bluuebunny/crossref_metadata_2025 huggingface dataset:
On the Mechanism of Membrane Damage by Complement: The Effect of Length and Unsaturation of the Acyl Chains in Liposomal Bilayers and the Effect of Cholesterol Concentration in Sheep Erythrocyte and Liposomal Membranes
Authors: Shin Moon L. , Paznekas William A. , Mayer Manfred M.
Published: December 2022

As shown previously, the action of activated complement (C) on a lipid bilayer produces two distinct effects, namely, formation of transmembrane channels and transfer of phospholipid from the bilayer to the aqueous phase. In the present work, we show that both phenomena are affected by the structure of the membrane lipids. Increasing acyl chain length of lecithin liposomes containing cholesterol was found to decrease release of 86Rb+ by antibody (Ab) and C or by purified C5b,6 + C7 + C8 + C9. These changes in 86Rb+ release suggest that channel formation is affected by membrane thickness. Similarly, release of 14C-PC by Ab + C diminished with increasing acyl chain length.

Release of 86Rb+ from egg lecithin liposomes by Ab + C was found to diminish with increasing cholesterol concentration. On the other hand, higher release of 14C-PC was observed with increasing cholesterol concentration. Cholesterol itself was not released by Ab + C.

The effect of membrane cholesterol was also studied with sheep erythrocytes by incubation of the cells with a cholesterol-enriched medium. In accord with the results obtained with liposomes, the susceptibility of erythrocytes to hemolysis by C5b-9 was found to diminish with increasing cholesterol enrichment of the membrane, which indicates that cholesterol affects channel formation.

The effect of unsaturation was tested by treatment of C18:0 or C18:1 lecithin liposomes containing 43% molar cholesterol with Ab + C. 86Rb+ release was decreased, whereas 14C-PC release was slightly increased when unsaturated chains were present. Membrane fluidity is not believed to be the sole controlling factor since the changes in this parameter by varying cholesterol or by unsaturation did not correlate with the effects on marker release.
Two Distinct Populations of Suppressor Cells in the Spleens of Mice Bearing Methylcholanthrene-Induced Tumors
Authors: Pope B. L. , Whitney R. B. , Levy J. G.
Published: December 2022

The suppressor cells found in the spleens of mice bearing advanced tumors could be divided into two different populations by centrifugation on Hypaque-Ficoll of specific gravity 1.08. The cells in the light fraction suppressed the in vitro PFC response to SRBC and DNP-LPS. They appeared to be of the macrophage/monocyte line since they were adherent to plastic and nylon wool but were not removed by anti-BA-ϑ serum plus complement (C). They did not require DNA synthesis for activity since they were not inactivated by mitomycin C. The cells in the dense fraction suppressed only the response to SRBC. They may have been T cells since they were not adherent to plastic or nylon wool and their activity was reduced by treatment with anti-BA-ϑ serum plus C. They did require DNA synthesis since their activity was removed by treatment with mitomycin C. Suppression of normal lymphocyte function could be affected when splenic suppressor cells were separated from the normal population by a cell-impermeable membrane. Suppression occurred when the two populations were separated by a 0.8-µ Nuclepore membrane or by regular dialysis tubing but not when low porosity dialysis buting (m.w. <3,500 retained) was used. The suppressor cell in this system was nonadherent to plastic, partially adherent to nylon-wool, not removed by carbonyl iron and magnet, and pelletted to the bottom of a Hypaque-Ficoll gradient.
Cell Types Involved in Cytotoxicity Induced by Heated Cells and Inhibition of This Cytotoxicity by Anti-Human B Cell Sera
Authors: Kasakura Shinpei
Published: December 2022

We have studied the induction of cytotoxic activity in human peripheral blood lymphocytes by heated allogeneic cells. By separating T and B cells from the responder and stimulator cell populations we found that cytotoxic cells are generated in responder T cell populations by both T and the B stimulator cells.

Rabbit antisera to a membrane glycoprotein complex (33,000 and 27,000 m.w. by SDS-gel electrophoresis) isolated from a human B cell line were utilized to explore further the nature of the effector cells in this type of cytotoxicity. This antiserum, present during the 6-day-culture period, blocked generation of cytotoxic effector cells. Depletion of cells bearing the B cell antigen from the responder cell population by anti-B cell serum and complement (C) eliminated cytotoxicity. Furthermore, heated cell-induced cytotoxicity was blocked by simply pretreating the responder or the stimulator cell populations with anti-B cell serum in the absence of C. Apparently the human lymphocyte that functions as the effector cell in heated cell-induced cytotoxicity bears the Ia-like antigen that might be important in triggering this type of cytotoxicity.
Isolation and Analysis of H-2 and Ia Alloantigens from Wild Mouse Strains
Authors: Cullen Susan E. , Hansen Ted H. , Klein Jan
Published: December 2022

Immunoprecipitation and SDS-PAGE analysis were used to examine H-2 and Ia antigens from mouse strains with wild-derived MHC haplotypes. Antisera raised against the wild-derived strains contained anti-H-2 and anti-Ia antibodies which precipitated antigen molecules readily distinguishable by a single assay procedure. The antibodies to wild strains were cross-reactive with standard laboratory haplotypes. Evidence supporting the similar genetic organization of wild and standard haplotypes was found, including isolation of separate H-2K and H-2D molecules from a wild-derived strain, and isolation of two separate Ia molecules from a wild-derived strain.
Light Chain Diversity of Murine Anti-Streptococcal Antibodies: IgCH-Linked Effects on L Chain Expression
Authors: Perlmutter Roger M. , Briles David E. , Greve Jeffrey M. , Davie Joseph M.
Published: December 2022

