bluuebunny/crossref_metadata_2025_split huggingface dataset:
Use of a Monoclonal Antibody (W6/32) in Structural Studies of HLA-A,B,C Antigens
Authors: Parham Peter , Barnstable Colin J. , Bodmer Walter F. 
Published: December 2022

Abstract
The experiments reported here show that W6/32 antigenic activity is retained in papain-solubilized HLA antigens and that the W6/32 antibody provides a useful standard reagent to detect and assay HLA-A,B,C antigens. The W6/32 antibody was purified and used to construct an immunoaffinity column. Soluble HLA-A,B,C antigens from papain or detergent-treated membranes could be highly purified in a single step with this column and serologic analysis showed that HLA-A,B and C antigens were bound to the column. Thus, the W6/32 antigenic determinant is present on gene products of all three loci.
The amount of HLA-A,B,C antigens on the surface of human B cell lines and peripheral blood lymphocytes was measured with W6/32 antibody. B cell lines expressed, on average, 9 times as much cell surface HLA-A,B,C antigens as peripheral lymphocytes, although there was appreciable variation within each group. The B cell line Bri 8, for example, expressed 1.5 × 106 W6/32 antigenic sites, and by inference, HLA-A,B,C molecules per cell. Equal amounts of β2m and HLA-A,B,C chain were found on both cell types.
The isolated HLA-A chain from intact 125I-HLA-A2 antigens weakly bound to W6/32 antibody in contrast to 125I-β2-microglobulin (β2m) isolated from the same preparation of HLA-A2 antigens that showed no demonstrable binding. when an excess of cold β2m was added to the isolated 125I-HLA-A2 chain, the binding to W6/32 antibody was considerably enhanced. These results suggest that the W6/32 antigenic determinant involves only amino acids of the HLA-A,B,C chain and is a product of their three dimensional configuration. Stable maintenance of this configuration appears to be dependent on the association of the HLA-A,B,C chain with β2m.
Studies on in vivo Priming of the TNP-Reactive Cytotoxic Effector Cell System
Authors: Fujiwara Hiromi , Levy Robert B. , Shearer Gene M. , Terry William D. 
Published: December 2022

Abstract
A comparison was made of the effects of i.v. inoculation of trinitrophenyl-(TNP) conjugated syngeneic cells on the subsequent in vitro generation of TNP-reactive effector cell activity and on the in vivo development of TNP-contact sensitivity. The administration of syngeneic TNP-conjugated spleen cells before 2,4,6-trinitro-1-chlorobenzene (TNCB) painting abolished the capability of animals to develop TNP-contact sensitivity. In contrast, the same treatment resulted in an appreciable augmentation in the generation of TNP-reactive cytotoxic effector cell activity as measured by subsequent in vitro sensitization with TNP-conjugated cells. The possible mechanisms by which enhanced TNP-reactive cytotoxic effector cell activity was elicited under conditions identical to those that induced unresponsiveness for TNP-contact sensitivity are discussed.
Activation of Human Natural Killer Cells Cytotoxic for Human Leukemia Cells by Purified Interferon
Authors: Zarling Joyce M. , Eskra Linda , Borden Ernest C. , Horoszewicz Julius , Carter William A. 
Published: December 2022

Abstract
Interferon (IF) inducers and IF-containing preparations of leukocyte origin have recently been reported to augment natural killer (NK) cell activity. Our studies were undertaken to determine whether highly purified human fibroblast interferon (HFIF), induced by polyinosinic: polycytidylic acid (poly I:C), would augment human NK cell activity against an NK-sensitive leukemia cell line, K562, and, more importantly, to determine whether HFIF would activate normal lymphocytes to lyse human leukemia cells that have not been adapted to tissue culture or established as cell lines. Pretreatment of peripheral blood mononuclear cells for 1 to 16 hr with purified HFIF (150 to 300 units/ml) caused 2.5- to 5-fold increases in cytotoxicity against the K562 cells. Both native and HFIF-augmented cytotoxicities were mediated by FcR+ lymphocytes (lymphocytes expressing receptors for the Fc portion of IgG). Removal of FcR+ lymphocytes, but not monocytes, prevented HFIF from augmenting cytotoxicity, suggesting that HFIF activates preexisting FcR+ NK cells and that monocytes are not required for this effect. Our findings that NK cell activity is augmented by poly I:C-induced fibroblast IF, which lacks lymphokines and viral products present in virus-induced leukocyte IF preparations previously used, together with the fact that our IF preparation was more highly purified than any previously used for such studies, strongly suggest that the IF protein itself augments NK activity.
HFIF-treated cells from some individuals lysed many patients' leukemia cells; however, the HFIF-treated cells were not cytotoxic for autologous or allogeneic normal lymphocytes or concanavalin A blasts. FcR+ cells were required for the HFIF-induced cytotoxicity against the patients' leukemia cells. These results thus demonstrate that not only leukemia cell lines, but also human leukemia cells that have not been adapted to tissue culture or established as cell lines, can be lysed by IF-activated human NK cells.
Studies on the Terminal Stages of Immune Hemolysis
Authors: Boyle Michael D. P. , Borsos Tibor 
Published: December 2022

