bluuebunny/crossref_metadata_2025 huggingface dataset:
Stimulation of Mitogenic Responses in Human Peripheral Blood Lymphocytes by Lipopolysaccharide: Serum and T Helper Cell Requirements
Authors: Miller Richard A. , Gartner Suzanne , Kaplan Henry S.
Published: December 2022

Stimulation of human peripheral blood lymphocytes by lipopolysaccharide (LPS) was studied by the incorporation of 3H-thymidine. Peak stimulation occurred at 7 to 9 days over a broad range of LPS concentrations. Both Escherichia coli and Salmonella typhimurium LPS were effective mitogens with S. typhimurium having slightly higher activity. There was a strict serum requirement; pooled fresh frozen human serum was found to best support stimulation. In fetal calf serum, LPS caused a reduction in culture-induced stimulation. Cell separation procedures were employed in order to study the nature of the responding cell population. It was found that only non-T cells were stimulated by LPS, but in order for maximal stimulation to occur there was a requirement for helper T cells.
T Cell Regulation of Specific B Cell Responses
Authors: Campos-Neto Antonio , Levine Herbert , Schlossman Stuart F.
Published: December 2022

The role of T cells in the generation of specific antibodies to antigens under Ir gene control was investigated in strain 2 guinea pigs. The specificity of anti-α,Dnp-Lys9 antibody was studied in guinea pigs with intact immune responses and in strain 2 guinea pigs deprived of T cells by adult thymectomy, lethal irradiation, and reconstitution with syngeneic bone marrow cells. Conventional strain 2 guinea pigs developed both T cell responses and highly specific anti-α,Dnp-Lys9 antibody. In contrast, strain 2 “B” guinea pigs failed to develop delayed hypersensitivity and in vitro T cell responses. Nevertheless, these “B” guinea pigs could produce anti-Dnp antibody of restricted heterogeneity and in quantities comparable to those produced by the conventional strain 2 guinea pigs. It was shown that anti-α,Dnp-Lys9 antibody produced in strain 2 “B” guinea pigs lacking specific T cell responses could not distinguish α,Dnp-Lys9 from dinitrophenol. Similar antibodies are produced in strain 13 guinea pigs that are poly-L-lysine, Ir gene negative. Reconstitution of strain 2 “B” guinea pigs with normal T cells enabled them to develop both cell-mediated immune responses and antibody of highly discriminatory power. These findings support the notion that in the absence of specific T cells, comparably specific B cell antibody responses cannot be generated, and perhaps more importantly, that specific T cells are required to facilitate the amplification of unique B cell clones.
Natural Cell-Mediated Cytotoxicity in Guinea Pigs: Properties and Specificity of Natural Killer Cells
Authors: Altman Amnon , Rapp Herbert J.
Published: December 2022

Lymphoid cells of guinea pigs were found to be naturally cytotoxic against a variety of target cells. Natural killer (NK) cell activity was high in peripheral blood and spleen, low in lymph nodes and peritoneal exudates, and absent in the thymus. Significant activity was found between the ages of 3 and 40 weeks, and strain 2 guinea pigs had a higher NK activity than strain 13 guinea pigs. Peripheral blood NK cells were sensitive to treatment with ammonium chloride solution, whereas splenic NK cells were resistant to the same treatment. The NK cells were found to be nonadherent to nylon wool and plastic and were enriched among cells bearing receptors for the Fc portion of IgG. The NK activity of guinea pig lymphocytes was selective in that only some, but not all, of the target cells used were susceptible to the cytotoxic effector cells. Susceptible target cells included syngeneic, allogeneic, and xenogeneic tumor cell lines, especially the human myeloid cell line K562, and some passages of primary syngeneic fibroblast cultures. A positive correlation was found between the direct susceptibility of different targets to NK cells and their ability to inhibit competitively the killing of K562 target cells. The NK cells in guinea pigs appear to be similar to human NK cells in several aspects and may, therefore, serve as a useful model for studying certain aspects of human NK activity.
Differences in Kinetic Parameters of Lysis by Syngeneic and Allogeneic T Lymphocytes Immune to the Same Tumor
Authors: Huber S. A. , Lucas Z. J.
Published: December 2022

Kinetic parameters of cell-mediated cytotoxicity (CMC) to the 13762A rat mammary adenocarcinoma were studied by using immune syngeneic (Fischer) and allogeneic (Wistar Furth) spleen cells. Treating the spleen cells with rabbit anti-T cell serum and complement eliminated cytotoxicity. Both types of cytolytic cells bound to tumor cells in Ca2+-free medium but required Ca2+ to lyse them. Cytochalasin A did not interfere with the lysis of target cells already bound to cytolytic cells but it did prevent attachment of cytolytic cells to new targets. Cytolysis of the tumor occurred in three sequential steps: 1) binding of the lymphocytes to the target; 2) production of a lytic lesion in the target by the lymphocyte; and 3) lysis of the target. Allogeneic and in vitro augmented syngeneic lymphocytes required about 30 min for maximum binding to targets compared to 4 hr for syngeneic cells sensitized in vivo. Induction of the lytic lesion required 1 hr for the allogeneic cells and up to 8 hr for both types of sensitized syngeneic cells. Target cell death was slightly faster after incubation with allogeneic (8 hr) than with syngeneic (12 hr) cells. Pretreatment of the tumor cells with mitomycin C or actinomycin D did not enhance target lysis. Despite the many similarities between allogeneic and syngeneic effector cells, the increased time required for induction of the lytic lesion by syngeneic effector cells might indicate employment of a different lytic mechanism or a lower lytic efficiency compared to allogeneic cells.
Enhanced Intracellular cGMP Concentrations and Lectin-Induced Lymphocyte Transformation
Authors: Atkinson John P. , Kelly James P. , Weiss Avery , Wedner H. James , Parker Charles W.
Published: December 2022