L chains derived from anti-group A streptococcal carbohydrate antibodies raised in A/J, BALB/cJ, C57BL/6J, CB-20, BAB-14, and CAL-20 mice were examined by isoelectric focusing. Multiple strain-associated differences in the degree and frequency of expression of particular L chain spectrotypes were observed. Analysis of L chain-focusing patterns in allotype-congenic mice revealed that IgCH-linked genes can have profound effects on the L chain phenotypes expressed by strains with identical L chain genotypes. Lastly, the overall spectrotypic diversity of L chains from anti-GAC antibodies appears to be less extensive than the diversity of the antibodies from which these L chains derive, documented by similar techniques. These results are interpreted in light of the significance of combinatorial diversity in generating antibody heterogeneity.
Identification and Characterization of a New Subset of Rat T Lymphocytes
Authors: Kierszenbaum Felipe , Budzko Delia B.
Published: December 2022

Subpopulations of rat T lymphocytes described in this paper are defined by their susceptibility to the selective cytotoxic effects of normal guinea pig serum (GPS). These cells were present in relatively large proportions in the bone marrow and thymus and in very small percentages in the spleen and lymph nodes. Tissue distribution of GPS-sensitive lymphocytes was similar in three inbred strains and, therefore, was characteristic of the species rather than a single strain. Adherent peritoneal cells were largely insensitive to GPS. The partial cytotoxicity of GPS on rat lymphoid cells contrasted with the indiscriminate killing capacity of normal sera from humans, rabbits, and chickens, which affected over 98% of the cells regardless of organ location. Variations of the number of cells used in otherwise identical GPS-cytotoxicity assays did not significantly alter the proportion of either thymus or bone marrow cells killed. This suggests a selective sensitivity by discrete subclasses of lymphocytes present in these organs. GPS-sensitive lymphocytes occurred in rat bone marrow from birth and constituted 30 to 50% of the nucleated cells. The percentage of these cells decreased after 5 weeks of age to 15 to 30%, with most value being around 20%. Cells susceptible to GPS cytotoxicity were absent in the thymus of 35-day-old or younger rats but were readily detectable in 8-week-old animals, suggesting that GPS-reactivity defines a thymus cell surface-differentiation component. The subset of the thymus cell expressing this marker was essential for responsiveness to either PHA or Con A. Prior killing of bone marrow cells with GPS left no cells to be killed specifically by rabbit anti-rat Thy 1 IgG plus complement (C) free of nonspecific cytotoxicity. Initial killing of bone marrow cells with anti-Thy 1 and C consistently eliminated the GPS-sensitive cells. These results indicate that in the rat bone marrow, unlike in the thymus, GPS-sensitive and Thy 1-bearing lymphocytes constitute the same cell subpopulation. GPS is capable of “recognizing” and killing subpopulations of rat T lymphocytes, a property that can be used to characterize further these cells on a functional basis.
The Control of Autoreactivity
Authors: Osband Michael , Parkman Robertson
Published: December 2022

To determine if autocytotoxic lymphocytes were naturally occurring in murine spleens, C57BL/6 spleen cells were fractionated on discontinuous BSA gradients. Autocytotoxicity was assessed in vitro (cytotoxicity of C57BL/6 fibroblasts) and in vivo (splenomegaly and popliteal node enlargement in C57BL/6 mice). A medium density subpopulation of lymphocytes was shown to be autocytotoxic and to be similar to autoreactive lymphocytes produced by the in vitro “sensitization” of splenic lymphocytes on syngeneic fibroblasts monolayers. The naturally occurring autocytotoxic lymphocytes express no detectable Θ antigen, did not adhere to nylon, but did have an Fc receptor. In recombination experiments, BSA-fractionated lymphocytes were incubated with autocytotoxic and “sensitized” lymphocytes. A light density subpopulation was shown to suppress both autoreactive lymphocyte subpopulations at a 1:50 ratio. The suppressor cells were nylon nonadherent T lymphocytes. The lack of autoreactivity of unfractionated murine spleen cells is due to the concomitant presence of autocytotoxic and suppressor lymphocytes. If suppressor lymphocytes are selectively removed in vitro, the reactivity of autocytotoxic lymphocytes can be detected.
IgE-B Cell-Generating Factor from Lymph Node Cells of Rats Infected with Nippostrongylus Brasiliensis
Authors: Urban Joseph F. , Ishizaka Kimishige
Published: December 2022

Cell-free culture supernatants (CFS) of mesenteric lymph node cells from Nippostrongylus brasiliensis-infected rats specifically increase the proportion of IgE-bearing cells in normal rat bone marrow cultures without changing the proportion of IgM-bearing cells and IgG2a-bearing cells. Virtually all of the IgE-bearing cells appearing in the culture carried IgM determinants on their surface. Precursors of IgE-bearing cells in normal bone marrow cells were removed by depletion of immunoglobulin-bearing cells. These findings indicate that IgE-B cell-generating factor converts IgM-bearing virgin B cells to IgM-IgE double bearing cells. Pretreatment of normal bone marrow cells with mitomycin C failed to inhibit the generation of IgE-bearing cells by the soluble factor. Indeed, most of IgE-bearing cells generated in cultures of mitomycin C-treated bone marrow cells did not contain 3H-thymidine, which was present during the entire culture period. It appears that DNA synthesis is not required for the differentiation of IgM-bearing cells to IgE-IgM double bearing cells. However, the appearance of IgE-bearing cells in cultures of normal bone marrow cells by CFS was abated by 0.5 µg/ml of actinomycin D or by pretreatment of the cells with pactamyicn. These findings suggest that the observed differentiation involves some process dependent on transcription of DNA and protein synthesis.
The Induction of Hapten-Specific Immunologic Tolerance and Immunity in B Lymphocytes
Authors: Fidler John M.
Published: December 2022