Abstract
The inhibitory effects of 0.1 M EDTA on the lysis of E* prepared by incubating EA with whole GPC was studied. At high end point lysis (&gt;70%) 0.1 M EDTA failed to prevent hemoglobin release whereas at lower end point (&lt;60%) 0.1 M EDTA was effective. In all cases hemoglobin release was inhibited by 25% BSA. When E* were prepared by incubating EAC1-8 with C9, similar results were obtained. In this system the difference in the ability of 0.1 M EDTA to inhibit hemoglobin release at high or low end point lysis could not be correlated with the number of lesions/cell but appeared to be related to the C9 to SAC1-8 ratio. With limiting SAC1-8 and excess C9, E* were produced from which hemoglobin release could not be prevented by 0.1 M EDTA whereas at lower C9 to SAC1-8 ratios hemoglobin release was prevented by 0.1 M EDTA. These differences most probably reflect functionally different sized transmembrane channels that were produced at different C9 to SAC1-8 ratios.
Studies on the Terminal Stages of Immune Hemolysis
Authors: Boyle Michael D. P. , Gee Adrian P. , Borsos Tibor 
Published: December 2022

Abstract
We have previously shown that 0.1 M EDTA could be used to distinguish functionally different transmembrane channels produced during complement-(C) mediated hemolysis of E. In this paper we have studied the ability of sugars of varying Stokes' radii to prevent hemoglobin release from E* intermediates whose lysis was inhibitable or not inhibitable by EDTA. On the basis of these experiments we propose that the inhibition of E* transformation by high molarity EDTA occurs by virtue of the size of the EDTA molecule in solution. Studies on the effect of EDTA on red cell lysis induced by polyene antibiotics that form transmembrane channels of a defined size support this conclusion. The results of these experiments were interpreted to mean: 1) The EDTA inhibitable lesion on E* has a smaller effective radius than the noninhibitable lesion; 2) the effective radius of the smallest lesion that yields a lytic site was less than 3.6 Å; 3) the lesions produced in the red cell membrane by C are not uniform but vary in size depending on the C9 to SACI-8 ratio used to produce E*.
Latent Lymphokines: Isolation of Guinea Pig Latent Lymphocyte-Derived Chemotactic Factor for Monocytes. Its Activation by Trypsin and a Soluble Factor from Macrophages
Authors: Postlethwaite Arnold E. , Kang Andrew H. 
Published: December 2022

Abstract
Guinea pig lymphocytes when depleted of macrophages and stimulated by the T cell mitogen phytohemagglutinin produce a latent form of lymphocyte-derived chemotactic factor for monocytes (LDCF-M). Latent LDCF-M is also produced when immune lymphocytes are stimulated in vitro with specific antigen, horseradish peroxidase. Latent LDCF-M from both sources can be activated and converted to “classical” LDCF-M by trypsin and by a soluble factor obtained from sonicated macrophages. These observations suggest that macrophages may modulate lymphokine activities in vivo by releasing soluble factors that convert inactive latent lymphokines to biologically active substances.
Specificity and Suppressor Function of Human T Cells Responsive to Autologous Non-T Cells
Authors: Sakane Tsuyoshi , Green Ira 
Published: December 2022