On the basis of recent reports that sodium azide, ascorbic acid, nitrosamines, and nitrosoguanidines elevated cGMP levels in some tissues, we examined their effects on cGMP metabolism in human lymphocytes. Sodium nitroprusside (NP), a nitrosoguanidine (MNNG), sodium azide, and ascorbic acid augmented intracellular cGMP concentrations many-fold in the presence or absence of calcium in the extracellular milieu. The response was usually maximal by 30 sec, persisted for at least 1 hr, and occurred in platelet-, granulocyte-, and monocyte-free preparations composed of greater than 90% T lymphocytes. In broken cell preparations these same agents augmented guanylate cyclase activity but had no effect on cAMP or cGMP phosphodiesterase. The magnitude of the increase in cyclase activity was in most cases proportional to the enhancement produced in intact cells, although sodium azide was a better stimulator of guanylate cyclase in broken cell preparations, and ascorbic acid in intact cells. With these agents as positive controls and four separate methods of cyclic nucleotide extraction before radioimmunoassay, significant and reproducible increases in cGMP were not demonstrable with Con A or E-PHA over a wide concentration range. The cGMP agonists did not in themselves produce lymphocyte mitogenesis or modulate E-PHA or Con A-induced transformation at optimal mitogenic concentrations. At supraoptimal concentrations of E-PHA or Con A (>25 µg/ml), NP increased thymidine incorporation and cell number. Cyclic GMP and its analogs alone neither enhanced thymidine incorporation nor modulated mitogen-induced lectin stimulation except for cGMP itself (1 and 0.1 mM) which increased 3H-thymidine transport but not cell number. These data do not support the concept of cGMP being a critical second messenger or having a permissive, positive regulatory role in lectin-induced transformation in human lymphocytes.
Binding Constants of Dextrans and Isomaltose Oligosaccharides to Dextran-Specific Myeloma Proteins Determined by Affinity Electrophoresis
Authors: Takeo Kazusuke , Kabat Elvin A.
Published: December 2022

In polyacrylamide electrophoresis, mobilities of dextran-specific myeloma proteins W3129 and QUPC52 were retarded when dextrans were added to the separating gel. Retardation by dextran was reversed by adding isomaltose oligosaccharides to the gel. From measurements of the extent of retardation by dextran or of reversal by hapten as a function of concentration, association constants of dextrans (Ka) and of oligosaccharides (Kia) were determined.

With W3129, NRRL dextrans B 742 FR.S, B 742 FR.L, N279, B 1355 FR.S, B 1399, B 1299 FR.S, and NRC fractions 6, 7, and 8 have Kα values ranging from 2 to 6 × 104 ml/g. With QUPC52, dextrans in general show lower Ka than with W3129; N279, NRC fractions 6, 7, and 8, and B 1399 have Ka values ranging from 1 to 7 × 104 ml/g, whereas B-742 FR.S, B-1299 FR.S, and B-1355 FR.S have Ka values ranging from 3 to 6 × 102 ml/g. Dextran D3, a completely linear α(1 → 6)-linked glucosyl chain having a m.w. of 36,500 has a relative high affinity for QUPC52 (Ka = 2.3 × 103 ml/g or 8.3 × 104 M-1), although it does not show any affinity for W3129. This result confirms that myeloma protein W3129 is specific for a terminal chain of α(1 → 6)-linked glucosyl residues, whereas QUPC52 reacts with the nonterminal portion of α(1 → 6)-linked glucosyl chains. Kia values obtained coincided with those by other methods reported earlier by Cisar et al. With W3129, affinity increases with chain length up to isomaltopentaose (IM5) with the highest affinity Kia = 6.7 × 104 M-1; Kia of isomaltohexaose (IM6) and isomaltoheptaose (IM7) being 6.3 × 104 and 2.3 × 104 ml/g, respectively. With QUPC52, affinity increases as the number of residues in the glucosyl chain increases, reaching an upper limit with IM6 and IM7, 1.2 and 1.4 × 104 M-1.

This method of affinity electrophoresis is useful for obtaining binding constants ranging from 102 to 106 M-1. It may be performed on antibodies, lectins, and other binding proteins by using not only the purified proteins but also crude protein mixtures such as the ascitic fluid used in this study.
Differential Protective Effects of Immune Lymphoid Cells against Transplanted Line Ib Leukemia and Immune Polioencephalomyelitis
Authors: Duffey Paul S. , Lukasewycz Omelan A. , Olson David S. , Murphy W. H.
Published: December 2022

The capacity of immune cells obtained from the major lymphoid compartments to protect C58 mice from transplanted line Ib leukemia, and from an age-dependent autoimmune CNS disease (immune polioencephalomyelitis = IPE) elicited by immunizing old C58 mice with inactivated Ib cells was quantified. Cells used for comparative adoptive protection tests were harvested from the major lymphoid compartments 14 to 15 days after young C58 mice were immunized with inactivated Ib cell preparations. Regression curves were plotted from survival data and the log10PD50 values were determined. Immune spleen (ISC) and peritoneal cells (IPEC) were significantly more protective against transplanted Ib cells than immune lymph node (ILNC), thymic (ITC), and marrow cells (IMC). In contrast, IPEC and IMC were not protective against IPE and ITC were only marginally protective. ILNC afforded significant protection to transplantable leukemia but were only marginally protective to IPE. When ISC were treated with anti-thy 1.2 serum and complement, protection against transplanted leukemia and IPE was reduced > 99%. When donors of immune lymphoid cells were treated with 12.5 mg of cortisone acetate daily for 2 days before lymphoid cells were harvested, protection against transplanted Ib cells by ISC was reduced by approximately 90% whereas protection against IPE was totally eliminated. Considered together, these results indicate that the protective mechanisms to transplantable leukemia and IPE differ significantly in the same indicator mouse strain.
Activation of Human B Lymphocytes
Authors: Dimitriu Alexandra , Fauci Anthony S.
Published: December 2022

