bluuebunny/crossref_metadata_2025 huggingface dataset:
Destruction of Tumor Cells by BCG-Activated Alveolar Macrophages
Authors: Zwilling Bruce S. , Campolito Laura B.
Published: December 2022

Alveolar macrophages obtained from Syrian golden hamsters were tested for their ability to destroy tumor cells. Only macrophages obtained from BCG immune animals rechallenged intratracheally with BCG five days before assay exhibited cytotoxic activity. Maximum destruction of tumor cells occurred after 5 days of incubation. Immunologic activation of macrophages was required to attain cytotoxic alveolar macrophages. Induction of inflammatory lung exudates by a variety of nonspecific irritants did not result in tumor cell destruction by macrophages. These observations may prove useful in designing an approach for immunotherapy of lung cancer.
Immune Responses of Inbred Guinea Pigs to the Sequential Polymer Poly(L-Tyr-L-Glu-L-Ala-Gly): Studies with the Oligomers of the Polymers
Authors: Lai Chang-Hai , Maurer Paul H.
Published: December 2022

Synthetic known sequence polypeptides poly(Tyrosine-Glutamic acid-Alanine-Glycine) (T-G-A-Gly)n were found to be very immunogenic in responder inbred guinea pigs. Two and one-half micrograms were enough to elicit both humoral and cellular responses. Only the α-helical oligomers were immunogenic and were able to inhibit the homologous antigen-antibody reactions. The random polymers of comparable amino acid composition, i.e., poly(glutamic acid60alanine40) (GA), poly(glutamic acid50 tyrosine50) (GT), poly(glutamic acid50alanine30tyrosine10) (GAT10), did not inhibit. the antibodies against (T-G-A-Gly)n did not bind to the closely related known sequence polymer poly tyrosine-alanine-glutamic acid-glycine) (T-A-G-Gly)n or to the above random polymers. It is thus concluded that antibodies against (T-G-A-Gly)n are directed against conformational determinats.
Structural Evidence for an Unblocked VHI Sub-Subgroup of Human Heavy Chains
Authors: Hopper John E. , Brahn Ernest
Published: December 2022

The N-terminal amino acid sequence of an unblocked human heavy chain of an IgGκ paraprotein from patient Tho is reported. This unblocked VH sequence belongs to the VHI subgroup and has striking structural homology to an unblocked VHI sequence previously reported for the IgGκ paraprotein isolated from the serum of patient Bro. Direct sequence comparison of the Tho and Bro proteins reveals complete structural identity in 25 of the N-terminal 27 residues, with unique amino acid substitutions shared at the N-terminus and positions 16, 18, and 24. This remarkable sequence homology suggests that the two VH sequences represent examples of an unblocked VHI sub-subgroup.

The Tho and Bro sequences possess an ala-glu-val basic triplet at positions 9 to 11 in common with the other previously reported VHI proteins which help to further establish this sequence triplet as a recognizable VHI subgroup-specific region. Moreover, in this regard, the unblocked sequences of Tho and Bro emphasize the importance of extending an unblocked VHI sequence beyond positions 9 to 11 before a subgroup assignment can be determined.
Antibody Response to Simian Virus 40 Tumor Antigen in Nude Mice Reconstituted with T Cells
Authors: Cicurel Lucienne , Croce Carlo M.
Published: December 2022

Athymic BALB/c nude mice (nu/nu) fail to generate circulating antibodies to simian virus 40 (SV40) tumor (T) antigen when immunized with SV40-transformed mouse cells or with T antigen positive somatic cell hybrids derived from SV40-transformed human and normal mouse parental cells. However, normal BALB/c mice readily produce antibodies to SV40 T antigen. When nude mice were reconstituted with normal syngeneic T lymphocytes from spleen or thymus source, the humoral immune responsiveness to SV40 T antigen was restored.
In Vitro Sensitization of Human Lymphocytes Against Histiocytic Lymphoma Cell Lines
Authors: Treves Abraham J. , Feldman Michael , Kaplan Henry S.
Published: December 2022