Abstract
Human T cells proliferate in response to autologous non-T cells. This reaction, called the autologous MLR has been shown to have both immunologic specificity and memory. Our present study first demonstrates that specific secondary responses can be demonstrated in the autologous MLR. A BUdR and light technique was then used to ablate cells proliferating in response to either autologous or allogeneic non-T cells. The removal of T cells responsive to autologous non-T cells left the responsiveness to allogeneic T cells relatively intact, and conversely the removal of T cells responsive to allogeneic non-T cells left the responsiveness to autologous non-T cells relatively intact. Thus, the T cells responsive to autologous non-T cells and those responsive to allogeneic non-T cells are largely separate cell populations.
Since the lymphocytes from patients with SLE fail to demonstrate a normal autologous MLR and also have a defect in the development of Con A-induced suppressor cells, we investigated whether the T cell population responsive in the autologous MLR was particularly enriched in cells capable of developing suppressor capacity. A BUdR and light treatment was used to remove T cells responsive to either autologous or allogeneic non-T cells. The remaining cells were then activated by Con A, and their subsequent suppressor activity was measured. Removal of T cells proliferating in response to autologous non-T cells produced significantly greater failure of suppressor cell development than did the removal of T cells responsive to allogeneic non-T cells. Thus, T cells capable of responding to autologous non-T cells are particularly enriched in cells capable of becoming suppressor cells.
Human Monocyte, Lymphocyte, and Granulocyte Antibody-Dependent Cell-Mediated Cytotoxicity Toward Tumor Cells
Authors: Levy Paul C. , Shaw George M. , LoBuglio Albert F. 
Published: December 2022

Abstract
Previous investigations of antibody-dependent cell-mediated cytotoxicity (ADCC) by human leukocytes indicated that lymphocytes and granulocytes, but not monocytes, lysed tumor cell targets. We have recently shown that monocytes, too, can carry out ADCC toward at least three allogeneic tumor cell lines. This paper examines the characteristics of ADCC by all three Fc receptor-bearing human leukocytes (monocytes, lymphocytes, and granulocytes) toward the same antibody-coated 51Cr-labeled tumor target cells. For each leukocyte effector, ADCC was found to be temperature dependent and rapid, with most target cell lysis occurring within 4 hr of incubation. The extent of target cell destruction was positively related to the effector cell/target cell ratio and to the degree of antibody sensitization of the target cells. ADCC was clearly antibody dependent since the cytotoxic event did not involve nonantibody-coated “innocent bystander” target cells and was inhibited by staphylococcal Protein A and nonspecific monomeric immunoglobulin. Granulocyte ADCC required considerably larger amounts of target cell-bound antibody than did ADCC by lymphocytes and monocytes. Conversely, larger amounts of monomeric immunoglobulin were required to inhibit granulocyte ADCC as compared to monocyte and lymphocyte ADCC in experiments utilizing identical antibody-coated target cells. In innocent by-stander experiments, no evidence for a soluble mediator of target cell lysis was found for any of the effector leukocytes. We suggest that this experimental system which involves three Fc receptor-bearing human leukocyte populations and identical antibody-coated target cell preparations provides a new and unique method for studying this cell-mediated cytotoxic event.
The Role of H-2-Linked Genes in Helper T Cell Function
Authors: Swierkosz James E. , Marrack Philippa , Kappler John W. 
Published: December 2022

Abstract
We have studied the H-2 genes that control interactions between antigen-presenting macrophages (Mø) and helper T cells both during and after priming. B6AF1 (H-2b × H-2a) mice were initially primed in vivo with keyhole limpet hemocyanin (KLH)-pulsed Mø of either parental H-2 type, and their T cells were tested for helper activity in the in vitro anti-trinitrophenol (TNP) response to TNP-KLH. In each case, F1 T cells cooperated well only with B cells and Mø having the same K-I-A subregion as the Mø used for priming.
In a second approach, F1 mice were instead primed with free antigen, and T cells enriched in specific helper function were then isolated in vitro by adherence to antigen-pulsed parental Mø. The F1 T cells isolated in this manner exhibited enriched helper activity only when tested with B cells and Mø that had the K-I-A region type of the Mø used for isolation. With this technique, the same genetic restrictions were also found for responses to sheep red blood cells (SRBC).
A direct role for I-region genes in this system was established by the ability of anti-Ia alloantisera to specifically inhibit the enrichment of helper T cells on Mø monolayers. Taken together these results pinpoint I-A as the relevant subregion involved in the associative recognition of these complex antigens by helper T cells.
Autoimmune Cell-Mediated Responses: Spontaneous Responses to Syngeneic Cells in NZB/NZW F1 Mice
Authors: Senitzer David , Cafruny William , Raeder Roberta , Freimer Earl H. 
Published: December 2022