Selective effects of in vitro azathioprine (AZ) on subpopulations of human lymphocytes involved in the pokeweed mitogen (PWM)-induced plaque forming cell (PFC) response against sheep red blood cells (SRBC) have been described. First, it was found that the responding B cell itself was very sensitive to AZ as concentrations as low as 0.01 µg/ml markedly suppressed the PWM-induced PFC responses of unfractionated peripheral blood mononuclear (MN) cell suspensions. The fact that AZ suppresses the PFC response of B cells only when added at the beginning of the cultures suggests that the drug interferes with some early events involved in B cell triggering by PWM. Furthermore, it was demonstrated that higher concentrations of AZ, i.e., 1 and 10 µg/ml selectively eliminated certain regulatory subsets of cells known to act as suppressors of this polyclonally induced PFC response. Preincubation of a) T cell-enriched (TCE) suspensions of cells, b) concanavalin A (Con A)-generated suppressor cells, and c) naturally occurring suppressor cells from nonresponder individuals with 1 and 10 µg/ml of AZ before co-culturing them with fresh MN cells in the presence of PWM caused a differential suppression of suppressor cell function and a selection for a pure helper effect with a resulting increase in B cell responses. Finally, regulatory helper cells were quite resistant to the in vitro effects of this drug as they still expressed their helper activity after pretreatment with 10 µg/ml AZ. Thus, this study demonstrated that B cells are highly sensitive, suppressor cells are less sensitive, and helper T cells are quite resistant to the in vitro effects of AZ.
Antigen-Induced Murine B Cell Lymphomas
Authors: Haughton Geoffrey , Lanier Lewis L. , Babcock George F. , Lynes Michael A.
Published: December 2022

In the accompanying report, we have described the characterization of two unusual murine B cell lymphomas, CH1 and CH2. A heterologous antiserum, which we refer to as “anti-idiotype” serum, has been raised to the detergent-solubilized surface immunoglobulin of CH1. The following criteria have established that this antiserum is specific for the CH1 tumor and that it reacts with V region determinants of the tumor surface IgM: 1) the antiserum reacts with CH1 tumor cells, but not normal mouse lymphoid cells or CH2 tumor cells, in indirect immunofluorescence and C-dependent cytotoxicity testing, 2) capping with the anti-idiotype serum removes all or most of the tumor surface Ig, 3) the antiserum forms a single band of precipitation against serum from CH1 tumor-bearing mice, when tested by double diffusion precipitin analysis, and 4) a single band of precipitation is formed in the electrophoretic migration position of IgM when the anti-idiotype antiserum is tested against serum from CH1 tumor-bearing mice in immunoelectrophoresis. Furthermore, we have demonstrated that this antiserum is useful in monitoring tumor growth and is a potent immunotherapeutic agent. Specifically, 50% of mice injected with a lethal tumor inoculum and given a small dose of anti-idiotype serum 2 days later remain tumor free, whereas all tumor-challenged control mice died within 30 days.
Genetic Control of the Immune Response to the H-2Dk Private Specificity, H-2.32
Authors: Shinohara Nobukata , Hansen Ted H. , Schurko Elizabeth , McKenzie Ian F. C. , Sachs David H.
Published: December 2022

B10.AKM mice (H-2m) when immunized with H-2k cells showed very low cytotoxic antibody responses to the H-2Dk private specificity H-2.32, whereas AKR.M and (AKR.M × B10.AKM)F1 mice that possess the same H-2m haplotype mounted reasonable anti-H-2.32 antibody responses. The genetic nature of the non-H-2 linked gene(s) controlling the anti-H-2.32 response was analyzed on the backcross progeny raised between (AKR.M × B10.AKM)F1 and B10.AKM mice. The anti-H-2.32 antibody response was found to be predominantly controlled by a single locus. This locus segregated independently of the Ig heavy chain locus, the Ly2 locus, and the Mls locus. Despite the observed difference in antibody production, no significant differences between AKR.M and B10.AKM mice were detected in induction of H-2Dk-specific killer T cells. Thus, the defect in the response of B10.AKM mice to H-2.32 can be detected at the level of B cell function and is controlled by a single non-H-2-linked genetic locus, but is not attributable to genes linked to the major immunoglobulin structural genes nor to the Mls locus.
Allergy to Insect Stings
Authors: Sobotka Anne K. , Adkinson N. Franklin , Valentine Martin D. , Lichtenstein Lawrence M.
Published: December 2022

Radioallergosorbent tests (RAST(s)) have been developed and assessed for the diagnosis of insect hypersensitivity by using a purified allergen from honeybee venom, phospholipase A, and crude yellow jacket venom. Sera from 193 patients positive both by history and skin test to one of these insects were compared with various groups of control sera. Eighty percent of sera from skin test-positive patients were RAST positive; positive RAST were found in 16% of sera tested from skin test-negative patients. A highly positive RAST correlates well with a positive skin test and clinical sensitivity, but serum IgE is not measurable in many patients with mast cell or basophil bound antibody. Since biologically important reactions of antigen with IgE require that the antibody be cell bound, skin testing would be preferred to RAST if one were limited to a single test for the diagnosis of insect allergy.
Evidence for a Silent or Null Gene in Hereditary C2 Deficiency
Authors: Pariser Kenneth M. , Raum Donald , Berkman Eugene M. , Alper Chester A. , Agnello Vincent
Published: December 2022