Peripheral blood lymphocytes from normal donors expressed spontaneous cytotoxic activity against human diffuse histiocytic lymphoma cell lines. In the unfractionated state, they could not be further sensitized in vitro against these cell lines. By applying cell separation techniques before culture, subpopulations of lymphocytes were obtained which could be sensitized in vitro and manifested cytotoxic activity against human histiocytic lymphoma cells. Three methods of separation were found effective: E rosette enrichment; elimination of Fc receptor positive cells; and removal of nylon wool adherent cells. Under these conditions, cross-reactive cytotoxicity was observed against non-neoplastic lymphoblastoid cell lines, but not against normal lymphocytes.
The Presence of Immunoregulatory Cells in Chicken Thymus: Function in B and T Cell Responses
Authors: Moticka Edward J.
Published: December 2022

The presence of immunoregulatory cells in chicken thymus was studied by using several different systems. Chickens injected with large numbers of syngeneic thymocytes were tested for their ability to produce antibody to heterologous red cells. Similar chickens were studied for their ability to reject allogeneic skin grafts. In separate studies, mixtures of thymocytes with spleen cells or with peripheral blood leukocytes were assayed for their ability to respond to PHA or to produce a graft-vs-host reaction in embryonic chicks. These studies indicated that immunoregulatory cells exist in chicken thymus, which displays both helper and suppressor activity. The suppressor cells were more prevalent or more easily detectable in young birds and in chickens with intact bursas. The helper function of thymocytes was seen to better advantage with cells derived from older animals and from bursectomized donors.
The Differential Effects of 6-MP and Cyclophosphamide on Immunoglobulin Deposition in the Skin and Kidney of NZB/NZW F1 Mice
Authors: Gilliam James N. , Hurd Eric R.
Published: December 2022

The effects of cyclophosphamide (CY) and 6-mercaptopurine (6-MP) on the accumulation of subepidermal Ig and glomerular Ig in the female NZB/NZW F1 (B/W) mouse were compared. Five-month old female mice, given daily i.p. injections of CY or 6-MP, and untreated control mice were killed after 2 months and specimens obtained for direct immunofluorescent staining of skin and kidney. In addition, electron microscopic examination of the kidneys was performed. Subepidermal Ig was present in 30% of the untreated animals, in none of the CY-treated, and in 83% of the 6-MP treated animals. Glomerular Ig was observed in 81% of the untreated animals, 40% of the CY-treated animals, and 85% of the 6-MP treated animals. ANA tests were positive in 83% of the untreated, 29% of the CY-treated, and 94% of the 6-MP treated animals. All differences between the CY- and 6-MP-treated groups were statistically significant. Electron microscopy of the kidneys demonstrated a marked reduction of electron dense deposits in the CY-treated animals but no reduction was seen in the 6-MP treated animals. These findings showed: 1) CY markedly reduced subepidermal Ig, glomerular Ig, and ANA; and 2) 6-MP increased subepidermal Ig whereas glomerular Ig and ANA remained essentially unchanged. These observations indicate that CY not only appears to be more effective than 6-MP in the treatment of B/W immune complex disease but also that 6-MP enhances Ig deposition in the skin.
Complement Activation in Semisolid Media: Insolubilization of Alternative Pathway Convertases in Agar Gels with C3 Nephritic Factor-Containing Sera
Authors: Arnaout Mohammed Amin , Davis Alvin E. , Rosen Fred S. , Alper Chester A.
Published: December 2022

It was previously shown that when normal human serum and purified properdin or serum containing nephritic factor were allowed to diffuse toward each other in agar, a stainable precipitin line formed only in the presence of an intact alternative pathway. In the present study we have shown that when guinea pig erythrocytes were incorporated in agar and normal human serum was allowed to diffuse toward C3 nephritic factor-containing serum, a line of hemolysis appeared that coincided with the stainable line. The line of hemolysis only formed in the presence of C3, factor B and Mg++, as does the properdin or C3 nephritic factor-induced stainable line. When C2-deficient serum was incorporated in agar with guinea pig erythrocytes and serum samples containing nephritic factor were applied in wells, rings of lysis developed, the areas of which correlated significantly with nephritic factor activity. When partially purified nephritic factor (contaminated only with IgG) or serum containing nephritic factor was subjected to electrophoresis in agar-guinea pig erythrocyte gels and overlaid with normal human serum, bands of lysis developed that required factor B and C3. Lysis of guinea pig erythrocytes in this system appears to be due to insolubilization of alternative pathway convertases with activation of the membrane attack unit C5–C9.
Analysis of the Diversity of Murine Antibodies to Dextran B1355
Authors: Hansburg Daniel , Briles David E. , Davie Joseph M.
Published: December 2022

