bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibody-Induced Suppression of the Fifth Component of Complement in Mice
Authors: Goldman John N. , Goldman Margaret B.
Published: December 2022

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

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

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 (>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

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

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

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

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<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 > IgE BA > 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<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.
Immunologic Rejection of Mammary Adenocarcinoma (TA3-St) in C57BL/6 Mice: Participation of Neutrophils and Activated Macrophages with Fibrin Formation
Authors: Dvorak Ann M. , Connell Anne B. , Proppe Karl , Dvorak Harold F.
Published: December 2022

The ultrastructural cellular kinetics of an in vivo model of immunologic rejection was studied with the strain-specific TA3-St mammary adenocarcinoma in C57BL/6 mice. The peritoneal cavity of tumor-challenged mice had numerous neutrophils, a few lymphocytes, and normal macrophages at early times after challenge. Neutrophils formed contacts with viable tumor cells, remained viable, nonphagocytic, and did not extrude their cytoplasmic granules. Tumor cells so contacted showed morphologic evidence of cell injury followed by cell death. Later times after tumor challenge showed numerous activated macrophages rapidly phagocytosing tumor debris resulting in complete elimination of tumor at 24 hr. These macrophage aggregates had numerous elongated interdigitated villi and a prominent fibrin meshwork associated with their processes and cell surfaces.

The findings in this study demonstrate the morphologic participation of neutrophils in tumor rejection with the later participation of phagocytic, activated macrophages whose surfaces were covered with fibrin. The mechanism for attraction and binding of neutrophils to target tumor cells is not known. Possible similarities to the recently described role for neutrophils in antibody-dependent cellular cytotoxicity should be considered. The mechanism of tumor killing by neutrophils was not apparent by ultrastructural analysis since extrusion of cytoplasmic granules was not present. Release of potent cytotoxic materials from these cells by other anatomic routes is postulated. Activated macrophages participated primarily as scavengers of dead tumor. The significance of their surface-associated fibrin meshwork requires further study.
Isolation and Characterization of Naturally Occurring Subclasses of Human Peripheral Blood T Cells with Regulatory Functions
Authors: Strelkauskas Anthony J. , Schauf Victoria , Wilson Barry S. , Chess Leonard , Schlossman Stuart F.
Published: December 2022

By utilizing naturally occurring autoimmune antibodies from patients with juvenile rheumatoid arthritis, we have isolated and functionally characterized two unique subpopulations of T cells. JRA + T cells, i.e., those identified by sera from these patients, react poorly in response to allogeneic cells, respond to Con A but not PHA, and do not help in the synthesis and secretion of Ig by B cells. In contrast, JRA- T cells, i.e., those not identified by sera from these patients, respond very well to allogeneic cells, proliferate well in response to PHA but not Con A, and more interestingly, can greatly enhance the secretion of Ig by B cells.
Suppressor Cell Activity in Tumor-Bearing Mice
Authors: Elgert Klaus D. , Farrar William L.
Published: December 2022

Spleen cells from mice bearing methylcholanthrene-induced fibrosarcomas impaired mitogen responses of normal syngeneic lymphocytes. Nylon wool column and other depletion techniques were utilized to characterize the cellular source of suppressive activity in tumor-bearing host (TBH) spleens. Evidence is presented for two distinct suppressor cell systems operating in the spleens, but not lymph nodes, of BALB/c mice bearing transplanted tumors. Spleens from TBH were shown to have greatly increased numbers of macrophages over their normal counterparts. TBH macrophages were observed to have suppressive activity at low in vitro concentrations. Anti-Thy 1 serum treatment of TBH macrophages abrogated low dose inhibition but not suppression due to high numbers of macrophages. No functional difference was detected between anti-Thy 1 serum-treated TBH and normal splenic macrophages. In a macrophage-depleted culture system, mildly nylon wool adherent, anti-Thy 1 serum, and hydrocortisone succinate-sensitive suppressor cells could be detected. Soluble supernatant products of TBH spleen and thymus cells were also found to inhibit in vitro mitogen responses, whereas TBH macrophages and lymph node cells demonstrated no soluble suppressive activity. The major source of soluble inhibitor of DNA synthesis (IDS) seems to be an anti-Thy 1 serum, hydrocortisone-sensitive population.
Imbalances in T Cell Subpopulations in Multiple Sclerosis Patients
Authors: Santoli Daniela , Moretta Lorenzo , Lisak Robert , Gilden Donald , Koprowski Hilary
Published: December 2022