Hereditary deficiency of the second component of complement (C) was the first human inherited C component deficiency state described (1). It also appears to be the most common C component deficiency thus far studied with a gene frequency of 1% (2, 3). The genetic basis for this deficiency is not as yet known. In contrast, the basis for the genetic deficiencies of C3 and C6, which occur much less frequently, have been established. These deficiencies have been shown to be due to inheritance of silent or null genes allelic with the structural genes (4, 5). The approach used in those studies was to identify families with the deficiency that had informative electrophoretic variants of the proteins. Study of the inheritance of the structural variants and the serum levels of C components in such families provided evidence for the silent or null gene mechanism.
Identity of C3- and C5-Receptors on Lymphoid Cells
Authors: Landen Beate , Dierich Manfred P.
Published: December 2022

Tannic acid-treated SRBC, incubated with increasing concentrations of C5 (Etan-C5) can be attached to C3 receptor-carrying (Raji) cells. This binding is dependent on the amount of C5 on Etan-C5 and can be inhibited by pretreatment of the Raji cells with either C5 or C3. Similar inhibition by soluble C3 and C5, respectively, is obtained for the interaction between Raji cells and Etan-C3. In addition, the immune adherence reaction between Ehum and EAC1423b could be blocked by previous treatment of Ehum with C5 or with C3. These results suggest the presence of binding sites for C5 on lymphoid cells and their identity with C3 receptors.
T Cell Helper Defect in Patients with Chronic Lymphocytic Leukemia
Authors: Chiorazzi N. , Fu S. M. , Montazeri Ghodrat , Kunkel H. G. , Rai Kanti , Gee T.
Published: December 2022

Purified peripheral blood T lymphocytes from normal donors were shown to help allogeneic tonsillar B cells to differentiate and secrete specific anti-SRBC antibody in vitro in a plaque-forming assay. Utilizing this system, a comparison was made between the allogeneic helper activity generated by the T cells of normal individuals and patients with various disease states. Allogeneic helper activity was absent when T lymphocytes from patients with CLL were used. Conversely, relatively normal allogeneic helper function was provided by T cells of patients with a variety of other disorders studied. Thus, a functional deficiency was identified in CLL patients in the subpopulation of regulatory T cells responsible for providing helper activity in allogeneic interactions.
Properties of Murine Leukemia-Associated Suppressor Cells
Authors: Grinwich Kazimiera D. , Alexander Thomas S. , Cerny Jan
Published: December 2022

The spleens of BALB/c mice bearing leukemia induced with Moloney leukemia virus contain cells capable of inhibiting the antibody response of normal immunocompetent lymphocytes in Marbrook culture vessels. The immunosuppression requires contact between the cells and appears to be mediated by leukemia-associated suppressor cells rather than by the leukemic cells themselves. Thymus-dependent antibody responses to sheep red blood cells (RBC), horse RBC or trinitrophenol coupled to ovalbumin (TNP-OVA) were inhibited proportionally to the number of suppressor cells added. In contrast, thymus-independent responses to Vibrio cholerae lipopolysaccharide and to TNP coupled to Ficoll were either unaffected or suppressed only with a high number of suppressor cells. The leukemia-associated suppressor cells from BALB/c mice (H-2d) inhibited the anti-sheep RBC response of syngeneic lymphocytes and of allogeneic DBA/2 lymphocytes bearing the H-2d haplotype but failed to inhibit lymphocytes from other allogeneic strains DBA/1 (H-2q), C57BR (H-2k), A (H-2a), and C57BL/10 (H-2b). Experiments with lymphocytes from congenic strains carrying various H-2 gene complexes on the B10 genetic background confirmed that the suppressor effect is restricted by genes in the H-2 complex. Further mapping suggested that the suppression requires identity in the K region between the suppressor and the responder cells. The inability to suppress the allogeneic, H-2 different lymphocytes was retained even when a majority of the leukemic, virus-producing cells were removed from the spleen cell suspension by using a specific cytotoxic antiserum. There was no correlation between permissivity of lymphocytes to infection by the Moloney leukemia virus in vitro and between their inhibition by the suppressor cells. Thus, the nonspecific (polyclonal) leukemia-associated suppressor cells preferentially inhibit helper T cells and the suppression involves a specific interaction with product(s) of K region genes on the surface of the responder cells.
Inhibition of Lymphocyte Blastogenesis by C3c and C3d
Authors: Schenkein Harvey A. , Genco Robert J.
Published: December 2022

The C3 cleavage products C3c and C3d were tested for their ability to alter the immunoproliferative response of human peripheral mononuclear cells to the antigens SLO and SK-SD, and to the mitogens PHA and PWM. It was found that both C3c (30 to 120 µg/ml) and C3d (10 to 40 µg/ml) inhibited lymphocyte blastogenesis in the presence on antigens but not mitogens, when cells were cultured in either autologous plasma or FCS. Similarly, the response to antigens of cell populations enriched for T lymphocytes was inhibited, whereas the response to optimal or suboptimal doses of mitogens was unaffected. When nonadherent (NA) cells were reconstituted with increasing numbers of adherent (AD) cells to potentiate the proliferative response of NA cells to the antigen SLO, the addition of either C3c or C3d abolished the potentiation of the response at low levels of reconstitution. However, at a given dose of C3c or C3d, addition of excess AD cells could restore the proliferative response. These results suggest that both C3c and C3d can inhibit T cell proliferation in response to antigen and that they may act at the level of the monocyte-T lymphocyte interaction to modulate cellular immune responses.
Immunoadjuvant Treatment of Primary Grafted and Spontaneous AKR-Leukemia
Authors: Olsson Lennart , Ebbesen Peter
Published: December 2022