We have developed radioimmunoassays that detect idiotypic (variable region) differences among the α(1 → 3) dextran-binding myeloma proteins U102, J558, and M104 as well as an assay that detects variable region determinants common to all three proteins. Using these assays, we have examined 7S and 19S anti-α(1 → 3) dextran antibodies induced in five murine strains of the a1 IgCH linkage group and the recombinant strain BAB/14. All idiotypes were expressed in both 19S and 7S antibodies from all strains, but with considerable strain-specific variability in penetrance. In two strains, one additional type of antibody, which lacked all four idiotypic determinants, generally constituted the bulk of total anti-α(1 → 3) dextran antibodies.
Genetic Control of Immune Response to Protein Antigens
Authors: Passos Hélcio C. , Siqueira Maria , Reis Moema H. , Ferreira Vera C. Annes , Ibanez Olga M. , Sant'Anna Osvaldo A. , Biozzi Guido
Published: December 2022

A high (H/p) and a low (L/p) responder line of mice have been produced by selective breeding for the character “quantitative antibody responsiveness” to BSA and RgG. These two Ag were administered at optimal immunizing doses and alternated at each generation in order to avoid the interference of maternally transmitted antibodies.

H/p and L/p diverged progressively during the first seven generations of selective breeding, suggesting a polygenic regulation of the character investigated. The maximal interline separation (RT) was reached after seven consecutive generations of selective breeding (F7). The continuation of the selection until the F15 generation did not significantly modify the RT value of F7. At the maximal interline separation (F7 to F15) the titer of antibody measured in terms of passive hemagglutinins was about 250 times higher in H/p than in L/p. The decrease of phenotypic variance during the selective breeding shows that 76% of the phenotypic variance of the foundation population (F0) is due to genetic factors: VG, and that 24% is produced by environmental effects: VE.

The heritability of the character (h2) measured by the linear regression between the selection differential (S) and the response to selection (R) during the first seven generations is 0.22 ± 0.06. A provisional estimation that needs to be confirmed by experiments under way indicated that the character investigated may be controlled by two or three independent loci.
Serologic Identification of Hemopoietic Progenitor Cell Antigens Common to Mouse and Man
Authors: Krogsrud Richard L. , Bain Jerald , Price Gerald B.
Published: December 2022

An immunologic approach to the identification of human hemopoietic stem cell antigens has yielded two types of antisera. Both recognize antigens on hemopoietic progenitors which are identical or cross-reactive with determinants on sperm, brain, and, across the species barrier, with murine hemopoietic stem cells (CFU-S). The first antiserum was the result of an autoimmune reaction in a post-vasectomy patient. The second was produced in rabbits after injection of human brain cells.
Specific Binding of T Lymphocytes to Macrophages
Authors: Ben-Sasson Shlomo Z. , Lipscomb Mary F. , Tucker Thomas F. , Uhr Jonathan W.
Published: December 2022

Peritoneal exudate lymphocytes from immune guinea pigs that bind in vitro to autologous antigen-pulsed macrophages were allowed to proliferate for 1 week to give a population markedly enriched in antigen-specific T cells. This enriched population was then studied with regard to its binding to fresh autologous antigen-pulsed macrophages. Specific binding was not inhibited by a large excess of antigen in the media (5000-fold greater than the amount of antigen associated with the macrophages) either soluble or bound to Sepharose beads, or by coating the antigen-pulsed macrophages with antibody to the exogenous antigen, by reacting a second layer of antibody to the heterologous antibody, or by haptenating the antigen and treating the hapten-antigen macrophage complex with excess anti-hapten antibody.