In vivo administration of trinitrobenzenesulfonic acid (TNBS) induces a state of immunologic tolerance to the trinitrophenyl (TNP) hapten in the bone marrow-derived (bursal equivalent, B) lymphocytes of mice. An in vitro culture system utilized to generate an antibody forming cell (AFC) response was characterized and used to evaluate the unresponsive state. The tolerance induced by TNBS injection was TNP-specific and dose-dependent, and was present in response to both the thymic-derived (T) cell independent (TI) antigen TNP-lipopolysaccharide (TNP-LPS) and a T cell dependent (TD) form of TNP (TNP-sheep erythrocytes, TNP-SRC). Unresponsiveness persisted for over 8 weeks in response to either antigen.

Experiments were conducted to determine the mechanism of TNBS-induced immunologic tolerance with emphasis upon differentiation between B cell deletion and blockade of surface immunoglobulin (sIg) receptors by tolerogen. Tolerant and normal spleen cells responded independently to TNP-LPS when co-cultured, indicating that the observed unresponsiveness in TNBS-treated spleen cells was not due to suppression of TNP-specific B lymphocytes by other cells in the preparation. Used at polyclonal B cell-activating concentrations, TNP-LPS did not reverse the tolerant state. Additionally, unconjugated LPS in either serum-containing or serum-free cultures did not break tolerance. Because polyclonal B cell activators do not act in triggering through sIg receptors, the failure of a polyclonal stimulator to reverse the tolerant state suggests that blockade of TNP-specific surface receptors is not the mechanism of tolerance. Tolerant spleen cells did not reduce the number of TNP-specific plaque forming cells (PFC) produced upon stimulation of normal cells by a wide range of LPS doses, supporting the conclusion that suppressive cells or factors are absent. Antigen-free incubation of tolerant spleen cells for 24 hr and washing at 4°C did not abrogate the unresponsive state when followed by challenge with either TNP-LPS or TNP-SRC. Washing at elevated temperatures after pre-culturing in order to avoid possible reassociation of sIg-bound TNP conjugates was equally unsuccessful in reversing tolerance. Thus, antigen-free precultivation where unblocking or turnover and resynthesis of sIg receptors should have taken place yielded no evidence supporting receptor blockade as the basis of TNBS-induced tolerance. Treatment with the proteolytic enzyme pronase, in order to strip potential tolerogen from the cell surface, failed to reverse the unresponsive state. Incubation with anti-TNP or anti-Ig antibody under conditions designed to allow capping and removal of surface-bound trinitrophenylated moieties or sIg-bound tolerogen also revealed a stable tolerant state.

These results and evidence presented previously support the conclusion that TNBS induces tolerance in vivo through a deletion of TNP-specific B lymphocytes.
Dextran Alters Induction of Cell-Mediated Hypersensitivity
Authors: Alevy Y. G. , Battisto J. R.
Published: December 2022

Administering dextran to 6- to 8-week-old CBA/J mice just before applying the initial sensitizing dose of a hapten caused induction of heightened delayed-type responsivity to the hapten. Measured by an isotope localization method, the cell-mediated hypersensitivity engendered by a hapten, picryl chloride, was twice as great in animals receiving dextran than in control animals given the saline vehicle. Optimal enhancement was achieved when dextran administration preceded hapten by 2 hr. Furthermore, the augmentation was observed at the first test after the initial sensitization period but not after repetitive dermal applications of the hapten had been made over a prolonged time interval. In the latter instance diminution of sensitivity was observed.

Washed lymphoid cells, taken from animals 2 hr after treatment with dextran, transferred to x-irradiated syngeneic recipient animals the heightened capacity to become sensitized. Such lymphoid cells retained the ability to transfer the heightened capacity to become sensitized despite removal of macrophages by plastic-adherence.

Although dextran is known to cause murine T cells to release factors that increase responses of other T cells to mitogens, it is not known whether dextran affects that subpopulation of T cells directly mediating contact sensitivity or whether its action is upon a separate subpopulation of T cells concerned with immuno-regulation.
Suppressor T Cell Mechanisms in Contact Sensitivity
Authors: Miller Stephen D. , Sy Man-Sun , Claman Henry N.
Published: December 2022

We investigated mechanisms of suppression of contact sensitivity to DNFB in mice. In particular, we studied the suppression produced by suppressor T cells which had been induced by injection of syngeneic DNP-modified lymphoid cells (syninduced Ts). Syninduced Ts were found by several criteria to inhibit the efferent limb of sensitivity by inhibiting the expression of LN cells from DNFB-sensitized animals. First, syninduced Ts can suppress previously sensitized recipients in an antigen-specific manner. Second, co-transferred syninduced Ts could block the passive transfer of contact sensitivity by DNFB-immune LN cells given to normal recipients. This inhibition was shown to be dose dependent, antigen specific, and non-MHC restricted. Third, in reverse transfers of sensitized cells into tolerant recipients, only those recipients containing active syninduced Ts could inhibit the expression of sensitivity by the passively transferred immune T cells. In contrast, syninduced Ts were incapable of inhibiting the afferent limb of the response as assessed by the normal, post-sensitization DNA synthetic response of draining LN cells in Ts recipient mice. Syninduced Ts were not inhibitable by prior treatment with anti-TNP antiserum, indicating a lack of surface-associated tolerogen. The locus of suppression was most clearly shown in experiments where mice received syninduced Ts at the same time they were sensitized. These mice were themselves unresponsive, but did contain immune lymphoid cells able to express contact sensitivity after transfer to normal recipients followed by DNFB challenge. The data suggest that the suppressive environment allows the development of a normal complement of DNFB-specific immune T cells, but inhibits the expression of this sensitivity. The data are discussed in terms of current models of suppression, and are compared with mechanisms of suppression in other contact sensitivity models.
Loss of Self-Tolerance to Single-Stranded Deoxribonucleic Acid (sDNA) in Vitro
Authors: Roder John C. , Bell David A. , Singhal Sharwan K.
Published: December 2022