AKR mice grafted with spontaneous AKR leukemia cells and treated with BCG may become leukemic due to graft leukemia or spontaneous leukemia, may die in a wasting-like syndrome, or may appear cured from leukemia and be without signs of wasting-like syndrome. Lymphoid spleen cells from these groups of mice were used at various times from cytotoxicity against AKR leukemia cells, against nonmalignant AKR fibroblasts, and against nonmalignant BALB/c fibroblasts. Spleen cells from cured mice and wasting-like mice were cytotoxic to AKR leukemia cells. The cytotoxic activity could be enhanced by depleting the lymphoid cells of T lymphocytes. AKR leukemia cytotoxic spleen cells were invariably also cytotoxic to AKR fibroblasts, but never to BALB/c fibroblasts. Quantitative differences in sensitivity to cytotoxic attack could be demonstrated between AKR leukemia cells and AKR fibroblasts since the former cell type was about 1000 times more sensitive to cytotoxic attack as compared to AKR-fibroblasts. The insensitivity of BALB/c fibroblasts to cytotoxic attack seemed to be a qualitative characteristic. Addition of lymphoid cells from cured AKR mice inhibited the cytotoxic activity of otherwise cytotoxic cells, thus indicating some suppressor activity in cured mice. It is suggested that successful anti-neoplastic immunoadjuvant treatment depends on a quantitative difference between malignant and nonmalignant cells with respect to cytotoxic attack of self-directed cytotoxic cells.
Activation of the Classical Pathway of Complement by the C3NeF-Stabilized Cell-Bound Amplification Convertase
Authors: Daha Mohamed R. , van Es Leendert A.
Published: December 2022

C3 nephritic factor (C3NeF̄) has been shown to be composed of two heavy and two light chains, like IgG; in addition it shares antigenic determinants with IgG. C3NeF, purified from the sera of eight patients by incorporation of C3NeF into the stabilized fluid phase amplification C3 convertase, C3b,Bb(C3NeF), followed by its release after decay of convertase function, was investigated for its ability to bind 125I-C1q and to activate 125I-C1. It was found that although fluid phase C3b,Bb(C3NeF) is fully capable of binding 125I-C1q, it is not able to activate 125I-C1 even at concentrations of 1.3 × 1012 C3b,Bb(C3NeF) complexes/ml. On the other hand, cell-bound C3b,Bb(C3NeF) is capable of both binding 125I-C1q and activating 125I-C1. This discrepancy between fluid phase and cell-bound C3b,Bb(C3NeF) was found for C3NeF preparations from eight different patients and therefore seems to apply to all C3NeF preparations.
Heavy Chain Variable Region Allotypic Subspecificities of Rabbit Immunoglobulins
Authors: Horng Wayne J. , Knight Katherine L. , Dray Sheldon
Published: December 2022

An a1a2 rabbit (P286-3), neonatally suppressed for the expression of the a1 allotype, was immunized with autologous a1 IgG at 2 months of age. Both auto anti-a1 Ab and a1 IgG molecules were found in the serum of this rabbit after the auto-immunization. The auto anti-a1 Ab and the IgG from the auto anti-a1 Ab-depleted serum were isolated. Of the previously defined a1-AB, a1-AC, and a1-AD Ig subpopulations, the a1 IgG in the IgG preparation from rabbit P286-3 were all of the a1-AB Ig subpopulation. The auto anti-a1 Ab from rabbit P286-3 did not react with the a1-A, a1-B, and a1-C allotypic subspecificities; thus, it was presumably specific for the a1-D allotypic subspecificity. Thus, the a1-AB Ig subpopulation escaped from allotype suppression in rabbit P286-3, whereas the a1-AC and a1-AD Ig subpopulations remained suppressed. The a1-AD Ig subpopulation will probably remain suppressed for a long time and perhaps permanently since rabbit P286-3 has produced circulating auto-Ab specific for the a1-D allotypic subspecificity. These results indicate that the a1 Ig subpopulations are synthesized by distinct clones of lymphocytes under separate control.
Some Quantitative Aspects of the Passive Sensitization of Human Basophils
Authors: Goldstein Byron , Dembo Micah , Malveaux Floyd J.
Published: December 2022

We review the theory for the binding of IgE to Fc receptors on basophil surfaces. We then use the theory to analyze binding experiments of Malveaux et al. and T. Ishizaka on the passive sensitization of basophils from a patient with chronic myelocytic leukemia and 75% basophilia. From their data we determine that the forward rate constant for the binding of human IgE to receptors on these human basophils is (3.0 ± 1.0) × 104 M-1 sec-1.
Intestinal Goblet Cell Mucus Release
Authors: Lake Alan M. , Bloch Kurt J. , Neutra Marian R. , Walker W. Allan
Published: December 2022

We previously reported that the infusion of certain soluble immune complexes stimulated mucus release from the rat small intestine in vivo. The present studies sought to evaluate the response of the intestine of normal and immunized rats to the infusion of antigen alone. One hour after the intraduodenal infusion of antigen, small intestinal washings were obtained and analyzed for the presence of 35S-labeled, high m.w. glycoprotein of goblet cell origin. The amount of goblet cell glycoprotein released was estimated from the radioactivity present in the void volume of a Sepharose 4B gel filtration column.

