bluuebunny/crossref_metadata_2025 huggingface dataset:
Bacterial lipopolysaccharide (endotoxin) interferes with the induction of tolerance and primes thymus-derived lymphocytes.
Authors: Parks D E , Walker S M , Weigle W O
Published: December 2022

The injection of deaggregated human gamma-globulin (DHGG) into mice results in the establishment of specific immunologic unresponsiveness in T and B lymphocytes. However, the additional injection of bacterial lipopolysaccharide (LPS) 3 hr later converts the tolerogenic DHGG into an immunogen for both helper T and B cells. This generation of immunocompetence to HGG during interference with tolerance induction by LPS is demonstrated by both a primary plaque-forming cell response to DHGG + LPS and a secondary antibody response to a subsequent antigenic challenge. The secondary response to antigen stimulation in the B cells is, however, T cell dependent. Interference with tolerance induction in antigen-specific helper T cells can be demonstrated by challenge with the DNP-HGG hapten-carrier conjugate or by adoptive cell transfer with primed B cells and subsequent challenge with immunogenic HGG. Furthermore, helper T cells in the spleen and thymus become primed by DHGG + LPS as evidenced by their cooperation with primed B cells in the adoptive transfer assay. In addition to its interference with tolerance induction, LPS injected with either tolerogenic or immunogenic HGG also facilitates priming of helper T cells for the response to DNP-HGG. These data indicate that exposure of antigen-specific helper T and B cells to antigen alone results in the induction of tolerance, whereas induction of immunocompetence in these cells requires at least 2 signals, 1 provided by interaction with antigen and a 2nd delivered by a nonspecific stimulus similar to that provided by LPS in the present experiments.
Antigens of Ambrosia elatior (short ragweed) pollen. I. Crossed immunoelectrophoretic analyses.
Authors: Løwenstein H , Marsh D G
Published: December 2022

A crude extract of short ragweed pollen was obtained by extraction, centrifugation, dialysis and freeze-drying. Crossed immunoelectrophoresis, with a high-titer purified rabbit antibody fraction, revealed that the extract contained at least 52 antigens, of which 33 migrated toward the anode, 16 toward the cathode, and 3 toward both the anode and cathode. The electrophoretic precipitation pattern indicated a great deal of electrophoretic heterogeneity in several of the antigens. The overall distribution of apparent m.w. for 35 of the 52 antigens and the approximate m.w. for 19 identified antigens were determined by a combination of gel filtration and immunochemical analysis. Among antigens with m.w. weights less than or equal to 10,000, 8 of 10 were basic; on the other hand, all 5 identified antigens with m.w. above 40,000 were acidic. The apparent pI distribution for 30 antigens, and more precise pI values for 20 of 52 antigens, were determined by means of sucrose-gradient isoelectric focusing. The data obtained provide a reproducible reference system of considerable use in ragweed allergen chemistry and enable one to predict conditions for isolation of ragweed antigens.
Genetic influences on the augmentation of natural killer (NK) cells during murine cytomegalovirus infection: correlation with patterns of resistance.
Authors: Bancroft G J , Shellam G R , Chalmer J E
Published: December 2022

Cytotoxic activity attributable to natural killer (NK) cells was augmented in the spleens of mice infected with murine cytomegalovirus (MCMV). Stimulation was observed as early as 10 hr after virus administration but was dependent upon the dose used, and in particular, the host genotype. Effector cells were capable of killing a range of tumor cell targets as well as normal syngeneic thymocytes, and were induced in nude (nu/nu) and heterozygous (nu/+) mice. A significant correlation existed between resistance to the lethal effects of MCMV infection and the degree of NK cell augmentation by this virus in 10 of 11 strains examined. Non-H-2 related differences in activity were observed, but a clear association between high NK cell responses and the possession of a particular H-2 haplotype was not apparent. Beige mutant C57BL/6J mice, previously reported as defective in NK cell function were susceptible to MCMV and expressed low levels of cytotoxicity during infection. Thus, the genetically controlled stimulation of NK cells in the early stages of infection with MCMV may contribute to the genotype related patterns of resistance seen with this virus.
Induction of IgG production in a human monoclonal B lymphoblastoid cell line by a B cell-specific monoclonal antibody.
Authors: Miki Y , Kishi H , Muraguchi A , Maruyama S , Kishimoto S , Yamamura Y , Wang C Y , Kishimoto T
Published: December 2022

IgG production was induced in a human B blastoid cell line, CESS, by stimulation with a human B cell-specific monoclonal antibody (IIIA4). The number of IgG-producing cells reached its maximal level within 48 hr after incubation with IIIA4. IgG-producing cells induced by an optimal concentration of the antibody was about one-half as many as those induced with TRF, but higher concentrations of the antibody were rather inhibitory. Simultaneous addition of an optimal concentration of the antibody with TRF partially inhibited TRF-induced IgG production in CESS cells, but higher concentrations of the antibody did not block TRF-induced IgG production. TRF activity was completely absorbed with IIIA4-coated CESS cells as well as nontreated CESS cells. These results suggested that the membrane antigen recognized by IIIA4 was not the acceptor molecule for TRF. IIIA4 did not show any inhibitory effect on PWM-induced proliferation of normal B cells, but completely inhibited PWM-induced Ig production of normal B cells.
Regeneration of T cell subpopulations after bone marrow transplantation: cytomegalovirus infection and lymphoid subset imbalance.
Authors: Schroff R W , Gale R P , Fahey J L
Published: December 2022

