bluuebunny/crossref_metadata_2025_split huggingface dataset:
Observations on the Number of Immunoglobulin-Bearing Lymphocytes in Human Peripheral Blood with the Mixed Antiglobulin-Rosetting Reaction and Direct Immunofluorescence
Authors: Haegert David G. 
Published: December 2022

Abstract
Human peripheral blood lymphocytes were examined for the presence of membrane-associated immunoglobulin (Ig) by using direct immunofluorescence (DIF) and the mixed antiglobulin-rosetting reaction (MARR); to compare the two tests identical fluorescein-conjugated antiglobulin reagents were used both for fluorescent staining and for the rosetting reaction. The MARR was unaffected by incubation of lymphocytes in serum-free medium at 37°C or in acetate buffer (pH 4.0) before the actual rosetting procedure, whereas these conditions of preincubation reduced the number of fluorescent staining lymphocytes. Further, the MARR (in contrast to DIF) gave essentially the same results whether the fluorescent polyvalent antiglobulin reagent was IgG or a derived F(ab′)2 preparation. Evidence is presented that indicates that unlike DIF, the MARR demonstrates only membrane-incorporated Ig.
With fluorescein-conjugated anti-µ, anti-κ, anti-λ, and polyvalent antiglobulin reagents, the MARR was found to be more sensitive than DIF under the present test conditions, and approximately 20% of all peripheral lymphocytes were found to be Ig-bearing with the MARR. It should be stressed, however, that although the MARR appears to have been performed optimally, DIF was performed by using methodology that is widely available but may be suboptimal.
Of potential importance is the finding that summation of the number of sheep erythrocyte- (E) rosetting and mixed antiglobulin-rosetting lymphocytes approximated 100%, yet only 3% of lymphocytes in T-enriched preparations formed mixed antiglobulin rosettes. These findings suggest that E rosette-negative, DIF-negative null cells are B cells that have membrane-incorporated Ig that is not easily detectable by DIF. This interpretation is discussed with regard to possible technical difficulties with the MARR and with respect to other data that indicate the presence of a third lymphocyte population distinct from T and B lymphocytes.
Studies on the Mechanism of Action of Guinea Pig Lymphotoxin
Authors: Okamoto Mitsuhiro , Mayer Manfred M. 
Published: December 2022

Abstract
In the previous paper it was suggested that the primary action of guinea pig lymphotoxin (LT) involved creation of ionic imbalances within the target L cells. The nature of these ionic disturbances is explored in this paper. The exogenous addition of CaCl2, but not KCl or NaCl, inhibited the cytotoxic action of LT. Cellular uptake rates of 45Ca++, but not 86Rb+, increased in LT-damaged L cells. The factor responsible for increasing the 45Ca++ uptake rate cochromatographed on a hydroxyapatite column with the cytotoxic activity of LT. Ouabain prevented the LT-mediated lysis and, concomitantly, depressed the LT-induced increase of 45Ca++ uptake rate. The LT-damaged L cells excluded trypan blue to the same extent as the normal cells. The addition of LT to an LT-resistant L cell mutant affected neither the 45Ca++ uptake rate nor the viability. From these observations, damage to the calcium transport system in the L cell plasma membrane is proposed as a mechanism of LT action.
Immunologic Responses to a Murine Mammary Adenocarcinoma: in Vitro Production of Specific Killer Cells Is Dependent on Active T Lymphocytes
Authors: Yamamura Yasuhiro 
Published: December 2022

Abstract
The mode of production of specifically armed monocytic killer cells was investigated with the T1699 mammary adenocarcinoma in syngeneic DBA/2 mice. After overnight in vitro incubation of cells from the spleen but not from the lymph nodes, blood, or from the peritoneal cavity produced specific killer cells. The activation of spleen cells was inhibited by pretreatment with anti-ϑ serum and C; however, already activated specific killer cells were not sensitive to the same treatment. Removal of phagocytic cells did not significantly affect the cytotoxicity of the splenic killer cells whereas removal of rayon-wool adherent cells greatly reduced both the total cytotoxicity, and to a lesser extent, the cytotoxicity indices. Overnight co-cultivation of normal peritoneal-exudate cells with the lymph node cells from tumor-bearers, although neither class of cells alone was cytotoxic to T1699 cells in vitro, produced specific monocytic killer cells, through steps dependent on active T lymphocyte functions. Culture supernatants of tumor-bearer's spleen cells also contained factor(s) which induced cytotoxicity mediated by normal peritoneal-exudate cells against T1699 cells in vitro; and the production of the factor(s) was also inhibited by pretreatment of the spleen cells with anti-ϑ serum but not by anti-mouse IgG or anti-mouse whole γ-globulins serum and C.
Circulating and Mitogen-Induced Immunoglobulin-Secreting Cells in Human Peripheral Blood: Evaluation by a Modified Reverse Hemolytic Plaque Assay
Authors: Ginsburg William W. , Finkelman Fred D. , Lipsky Peter E. 
Published: December 2022