The spleens of many normal and autoimmune-susceptible strains of mice mount a specific, IgM, anti-sDNA plaque-forming cell (PFC) response in vitro in the absence of an exogenous source of antigen. This response was not related solely to levels of xenotropic or ecotropic virus and was generated from a small number of precursor cells capable of binding sDNA. Small numbers of anti-sDNA PFC were also apparent in serum-free medium and the response of low-responder strains could be augmented with pokeweed mitogen. T cells and macrophages were not essential and cell division was required early in culture to obtain a peak response on day 5. These results suggest that autoantigen-sensitive cells may escape normal regulatory mechanisms in vitro and differentiate into clones of autoantibody secreting cells.
Oxygen Immunosuppression: Modification of Experimental Allergic Encephalomyelitis in Rodents
Authors: Warren Joel , Sacksteder Miriam R. , Thuning Claire Ann
Published: December 2022

Using protocols that incorporated double blind examination of animals sensitized to CNS antigen, we confirmed and amplified earlier findings of the complete suppression of EAE in rodents by hyperbaric oxygen. The effects of O2 were related to the gas pressure and duration of treatment. The development of paralytic disease was prevented for 34 days after sensitization (the longest interval studied). Compressed air or normobaric O2 administered under similar conditions did not modify the course of illness. Within 7 to 10 days after the discontinuance of oxygen therapy the majority of treated guinea pigs developed typical signs of EAE with characteristic lesions in the CNS. The relapses occurred sooner in the Lewis rat. The development of the delayed hypersensitivity reaction to myelin basic protein and to tuberculin is also suppressed by O2 therapy indicating that its effects upon autoimmune encephalomyelitis involves fundamental alterations of the cellular components of the immune response, some or all of which are reversible.
Histamine Release Due to Bivalent Penicilloyl Haptens Control by the Number of Cross-Linked IgE Antibodies on the Basophil Plasma Membrane
Authors: Dembo Micah , Goldstein Byron , Sobotka Anne K. , Lichtenstein Lawrence M.
Published: December 2022

We describe the characteristics of in vitro histamine release from human basophils passively sensitized with serum from a penicillin-allergic individual. The histamine release is induced by a synthetic bivalent hapten, bis benzylpenicilloyl 1,6 diaminohexane (BPO)2. We present data on the effect of a monovalent hapten, benzylpenicilloyl formyl-l-lysine (BPO)1, on the histamine release. We also examine how histamine release depends on the concentration of serum used for passive sensitization, the source of cells used for passive sensitization, and the time allowed for histamine release.

We interpret these experiments in terms of a theory of equilibrium binding of bivalent haptens to cell surface antibody that is presented in the previous paper. The results are consistent with the idea that the amount of histamine release is controlled by the number of cross-linked IgE molecules on the cell surface. In particular, the histamine dose-response curve rises because cross-links rise, has a maximum because the cross-links are a maximum, and falls because the cross-links fall.
Regulation of the Autoimmune Plaque-Forming Cell Response to Single-Strand DNA (sDNA) in Vitro
Authors: Roder John C. , Bell David A. , Singhal Sharwan K.
Published: December 2022

We have shown that young autoimmune and normal strain mice possess autoantigen-sensitive cells potentially capable of producing anti-sDNA autoantibody in the absence of normal regulatory mechanisms in vitro. In certain strains such as B/W mice, these regulatory mechanisms presumably break down with increasing age, and autoimmunity develops. These regulatory mechanisms might consist of sDNA, T cells, or some combination of these since both of these agents suppressed the anti-sDNA PFC response in vitro. The sDNA may have inhibited PFC development by a receptor blockade mechanism since i) spleen cells pulsed with sDNA for short periods and then washed were suppressed after 5 days of culture; ii) treatment of these blocked cells with trypsin and DNase I restored the anti-sDNA response; iii) the PFC remaining in partially blocked cultures were of lower avidity than PFC in unblocked cultures; and iv) the target of sDNA may be a B cell.