Abstract
The effect of various stimulator cell populations on B/W mouse spleen cell cultures was analyzed by measuring 3H-thymidine incorporation. The data show that syngeneic heart and kidney cells induce strong stimulation of DNA synthesis in B/W spleen cells. In contrast, syngeneic liver cells are not stimulatory. Autologous stimulation of adult B/W spleen cells by heart and kidney cells was also observed. BALB/c heart and kidney cells were stimulatory for B/W spleen cells, although BALB/c spleen cells were only slightly stimulated by B/W or BALB/c heart and kidney cells.
The responses to syngeneic heart and kidney cells were observed to peak at 7 to 8 days of culture. The magnitude of the observed responses did not appear to correlate with the age or sex of the B/W spleen cells used, and the ability to response to heart and kidney cells with large increases in DNA synthesis occurred at a very early age. Spleen cells from old B/W mice lose the ability to become stimulated by concanavalin A, although no concomitant loss of reactivity to syngeneic heart or kidney cells is observed. The presence of θ-bearing lymphocytes in B/W spleen cell populations is not required during culture initiation for the stimulation of DNA synthesis by syngeneic heart or kidney cells. However, during the peak of DNA synthesis, the majority of responder spleen cells carry θ-antigen.
We postulate that antigenic determinants present on heart and kidney cells induce in vitro cell-mediated immune responses of an autoimmune nature in B/W spleen cell populations. The results are in aggrement with an early defect in immunologic regulation that persists throughout the life of the B/W mouse.
Cloning of B Cells Positive or Negative for Surface IgD
Authors: Layton Judith E. , Teale Judy M. , Nossal G. J. V. 
Published: December 2022

Abstract
As in the accompanying paper, spleen cells from 2½-week-old mice, containing equal proportions of sIgD+ and sIgD- B cells were separated in the fluorescence-activated cell sorter into sIgD+ and sIgD- populations. The activity of each population and the effect of tolerance induction were measured in a T-dependent B cell-cloning assay: the splenic focus assay of Klinman and colleagues. The anti-DNP precursor frequency was found to be essentially the same in each population when calculated on a per B cell basis. In addition, the response from the sIgD- population was unlikely to be from cells that had acquired sIgD during the culture period, since the presence of anti-δ serum in the cultures did not inhibit the response from this population. The isotype of the antibody produced by responding clones was analyzed and the results showed that the sIgD- population produced a slightly higher proportion of clones secreting IgM only, and a lower proportion of IgG-secreting clones, than the sIgD+ population. Tolerance induction in the separated populations indicated that the sIgD- subset was only marginally susceptible to tolerogenesis whereas the sIgD+ subset was resistant. Furthermore, if the expected contribution of the highly tolerizable sIg- cells contained in the sIgD- subset was subtracted, the degree of tolerance was reduced to an insignificant level. Thus, the response to a T-dependent antigen is not restricted to the sIgD+ subset of cells and resistance to tolerogenesis does not appear to be caused by acquisition of sIgD, even when a T-dependent challenge antigen is used. The results are discussed in the light of other work on the function of IgD.
Nature of T Lymphocyte Recognition of Macrophage-Associated Antigens
Authors: Thomas David W. , Meltz Susan K. , Wilner George D. 
Published: December 2022