Abnormal proportions of a distinct T cell subpopulation able to bind IgG immune complexes (T.G cells) were found in peripheral blood samples from patients with MS. About 50% of the patients examined had an overabundance of T.G cells. The possible role of these cells in the pathogenesis of MS is considered.
The Modulation of the Alternative Pathway of Complement in C2-Deficient Human Serum by Changes in Concentration of the Component and Control Proteins
Authors: Nydegger Urs E. , Fearon Douglas T. , Austen K. Frank
Published: December 2022

The opposing actions of component and regulatory proteins of the alternative pathway of C on activation of the sequence were demonstrated in C2-deficient serum utilizing two initiating particles and incremental additions of proteins purified to homogeneity. Activation of the alternative pathway in C2-deficient serum by zymosan was assessed by measurement of the inactivation of C3 and B, while activation by rabbit erythrocytes (ER) was measured by their lysis. Participation of the classical C-component sequence was excluded by the presence of Mg-EGTA and by the failure of reconstitution of C2 to have a measurable effect. Incremental additions of either purified C3bINA or β1H, which increased the serum concentrations of the proteins by 43% to 175% or 15% to 30%, respectively, produced dose-related suppression of zymosan-induced B and C3 inactivation throughout the 45 min period of observation. Additions of either C3 or B to C2-deficient serum so as to increase their concentrations by 35% reversed the combined 50%-inhibitory effect of added C3bINA and β1H on the zymosan-induced inactivation of C3 and B. Dose-dependent inhibition of the rate and extent of lysis of ER in C2-deficient serum was obtained with increases of C3bINA concentrations of 25 to 100% and of β1H concentrations of 12.5 to 50%. As little as a 25% increase in the concentration of B increased the rate of lysis of ER in C2-deficient serum and partially reversed the inhibitory effect of the combined addition of C3bINA and β1H. Thus, the disequilibrium between component and control proteins of the alternative pathway created by the surface of an activating particle, that permits transition to the amplification phase of C3 cleavage, can be modulated by alterations in the concentrations of these proteins. Modest increases in the concentrations of C3bINA and β1H served to maintain balance, whereas comparable augmentations in the concentrations of C3 and B facilitated activation by the initiating particle.
Isolation and Characterization of a Rabbit T Lymphocyte-Specific Antigen
Authors: Navran James E. , Archer Stanley J.
Published: December 2022

Rabbit T lymphocytes may be differentiated from B lymphocytes by the presence of a T lymphocyte-specific surface antigen. This unique antigen has been extracted from the plasma membrane of rabbit thymocytes by 3 M KCl. The presence of the antigen in the membrane extract was demonstrated by inhibition of cytotoxicity with goat anti-rabbit T cell serum (ATS). The crude membrane extract was fractionated by gel electrophoresis and the fractions containing the T cell antigen identified by inhibition of cytotoxic ATS and by passive hemagglutination. The purified T cell antigen was found to have a m.w. of approximately 12,000 and contained approximately 2.5% carbohydrate. Evidence was also obtained to suggest that the rabbit T cell antigen exists in multiple forms, each having the same m.w. but exhibiting different electrophoretic characteristics.
Immunologic Factors Influencing the Intra-tumor Localization of ADCC Effector Cells
Authors: Haskill Stephen , Parthenais Elaine
Published: December 2022

The role of cellular and humoral immunity in the localization of blood-borne, bone marrow-derived ADCC effector cells into the T1699 mammary adenocarcinoma was investigated. Administration of ALG before tumor inoculation caused total immunosuppression and resulted in minimal in situ inflammation. ALG treatment started at the time of tumor inoculation suppressed the delayed hypersensitivity response below a detectable level but permitted a significant antibody response. Under these circumstances, the localization of ADCC effector cells into the tumors appeared normal. Similarly, administration of ALG at later stages of tumor growth, where ALG acts as an anti-inflammatory agent, did not interfere with the normal infiltration of ADCC effector cells in situ, although the delayed hypersensitivity response was totally suppressed. When melphalan treatment was used to produce tumor-bearing mice with an intact delayed hypersensitivity response but devoid of a significant antibody response, the drug-treated animals were found to have high levels of ADCC effector cells in situ. These results demonstrate that although the in situ inflammatory reaction appears to be immunologically dependent, neither the delayed hypersensitivity nor the antibody response is solely responsible for the localization of ADCC effector cells in the T1699 mammary tumor.
Differential Cytotoxic Effect of Natural Thymocytotoxic Autoantibody of NZB Mice on Functional Subsets of T Cells
Authors: Shirai Toshikazu , Hayakawa Kyoko , Okumura Ko , Tada Tomio
Published: December 2022