Abstract
The number of immunoglobulin-secreting cells (ISC) found circulating in normal human peripheral blood and the number generated after in vitro stimulation with pokeweed mitogen (PWM) were assessed by using a modified reverse hemolytic plaque assay (RHPA). This technique involves incubation of the lymphocyte population to be tested in agarose with sheep erythrocytes coated with antibody to human immunoglobulin (Ig). The subsequent addition of anti-Ig-developing antiserum and C results in the formation of hemolytic plaques around active Ig-secreting cells. The RHPA allows for quantitation of total ISC as well as cells individually secreting IgM, IgG, or IgA. Freshly prepared human peripheral blood mononuclear cells (PBM) from 15 individuals were assayed for ISC and found to contain a mean of 747 ± 364 ISC/106 PBM, of which a mean of 445 secreted IgG, 164 IgM, and 239 IgA. In vitro culture of PBM with PWM resulted in the generation of 10-fold more ISC than were found in unstimulated cultures (9353 vs 925). Increased numbers of ISC were evident after 3 to 4 days of incubation and reached a peak after 5 to 7 days. There was great variation in the number of ISC generated in response to PWM in cultures of PBM obtained from different individuals. Moreover, PWM induced a predominance of IgM-secreting cells in cultures of PBM from some individuals, whereas stimulating predominantly IgG-secreting cells in others. The predominant Ig isotype of the ISC generated in response to PWM in cultures of PBM from different individuals was found to vary with the race and sex of the donor.
Evidence for Multiple Clones of Cytotoxic T Cells Responding to Antigenic Determinants on the Same Molecule
Authors: Geib Roy , Chiang Chinlee , Klein Jan 
Published: December 2022

Summary
Hzl lymphocytes stimulated in the CML assay by B6 cells kill not only B6 but also M505 targets. (Hzl and M505 are strains that differ from B6 by recent mutations that occurred in the H-2Kb gene.) When the Hzl anti-B6 lymphocytes are adsorbed on M505 monolayer, the nonadherent cells still react with B6 but not with M505 targets. This result suggests that different antigenic determinants carried by the same H-2 molecule stimulate different T cell clones. The implication of this finding is that probably large numbers of T cell clones are stimulated by a foreign H-2 haplotype coding for a great number of antigenic determinants.
Thermal Inactivation of Hepatitis B Surface Antigen
Authors: De Flora Silvio 
Published: December 2022

Abstract
Thermal inactivation of hepatitis B surface antigen (HBs Ag) was investigated under various experimental conditions by means of a quantitative radioimmunoassay. No decrease of HBs Ag immunologic reactivity could be detected at -80°C, up to 24 months of storage, whereas at -20 and 4°C survival of serum HBs Ag after 6 months was 71.4 and 48.7%, respectively. Freezing and thawing of serum affected the activity of the antigen slightly (60.6% after 26 cycles in 6 months). The loss of activity of purified HBs Ag in phosphate buffer at 20, 37, 44, 56, 70, and 98°C was linear with respect to time, and the half-life of inactivation ranged from 6 min at 98°C to 52 days, 18 hr, and 12 min at 20°C. Over this range of temperature, the velocity constants for inactivation (k) showed a linear relationship with the reciprocal of absolute temperature, and the energy (ΔH) and the entropy (ΔS) of activation of the process were 23.382 kcal/mole and -8.468 cal/mole/deg, respectively. No residual activity was detected after 15 min at 121°C (1 Atm). At neutral pH an increasing protective effect was afforded by deionized water, 0.15 M NaCl, 0.067 phosphate mixture, 0.1 M citric acid-0.2 M sodium phosphate dibasic buffer and human serum. Heat-stability of HBs Ag was significantly reduced by modifying the pH of the medium whereas addition of 1 M MgCl2 had a considerable stabilizing effect. Survival of HBs Ag was affected by contamination with a variety of microbial strains. Traces of the antigen dried on a glass surface could be recovered after 3 and 4 weeks of exposure at room temperature (21 ± 2°C) to ambient air. Subtypes adw and ayw were destroyed to the same extent by heat exposure, and no selective inactivation of a, d, and y subdeterminants was observed.
Comparison of Radioimmunoassay Techniques in the Detection of IgE and IgG Antibody Activity against Aspergillus Fumigatus Antigens
Authors: Patterson Roy , Wang Josephine L. F. , Roberts Mary , Zeiss C. Raymond 
Published: December 2022