Immune reconstitution was studied serially by using T lymphocyte cell surface differentiation antigens in 37 individuals receiving bone marrow transplants. Antigen expression was assessed by immunofluorescence analysis using monoclonal antibodies to T lymphocytes including Leu-3, which defines a T lymphocyte subpopulation in healthy individuals with T helper or inducer activity (L3+), and Leu-2, which defines a T lymphocyte subpopulation in healthy individuals with T helper or inducer activity (L3+), and Leu-2, which defines a T lymphocyte subpopulation with suppressor or cytotoxic activity (L2+). These studies demonstrated that the L2+ subpopulation regenerated more rapidly after transplant than did the L3+ subpopulation. Imbalances between these two T lymphocyte subpopulations, indicated by a decreased L3/L2 ratio, persisted for periods up to 12 mo post-transplant. Expression of a cell surface antigen associated with immature lymphocytes (OKT-10), and of HLA-DR (Ia-like) antigens was markedly increased during the post-transplant period. HLA-DR antigen expression did not appear related to immune activation in that increased reactivity was not detected with a monoclonal antibody (anti-TAC) specific for activated T cells. These observations in bone marrow transplant recipients and other disorders characterized by lymphoid restoration or immaturity indicate that inversion of the normal L3/L2 ratio and increased expression of OKT-10 and HLA-DR antigens may be features of a regenerating immune system. Furthermore, serial observation of individual patients indicated that infection with cytomegalovirus was associated with a progressive decrease in the L3/L2 ratio.
Abnormal natural killer cell activity in systemic lupus erythematosus: an intrinsic defect in the lytic event.
Authors: Katz P , Zaytoun A M , Lee J H , Panush R S , Longley S
Published: December 2022

The natural killer (NK) cell activity in fifteen systemic lupus erythematosus (SLE) patients was investigated by employing 51-chromium- (51Cr) release microcytotoxicity and single cell cytotoxicity assays against K562 target cells. Although the SLE patients as a group had depressed NK function in the 51Cr-release assay compared to normal subjects (p less than 0.005), those with clinically active disease displayed the greatest impairment in this activity (p less than 0.001). Active SLE patients were deficient in overall NK activity (Vmax) (p less than 0.005) but had normal percentages of potentially cytotoxic target binding cells (TBC). These TBC, however, were unable to normally kill bound target cells (p less than 0.01), which is indicative of a deficiency of "active" NK cells (p less than 0.005). Those NK cells with intact cytotoxic capabilities could "recycle" and repeat the lytic sequence normally. Exposure of normal lymphocytes to SLE sera did not impair any phase of NK function. These studies indicate that defective NK activity in SLE is secondary to an abnormality in the lytic event itself and is not due to a deficiency of NK cells, an abnormality in target binding, or an inability of NK cells to lyse multiple targets. Additionally, serum factors do not appear to play a major etiologic role in the cytotoxic abnormalities of these patients.
Cyclosporin receptors on human lymphocytes.
Authors: Ryffel B , Götz U , Heuberger B
Published: December 2022

Utilizing (3H) cyclosporin C (3H CS-C), a dihydroderivative of cyclosporin A (CS-A), an assay for cyclosporin receptors on human peripheral blood lymphocytes was developed. The specific binding of (3H) CS-C was saturable, time-dependent, and reversible. A Kd of 1.2 x 10(-7) M and a maximum binding capacity (Bmax) of 7 pmol/10(6) cells was calculated from equilibrium binding studies. A Scatchard analysis confirmed a single population of high affinity binding sites and about 7 x 10(5) sites/cell. Kinetic analysis of specific binding at 37 degrees C yielded a pseudo-first order rate association constant of 1.1 x 10(6) M-1 min-1 and a zero order dissociation rate constant of 0.19 min-1. The Kd 1.7 x 10(-7) M calculated from kinetic studies agreed well with the value of 1.2 x 10(-7) M determined in equilibrium binding studies. Regarding the specificity of binding, (3H) CS-C binding to lymphocytes was inhibited with CS-A and CS-C; however, no inhibition with the biologically inert keto-CS-A was observed. Mitogens and various growth factors did not compete for the binding site when added simultaneously with the radiolabel. However, pretreatment with T cell mitogens such as PHA, Con A, or the OKT3 antibody reduced (3H) CS-C binding significantly. These results suggest that the binding site for CS-A is closely associated with the mitogenic receptor on lymphocytes.
Cord IgM antibody specific for human killer T cells: T lymphocytotoxic human fetal antibody (TLFA) recognizing maternal killer T cells proliferating in the presence of interleukin 2.
Authors: Miyagawa Y , Komiyama A , Akabane T , Uehara Y , Yano A
Published: December 2022

T lymphocytotoxic human fetal antibody- (TLFA) containing cord IgM (TLFA IgM) was purified from cord blood. The proliferation of maternal mixed lymphocyte culture-(MLC) induced and long-term cultured T cells (LCT) specific to the paternal non-T cells in the presence of interleukin 2 (IL 2) was inhibited by treatment with the respective TLFA IgM plus complement (C). Sixty-six to 78% of Leu-2a-positive maternal LCT were stained with the respective TLFA IgM, as determined by anti-Leu antibodies and TLFA IgM double-staining. Moreover, TLFA IgM plus C treatment reduced the killer activity of the respective maternal LCT against the PHA-induced paternal target cells. Therefore, TLFA-positive killer T cells of maternal LCT belong to certain killer T cell subsets of Leu-2a-positive pan-killer T cells, and TLFA contains a potential antibody against maternal killer T cells.
Dissection of immunoregulatory subpopulations of T lymphocytes within the helper and suppressor sublineages in man.
Authors: Gatenby P A , Kansas G S , Xian C Y , Evans R L , Engleman E G
Published: December 2022