Results of treating antigen-pulsed macrophages with the proteolytic enzymes trypsin and pronase indicate that exogenous antigen is on the macrophage surface, but the experiments failed to prove that the removable antigen is essential for binding. The simplest interpretation of these results is that the T cell recptor is not specific for native exogenous antigen.
Immunochemical Characterization of IgM in Human Intestinal Fluids
Authors: Richman Lee K. , Brown William R.
Published: December 2022

We have investigated the immunochemical characteristics of IgM in intestinal fluids from healthy human subjects. The concentration of IgM was measured by double-antibody radioimmunoassay. The molecular size of IgM was evaluated by rate zonal sedimentation on sucrose gradients. The presence of SC on IgM was determined by solid-phase immunosorption of intestinal fluid samples with anti-µ chain-Sepharose followed by reaction of the immunosorbent with radioiodinated anti-SC antibodies. Confirmation of this method was obtained by differential immunosorption of pooled density gradient IgM fractions with anti-SC or nonimmune rabbit gamma globulin-Sepharose. The stability of 131I-IgM-125I-SC complexes was evaluated by incubation in intestinal fluid followed by density gradient ultracentrifugation.

The mean concentration of IgM in small intestinal fluid was 1.53 mg/dl. IgM molecules sedimenting at 19S, 10S, 7S, and less than 3.5S were identified. SC was present on IgM in a minority of specimens and was found to be associated solely with 19S species of IgM. SC was rapidly cleaved from IgM-SC complexes by incubation in intestinal fluid at 37°C. The data are consistent with a model for IgM secretion in the human intestine in which IgM is secreted complexed with SC but is readily degraded within the intestinal lumen.
Further Purification and Characterization of a Circulating Antigen in Schistosomiasis
Authors: Nash Theodore E. , Nasir-Ud-Din  , Jeanloz Roger W.
Published: December 2022

Previous studies showed that an antigen found in the circulation of animals heavily infected with Schistosoma mansoni was extracted in a trichloroacetic acid solublechloroform insoluble fraction (TCA-S-C) of adult worms. Antigenic activity was destroyed by periodate treatment but remained unaltered after treatment with proteolytic enzymes, DNase, RNase, and lyophilization. In the present study, chromatography of TCA-S-C on a DEAE cellulose column revealed six substances, one of which was antigenic. After electrophoresis in agarose antigenic activity corresponded to a slower moving, toluidine blue-staining material. A faster moving, toluidine blue-staining substance seems to be responsible for the large 260 nm. absorbing peak. Analysis of a fraction containing only antigen revealed a large amount of carbohydrate, primarily N-acetylglucosamine and D-glucuronic acid but also galactase, glucose, N-acetylglucosamine, and trace amounts of other sugars. Amino acids accounted for about 11% of the weight of the antigen. The antigen appears to be a proteoglycan.
Dextran Abrogates Stimulation of IgG Antibody Synthesis by Allogeneic Effect Factor
Authors: Yen Belinda , Battisto Jack R.
Published: December 2022

The influence of dextran upon in vitro immunization of unfractionated splenic cells and of purified splenic B cells against sheep erythrocytes has been studied with particular emphasis upon the relationship of dextran to the augmentation of antibody synthesis by allogeneic effect factor (AEF). Dextran enhanced the number of SRBC-specific IgM PFC when added to unfractionated splenic cells undergoing a primary immune response in vitro. When it was added to purified splenic B cells no such effect was observed.

The established capacity of AEF to facilitate antibody synthesis by splenic B cells was found to be markedly altered by addition of dextran to the cultures. Although the capacity of AEF to increase the number of IgM PFC remained virtually unchanged, the enhancement of IgG PFC brought about by AEF was abrogated. These observations suggest that either two separate molecules may be responsible for the activities of AEF or that one molecule possesses two separable functions: one increases IgM PFC and another stimulates IgG PFC. Data are given that indicate the activity of the latter may be interfered with by dextran at the B cell surface membrane.

We hypothesize that the receptor for AEF on the IgG-producing B cell may be, in part, structurally related to dextran (i.e., a polysaccharide molecule with α-1,6 glucosidic linkages) and for this reason may interfere with reception of an AEF signal by B cells. If this is the case, then antigens that cause the appearance of IgM antibodies but not IgG antibodies may be incorrectly regarded as “T cell-independent antigens” primarily because they intercept the latter activity of AEF.
Functional Analysis of B Cell subpopulations: Responsiveness of Phenotypically Characterized Cr+ and CR- B Cell Subsets to a T-Dependent Antigen in Vitro
Authors: Simon Markus M. , Hämmerling Ulrich
Published: December 2022

B lymphocytes were separated by velocity sedimentation on the basis of receptors for activated complement (C3). The C receptor-positive (CR+) and C receptor-negative (CR-) subpopulations were assayed for in vitro 19S PFC responses to the trinitrophenyl group coupled to burro erythrocytes (TNP-BRBC). Both B cell subsets are equally able to respond to the antigen in the presence of either unprimed or carrier-primed T cells. The antibody responses of either CR+ or CR- B cells were not dependent on the presence of adherent spleen cells in cultures supplemented with 2-mercaptoethanol, but addition of peritoneal macrophages slightly increased the number of PFC in both populations.