Abstract
Radioimmunoassays suitable for detection of antibodies against mixtures of antigens were used to provide quantitative estimation of antibody activity against Aspergillus fumigatus (Af) antigens. Sera from subjects with allergic bronchopulmonary aspergillosis, aspergillomas, and asthma with immediate-type cutaneous reactivity were studied. With 125I IgE and 125I IgG, a solid phase polystyrene tube multilayered type radioimmunoassay detected IgE and IgG antibodies in the same serum samples. With 125I Af, a solid phase polystyrene tube and a double antibody assay detected IgE and IgG antibodies, respectively, in the same serum samples. We conclude that either type of assay may be used for detection of antibody activity and that demonstration of elevated IgE and IgG antibody levels in the same serum may be useful in characterization and differentiation of disease states in which Af is involved.
Mechanism of Nonspecific Macrophage-Mediated Cytotoxicity: Evidence for Lack of Dependence upon Oxygen
Authors: Sorrell Tania C. , Lehrer Robert I. , Cline Martin J. 
Published: December 2022

Abstract
Peritoneal macrophages elicited in C3H/HJ mice by the i.p. injection of Corynebacterium parvum were cytotoxic to allogeneic virus-transformed fibroblasts in vitro. Cytotoxicity was demonstrated in a morphologic (plaque) assay, and quantitated by measuring macrophage-mediated inhibition of incorporation of 3H-thymidine by the target cells. The cytotoxic effect was well established by 6 hr of macrophage-fibroblast interaction, and was retained in cultures from which the supernatant was removed before the addition of 3H-thymidine. Cytotoxic activity of macrophages diminished rapidly after 22 hr of cultivation in vitro. Maximal cytotoxic effect could be prolonged by addition of C. parvum, 50 µg/ml, to macrophage monolayers preincubated in vitro for 22 hr. It could neither be retained nor regenerated when C. parvum was added to monolayers greater than 22-hr old.
C. parvum-activated macrophages, grown under anaerobic conditions for 8 hr, retained the ability to phagocytize heat-killed Candida albicans and to exclude trypan blue dye. There was a small but significant reduction in the ability of macrophages to inhibit 3H-thymidine incorporation by target fibroblasts under anaerobic conditions.
The cytotoxic effect of activated macrophages in air was not altered by the presence of catalase and was enhanced by enzymatically active superoxide dismutase. We conclude that the processes involved in macrophage-mediated cytotoxicity against allogeneic fibroblasts in this system are largely independent of oxygen.
Antibody-Induced Suppression of the Fifth Component of Complement in Mice
Authors: Goldman John N. , Goldman Margaret B. 
Published: December 2022

Abstract
We have shown that administration of anti-C5 antibody to newborn mice heterozygous for C5 deficiency can suppress C5 levels for prolonged periods. This is analogous to antibody-induced immunoglobulin allotype and idiotype suppression. Only certain sources of anti-C5 are effective in suppression. Of the three antisera extensively tested in (SWR × RIII)F1 hybrids, A/He anti-RIII was the most effective, SWR anti-RIII was less effective, and SWR anti-DBA/1 was ineffective. Only certain strain combinations of F1 hybrids are susceptible to C5 suppression. C5 suppression was seen with (SWR × RIII)F1 hybrids but not (SWR × DBA/1)F1 hybrids. Kinetics of C5 suppression suggest a requirement for an active, ongoing process in the maintenance of low C5 levels.
A Cytotoxic Proteinase Isolated from Human Lymphocytes
Authors: Hatcher Victor B. , Oberman Martin S. , Lazarus Gerald S. , Grayzel Arthur I. 
Published: December 2022