The helper/inducer (Leu-3) and suppressor/cytotoxic (Leu-2) sublineages of human peripheral blood T cells can both the subdivided into functionally distinct subsets with anti-Leu-8, a new monoclonal antibody that identifies 75 +/- 10% of the Leu-3 cells and 60 +/- 10% of the Leu-2 cells. Using the autologous MLR as a T cell-dependent stimulus for immunoglobulin synthesis, we have shown that the major helper effect for antibody formation lies within the numerically minor Leu-3,8- subset. In addition, neither Leu-2,8- cells alone suppress Leu-3-induced immunoglobulin synthesis, but in combination these subsets are markedly inhibitory. These results indicate that at least two phenotypically distinct cell types of suppressor lineage interact to produce suppression of an immune response in man.
A monoclonal antibody specific for the Thomsen-Friedenreich cryptic T antigen.
Authors: Rahman A F , Longenecker B M
Published: December 2022

A monoclonal antibody (49H.24) is described that was made after immunization of BALB/c mice with human neuraminidase-treated erythrocytes (NE-RBC). 49H.24 is an IgM and reacts with NE-RBC but not untreated, normal human RBC. As little as 100 pg of antibody protein produced detectable direct agglutination of, or binding to, NE-RBC. The fine specificity of 49H.24 was determined by using a series of synthetic sugar haptens as inhibitors of agglutination or binding of 49H.24 to NE-RBC. Only synthetic T hapten (beta Gal(1 leads to 3)alpha GalNAc) produced complete inhibition of agglutination or binding, and no inhibition was produced by several other closely related haptens. Synthetic T hapten-coated silica beads (Synsorb) were used to affinity purify 49H.24. Affinity-purified antibody was radiolabeled with 125I and used in a sensitive competition assay to detect natural T antigen associated with cell membranes or in soluble form. The potential use of this or similar monoclonal antibodies as probes for an important human tumor marker is discussed.
A rapid simple method of basophil purification by density centrifugation on Percoll.
Authors: Raghuprasad P K
Published: December 2022

This study describes simple and relatively rapid methods of purifying basophils from human peripheral blood. Our initial studies defined the densities of basophils in several donors and helped develop strategies for selectively concentrating basophils by isopyknic centrifugation over Percoll. We layered anticoagulated blood over Percoll of appropriate density, centrifuged these for 10 min, and collected the cells held in Percoll separately from the overlying band of mononuclear cells. Most of the basophils were distributed between densities of 1.072 and 1.078 g/ml, with a peak at approximately 1.074 to 1.076 g/ml. Basophil purity of 24.4 +/- 4% (range 5.6 to 50%) was achieved in a Percoll gradient of 1.076 g/ml due to both the increased numbers of basophils and the low numbers of mononuclear cells at this gradient. The cells isolated by this method are viable, appear morphologically normal, and release histamine to various stimuli in a normal manner. A second centrifugation of the cells obtained from this density over appropriate discontinuous gradients of Percoll leads to further enhancement of basophil purity. The main limitation of this technique appears to be the relatively low yield (percent of recovery) of the basophil (less than or equal to 20% for the two-step procedures).
Experimental murine hypersensitivity pneumonitis: multigenic control and influence by genes within the I-B subregion of the H-2 complex.
Authors: Wilson B D , Sternick J L , Yoshizawa Y , Katzenstein A L , Moore V L
Published: December 2022

An animal model of hypersensitivity pneumonitis was developed in the mouse by immunization with pigeon dropping extract (PDE) in complete Freund's adjuvant, followed by daily aerosol challenge with soluble specific antigen. C3H/He (H-2k) mice developed intense diffuse interstitial lung disease and were classified as high responders. On the other hand C57BL/10 (H-2b) mice developed only mild perivascular infiltration. Moreover, although high-responder C3H/He mice developed delayed hypersensitivity to PDE, low-responder C57BL/10 mice failed to develop this reactivity. Breeding studies in F1 and F2 progeny of C3H/He high responders and C57BL/10 low responders showed that responsiveness was multigenic. B10.BR (H-2k) mice also developed intense PDE-induced diffuse interstitial lung disease, suggesting that genes within the H-2 complex influence the development of pulmonary inflammation. On the other hand, C3H.SW (H-2b) mice were also high responders, possibly because of non-H-2 genes in the C3H/He background that influenced responsiveness. Studies in H-2 recombinant strains with C57BL/10 backgrounds mapped the H-2-linked genes that influence responsiveness within the I-B subregion.
Schistosoma mansoni surface antigen defined by a rat monoclonal IgG2a.
Authors: Dissous C , Grzych J M , Capron A
Published: December 2022