CR- B cell populations obtained by two different methods [i.e., passage of spleen cells through a column of C3-coated Sephadex beads (CR-col) versus velocity sedimentation of EAC-rosetted spleen cells (CR-sed)] differed in their composition. The first method appears to select nonspecifically for relatively less Ia+ and more Ig+ cells than the second method. This enrichment for less mature B cells is reflected in the response characteristics toward TNP, since the CR-col cells needed a higher proportion of helper cells as compared with CR-sed cells for eliciting equivalent numbers of anti-TNP plaques. The results suggest that CR+ and CR- B subpopulations show quantitative rather than qualitative differences in their response characteristics to a T-dependent antigen.
Synthesis and Characterization of Membrane Immunoglobulin, Ia, and H-2 Molecules of Thy 1+ AKR/J Lymphomas
Authors: Schwartz Benjamin D. , Campbell Pamela S. , Asofsky Richard , Greenberg Robert S. , Zatz Marion M.
Published: December 2022

Three AKR lymphomas displaying B cell and T cell characteristics have been described. Because of the proclivity of normal AKR/J mice to develop T cell lymphomas, and the rarity of lymphomas with dual characteristics, the B cell markers of these tumors were studied more intensively. Fluorescence data with class-specific anti-immunoglobulin reagents demonstrated that the tumor cells stained only with class-specific anti-IgM reagents. Because of the possibility that the surface Ig was passively acquired and of reports that certain anti-µ-chain sera react with “IgT”, chemical characterization of the immunoglobulin molecules was performed. Using 3H-leucine internal labeling, we showed that all three tumor lines synthesized the immunoglobulin found on their surface, and that the immunoglobulin had the chemical and immunologic characteristics most typical of monomeric surface IgM, and was composed of µ-chains and light chains. The Ia antigens found on these cells were also examined. These antigens were also synthesized by the cells and were present in the same molecular form and in the same approximate quantity as Ia antigens on normal spleen cells.
Fc Receptor Heterogeneity: Immunofluorescent Studies of B, T, and “Third Population” Lymphocytes in Human Blood with Rabbit IgG b4/Anti-b4 Complexes
Authors: Winfield John B. , Lobo Peter I. , Hamilton Maurice E.
Published: December 2022

IgG Fc receptors on human peripheral blood lymphocytes (PBL) were characterized by immunofluorescence studies with defined rabbit IgG b4 allotype/anti-allotype complexes. Three discrete types of Fc receptor-bearing cells, totaling ∼33% of PBL, were identified. Fc receptors of the three types differed in their sensitivity to trypsin and in either absolute or localized density (topography) as determined by variable requirements for anti-IgG cross-linking in order to visualize bound complexes microscopically. The question of additional heterogeneity related to differences in individual Fc receptor affinity for complexed IgG was not approached in this study. Ten to 15% of PBL had pronase-sensitive, trypsin-resistant Fc receptors readily detected by direct immunofluorescence by using large fluorescein-conjugated complexes prepared near equivalence. Double-label and lymphocyte fractionation experiments established this population to be largely distinct from surface IgM + B cells and T cells, and identical to EARipley rosette-forming cells. Approximately 50% of surface IgM + B cells and ∼10% of T cells had lower density Fc receptors identified by indirect immunofluorescence with small complexes prepared in antigen excess or by cross-linking fluorescein-conjugated complexes with anti-rabbit IgG anti-serum. An additional ∼15% peripheral T and B cells had very low density Fc receptors detectable by complexing the IgG on the cell surface by sequential incubations of cells with b4 IgG and anti-b4. Fc receptors on B and T cells were sensitive to both pronase and trypsin digestion.