The release of goblet cell mucus was enhanced by antigen stimulation in orally immunized animals. The discharge of goblet cell mucus was not increased after antigen infusion in animals immunized by the i.p. route despite the induction of high levels of serum antibody. The inability to demonstrate release of mucus after antigen challenge in systemically immunized rats suggests that the amount or the type(s) of antibody required at the mucosal surface is produced only after oral immunization.
Activated T Cells in the Synovial Fluid of Arthritic Patients: Characterization and Comparison with in Vitro Activated Humand and Murine T Cells in Cooperation with Monocytes in Cytotoxicity
Authors: Galili Uri , Rosenthal Lili , Galili Naomi , Klein Eva
Published: December 2022

Lymphocytes were separated from the synovial fluid of 34 arthritic patients. The majority of lymphocytes rosetted with SRBC and were thus of T origin. A proportion of these cells were activated as indicated by their high 3H-thymidine incorporation in vitro, the formation of “stable” E rosettes, and their ability to attach to autologous monocytes and to human cells. Such attached activated T cells enhanced the cytotoxic activity of monocytes. Human and murine-activated T cells generated in mixed lymphocyte cultures also attached to cell lines of the same species, and after their attachment they enhanced the cytotoxic activity of monocytes and natural killer cells. It is suggested that one of the possible roles of activated T cells in immunologically damaged tissues is the attraction of circulating nonspecific killer cells to the site of the response.
Monocyte Dependence of Pokeweed Mitogen-Induced Differentiation of Immunoglobulin-Secreting Cells from Human Peripheral Blood Mononuclear Cells
Authors: Rosenberg Stuart A. , Lipsky Peter E.
Published: December 2022

Human peripheral blood mononuclear cells (PBM) lost the capacity to generate immunoglobulin-secreting cells (ISC) in response to pokeweed mitogen (PWM) when depleted of adherent cells (AC). The diminished responsiveness of the nonadherent cells (NAC) could not be ascribed to cell death, altered PWM dose response characteristics, or a change in the length of incubation required to generate a response. Supplementation with autologous or homologous AC, but not 2-mercaptoethanol, restored the capacity of NAC to generate ISC after PWM stimulation. By standard criteria AC were found to contain 85 to 90% monocytes. Furthermore, the monocytes and not the few lymphocytes contaminating the AC were responsible for restoring PWM responsiveness to the NAC. PWM-induced DNA synthesis of NAC also was markedly reduced compared to PBM. Again, supplementation with monocytes restored responsiveness to NAC. The monocyte dependence of PWM-induced proliferation and generation of ISC was most apparent when cultural conditions were employed that limited cell-to-cell interaction.
Lack of Responsiveness of C3H/HeJ Macrophages to Lipopolysaccharide: The Cellular Basis of LPS-Stimulated Metabolism
Authors: Ryan J. L. , Glode L. M. , Rosenstreich D. L.
Published: December 2022

The rate of glucose utilization has been used as a measure of LPS-induced activation of cultures of C3H/HeN and C3H/HeJ spleen cells, peritoneal cells, and purified peritoneal adherent cells. Peritoneal cells utilized 40 to 60 times more glucose than did spleen cells and purified adherent monolayers were more active than mixed peritoneal cells, suggesting that only macrophage metabolism was being measured. The cell preparations for C3H/HeJ mice were not activated by Escherichia coli K235 LPS prepared by extensive phenol extraction, whereas C3H/HeN cells were activated by that LPS. Cells from both strains were activated by a commercially obtained E. coli 0111:B4 LPS and butanol-extracted K235 LPS.

The addition of 10% C3H/HeN spleen cells to C3H/HeJ peritoneal cells resulted in a marked enhancement of glucose utilization. These findings suggest that LPS-induced enhancement of macrophage metabolism occurs both by direct action of LPS on macrophages as well as indirectly through activated lymphocytes.
T-T Cell Synergy in the in Vitro Generation of Secondary Cell-Mediated Cytotoxicity Against Syngeneic SV-40 Transformed Cells
Authors: Glaser Moshe
Published: December 2022

The concept of T-T cell interaction that was first suggested during cell-mediated immune response to alloantigens was evaluated in vitro in a syngeneic tumor system. Combinations of lymph node and thymus cells from mice immune against SV-40 transformed cells were shown to exhibit synergism with respect to in vitro generation of secondary cytotoxic effector cells against the corresponding target cells. That is, the activity exhibited by combinations of lymph node and thymus cells was greater than that given by equal numbers of the two cell types cultured separately. With respect to the kinetics and magnitude of effector cell generation, the response occurring in synergizing cultures closely resembled that of optimal numbers of lymph node cells. In addition, the antigen specificity of effector cells generated by synergizing cultures was similar to that of effectors derived from cultures containing optimal numbers of responding lymph node cells. Pretreatment of either lymph node or thymus cells with mitomycin C or x-ray irradiation completely abolished their synergistic interaction. The lymph node and thymus cell populations participating in synergy were found to be thymus dependent. since we have previously shown that similar T-T cell interaction occurs in rejection of a syngeneic SV-40 induced sarcoma in mice, it is proposed that the in vitro generation of secondary cytotoxicity provides an in vitro model for the study of in vivo thymus cell interactions in response to tumor-associated antigens. These results suggest that we may be dealing with the same or similar T cell populations described before as displaying synergy in response to alloantigens in the graft-vs-host, mixed lymphocyte, and cell-mediated cytotoxicity reactions.
Human T Lymphocyte Subpopulations: Studies of the Mechanism by Which T Cells Bearing Fc Receptors for IgG Suppress T-Dependent B Cell Differentiation Induced by Pokeweed Mitogen
Authors: Moretta Lorenzo , Mingari Maria Cristina , Moretta Alessandro , Cooper Max D.
Published: December 2022