A schistosomula surface antigen of 38,000 daltons has been isolated by using a rat monoclonal IgG2a that has been shown to confer protection against S. mansoni infection by passive transfer in rats. This antigen is one of the previously characterized surface proteins reacting with sera from various infected hosts, including rat, mouse, monkey, and human. Studies on parasites of different developmental stages indicated the presence of this antigen on cercariae and skin-derived or mechanically transformed schistosomula.
Tumor-specific antigen of murine T-lymphoma defined with monoclonal antibody.
Authors: Allison J P , McIntyre B W , Bloch D
Published: December 2022

A panel of hybridomas was constructed by fusion of P3X63Ag8 myeloma cells with spleen cells from a BALB/c mouse that had been immunized with a C57BL/Ka x-ray-induced lymphoma, C6XL. One of forty-three hybridomas secreting antibodies reactive with the tumor cells was found to be unreactive with normal spleen cells in a radioimmunometric assay. This antibody, designated 124-40, was unreactive with normal adult thymus, spleen, lymph node, or bone marrow cells, or with fetal spleen or thymus cells in radioimmunometric or radioimmunoprecipitation assays. Flow microfluorometric analysis of these nonmalignant lymphoid cells failed to reveal subpopulations reactive with MAb 124-40. The antibody was highly specific for the lymphoma cells used for immunization and did not react with a panel of other spontaneous or x-ray-induced or chemically induced lymphomas. The antigen reactive with MAb 124-40 was isolated by radioimmunoprecipitation and found to be a glycoprotein composed of disulfide-bonded subunits of 39,000 m.w. and 41,000 m.w. A cell surface component of similar structure, but not reactive with MAb 124-40 could be detected by two-dimensional electrophoresis in extracts of purified T cells, but not B cells. These results suggest that the apparently individually specific lymphoma antigen reactive with MAb 124-40 might be a clonally expressed epitope carried by a T cell surface component.
The Effect of Mechanical Obstruction of the Hepatic Veins Upon the Outflow of Lymph from the Thoracic Duct
Authors: Simonds J. P. , Brandes W. W.
Published: December 2022

Simple mechanical obstruction of the hepatic veins in otherwise normal dogs increases the outflow of lymph from the thoracic duct from 2.4 to 6.2 times, with an average increase of 5.2 times, the normal. This increase in the outflow of lymph is almost identical with the average increase (5.45 times) observed by Petersen, Jaffé, Levinson and Hughs in anaphylactic shock, but was much less than that recorded by them in peptone shock. Since it has been previously shown by us that little or no blood escapes from the liver during the height of anaphylactic and peptone shock it is suggested (a) that the engorgement of the liver due to obstruction to outflow of blood may be an important factor in the increased output of lymph from the thoracic duct in these two types of shock; (b) that this may be the chief cause in anaphylactic shock; and (c) that there is an additional factor, probably increased permeability of the sinusoidal endothelium, in peptone shock. It is further suggested that the substance produced when the homologous serum passes through the liver of a sensitized dog is not entirely identical in its action with peptone. It at least seems to lack the powerful direct lymphogogic effect of peptone.
Production of Staphylococcus Antitoxin in Horses
Authors: Parker Julia T. , Banzhaf Edwin J.
Published: December 2022

It has been shown by one of us (1, 2) that most strains of Staphylococcus aureus from pathogenic sources produce a poison which in 0.1 cc. amounts causes marked necrosis when injected into the skin of rabbits. At the time of writing these papers we were not aware that von Lingelsheim (3) as early as 1900 described experiments in which he inoculated subcutaneously 8 to 10 cc. of sterile filtrates of broth cultures of Staphylococcus aureus and obtained marked inflammatory changes and sometimes necrosis. More recently, Neisser and Wechsberg (4) confirmed these findings with smaller injections (0.2 to 0.3 cc.) of toxic filtrates. These observers found as we did that this necrotizing poison was destroyed when heated to 58° for one hour.

On further analysis, we showed this poison to be an exotoxin as the serum of rabbits immunized thereto neutralized it in multiple quantities.
Studies in Hypersensitiveness
Authors: Brown Aaron
Published: December 2022

Thirty-one late hay fever subjects were tested with extracts of high and low ragweed pollens, and each one was found to be equally sensitive to the two extracts. One group of these individuals were given the usual series of prophylactic injections of one of the extracts, and the remainder were similarly treated with the other extract. After a maximal tolerance had been established to the one extract, an equivalent or larger quantity of the other extract was injected. This change from one extract to the other was not followed in any case by a constitutional reaction, which must have been expected if the atopen of the two pollens were not identical.
Studies on the Serologic Classification of B. Botulinus
Authors: Gunnison J. B. , Schoenholz P.
Published: December 2022

The results of the complement fixation tests fully confirm those obtained by Schoenholz and Meyer (1) by means of the agglutination reaction. This study has given no indication of the need for the creation of new groups for any further revision of the classification. By means of these reactions, eleven of the inagglutinable strains have been classified, and seven strains have been shown to be ultraspecific. No strain which had been definitely assigned to a group has mutated to another in the course of three to four years of artificial cultivation. This would indicate that the serologicic groups possess a marked degree of permanence. In view of these results, the final serologic classification may be stated as follows. There are seven groups of B. botulinus types A and B. The type A strains are divided into three groups, of which the third is composed of ultraspecific strains. The type B strains fall into three specific groups and the seventh group includes the ultraspecific strains of this type.
Antigenic Properties and Virulence Among Mammalian Tubercle Bacilli
Authors: Mudd Stuart , Fürth Jacob
Published: December 2022

The human and bovine varieties of Mycobacterium tuberculosis, although possessing characteristic differences in pathogenic and cultural characters, have been shown to be antigenically similar. Nevertheless it has seemed of interest to select certain human and bovine strains which in respect to virulence are atypical and subject these to serological study by a greater diversity of method than has been brought to bear on this problem heretofore.