The heterogeneity of IgG Fc receptors on different lymphocyte subpopulations as defined by these experiments may be of relevance for further analysis of normal and abnormal immune function.
Modulation of Fc-Mediated Phagocytosis by Cyclic AMP and Insulin in a Macrophage-Like Cell Line
Authors: Muschel Ruth J. , Rosen Neal , Rosen Ora M. , Bloom Barry R.
Published: December 2022

Studies on genetically selected mutants in phagocytosis of E[IgG] indicated that the defect in some mutants could be corrected by addition of 8 Br-cAMP, and suggested that cyclic AMP may be involved in the mechanism of phagocytosis through the Fc receptor. In order to elucidate the role of cyclic AMP in phagocytosis in the parental, nonmutant macrophage-like cell line, J774.2, it was necessary to employ restrictive conditions which rendered phagocytosis suboptimal. When J774.2 cells were cultured in nontissue culture Petri dishes phagocytosis was markedly reduced. Addition of 8 Br-cAMP or inducers of intracellular cyclic AMP such as isoproterenol restored the phagocytic ability of these cells. Similarly, treatment of the parental J774.2 cells with insulin reduced the level of phagocytosis, and once again this suppression could be corrected by addition of 8 Br-cAMP. In no case did AMP mimic the effects of 8 Br-cAMP. The effect of cyclic AMP action in this system was not instantaneous, but rather reached optimal levels at 5 to 10 hr, suggesting that cyclic AMP is not the immediate signal for phagocytosis. The genetic analysis of macrophage variants may provide a useful model for studies on the mechanisms of phagocytosis, and also the effects of insulin and cyclic AMP on an easily measurable biologic function in a specialized cell type.
B Memory Cells in the Thymus: Part of the Pool of Potentially Circulating Memory Cells
Authors: Benner Robert , van Oudenaren Adrianus , de Ruiter Hans
Published: December 2022

Immunization of mice with sheep red blood cells (SRBC) or Escherichia coli lipopolysaccharide (LPS) induces the appearance of B memory cells in the thymus. In this paper the origin of these B memory cells was investigated. Therefore, mice primed with either SRBC or LPS 6 months previously and nonprimed mice were joined for parabiosis. Four weeks later the parabiotic mice were separated from each other. Another 3 weeks later thymus cells from the primed and nonprimed mice were transferred separately into lethally irradiated mice in order to determine the adoptive PFC response. It was found that the 4-week period of parabiosis could account for the appearance of a distinct population of B memory cells in the thymus of the nonprimed mice. This result suggests that the B memory cells which appear in the thymus belong to the pool of potentially circulating memory cells.
Functional Differentiation of B Lymphocytes in Congenital Agammaglobulinemia
Authors: Dosch Hans-Michael , Percy Maire E. , Gelfand Erwin W.
Published: December 2022

Despite the absence of B lymphocytes, peripheral blood lymphocytes (PBL) from four of five patients with congenital agammaglobulinemia (cAγ) generated a specific hemolytic plaque-forming cell (HcPFC) response in vitro to sheep red blood cells and ovalbumin. The kinetics, antigenic, and cellular requirements were similar to normals, but significantly less HcPFC were found in patient cultures. Normal but not patient HcPFC-precursor cells were inactivated by treatment with anti-µ antisera whereas generated HcPFC in both controls and patients were sensitive to treatment with anti-µ. Pokeweed mitogen (PWM) and dextran sulfate (DxS) enhanced the HcPFC-response of normal PBL; cAγ-cells were unresponsive to DxS and, in the presence of PWM, the development of HcPFC was inhibited. These findings indicate the presence of B lymphocyte precursors in the majority of patients with cAγ investigated.
Modulation of B Cell Responses by Glycolipid Released from Antigen-Stimulated T Cells
Authors: Esselman Walter J. , Miller Harold C.
Published: December 2022