Abstract
A proteinase active at physiologic pH was isolated from unstimulated human peripheral blood lymphocytes with gel filtration and affinity chromatography. The proteinase with a molecular mass of approximately 30,000 daltons was completely inhibited by diisopropylfluoro-phosphate (DFP) and soybean trypsin inhibitor (STI). Incubation of the lymphocyte enzyme with 3H-proline labeled T24 human bladder carcinoma cells resulted in significant cytotoxicity of the target cells. Cytotoxicity was only observed with much higher concentrations of trypsin. Similar results were obtained with a 51Cr release assay. This investigation demonstrates that unstimulated human peripheral blood lymphocytes contain a cytotoxic proteinase capable of killing pre-labeled target cells. The proteinase may be involved in lymphocytemediated cytotoxicity.
Kinetics of CTL Induction by Concanavalin A
Authors: Tartof David , Fitch Frank W. 
Published: December 2022

Abstract
Induction of maximal CTL activity was achieved within 12 hr of exposure to Con A in vitro in various mouse lymphoid cell populations. These included spleen cells from normal unsensitized mice, spleen cells from mice previously immunized with alloantigen, and mouse spleen cells exposed to alloantigen in long-term mixed leukocyte culture (LTMLC). Although induction of maximal incorporation of tritiated thymidine was accomplished within this same period in the cells obtained from LTMLC, a much longer period of Con A exposure (&gt;24 hr) was required for freshly prepared spleen cells from normal or previously immunized mice. These findings indicate that the increased tritiated thymidine uptake induced in freshly prepared spleen cells on continued exposure to Con A beyond 12 hr is not associated with the development of cytolytic activity, and that it probably represents stimulation of subpopulations no longer present in the LTMLC population where positive selection for cells responsive to cellular alloantigens has taken place.
Role of the Classical and Alternative Complement Pathways in Chemotaxis and Opsonization: Studies of Human Serum Deficient in C4
Authors: Clark Robert A. , Klebanoff Seymour J. 
Published: December 2022

Abstract
The roles of the classical and alternative C pathways in the formation of chemotactic factors and opsonins in human serum were evaluated by comparative studies on normal serum and serum from a patient with a nonfunctional classical pathway resulting from an inherited absence of C4. Neutrophil chemotactic activity was determined in serum activated by zymosan, bacterial endotoxin, intact bacteria, and cobra venom factor, by using a 51Cr radioassay in Boyden chambers. Opsonization of zymosan by serum was determined by the iodination reaction with human neutrophils as the test cells. Chemotactic activity and opsonization were both markedly impaired in C4-deficient serum; these defects were most apparent when the serum concentration was low or the period of exposure to the activating agent was short. Normal chemotactic and opsonic activity was observed when either the serum concentration or the incubation time was increased with zymosan as the activating agent. In contrast, correction of the chemotactic defect was not achieved under similar conditions when endotoxin was the activating agent. The addition of purified C4 resulted in improvement or normalization of chemotactic and opsonic function. Similar defects were demonstrated in sera from an unrelated patient with inherited C4 deficiency and from a patient with absent C4 and C2. These observations point out the necessity for an intact classical pathway in order to achieve optimal formation of biologic mediators from the C system.
Opsonin-Independent Phagocytosis of Activators of the Alternative Complement Pathway by Human Monocytes
Authors: Czop Joyce K. , Fearon Douglas T. , Austen K. Frank 
Published: December 2022

Abstract
Activators of the alternative C pathway were ingested by human monocytes in a synthetic medium devoid of serum or proteins of the alternative C pathway. The rank order of the percentage of human monocytes ingesting rabbit, mouse C57B, and mouse CBA erythrocytes was the same as that observed for lysis of these erythrocytes by the alternative C pathway in C2-deficient human serum diluted in GVB-Mg-EGTA. Sheep and guinea pig erythrocytes, which were not ingested by human monocytes, were not lysed by the alternative C pathway. Although the capacity of human monocytes to ingest rabbit erythrocytes was consistent for a particular donor, there was a bimodal distribution among donors in terms of the percentage of monocytes involved. Zymosan ingestion, on the other hand, was observed with a high proportion of monocytes from all donors. The capacity of human monocytes to ingest zymosan particles and rabbit erythrocytes was largely diminished when the monocytes were pretreated with quantities of affinity-purified trypsin, which had little or no effect on their ability to phagocytose EA or even to bind E(IgM)C43b intermediates. Latex particles that did not activate the alternative pathway of C were ingested by human monocytes after treatment with a concentration of trypsin 10 times that required to reduce substantially ingestion of zymosan. The trypsin-sensitive mechanism of monocytes involved in the ingestion of zymosan and rabbit erythrocytes was regenerated over 48 hr during in vitro culture and is a result compatible with the restoration of function of a membrane protein. The capacity of human monocytes to ingest particulate activators of the alternative C pathway in the absence of surface-associated opsonizing proteins such as IgG or C3b by a trypsin-sensitive step represents a monocyte recognition mechanism specific for activating surfaces of the alternative C pathway.
Demonstration of Two Types of Helper T Cells for Different IgG Subclass Responses to Dinitrophenylated Flagellin Polymer
Authors: Minami Mutsuhiko , Usui Mitsuko , Kanno Tsunehiro , Tamura Noboru , Matuhasi Tyoku 
Published: December 2022