The antigenic properties of five strains of mammalian tubercle bacilli described as avirulent are in this communication compared with those of a human strain of low virulence, H37, and a bovine strain of high virulence, Bovine C. It is found that strains differing profoundly in virulence may show similar serological behavior even when tested with the aid of sensitive adsorption methods. Thus the avirulent strains B.C.G. and R1, and two cultures of H37 were not differentiated antigenically from the virulent strain Bovine C; on the other hand, the same methods served to differentiate Bovine III and the saprophytic “Koch” strain from the virulent organism.
Observations on the Skin Reaction Induced in Rabbits, Guinea Pigs and Goats by the Intracutaneous Injection of Dysentery Toxin
Authors: Jungeblut C. W.
Published: December 2022

Introduction. In 1923 Brokman (1) described an intracutaneous reaction in man occurring in a certain percentage of cases after the intradermal injection of a small amount of dysentery toxin (Shiga). A reaction of similar character had been noted before by Major and Nobel (2) in 1913. Brokman's later work, in which Przesmycki (3, 4) participated, proved that those individuals who gave a positive reaction had no specific antitoxin in their blood, while an appreciable amount could be detected in the serum of those who reacted negatively to the intracutaneous injection of the dysentery toxin. The relative frequency of positive reactions at different ages was subsequently studied by Brokman and Popowski (5). The findings of Brokman and his associates have only recently been confirmed by Zoeller and Lesbre (6). These authors, for the first time, injected also laboratory animals intracutaneously with dysentery toxin.
High frequency and nonrandom distribution of alloreactivity in T cell clones selected for recognition of foreign antigen in association with self class II molecules.
Authors: Ashwell J D , Chen C , Schwartz R H
Published: December 2022

Previous studies have demonstrated that a single T cell clone can respond to both a foreign antigen in the context of self major histocompatibility complex (MHC)-encoded molecules (self plus X) and to an allogeneic class I or class II molecule in the absence of antigen (non-self). We have used limiting dilution of T cells obtained from the draining lymph nodes of antigen-primed B10.A mice to establish a large number of T cell clones that recognize either GAT, pigeon cytochrome c, or sheep insulin in association with syngeneic antigen-presenting cells. Sixty-two antigen-specific T cell clones were assayed for their ability to proliferate in response to a panel of nine different allogeneic haplotypes. Of these, 38 (61%) responded to at least one allogeneic haplotype, and 15 of the 38 (39%) responded to more than one allogeneic stimulator. In addition, the patterns of alloreactivity varied with the immunizing antigen. The GAT-specific T cells had at least one responder to every haplotype tested, although H-2u-responsive T cell clones were the most common. In contrast, no pigeon cytochrome c-specific T cells responded to stimulators of the H-2u haplotype, but rather predominantly responded to H-2t4/H-2s and H-2i5/H-2b. Finally, sheep insulin-reactive T cell clones preferentially responded to H-2u stimulators, although stimulation by antigen-presenting cells of the H-2p and H-2q haplotypes was also common. A chi 2 analysis of the data demonstrated that the dependence of the pattern of alloreactivity observed upon the antigen used for immunization was statistically significant (p less than 0.01). The high frequency of alloreactivity found in antigen-specific T cell clones is discussed, as well as the implications that the antigen-dependent skewing of the distribution of alloreactivity have for a one-receptor model vs a two-receptor model of T cell recognition.
Requirements for activation of contrasuppressor T cells by type III pneumococcal polysaccharide.
Authors: Braley-Mullen H
Published: December 2022

Either S3-coupled spleen cells (S3-SC) or soluble S3 activates two populations of regulatory T cells, T suppressor cells (Ts) and contrasuppressor T cells (Tcs). The latter cells function to mask the activity of Ts in unfractionated T cell populations, so that Ts can be detected only after removal of Tcs. Activation of Tcs by S3 may be required for induction of an antibody response to S3. This is suggested by the findings that Tcs are activated only by immunogenic doses of S3, that Tcs are not detectable in the spleens of mice tolerant to S3, and that (CBA/N X BALB/c)F1 male (xid) mice, which are genetically unresponsive to S3, do not develop Tcs after immunization with S3. Moreover, the kinetics of activation of Tcs by S3 closely parallels the kinetics of the antibody response to S3. Tcs have no detectable activity in the absence of Ts, indicating that these cells do not function as amplifier or helper T cells.
Supernatant from a cloned helper T cell stimulates most small resting B cells to undergo increased I-A expression, blastogenesis, and progression through cell cycle.
Authors: Leclercq L , Cambier J C , Mishal Z , Julius M H , Theze J
Published: December 2022