The mechanism by which human T lymphocytes bearing receptors for IgG (TG cells) suppress B cell differentiation promoted by pokeweed mitogen (PWM) and helper TM cells was studied. In this model system, autologous TG cells were shown to suppress via soluble factors released without detectable cell proliferation. In order to release these factors, TG cells needed to interact with PWM in addition to their reaction with IgG immune complexes during the isolation procedure. Suppression occurred only when TG cells were added early to mixtures of helper T and B cells cultured in the presence of PWM. B lymphocytes appeared to be resistant to suppression under a variety of experimental conditions. B cells “preactivated” by helper cells and PWM differentiated into plasma cells without any further help and were resistant to suppression by TG cells. B lymphocytes re-isolated from 2-day cultures containing TM cells, TG cells, and PWM retained their capacity to differentiate into plasma cells when cultured with helper T cells and PWM. B cell differentiation induced by helper supernatants was resistant to activated TG cells or their soluble factors. When TM cells were preincubated with suppressor supernatants, their ability to induce B cell differentiation was reduced; in contrast, similar pretreatment of B cells had no effect on their differentiation. These results suggest that TG cells can suppress the induction of polyclonal B cell differentiation by acting indirectly on helper TM cells.
The Proliferative Response of Human Lymphocytes to Antigen Is Suppressed Preferentially by Lymphocytes Precultured with the same Antigen
Authors: Borkowsky William , Valentine Fred T.
Published: December 2022

Human blood lymphocytes activated in vitro with antigen to which the donor is reactive are capable of suppressing the secondary proliferative response of autochthonous fresh cells to antigen. Both antigen-specific and antigen-nonspecific suppression can be detected in each experiment. These suppressor cells act by decreasing the number of lymphocytes entering the proliferative response rather than by slowing or otherwise inhibiting ongoing proliferation. The suppressor cells must be added soon after fresh cells are stimulated with antigen to be effective, but the suppressor cells themselves need not proliferate to exert their effect. Suppressor cells are optimally effective when added in numbers equal to those of the responding population, but still exert a significant effect at one-eighth that number.
The Fate of IgE Bound to Rat Basophilic Leukemia Cells
Authors: Isersky Chaviva , Rivera John , Mims Sharon , Triche Timothy J.
Published: December 2022

The present study investigates the fate of the cell-bound IgE by using a well-characterized rat basophilic leukemia cell line and a purified IgE myeloma protein. Both histamine-releasing and nonreleasing cell lines were examined. In both cases, no evidence for cell-mediated IgE catabolism could be elicited. Both the dissociated IgE and the receptors remained intact for prolonged periods of time, as demonstrated by binding assays. Internalization and/or recycling of membrane-bound IgE could not be demonstrated by E. M. autoradiography. We found only limited time-dependent changes in accessibility to anti-IgE antibody, trypsin, or elution at low pH (2.9 to 3.1). A biphasic dissociation of cell-bound 125I-IgE during incubation in the presence of excess unlabeled IgE was reproducibly observed; the more slowly dissociated IgE was also less readily dissociated at pH 3.4. These studies lead us to conclude that, in vitro, IgE resides in a functional orientation on the surface of RBL-1 cells, for prolonged periods of time.
Detection and Characterization of Human Leukemia-Associated Antigens on Leukemic Cell Lines and Thymocytes
Authors: Yoshizaki Kazuyuki , Seon Ben K. , Barcos Maurice P. , Minowada Jun , Pressman David
Published: December 2022

The γG fractions of baboon and rabbit anti-MOLT-4 (a human leukemic T cell line) sera were extensively absorbed with glutaraldehyde-aggregated fetal calf serum and pools of normal human spleen cells and peripheral blood cells, both of which were derived from several different individuals. The spleen and peripheral blood cell preparations used contained normal T lymphocytes as well as B lymphocytes. The absorbed γG fractions were then further treated with several normal (nonmalignant) B cell lines that possessed HLA alloantigens known to be expressed on MOLT-4 cells. The resulting absorbed γG fractions of baboon (Abs-BαMOLT) and rabbit (Abs-RαMOLT) antisera were tested against thymocytes, leukemic T cell lines (MOLT-4, CCRF-CEM, CCRF-HSB-2, RPMI 8402) and a leukemic null cell line (NALM-1), various normal B cell lines, normal peripheral blood lymphocytes, normal spleen lymphocytes, and other control cells. Three different tests were used for both Abs-BαMOLT and Abs-RαMOLT: 1) A complement-dependent cytotoxicity test, 2) an 125I-antibody binding assay, and 3) a quantitative absorption-cytotoxicity test. Both the Abs-BαMOLT and Abs-RαMOLT that we used detected antigens common to the four leukemic T cell lines and to thymocytes and did not react with any of the control cells including normal T cells and cultured normal B cell lines. Abs-BαMOLT differed from Abs-RαMOLT in that only the former reacted with NALM-1, a leukemic null cell line, and only the latter reacted with an antigen(s) on MOLT-4 that was not detectable on thymocytes from certain donors. The Abs-RαMOLT was used for the isolation and partial characterization of the human thymus leukemia-associated antigens reported in the following paper.
Changes in Surface Immunoglobulin Isotypes on Purified Antigen-Binding Cells after Antigenic Stimulation
Authors: Kenny James J. , Kessler Steven W. , Ahmed Aftab , Ashman Robert F. , Scher Irwin
Published: December 2022

Antigen-binding cells (ABC) were isolated from 5-day immune and nonimmune mice via a two-step centrifugation procedure. Their surface immunoglobulins (sIg) were then stained with fluorescein-conjugated anti-Ig, anti-µ, and anti-γ-specific reagents and analyzed on the fluorescence-activated cell sorter (FACS), or labeled by lactoperoxidase-catalyzed surface iodination and examined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Nonimmune ABC, unfractionated and ABC-depleted lymphocytes had identical FACS profiles after anti-µ and anti-Ig staining. Surface radioiodination and SDS-PAGE analysis showed that these nonimmune ABC exhibited a greater amount of δ than µ on their surface with only a trace of γ. In contrast, immune ABC bore more µ than δ and 30 to 50% of these ABC also bore surface γ. Immune ABC also showed a reduced amount of total sIg compared to Fl-anti-Ig stained unfractionated or ABC-depleted lymphocytes.