Thymocytes and splenic T cells suppressed the anti-sDNA response but not the anti-SRBC response in vitro in a dose-dependent manner. The suppressive capacity of thymus cells did not decline with age in B/W mice. In addition, thymus cells activated by competing foreign antigens could also suppress the anti-sDNA response. The relationship between these modes of regulating autoreactivity remains to be investigated.
Effect of a Synthetic Adjuvant on the Induction of Primary Immune Responses in T Cell-Depleted Spleen Cultures
Authors: Watson James , Whitlock Cheryl
Published: December 2022

N-acetylmuramyl-l-alanyl-d-isoglutamine (muramyl dipeptide) stimulates in vitro primary immune responses to SRBC in T cell-depleted (nude) spleen cultures. The stimulation of immune responses by muramyl peptide was antigen dependent. A microculture system was used to compare the T cell-replacing activities of several structural analogues of muramyl dipeptide and to compare the activity of muramyl dipeptide to helper T cells. In a limiting dilution analysis with excess helper T cells or muramyl dipeptide, the frequency of B cell precursors that respond to SRBC was similar, ranging from 1.5 to 5 × 10-5. Decreasing the cell density in microcultures did not affect the efficiency of B cell precursor responses in the presence of muramyl dipeptide. Muramyl dipeptide was examined for mitogenic activity in spleen cell cultures. In serum-free medium, muramyl dipeptide stimulates slight (3-fold) increases in DNA synthetic activity. In medium supplemented with 5 to 20% fetal calf serum, muramyl dipeptide showed no significant mitogenic activity. There are a number of possible explanations for the T cell-replacing activity of muramyl dipeptide. The most likely is that muramyl dipeptide interacts directly with B cells to mimic the helper T cell signal in the inductive stimulus.
The Relationship Between Bacterial Endotoxin and Human B Cell-Activating Factor
Authors: Wood David D. , Cameron Patricia M.
Published: December 2022

The monokine, B cell-activating factor (BAF), has the property of stimulating murine B lymphocytes in the absence of T lymphocytes to make a humoral response in vitro to erythrocyte antigens. The factor is secreted by fresh human monocytes after stimulation with as little as 10-11 g/ml of bacterial endotoxin. It had previously been shown that commercial preparations of phytohemagglutinin stimulated BAF release, but subsequent analysis of this phenomenon showed that the BAF-inducing activity of the phytohemagglutinin preparations is resistant to heating (which reduces T cell mitogenicity) and is sensitive to the LPS inhibitor polymyxin B. Therefore, the BAF-releasing activity of the PHA preparations is probably due to contaminating LPS. Once it was realized that BAF release had only been observed in the presence of LPS, the question was raised whether BAF was a derivative of the added LPS. This seems unlikely since 1) BAF and LPS can be separated by ion exchange chromatography, 2) relative to its activity in the Limulus lysate test, BAF is at least 105 times more active than LPS as a B cell stimulator, 3) BAF is a poor mitogen compared to LPS, 4) BAF, but not LPS, loses activity in nonsupportive media, 5) the C3H/HeJ mouse responds to BAF but not LPS, and 6) polymyxin B inhibits LPS but not BAF. Finally, BAF is not a derivative of lipid A protein since lipid A protein (which resembles BAF by criteria 5 and 6) is not necessary to stimulate BAF secretion by monocytes.
Bacteria Induce Lymphokine Synthesis Polyclonally in Human B Lymphocytes
Authors: Räsänen Liisa , Karhumäki Eliisa , Arvilommi Heikki
Published: December 2022

We have studied the ability of various bacteria to stimulate human lymphocytes to produce leukocyte migration inhibitory factor (LIF). Mononuclear cells from adult and cord blood as well as purified T and B lymphocytes were stimulated with killed bacteria. The culture supernatants were tested for the presence of LIF by the agarose migration method. All nine bacterial strains tested activated unseparated mononuclear cells and B lymphocytes but not T cells to produce LIF. LIF was also present in cord blood cell cultures suggesting that the stimulation of lymphocytes was polyclonal rather than antigenic. Therefore, we propose that one of the physiologic functions of B lymphocyte lymphokines might be to form part of the nonspecific defense mechanisms against microbial invasion.
Allogeneic Restriction of Acquired Antimicrobial Resistance in the Rat
Authors: Jungi Thomas W. , McGregor D. D.
Published: December 2022

Allogeneic restriction of acquired antimicrobial resistance was demonstrated by infusing Listeria-immune thoracic duct lymphocytes (TDL) into syngeneic, semisyngeneic, or allogeneic recipients that had been infected i.v. with Listeria monocytogenes (LM). Syngeneic and semisyngeneic recipients enjoyed a high level of protection (3 to 5 log10 reduction in viable organisms within 64 hr, whereas allogeneic recipients were poorly protected (0.5 to 2.5 log10 reduction). Sharing one haplotype between TDL-donors and challenged recipients resulted in a 4- to 8-fold increase in the level of resistance transferred. Histoincompatibility reactions could be excluded as a cause of the observed allogeneic restriction. Therefore, it is likely that LM-immune TDL are unable to respond efficiently to the antigens of this organism in an allogeneic environment. But allogeneic immune TDL do confer a low level of protection that cannot be ascribed to B cell activity nor to GvH and/or HvG reactions. Evidence is provided that the modest protection afforded by allogeneic lymphocytes is conveyed by the same T cell subset that operates with much higher efficiency in genetically related rats.
Correlation of the Suppression of Mitogen Responsiveness and the Mixed Lymphocyte Reaction with the Proliferative Response to Viral Antigen of Splenic Lymphocytes from Cytomegalovirus-Infected Mice
Authors: Kelsey Douglas K. , Overall James C. , Glasgow Lowell A.
Published: December 2022