Helper T cell clone 52.3 supernatant (52.3 SN) was previously shown to be able to stimulate gradient-purified murine resting B cells in the absence of any additional stimulus. However, the proportion of cells that were accounting for the thymidine uptake and the Ig production was unknown. In this paper, we have studied induced changes that can be measured at the single cell level, and have thus determined the frequency of resting B cells that respond to 52.3 SN. Results indicate that 52.3 SN induces an increased I-A expression and a cell size enlargement on virtually all resting B cells. A significant proportion (30%) of these cells later becomes large blasts. Acridine orange staining revealed that in the presence of 52.3 SN a large fraction of the resting B cells undergoes the G0 to G1 transition. Furthermore, 52.3 SN is able to induce at least 20% of the cells to continue through the cell cycle into S phase as indicated by propidium iodide staining of DNA. Finally, a fraction of the 52.3 SN-stimulated cells differentiate to Ig-producing cells. Our present results suggest that resting B cells express functional receptors for some lymphokines and that these lymphokines can act in the absence of membrane Ig occupancy. Our findings further support the existence of a B cell-activating factor acting in a MHC-unrestricted manner and responsible for the entry of resting B cells into cell cycle. The relationship between this factor and other lymphokines is discussed.
Activation of cation transport by lymphokines in B cells without induction of DNA synthesis or immunoglobulin gene transcription.
Authors: Fotedar R , Severini A , Holowachuk E W , Shiozawa C S , Paetkau V , Kaplan J G , Diener E
Published: December 2022

We report on an experimental model that permitted us to evaluate the biologic relevance of membrane-associated biochemical events with respect to cell proliferation and maturation, each induced by distinct sets of signals. Antigen-affinity-enriched murine B cells cultured in the presence of a proliferative signal induced by LPS showed activation of Na+/K+ ATPase and enhanced the uptake of proline, followed by RNA, protein, and DNA synthesis, without the generation of antibody. Stimulation with both the proliferative signal(s) and the maturation signal(s) derived from lymphokines of an EL-4 thymoma induced B cells to proliferate and synthesize mRNA encoding mu-chain of IgM and to mature into IgM-secreting cells. Most important, the secretory product of EL-4, in the absence of LPS, activated Na+/K+ ATPase but failed to stimulate uptake of proline and synthesis of DNA or mu-specific mRNA. A similar response was observed in splenocytes depleted of T cells and in unfractionated spleen cells. Thus a component secreted by EL-4 can induce some of the early molecular events characteristic of the proliferative response but lacks the ability to initiate blast transformation and DNA synthesis.
Modulation of mouse anti-SRBC antibody responses by placental extracts. II. Antigen specificity and regulatory role of B and T cell populations affected by two distinct placental fractions.
Authors: Bobé P , Kanellopoulos-Langevin C , Bleux C , Voisin G A
Published: December 2022

Previous results from our group had shown that when CBA mice are primed to sheep red blood cells (SRBC) in the presence of various doses of placental extract (PE) (or liver extract [LE] as control), their spleen cells injected into normal syngeneic recipients have important immunoregulatory properties. Low doses of PE (0.25 to 4 mg per mouse) induce a marked decrease of the PFC response against SRBC in recipient animals. In contrast, higher doses of PE (8 to 13 mg per mouse) have a potentiating effect on the same response. LE is not different from a saline injection, at any dose. The suppressive and enhancing effects can be reproduced with two distinct placental fractions (PF) of 40 KD and 60 KD, respectively. In the present report, we have studied the requirement for an antigenic stimulation at the same time as the injection of PE, and the antigenic specificity of the subsequent immunoregulatory effects. In addition, we have analyzed the functional properties of the spleen cell populations affected by PE or placental fractions: surface Ig- cells mediate the suppressive effect due to low doses of PE or the 40-KD fraction, whereas surface Ig+ cells are responsible for the enhancing effect due to high doses of PE or the 60-KD fraction. These immunoregulatory activities do not appear to be related to the presence of Ig fragments in PF, because our results have shown that no Ig fragments can be detected in either PF. Surface Ig- T cell populations from spleen cells treated with the 40-KD fraction and antigen have been further separated into Lyt-2- and Lyt-2+ subpopulations. Our results showed that Lyt-2+ cells alone suppress the PFC response anti-SRBC in both normal and irradiated syngeneic recipients. Thus, the injection of a 40-KD PF in the presence of antigen activates splenic T suppressor cells capable of specifically regulating a secondary antibody response in vivo.
A phosphatidylinositol kinase in rat mast cell granules.
Authors: Kurosawa M , Parker C W
Published: December 2022

Intact granules were isolated from sonicated purified rat serosal mast cells on a Percoll gradient. The granules were shown to contain a highly active phosphatidylinositol kinase that catalyzes the formation of diphosphoinositide from endogenous phosphatidylinositol in the granule membrane. The enzyme requires ATP and Mg2+ or Mn2+ for activity; Ca2+, fluoride and cyclic AMP are inhibitory. The Km for ATP is 25 microM. The initial reaction is rapid, but the response ceases within a few minutes. A comparison of the rate of phosphorylation of intact and broken membrane granules suggests that the phosphorylation occurs on the outer (cytoplasmic) surface of the granules.
Association of human lambda light chain V/J/C segments: serologic analysis and primary structure of the lambda VI Bence Jones protein THO.
Authors: Ghiso J , Solomon A , Frangione B
Published: December 2022