Since the intensity of staining with Fl anti-µ did not change after immunization, the reduced total sIg and increased µ/δ on the immune ABC were attributable to loss of sIgD after antigenic stimulation of the ABC. This antigen-induced loss of sIgD could be simulated on non-immune ABC and on unfractionated spleen cells by proteolytic removal of the sIgD.
Slow Reacting Substance (SRS) from Ionophore A23187-Stimulated Peritoneal Mast Cells of the Normal Rat
Authors: Yecies Lewis D. , Wedner H. James , Johnson Stephen M. , Jakschik Barbara A. , Parker Charles W.
Published: December 2022

When rat peritoneal mast cells were exposed to the ionophore A23187, a principle was released that possessed the biologic properties of slow reacting substance (SRS) from various sources. The response was dose, time, and temperature dependent with no activity being demonstrated in unstimulated cells. Supporting evidence that the mast cell product was similar or identical to SRS obtained from other sources include: 1) appropriate differential bioassay profile, 2) resistance to lipolysis and proteolysis, 3) acid lability and base stability, 4) inactivation by limpet arylsulfatase, and 5) inhibition by low concentrations FPL 55712. These data demonstrate that the isolated rat peritoneal mast cell contains the biosynthetic capacity to produce a bioreactive substance with the properties of SRS.
C1q: Isolation from Human Serum in High Yield by Affinity Chromatography and Development of a Highly Sensitive Hemolytic Assay
Authors: Kolb William P. , Kolb Linda M. , Podack Eckhard R.
Published: December 2022

C1q, a subcomponent of the first component of complement, has been isolated from human serum in fully hemolytically active form by affinity column chromatography and gel filtration with Bio-Gel A-5M. The affinity column was prepared by covalent coupling of purified human IgG to CNBr-activated Sepharose 4B. Final yields of C1q ranged from 25 to 40% with 650- 890-fold purification based on recovery of hemolytic activity. The preparations were free of contaminating serum proteins as judged by SDS-polyacrylamide gel electrophoretic and immunochemical criteria. The final C1q preparations were also devoid of any demonstrable C1q-inhibitor activity.

A C1q-depleted reagent (C1qD) was obtained from the nonadsorbed protein containing fractions of the human IgG-Sepharose 4B affinity column and utilized in conjunction with sensitized sheep erythrocytes (EA) for the detection and quantitation of C1q hemolytic activity. Employing optimal quantities of C1qD in the hemolytic assay mixture, the highly purified C1q preparations contained 0.5 to 1 × 1013 effective molecules/mg and 0.5 to 1 × 1012 effective C1q molecules/ml of human serum. This assay would therefore reproducibly detect less than 1 ng of C1q hemolytic activity.
Orally Induced Tolerance
Authors: Dumont Allan E.
Published: December 2022

Mattingly and Waksman have recently reported clear-cut and convincing data demonstrating the formation of antigen-specific suppressor cells in gut-associated lymphoid tissue (GALT) after oral administration of antigen (1). To consider formation of suppressor cells in GALT as an independent process responsible for orally induced tolerance, however, is to ignore information specifically implicating the liver in the induction of this tolerance. Thus, the authors overlook the fact that direct intraportal injection of sheep erythrocytes (2), bovine γ-globulin (3), or picryl chloride (3) renders susceptible animals immunologically unresponsive to these materials, and that the tolerogenic effect of oral feeding of a simple chemical antigen is abolished by diverting portal flow from the liver (4).

The precise mechanism responsible for immunologic unresponsiveness after portal circulation of antigen to the liver is unknown but in several of the papers cited by Mattingly and Waksman, the following possibilities are considered: 1) the liver may sequester immunogenic fragments of antigen-antibody complexes and thus allow tolerogenic components to exert their effect (5); 2) intrahepatic portal lymphoid tissue may form IgA that joins with absorbed antigen to suppress immunologically active cells (6); 3) induction of suppressor cells may require access of antigen to liver (7).
Immunoprophylaxis of Respiratory Syncytial Virus Infection in the Infant Ferret
Authors: Suffin Stephen C. , Prince Gregory A. , Muck Katie B. , Porter David D.
Published: December 2022

Infant ferrets can be protected from respiratory syncytial virus challenge at 3 days of age by gestational infection of their mothers. Ferrets acquire their immunity to respiratory syncytial virus postpartum via immunizing products of lactation. The level of protection against viral replication correlates with the maternal serum neutralizing titer or a concomitant factor. Passive administration of adult ferret serum with a neutralizing titer of 1:1024 or greater, either i.p. or orally does not confer immunity. A nonantibody-mediated protective mechanism appears to play an important role in protecting the infant ferret from respiratory syncytial virus replication. Our findings allow the testing of the efficacy of future human vaccines before human clinical trial.
Isolation of Functional Subsets of Human Peripheral Blood Monocytes
Authors: Norris David A. , Morris Richard M. , Sanderson Richard J. , Kohler Peter F.
Published: December 2022

Monocytes were isolated by counterflow centrifugation of Ficoll-Hypaque separated peripheral blood mononuclear cells. The monocytes formed a bimodal volume distribution of “large” and “small” phagocytic esterase-positive, peroxidase-positive cells with peaks at 470 and 410 µ3, respectively. The large monocytes were predominately Fc receptor positive, and were able to lyse both sensitized human and chicken erythrocyte targets in ADCC assays, whereas the small monocytes were largely FcR negative and were inactive against sensitized human erythrocyte targets. However, ADCC against chicken erythrocyte targets was seen in some fractions containing small monocytes and was probably due to FcR+ lymphocytes (K cells) in those fractions. These experiments establish that monocytes are effectors of ADCC against both human and chicken erythrocyte targets and that the peripheral blood monocyte is heterogeneous in size, function, and surface receptor distribution.