The effect of the natural thymocytotoxic autoantibody (NTA) of NZB mice on the functional T cell subsets was studied in an in vitro secondary antibody response. Dinitrophenylated keyhole limpet hemocyanin (DNP-KLH)-primed BALB/c spleen cells were treated with appropriate dilutions of NTA and complement, and then stimulated with DNP-KLH. Whereas the cytotoxic treatment of DNP-KLH-primed spleen cells with a low dilution of NTA and complement slightly suppressed the anti-DNP antibody response, the treatment at a higher dilution resulted in a significant enhancement of the antibody response. The enhancement was found due to the selective elimination of a part of T cells including suppressor T cells, which were more sensitive to NTA than helper T cells. This was confirmed by an experiment in which KLH-primed spleen cells were treated with appropriate dilutions of NTA and complement and then co-cultured with DNP-KLH-primed B cells resulting in a marked enhancement of the anti-DNP antibody response. The treatment of KLH-primed spleen cells with NTA in the same manner resulted in the loss of activity of the extractable antigen-specific suppressive T cell factor (TsF). Both antigen-specific helper T cells and memory B cells were found to be highly resistant to the cytotoxicity of NTA. Fluorescence-activated cell sorter (FACS) analyses revealed that the cytotoxic treatment of splenic T cells with NTA with an appropriate dilution selectively eliminated the T cells bearing a high density of NTA-reactive antigen. These results suggest that NTA is responsible for the selective loss of the suppressor T cell function that appears early in life in New Zealand mice and plays, at least in part, a causative role in the development of autoimmune diseases.
Structure and Function of Immunoglobulin Domains
Authors: Barnett Foster Debora E. , Dorrington Keith J. , Painter Robert H.
Published: December 2022

The ability of various fragments of human myeloma IgG1 to inhibit rosette formation between human anti-D-coated human red blood cells and human polymorphonuclear leukocytes has been investigated. Although IgG and Fc showed a dose-dependent inhibition of rosettes at equimolar concentrations, neither of the fragments corresponding to the Cγ2 and Cγ3 homology regions obtained by acid-tryptic cleavage of Fc was able to inhibit rosette formation. The pepsin fragment of Fc, pFc′, which represents the complete Cγ3 domain, was also unable to prevent rosette formation. Reduction and alkylation of IgG or Fc markedly diminished cytophilic activity as measured by this system. These data indicate that the site in human IgG1 bound by granulocytes is dependent on the full quaternary structure of Fc, a requirement in marked contrast to that noted for binding by macrophages and monocytes.
Suppression of Polyclonal B Cell Activation in Scrapieinfected C3H/HeJ Mice
Authors: Garfin David E. , Stites Daniel P. , Zitnik Louise A. , Prusiner Stanley B.
Published: December 2022

A consistent modification in B lymphocyte activation has been observed 1 month after infection of C3H/HeJ mice with scrapie. The mitogenic response to lipopolysaccharide of splenocyte cultures from experimental mice was reduced 30 to 60% as compared to controls. This reduction in mitogen responsiveness was transient but coincided with the onset of detectable splenomegaly and with the reported recovery of maximum yield of infectious scrapie agent in the spleen. The DNA synthetic response to lipopolysaccharide stimulation of splenocytes from scrapie-infected C3H/HeJ mice was depressed relative to controls only between 20 and 40 days after intracerebral inoculation. At all other times, experimental and control responses were identical. Scrapieassociated decreases in mitogenesis were found whether the spleen cell cultures contained splenocytes from individual mice, splenocytes pooled from several mice, or gradient-purified mononuclear cells. The responses of C3H/HeJ splenocyte cultures to phytohemagglutinin or concanavalin A stimulation were unaffected by scrapie infection.
Regulation of IgE Antibody Production by Serum Molecules
Authors: Katz David H. , Tung Amar S.
Published: December 2022