Abstract
The IgG anti-DNP antibody response to DNP-POL was explored in mice in an adoptive transfer system. The adoptive secondary IgG anti-DNP response required the presence of T cells. The IgG anti-DNP response to DNP-POL could be obtained in the irradiated recipients transferred with DNP-primed B cells if the recipients had been primed with either carrier or unrelated antigens 5 days before the cell transfer. Passive transfer of DNP-primed B cells plus T cells activated by carrier resulted in a high IgG anti-DNP response to DNP-POL. Passive transfer of DNP-primed B cells plus either T cells activated by unrelated antigens or unactivated T cells also resulted in a low but appreciable IgG anti-DNP response. Moreover, there are qualitative differences among these IgG anti-DNP responses. The IgG1, IgG2a, and IgG2b responses to DNP-POL were induced only in the carrier-primed recipients given DNP-primed B cells. By contrast, the IgG3 response to DNP-POL could be induced in the KLH-primed recipients receiving DNP-primed B cells.
These results indicate that the IgG anti-DNP responses to DNP-POL are induced through at least two different pathways; one involves a collaboration between carrier-specific T cells and hapten-specific B cells, and the other involves an interaction between noncarrier-specific T cells and hapten-specific B cells.
Measurement of IgE Antibody by an Antigen-Binding Assay: Correlation with PK Activity and IgG and IgA Antibodies to Allergens
Authors: Platts-Mills T. A. E. , Snajdr M. J. , Ishizaka K. , Frankland A. W. 
Published: December 2022

Abstract
A quantitative radioimmunoassay for IgE antibodies (ab) is described. The assay measures IgE antigen-binding activity (BA) by using purified allergens radiolabeled with 125I, IgE myeloma serum as a source of carrier protein, and monospecific goat anti-IgE (anti-IgE) to precipitate the IgE. IgE BA for the Group I protein of rye grass pollen (Rye I) was measured in the serum of patients with grass pollen hay fever who had never received immunotherapy. Values for IgE BA were found to correlate well with radioallergosorbent technique (RAST) results (rs = 0.89, p&lt;0.001). By assaying IgE-rich fractions, pure IgG, and whole serum, we found a good correlation between IgE BA, Prausnitz-Küstner (PK) activity, and total IgE. A positive PK test could be elicited with 0.05 ml of dilutions containing as little as 1 or 2 units IgE BA for Rye I/ml. PK testing was more sensitive than either RAST or the antigen-binding assay. Estimates based on the quantity of antigen bound in antigen excess suggested that 1 unit IgE BA represented 0.1 ng IgE ab. With higher concentrations of antigen (up to 1 µg/ml), however, estimates suggested that 1 unit IgE BA represented as much as 0.4 ng IgE ab. These higher estimates appeared to correlate reasonably well with estimates based on RAST elution or RAST absorption. The assay for IgE BA was directly comparable with assays for IgG BA and IgA BA. Results of these assays on sera from the patients showed that IgG BA for Rye I was detectable in 25 of 25 IgE BA in 24 of 25, and IgA BA in 21 of 25. Quantitatively we found a characteristic pattern IgG BA &gt; IgE BA &gt; IgA BA (geometric mean values 862, 214, and 60.2 units/ml, respectively). In keeping with this there was a highly significant quantitative correlation between IgG BA and IgE BA (r = 0.72, p&lt;0.001). None of the sera from nonallergic persons contained detectable IgA or IgE BA for Rye I; however, 20% of these sera (10 of 48) contained IgG BA.