Lymphocyte function has been shown to be suppressed during cytomegalovirus infections of humans and mice. We compared nonspecific and specific lymphocyte reactions of splenic lymphocytes from murine cytomegalovirus (MCMV)-infected C3H adult mice during the acute and recovery phases of infection. The response to phytohemagglutinin or lipopolysaccharide was suppressed maximally 4 to 5 days after infection and returned to normal levels by 15 days. The mixed lymphocyte response was suppressed 2 to 10 days after infection and returned to normal by 15 days. In contrast, lymphocytes responded to MCMV-infected syngeneic mouse embryo fibroblasts (MEF) by day 2 of infection incorporating 52,183 ± 9866 counts per minute (cpm) of 3H-thymidine compared with 23,587 ± 3909 cpm with uninfected MEF. The response to infected MEF increased to 132,560 ± 27,739 cpm by day 10. The ability of splenic lymphocytes from infected mice to respond to MCMV-infected MEF was retained for at least 75 days after infection. Preliminary characterization of the cells responding in the MCMV-induced proliferation assay showed that they were sensitive to treatment with anti-θ serum and complement, but not to treatment with anti-mouse γ-globulin and complement. The development of a specific lymphocyte response to viral infected cells during MCMV infection at a time when the MLR and response to mitogens are suppressed suggests that viral induced immuno-suppression may result from the regulation of the immune response of the host rather than a direct effect of the virus.
Antibody-Dependent Lymphocytotoxicity: An Analysis of Effector Cell-Target Cell Interactions
Authors: Van Oers M. H. J. , De Goede R. E. Y. , Zeijlemaker W. P.
Published: December 2022

Antibody-dependent lymphocytotoxicity (ADL) was studied with human peripheral blood lymphocytes as effector cells and P815 mouse mastocytoma cells, sensitized with rabbit IgG antibodies, as target cells. Enzymesubstrate-like kinetics were used to describe ADL inhibition induced by two types of inhibitors. Human IgG, both native and heat-aggregated, proved to be a competitive inhibitor of ADL at the target cell level. Human peripheral blood monocytes inhibited ADL in an apparently irreversible fashion, without appreciable evidence for competition. The data obtained provide strong support for the validity of an enzyme-substrate-like mechanism of ADL. Moreover, our results indicate that in order to measure the lytic capacity properly, cell populations, well defined as to their composition, should be used.

In the second part of the study, it was investigated whether the two types of cells capable of lysing sensitized target cells, i.e., null cells and T cells, differed with respect to their affinity for the target cells. Application of enzyme-like kinetics revealed considerable differences in maximal killing rate between the two subsets of K cells. However, the parameter related to the affinity toward the target cells was found to be of the same magnitude for the two types of effector cells.
Activation of Human B Lymphocytes
Authors: Haynes Barton F. , Fauci Anthony S.
Published: December 2022

Heterogeneous subpopulations of peripheral blood (PB) mononuclear cells that are capable of being activated by concanavalin A (Con A) to suppress the pokeweed mitogen (PWM)-induced anti-sheep red blood cell (SRBC) direct plaque-forming cell (PFC) response of human B cells have been described. Removal of adherent cells by passage over a Sephadex G-10 column or by plating of PB mononuclear cells on plastic surfaces did not affect the generation of Con A-activated suppressor cells (SC) (p < 0.01). Separation of PB cells into T cell-enriched (TCE) and T cell-depleted (TCD) suspensions by SRBC rosetting procedures demonstrated maximal Con A-activated suppression in each of these subpopulations (p < 0.001). Separation of the TCE fraction into T cells that bear Fc receptors for IgG (T.G) and those that do not (T.G-depleted) demonstrated submaximal suppression in each group alone. The T.G-depleted subpopulation suppressed to 50 ± 22% of control whereas the T.G fraction obtained by 7S EA rosetting did not suppress (128 ± 66% of control). However, T.G-enriched cells obtained, without activating the 7S Fc receptor, by depleting TCE fractions of T cells bearing an Fc receptor for IgM (T.M-depleted) did suppress maximally. Finally, separation of unfractionated PB mononuclear cells into those cells that possess complement (C) receptors (CR positive) and those that do not (CR negative), demonstrated that there was significant Con A-activated suppression in the CR-negative fraction of cells (p < 0.01) whereas the Con A-activated CR-positive cell did not suppress (p > 0.2). Thus, these studies demonstrate that cells that are capable of being activated by Con A to suppress the PWM-induced PFC response of normal human PB mononuclear cells are found in both the TCE and TCD fractions. In the TEC suspensions, Con A SC are found in both IgG Fc receptor positive and negative fractions, although IgG Fc receptor negative cells were less potent suppressors than IgG Fc receptor positive cells. In the TCD suspension, Con A SC are nonadherent, E-rosette-negative, Cr-negative cells.
Sulfated Glycosaminoglycans of Guinea Pig Basophilic Leukocytes
Authors: Orenstein Neil S. , Galli Stephen J. , Dvorak Ann M. , Silbert Jeremiah E. , Dvorak Harold F.
Published: December 2022

Cytoplasmic granules of basophilic leukocytes stain metachromatically and have been thought to contain sulfated glycosaminoglycans, presumably heparin. To test this hypothesis, we identified the [35S]glycosaminoglycans synthesized by guinea pig blood basophils in culture and in vivo. Basophils isolated from guinea pig blood were cultured for 20 hr in F12 medium—10% guinea pig serum containing sodium [35S]sulfate. Alternatively, basophils were purified from animals receiving repeated i.v. injections of sodium [35S]sulfate. Glycosaminoglycans were isolated from these basophils after pronase digestion and identified by the use of selective glycosaminoglycan-degrading enzymes. Approximately 55% of the [35S]glycosaminoglycans was degraded by chondroitinase AC, indicating the presence of chondroitin sulfate; an additional 30 to 35% could be degraded by chondroitinase ABC, indicating that dermatan sulfate was also present. The 15% glycosaminoglycan remaining after chondroitinase ABC digestion was degraded by purified heparitinase (heparanase), which has no effect on authentic heparin but degrades heparan sulfate. Thus, the glycosaminoglycan content of guinea pig basophils is a mixture of chondroitin sulfate, dermatan sulfate, and smaller amounts of heparan sulfate. No heparin was detected.
Low Molecular Weight C1q-Precipitins in Hypocomplementemic Vasculitis-Urticaria Syndrome: Partial Purification and Characterization as Immunoglobulin
Authors: Marder Robert J. , Burch Frank X. , Schmid Frank R. , Zeiss Chester R. , Gewurz Henry
Published: December 2022