The complete amino acid sequence of the human monoclonal lambda VI light chain Bence Jones protein THO was determined. We have found it to have remarkable similarities to the previously sequenced lambda VI Bence Jones protein SUT. Immunochemical analyses demonstrated that both lambda VI chains belong to a V lambda VI sub-subgroup. The 98-residue V gene-encoded segments of proteins THO and SUT are closely homologous and are distinguished from other lambda VI chains by a one-residue deletion at the V-J recombination site. Proteins THO and SUT have identical 13-residue J segments and therefore are encoded by the same J lambda gene. Further, both proteins have identical 105-residue C regions that by sequence represent products of the C lambda 3 (Kern-, Oz+) gene. The primary structure and serologic properties of proteins THO and SUT imply at the protein level of association between certain types of V lambda, J lambda, and C lambda segments.
Changes in thymocyte proliferation at different stages of viral leukemogenesis in AKR mice.
Authors: O'Donnell P V , Traganos F
Published: December 2022

Proliferation in total populations of thymocytes from control AKR mice or AKR mice injected intrathymically with MCF 69L1 virus was measured by flow cytometry of acridine orange-stained cells. Cell sorting experiments showed that the majority subpopulations of small cortical and medullary thymocytes in control mice were noncycling and were predominantly in the Go phase of the cell cycle. Of the 15 to 20% cycling thymic lymphoblasts, approximately 50% were in the G1 phase, 35% were in the S phase, and 15% were in the G2 + M phases of the cell cycle. Cycling cells appeared to consist of a major subpopulation with low RNA content and a minority subpopulation with high RNA content. In virus-injected mice, no changes in cell cycling were observed at stage I of leukemogenesis (30 to 40 days postinjection), at which time infection of thymocytes by MCF virus is maximum and constant but no clonality is evident. Thus, MCF virus infection of thymocytes per se does not appear to alter cell proliferation. Increased cell cycling and a shift in cell cycle distribution to more cells in G1 was observed at stage II of leukemogenesis (50 to 60 days postinjection), at which time a clonally expanded cell population is known to emerge in thymuses of injected mice. Acridine orange staining resolved these novel cycling cells from subpopulations of normal thymic lymphoblasts on the basis of intermediate RNA content. The transition from stage II to stage III (50 to 60 days postinjection) was accompanied by the outgrowth of a major cycling population with a distinct, often increased, RNA content. As a result, the residual "normal" background of cycling cells often observed in stage II was markedly reduced or completely absent by stage III. Populations of cycling blasts from mice with frank leukemia differed from those at stage III by a variability in mean RNA content and in cell cycle distribution indicative of individual tumor heterogeneity. In addition, thymomas often contained multiple populations of cycling blasts that could be resolved by their discrete RNA distributions. Simultaneous staining of DNA and RNA by acridine orange appears particularly well-suited for studying a heterogeneous population of cycling and noncycling cells represented by mouse thymus. This method has permitted a rapid and quantitative analysis of cell cycle parameters at progressive stages of viral leukemogenesis in AKR mice.
Immune interferon modulation of in vitro murine anti-human T cell monoclonal antibody-mediated cytotoxicity.
Authors: Dillman R O , Johnson D E , Shawler D L
Published: December 2022

Limited antitumor effects have been achieved in clinical trials with murine monoclonal antibody T101, perhaps because of its limited ability to effect complement-mediated or cell-mediated cytotoxicity. We explored the effects of recombinant immune interferon on T101-mediated cytotoxicity in vitro. Interferon failed to enhance expression of the antigen detected by T101 on target cells, but it did increase Fc receptor binding of T101 and other IgG2A and IgG3 murine proteins, but not IgG1 or IgG2B. Preincubation of U937, HL60, and human mononuclear cells with 100 U of immune interferon for 48 hr, while T101 was preincubated with various T cell line targets or human CLL cells at 4 degrees C for 30 min before combining effectors and targets for 4 hr at 37 degrees C, resulted in cytotoxicity of 18 to 44% of maximum. Cytotoxicity in the absence of interferon or T101 was less than 5%. Unfortunately, rapid modulation of antigen-antibody when T101 was preincubated with targets at 37 degrees C prevented any increase in cytotoxicity under those conditions. We conclude that immune interferon can augment T101-mediated cytotoxicity in vitro, but it is unlikely that it would enhance T101-mediated cytotoxicity via complement or cell-mediated mechanisms in vivo.
Split tolerance induced by chick embryo thymic epithelium allografted to embryonic recipients.
Authors: Houssaint E , Torano A , Ivanyi J
Published: December 2022

To test the capacity of the epithelial component of the chick embryo thymus to induce tolerance to major histocompatibility complex (MHC) antigens, pre-colonized thymic rudiments were grafted into chick embryonic recipients. Semi-allogeneic or allogeneic transplantations were done between two lines of chickens histocompatible at the MHC locus. Approximately 10% of these thymic chimeras hatched and were studied 3 mo after hatching. Thymic grafts were not rejected by the allogeneic host. The tolerance of chimeric chickens to thymus donor MHC antigens was tested by using a skin graft rejection test and a graft-vs-host (GvH) assay. Chimeric chickens that received an MHC-incompatible thymic graft during the embryonic life tolerated skin graft with the MHC haplotype of the thymus donor. Nevertheless, the lymphocytes within the thymic graft, the host thymus, and the blood were tolerant to the host MHC antigens but were alloreactive in GvH reaction for the MHC antigens of the thymic graft type. These results suggest that the epithelial component of the thymus when taken before the starting of the colonization by hemopoietic precursors and grafted into an early chick embryonic host can induce a tolerance for the MHC determinants involved in allograft rejection but not in the GvH reaction.
Growth regulation of transformed T cells by nonactivated macrophages: the role of Ia expression.
Authors: Kedar I , Rosenberg Y J , Steinberg A D
Published: December 2022