Mice immunized with either 109 or 106 SRBC were used as a source of T suppressor (Ts) cells and T helper (Th) cells, respectively. T cells were prepared by purification on glass wool and nylon wool columns and were cultivated at 37°C for 3 days. The suppressive or helper effects of the conditioned medium obtained from the T cells were determined in spleen cell cultures by a Jerne-plaque assay. Addition of suppressor medium (Ts-M) to a spleen cell assay was modulatory in that it delayed the normal response time by several days. Thy-1 and GM1 ganglioside were associated with the modulatory substance because a normal response could be obtained by treatment of the Ts-M with anti-Thy-1.2 or anti-GM1 ganglioside sera. Addition of helper T cell medium (Th-M) significantly enhanced anti-SRBC PFC response but anti-Thy-1 or anti-GM1 ganglioside sera had no effect on the helper activity of Th-M. Glycolipids were isolated from Ts-M and Th-M, formulated into liposomes, and were added to PFC assay. A ganglioside isolated from Ts-M with the thin layer mobility (Rf) of brain GM1 ganglioside was found to inhibit anti-SRBC-plaques. Other glycolipids from Ts-M were not inhibitory and none of the glycolipids isolated from Th-M were inhibitory under our assay conditions. The modulatory activity of Ts-M and the isolated glycolipid was not antigen specific and could be neutralized with anti-Thy-1 or anti-GM1 sera. These findings demonstrate that the modulatory (or static) capacity of Ts-M isolated from cultures of partially purified T cells is associated with a cell product, probably a membrane complex, containing Thy-1 antigen and modulatory glycolipid.
Ia Antigens in Inbred Rats and Their Relationship to the MLR Phenotype
Authors: Radka Susan F. , Cramer Donald V. , Gill Thomas J.
Published: December 2022

Three different alloantisera were raised by using Ag-B/MLR disparate rats, and the cytotoxic activity remaining after absorption with erythrocytes to remove anti-Ag-B antibodies was examined. The alloantisera detected surface antigens present only on B cells and segregation studies demonstrated that the genes that code for these antigenic specificities were linked to the major histocompatibility complex. The reactivity of the alloantisera with splenic lymphocytes from a panel of strains representative of the currently known Ag-B groups showed that multiple specificities were present in two of the three antisera and that these specificities were shared by many inbred strains. The appropriate absorption studies showed, however, that each antiserum detected an unique specificity that was found only in those inbred strains that shared the same mixed lymphocyte reactivity (MLR) phenotype as the donor strain. The alloantiserum produced against the KGH strain inhibited the MLR reactions involving this strain only when it was used as the stimulating cell population. The antigens detected by the three alloantisera described here have the characteristics of Ia antigens, and they have tentatively been designated Ia.1 (ACI anti-KGH), Ia.3 (B3 anti-BN) and Ia.4 (MNR anti-DA).
Specific, Immunologic Regulation of Differentiation of Immunoglobulin Expression in MOPC-315 Cells during in Vivo Growth in Diffusion Chambers
Authors: Rohrer James W. , Lynch Richard G.
Published: December 2022

The cells of MOPC-315, a TNP-specific BALB/c plasmacytoma, differentiate during in vivo growth in diffusion chambers (DC) from nonsecreting lymphocytoid cells bearing cell surface immunoglobulin to immunoglobulin-secreting plasmacytes. To determine if such maturation is sensitive to host immunoregulatory signals, DC containing MOPC-315 cells and various TNP-immunogens were implanted i.p. in normal or carrier-primed BALB/c mice and harvested at various times thereafter. Tumor cell number, anti-TNP RFC, and anti-TNP PFC were monitored. Promotion and suppression of MOPC-315 cell growth and anti-TNP immunoglobulin expression were seen when DC containing MOPC-315 cells and TNP-SRBC were grown in mice primed with 4 × 108 and 4 × 106 SRBC, respectively, but not in normal or RRBC-primed mice. Conversely, 315 cells responded to TNP-RRBC only in RRBC-primed mice. Exposure of MOPC-315 cells to both SRBC and TNP-BALB/c RBC in SRBC-primed mice elicited neither help nor suppression. Likewise, no TNP-specific response was elicited by exposure of MOPC-315 cells to the T-independent immunogen DNP30-Ficoll. These results suggest that MOPC-315 cells: a) are responsive to inducible host carrier-specific soluble helper and suppressor factors, and b) provide a monoclonal antigen-binding population that may prove useful in studies of B cell regulation.
Basophil-Derived Platelet-Activating Factor (PAF) as an in Vivo Mediator of Acute Allergic Reactions: Demonstration of Specific Desensitization of Platelets to PAF during IgE-Induced Anaphylaxis in the Rabbit
Authors: Henson Peter M. , Pinckard R. Neal
Published: December 2022