A lupus-like syndrome involving chronic urticaria with cutaneous vasculitis, systemic symptoms, hypocomplementemia with preferential depletion of C1q, and low m.w. (7S) C1q-precipitins has recently been defined. The C1q-precipitin activity (C1q-p) seems to represent a diagnostic marker of the disease, but its chemical nature is not yet clear. We have partially purified and characterized C1q-p from the serum of two patients with this syndrome and compared its activity with the C1q-precipitating activity of aggregated human γ-globulin (AHGG), anti-C1q antibodies, and several polynucleotides including DNA and polyinosinic acid. C1q-p was found to partition with IgG during precipitation by ammonium sulfate and low ionic strength buffer as well as during column chromatography on DEAE-cellulose and G-200 Sephadex. Like AHGG, but in complete contrast to the polynucleotides, the C1q-precipitating activity of C1q-p was sensitive to pepsin, trypsin, and acidic conditions, but unaffected by DNAse or RNAse; the C1q-precipitating activity of anti-C1q antibody was not diminished by any of these procedures. Thus, C1q-p consists of γ-migrating protein of low m.w., and its C1q-precipitating activity is indistinguishable from that of AHGG. These results are consistent with the concept that C1q-p is comprised, at least in part, of IgG that binds C1q via the Fc portion of the molecule.
Surface Markers and Electron Microscopy of Human Blood L Cells: A Comparison with T and B Lymphocytes
Authors: Horwitz David A. , Niaudet Patrick , Greaves Melvyn F. , Dorling Jack , Deteix Patrice
Published: December 2022

Human blood L cells are a major subset of non-T, non-B (null) cells that lack easily detectable C3 receptors, but have high affinity Fc receptors for IgG. L cells have been purified by rosetting and immunoadherence procedures and characterized by immunofluorescence, histochemical and electron microscopic techniques. Purified L cells do not react with anti-monkey thymus serum (HuTLA) and most of them do not form rosettes with sheep or mouse erythrocytes even after the lymphocytes are treated with neuraminidase. Two major subsets of L cells are described. One has low density Ia-like surface antigens and this subset can be distinguished from B cells that have high density Ia-like surface markers. The other subset reacts with anti-myeloid serum, a reagent that characteristically stains an antigen whose density progressively increases with myeloid lineage maturation. L cells do not react with peroxidase or α-naphthyl esterase. They have ultrastructural features of lymphocytes although a deep cleft in the nucleus gives some a ‘monocytoid” appearance. These findings raise the possibility that some or most L cells are neither typical lymphocytes nor monocytes but are members of a separate hematopoietic lineage that occurpy an intermediate position in phylogeny and ontogeny.
Phagocytic Properties of Isolated Human Eosinophils
Authors: Grover William H. , Winkler Herbert H. , Normansell David E.
Published: December 2022

Viable eosinophils from normal human peripheral blood were separated by sedimentation through a nutrient-enriched linear gradient of sodium hypaque. Ingestion experiments with these cells showed that eosinophils would phagocytose latex beads coated with a mixture of allergen and corresponding allergic human serum. The stimulatory activity of the allergic serum correlated with the IgG fraction and not with those properties associated with IgE. Eosinophils isolated from allergic donors were found to be inferior in their phagocytic ability when compared with eosinophils from normal cell donors.
Immunity to Experimental Salmonella Infection: Studies on the Protective Capacity and Immunogenicity of Lipopolysaccharide, Acetone-Killed Cells, and Ribosome-Rich Extracts of Salmonella Typhimurium in C3H/HeJ and CD-1 Mice
Authors: Eisenstein Toby K. , Angerman Carl R.
Published: December 2022

Phenol-water-extracted lipopolysaccharide of Salmonella typhimurium, strain W118-2, was tested for protective capacity and immunogenicity in C3H/HeJ and CD-1 mice 21 days post vaccination. It was found that in C3H/HeJ mice the LPS was not protective and gave no anti-O titer, as measured by passive hemagglutination, whereas in CD-1 mice it was protective and immunogenic. In contrast, acetone-killed cells and a ribosomal vaccine prepared from the same strain of Salmonella were protective and resulted in an anti-O titer in both the C3H/HeJ and the CD-1 mice. It was shown, therefore, that LPS co-extracted with ribosomes, or present in whole cells, is more immunogenic for C3H/HeJ mice than phenol-water purified LPS. These observations imply that ribosome-rich extracts and acetone-killed cells contain substances, in addition to LPS, that modulate the immune response in C3H/HeJ mice. It was also observed that ribosomal vaccine prepared from a rough strain of S. typhimurium, strain TA1659, that lacks O antigens and makes LPS of the Rc type, did not protect C3H/HeJ or CD-1 mice against Salmonella infection. Mixtures of LPS with ribosomal vaccine derived from this mutant were also tested for their protective capacity. The results are discussed with regard to the antigens involved in immunity to Salmonella infection.
Immunochemical Characterization of Native Polysaccharides from Group B Streptococcus: The Relationship of the Type III and Group B Determinants
Authors: Kasper Dennis L. , Goroff Diana K. , Baker Carol J.
Published: December 2022