We studied the influence of unactivated mouse peritoneal macrophages on the proliferative capacity of a spontaneously transformed MRL-lpr/lpr T cell clone. Macrophages, 25%, induced a reduction in proliferative rate from 20% to 95% measured by [3H]thymidine incorporation and microscopic cytometry. MHC-compatible (H-2k) macrophages caused growth inhibition reciprocal to the amount of Ia expression on the macrophage. Thus, with increasing preculture of the macrophages there was both decreasing Ia and increasing suppression. H-2-incompatible macrophages had maximal inhibitory capacity without preincubation. Macrophages derived from the peritoneum of MRL-lpr/lpr mice were less suppressive than macrophages from other H-2k mice. In contrast to the case of activated macrophages in other studies, in the present system there was no killing of T cells, only reduction in proliferation. The inhibitory effect of the macrophages correlated with the spontaneous formation of rosettes between the macrophages and the T cell clone. The number of rosettes forming a single layer of T cells around the macrophages, but not the number of rosettes with multiple layers of cells, was reciprocally related to the amount of Ia expression. The results suggest that macrophages bear a surface structure that influences and modulates the growth of T cells.
Selective phagocytosis of gram-positive bacteria and interleukin 1-like factor production by a subpopulation of large granular lymphocytes.
Authors: Abo T , Sugawara S , Amenomori A , Itoh H , Rikiishi H , Moro I , Kumagai K
Published: December 2022

There has been a consensus that a large granular lymphocyte (LGL) population with natural killer (NK) function is nonadherent and nonphagocytic. However, a significant proportion of the nonadherent cells purified by the two-step depletion of adherent cells with a plastic surface and nylon wool columns engulfed Sta. aureus into their cytoplasm. These cells were morphologically identified as LGL in light and electron microscopies. Two-color immunofluorescence tests, furthermore, demonstrated that Leu-11+ LGL, Leu-11+7-, and Leu-11+7+, but not Leu-11-7+, phagocytosed Sta. aureus. Among the particles tested here, only Gram(+) bacteria were preferentially phagocytosed, whereas Gram(-) bacteria, other large-sized microbes (e.g., baker's yeast and Candida albicans), latex, silica, and carbonyl iron were not. LGL exhibited a substantial level of bactericidal activity against Sta. aureus, although the level was one third of that mediated by monocytes. When Gram(+) bacteria were incubated with nonadherent cells for 18 hr, significant amounts of interleukin 1 (IL 1)-like factors (or IL 1 itself) as well as interferon were detected in the supernatants. On the other hand, this incubation did not induce interleukin 2 (IL 2). The IL 1-like factor producer cells were demonstrated to be the low-density lymphocytes on Percoll separation and to have the Leu-11+ phenotype. The phagocytosis was suggested to be an important stimulus in producing IL 1-like factors from LGL. Thus, the treatment of cells with cytochalasin B, a microfilament disrupting agent, completely abrogated both phagocytosis and IL 1-like factor production. Some cell wall components of Gram(+) bacteria might be important to a recognition process of the phagocytosis, since the protoplasts of Sta. aureus, when prepared by the treatment of bacteria with lysostaphin, were no longer phagocytosed by LGL. The present results therefore identify an additional unique characteristic similar to, but not identical with, the myelomonocytic nature of Leu-11+ LGL.
Phenotype, frequency, and EBV responsiveness of human marrow B and pre-B cells.
Authors: Hibi T , Chan M A , Petsche D , Dosch H M
Published: December 2022

We have purified subpopulations of B lineage cells from human adult (rib) bone marrow by cell sorting and panning. Limiting dilution analysis was then used for a clonal analysis of cells able to secrete IgG, IgA, or IgM spontaneously or after infection with EBV. Nonproliferating, high rate IgG or IgA producers occurred at frequencies of about one per 1000 marrow mononuclear cells. Their frequency and Ig production was unaffected by EBV, and they appeared not to express EBNA after exposure to EBV. These cells were Ia+, B1+, and over 85% expressed sIg of the IgM/D (up to 75%) and/or IgG/A isotypes (40 to 60%). B cells committed to the secretion of IgM represent 2 to 10% of marrow B lymphocytes. They were found to be Ia+/B1+/B2+/CALLA- and C3b receptor (CR3)-cells, and most (greater than 90%) required infection with EBV and proliferation to develop into IgM-producing lymphocytes. Thirty to 40% of these cells did not express Ig (H or L chain) on their surface, and therefore resembled pre-B cells at the beginning of the 4- to 5-wk culture period. Proliferating pre-B cells from adult human marrow have been described, but their conversion into IgM-producing cells has not been formally demonstrated. Although EBV induces IgM production, the expression of EBNA, and several rounds of cell division in these cells, the induction of stable (greater than 5 wk) growth transformation represents a rare event in these pre-B cells: in several thousand limiting dilution wells, not a single culture of sIg-cells showed stable growth transformation. The dichotomy between EBV-induced high-rate IgM responses and absent growth transformation discriminates activation and transformation as distinct aspects of EBV-induced B cell "responses", and suggests that cellular properties play critical roles for viral transformation. We propose a model in which cellular target genes for transforming sequences in the EBV genome are transiently expressed during B cell differentiation.