The anaphylactic response to an i.v. injection of antigen into rabbits making only IgE antibody against the antigen was shown to be preceded by a disappearance of the metachromatic staining properties of the circulating basophils and was accompanied by marked but transient thrombocytopenia. The platelets which returned to the circulation 1 hr after the anaphylaxis were shown to be unresponsive to the secretion-inducing activity of basophil-derived platelet-activating factor (PAF) when compared with platelets examined before antigen challenge. By contrast, platelet responsiveness to other stimuli such as collagen, thrombin, and C3b was unchanged. The specific desensitization to PAF provides strong evidence for the action of this mediator on platelets in vivo during IgE-mediated anaphylaxis and provides a useful tool for detecting the effects of particular cell activators in inflammatory reactions.
The Relative Frequency of the Positive Dysentery-Skin Reaction at Different Ages
Authors: Brokman H. , Popowski S.
Published: December 2022

Conclusions

The skin-test carried out with the toxin of the Shiga-Kruse bacillus in 675 individuals of different ages, showed 51 per cent positive results in children up to the fourth years of age; 27 per cent between four and sixteen years; 59 per cent between sixteen and twenty-three years of age.
The Individuality of Semen, with Reference to its Property of Inhibiting Specifically Isohemoagglutination
Authors: Yamakami K.
Published: December 2022

In the course of an investigation, made for the purpose of determining whether the cells of various tissues can absorb the isohemagglutinins with the same degree of specificity as the erythrocytes, the very remarkable fact has been discovered in my laboratory by S. Shirai that human semen manifests a very marked inhibiting power upon isohemoagglutination, and, moreover, there is a fair degree of specificity in this inhibition.

Just in the like manner as the red blood cells of group I blood (Jansky) do not absorb the isohemoagglutinins, and consequently can not alter the agglutinating power of any serum, the semen produced by a man of group I blood has either no influence upon the agglutinating power of serum of any blood group, or its inhibiting influence is impartial upon all types of sera, if some inhibition occurs.
Smooth Muscle in Anaphylactic Guinea Pigs
Authors: Alexander H. L. , Holmes J. A. , Becke W. G.
Published: December 2022

Anaphylaxis is essentially a cellular phenomenon. Of the tissues that are known to participate in anaphylactic shock, that of smooth muscle has received most attention. Both Schultz (1) and Dale (2) by their classical experiment demonstrated that when smooth muscle of a sensitized animal was suspended in a bath of oxygenated Ringer's solution, it contracted when it came in contact with homologous antigen added to the bath. Likewise, Manwaring and his co-workers (3) showed that in living sensitized dogs, when certain organs containing smooth muscle were perfused with the antigen with which these animals had been injected, contraction occurred. Smooth muscle, also, has been shown to be responsible for many of the symptoms of anaphylactic shock. Examples of this are the classical asphyxia of guinea pigs from broncho-constriction, as described by Auer and Lewis (4), and the pulmonary symptoms in rabbits which Airila (5), Coca (6), and others have demonstrated to be due to vascular spasm.
Spontaneous Regression of Friend Virus-Induced Erythroleukemia
Authors: Marcelletti John , Furmanski Philip
Published: December 2022

The spontaneous regression of erythroleukemia induced by the RFV strain of Friend virus is a macrophage-dependent process. Functional suppression or elimination of the macrophage population in leukemic mice with silica, carrageenan, anti-macrophage serum, or trypan blue inhibited regression. Prior protection of the macrophages with PVNO allowed regression in silica or carrageenan-treated mice. Macrophage phagocytic activity was inhibited in about half the RFV-induced leukemic mice at 25 to 30 days post virus inoculation. Those animals with normal macrophages regressed, whereas those with inhibited macrophages did not. Progressor mice could be induced to regress by inoculation with normal syngeneic macrophages; other cell types were ineffective.

The inhibition of macrophage function in leukemic mice was the result of infection of the macrophages by virus. Removal of the infected cells by cytolysis with anti-virus antiserum and C restored the phagocytic activity of the population. Inhibited macrophages were less capable of responding to immobilized antigen-antibody complexes than normal macrophages, suggesting that the loss of function was due to a change in their Fc receptor.
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

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

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

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

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

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

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

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

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.