Three polysaccharides were purified from a type III strain of Group B Streptococcus grown under conditions varying in exposure to acid by washing the cells with an EDTA buffer or extracting with cold 10% TCA. Extraction of cells grown in a medium buffered with a pH titrator resulted in a polysaccharide antigen of 1.2 × 106 daltons having only type III serologic specificity. The antigen extracted with EDTA from cells grown in a medium where acid accumulated had a molecular size of approximately 600,000, and demonstrated both type III and group B serologic activity. The antigen extracted with TCA had a molecular size of approximately 500,000 and also reacted with type III and group B sera. Chemical studies revealed that all three polysaccharides contained varying concentrations of galactose, glucose, glucosamine, and sialic acid. Only the TCA antigen contained rhamnose. Alcohol fractionation of the TCA antigen allowed separation of type III from group B serologic reactivity. However, this method failed to separate the two determinants of the EDTA-non-pH titrated antigen. Other physical, chemical, and immunologic methods similarly failed to separate these two determinants. Mild hydrolysis of the EDTA-pH-titrated antigen exposed a determinant immunologically identical to the formamide-extracted group B antigen. These studies as well as electron microscopy and affinity chromatography demonstrated that two determinants exist on the native polysaccharide antigen of type III, Group B Streptococcus. One is the superficial capsular type specific determinant, which in the most native state masks the second determinant, a common group B determinant.

Quantitative precipitation inhibition studies confirmed that l-rhamnose was the immunodominant sugar for the formamide-extracted B antigen. No monosaccharide inhibited the cross-reactive determinant on the EDTA-non pH titrated antigen. Despite this observation, both the formamide-extracted B antigen and the EDTA-non pH titrated antigen were demonstrated to be precipitating antibodies of identical specificity.
Nature of the Antigenic Complex Recognized by T Lymphocytes
Authors: Thomas David W. , Shevach Ethan M.
Published: December 2022

In this paper we examined the effect of anti-TNP antibody on guinea pig T cell proliferation in response to TNP-modified macrophages in vitro. The addition of anti-TNP to TNP-modified macrophages immediately after conjugation inhibited their ability to stimulate TNP-specific T cell proliferation. This inhibition appeared to be specific for the TNP response since anti-TNP had no effect on the ability of TNP-modified macrophages pulsed with either PPD or TNP-Ova to stimulate efficient PPD or Ova T cell responses. On the other hand, anti-TNP had no effect on the TNP-specific response to TNP-modified macrophages that had been cultured overnight before addition to primed T cells or to macrophages which had been pulsed with TNP-Ova. We also demonstrated that the same TNP-specific T cell subpopulation responds to both freshly TNP-modified macrophages and overnight cultured TNP-modified macrophages. These results suggest that the relevant TNP-determinants recognized by T cells are not exposed on the macrophage surface and raise the possibility that macrophages must process membrane-conjugated TNP to create the immunogen recognized by T cells.
Opinions
Authors: North Robert J.
Published: December 2022

The term “activated macrophage” was introduced into the literature and employed extensively by Mackaness in the 1960s to describe the enhanced microbicidal activity of macrophages from animals with acquired immunity to infection with facultative, intracellular bacterial parasites. The notion that macrophages need to undergo functional modification in order to express resistance to infection with certain bacteria can be traced back to the writings of Metchnikoff at the turn of the century (1). It was subsequent studies of tuberculosis more than of any other infectious disease, however, that provided the evidence for the concept of a mechanism of cellular immunity based on the capacity of macrophages to acquire and to intrinsically express increased microbicidal mechanisms. The knowledge a) that a major visible aspect of the host response to tuberculosis consisted of the accumulation of macrophages at infective foci to form tubercles, b) that the tubercle bacillus is invariably engaged by and harbored inside macrophages, c) that secondary infection results in an accelerated rate of tubercle formation and in the destruction of the tubercle bacilli therein, and d) that all of these events proceed apparently without the participation of humoral antibody were sound reasons for postulating a mechanism of cellular immunity based on the intrinsic microbicidal powers of the macrophage.
Specificity of Lymphocyte-Mediated Cytotoxicity in Patients with Primary Intracranial Tumors
Authors: Levy Nelson L.
Published: December 2022

The specificity of a lymphocyte-mediated in vitro cytotoxic reaction was examined in a series of patients with primary intracranial tumors. Specific tumor-directed LMC was observed in 85% (35/41) of patients with gliomas and in 96% (24/25) of patients with nonglial tumors (meningiomas and acoustic neurilomas). The tumor specificity of these reactions was affirmed by testing the lymphocytes of each patient and the controls against nine to 12 different target cells. These target cells, each from an early passage culture, included both neoplastic and normal cells from the patient himself, as well as allogeneic tumor cells of types both related and unrelated to the patient's tumor. The specificity was further affirmed by showing that the LMC could be abolished by prior absorption of the lymphocytes on appropriate cell monolayers.

The direct assays, in concert with the absorption studies, showed that, in glioma patients, the LMC was directed against two different determinants. One was found on cells from all glial tumors, regardless of their degree of anaplasia, and was called the common glioma antigen (CGA). The other was expressed on anaplastic gliomas, melanomas, and fetal glial cells, but not on well-differentiated gliomas, normal adult glial cells, fetal fibroblasts, or other tumors. This distribution of the second antigen was believed due to the common origin of melanocytes and glial cells from the ectodermal cells of the embryonic neural tube, and it was thus called the glioembryonic antigen (GEA).