomized (ATx) or thymectomized, irradiated, and bone marrow reconstituted]. Grafts were tested for the presence of Ig+ and Thy 1+ cells and the response to phytomitogens and allogeneic cells. In normal and ATx recipients the grafts were partly populated with Thy 1+ cells of graft origin, which developed in situ from T cell precursors having migrated to the spleen before birth. The absence of the thymus (ATx recipients) did not prevent the appearance of Thy 1+ cells but impaired their functional maturation as assessed by Con A responses and mixed lymphocyte reaction. Such functions appeared to be sensitive to the humoral function of the thymus since they reappeared after grafting of a thymus gland in a cell-impermeable Millipore chamber. No T cell development was observed when neonatal spleens were grafted into T cell-deprived mice, suggesting that T cell precursors that preexisted in the spleen before grafting disappear in T cell-deprived mice whereas they persist in normal or ATx mice. One may thus hypothesize that the persistence and/or proliferation of T cell precursors in the neonatal spleen require the presence of a T cell environment, whereas its functional maturation is under the control of the humoral function of the thymus.