Abstract
The immune response of strain 2, strain 13, and (2 × 13)F1 guinea pigs was studied after immunization with human fibrinopeptide Bβ(hFPB), a 14 amino acid (Bβ1-14) thrombin-derived fragment of fibrinogen. Several weeks after immunization, strain 2 and (2 × 13)F1 antimals showed a positive anti-hFPB delayed-type hypersensitivity skin test and produced a dramatic hFPB-specific T lymphocyte proliferative response in vitro, as assessed by the incorporation of 3H-thymidine. Strain 13 animals immunized with hFPB did not produce an anti-hFPB skin test and showed little or no in vitro T cell proliferation with hFPB. Neither guinea pig strain produced detectable anti-hFPB antibody as determined by radioimmunoassay. Immune T cells were also tested for their ability to respond to several sequential synthetic homologues of hFPB. Strain 2 and (2 × 13)F1 T cells immunized with hFPB (Bβ1-14) were stimulated by fragments Bβ3–14, 5–14 and 7–14, but not by fragments Bβ1–13 and 9–14. In addition, strain 2 and strain 13 guinea pigs were immunized with the various homologues themselves and the immune T cells tested for responsiveness in vitro with hFPB and a battery of the fragments. Strain 2 T cells immunized with Bβ1–13 showed no proliferative response with Bβ1–14, 3–14, 5–14 and 7–14, and neglible stimulation with the homologous antigen Bβ1–13. After immunization with Bβ5–14 and 7–14, strain 2 T cells showed a proliferative response with Bβ1–14, 3–14, 5–14, and 7–14, but not with Bβ1–13. T cells from strain 2 animals immunized with B 9–14 were unresponsive with all homologues tested. T cells from strain 13 guinea pigs immunized with Bβ1–13 produced a dramatic proliferative response with Bβ1–13, but were unresponsive with Bβ1–14, 3–14, 5–14, and 7–14. Strain 13 animals immunized with Bβ5–14 and 9–14 showed no T cell response with any of the homologues tested, whereas Bβ7–14 immune strain 13 T cells produced a small response with Bβ7–14. These results are discussed with respect to T cell recognition of specific amino acid sequences of the fibrinopeptide molecule.
Ia Antigens in Mouse Skin Are Predominantly Expressed on Langerhans Cells
Authors: Tamaki Kunihiko , Stingl George , Gullino Marisa , Sachs David H. , Katz Stephen I. 
Published: December 2022

Abstract
We have investigated the expression of products of the mouse major histocompatibility complex (MHC) on BALB/c and A/J epidermal cells. By using reagents with specificity for various products of the MHC in an indirect immunofluorescence procedure, we found that H-2 antigens are expressed on the vast majority of epidermal cells. Ia antigens, by contrast, are present on only 2.4 to 6.9% of all epidermal cells. These Ia-bearing cells bear a receptor for the Fc portion of IgG and ultrastructurally exhibit the characteristics of Langerhans cells. Ia antigens on Langerhans cells are encoded for by at least the I-A and I-E/C subregions of the MHC.
Active Disassembly of the First Complement Component, C1̄, by C1̄ Inactivator
Authors: Ziccardi Robert J. , Cooper Neil R. 
Published: December 2022

Abstract
In order to clarify its mechanism of disassembly, the first component of human complement, C1, was reconstituted from C1q, 125I-C1r, and 131I-C1s. This radiolabeled C1 bound tightly to aggregated IgG covalently linked to Sepharose 4B. The subsequent addition of C1̄ inactivator (C1̄-In) led to the rapid release of equimolar quantitites of 125I-C1r̅ and 131I-C1s̅ from the agg IgG. SDS-PAGE analyses of such eluates showed that one C1̄-In molecule was associated with each C1r̅ polypeptide chain and another C1̄-In molecule was bound to each C1s̅ polypeptide chain. The released C1r̅, C1s̅, and C1̄-In co-sedimented in sucrose density gradients with a rate of 9S and monospecific anti-C1s̅ pelleted all three proteins. Therefore, C1r̅, C1s̅, and C1̄-In, in a molar ratio of 1:1:2, respectively, are released from C1̄ as a complex.
An analogous 9S complex containing C1r̅, C1s̅, and C1̄-In was generated in normal human serum after incubation with activators of the classical complement pathway. The C1r̅C1s̅(C1̄-In) complexes generated in both serum and the purified system were stable in the presence of EDTA.
A diffusion coefficient of 2.3 × 10-7 cm2/sec was determined for the C1r̅C1s̅(C1̄-In) complex from its behavior on gel filtration. An m.w. of 330,000 was then calculated from its sedimentation and diffusion coefficients. This m.w. is consistent with the value of 382,000 for a molecule of composition C1r̅C1s̅(C1̄-In)2 obtained by summing the weights of the subunits. These results indicate that C1-In efficiently disassembles C1̄, thereby releasing two C1r̅C1s̅(C1̄-In)2 complexes per C1̄ molecule.