IgE antibody production in mice of high and low IgE responder phenotypes, respectively, can be appreciably enhanced in magnitude after low dose whole body X-irradiation. Such enhanced responses, as well as adoptive secondary IgE responses, can be markedly suppressed by passive transfer of CFA-immune serum in low responder strains, but not in high responder strains. The studies presented here demonstrate that the suppressive activity of CFA-immune serum on IgE antibody production is strain specific. This is true even in reciprocal combinations of low IgE responder SJL and C57BL/6 mice, in which it was shown that serum capable of suppressing mice of the isologous strain was ineffective in diminishing IgE antibody production in the other low responder strain. Absence of suppressive activity in CFA-immune sera obtained from H-2 congenic donors which differed in their H-2 haplotypes while sharing many similarities in the background genome and, conversely, effective suppressive activity of H-2 congenic donor sera when H-2-identities between donor and recipient mice existed, strongly suggested a role, at least in part, of H-2 genes in dictating the strain specificity of such suppressive activity. Additional experiments provided evidence for a possible role of macrophages in catabolism of the active molecules in CFA-immune sera. These observations, together with those presented in the preceding paper, may provide valuable insight toward successful development of appropriate manipulations that could ultimately convert high IgE responder individuals into low responders.
Relationship of Expression of a Cell-Surface Antigen on Activated Murine Macrophages to Tumor Cell Cytotoxicity
Authors: Kaplan Alan M. , Bear Harry D. , Kirk Louise , Cummins Cecil , Mohanakumar T.
Published: December 2022

A cell surface antigen (AMøCSA) associated with activated macrophages was induced in vivo on murine peritoneal macrophages by pyran copolymer or Corynebacterium parvum but not by glycogen or thioglycollate. The antigen was detected by complement-dependent cytotoxicity by using an appropriately absorbed rabbit antiserum to the P388D1 macrophage cell line. Antigen was detectable on 90 to 100% of peritoneal macrophages 6 hr after i.p. inoculation of pyran or C. parvum and gradually declined to background levels, both with respect to the number of positive cells and the amount of antigen per cell, by 30 days after induction. Activated macrophages treated with pronase but not trypsin, neuraminidase, or β-galactosidase were no longer susceptible to lysis with AMøCSA-specific antiserum and C, but they could still be lysed with an antiserum directed to an organ-specific macrophage antigen shared by both normal and activated macrophages. Studies with four bacterial vaccines, Propionibacterium acnes, type I; two strains of P. acnes, type II, and P. granulosum have indicated that only two vaccines, P. acnes, type I and one strain of P. acnes, type II, could induce: a) regression of the MCA 2182 fibrosarcoma when inoculated intralesionally, b) splenomegaly, and c) peritoneal macrophages cytotoxic to tumor cells in vitro. The new antigen associated with activated macrophages was only detectable on macrophages activated by the two vaccines, P. acnes, types I and II, that induced functionally activated cells. These results are consistent with the induction of a new cell-surface antigen associated with activated but not with normal or elicited macrophages.
Genetic Control of the Immune Response to Phage fd
Authors: Kölsch Eckehart , Falkenberg Frank W.
Published: December 2022

The immune response to bacteriophage fd has been tested in congenic resistant strains of mice. B10 mice are high responders; B10.BR and B10.D2 mice are low responders. B10.A mice with the recombinant H-2a haplotype as well as heterozygous H-2k/d (B10.BR × B10.D2) F1 mice respond with high antibody titers, indicating the existence of two complementing Ir genes. From the results obtaiend with recombinant and F1 hybrid mice, Irfdα can be localized to the right of the crossover between the IE and IC subregions, Irfdα to the left of IB, presumably at IA. Complementation is observed with selected pairs of alleles only, suggesting α-β coupled complementation.

Low responsiveness of B10.A(2R) and (B10.A × B10.A(2R)) F1 mice is caused by a specific suppressor gene located at the Db subregion. This suppressor gene is active only with selected Irfdβ alleles.

In addition to genes in the H-2 complex, there are non-H-2 background genes modulating H-2 regulated anti-fd response.

B10.BR mice lack primary IgM and IgG antibodies, yet are able to mount a secondary IgG anti-fd response, giving evidence for intact memory formation in this low responder strain.

The combined data are most easily interpreted by assuming that both H-2 and non-H-2-genes modulate the effective immunogenic dose of antigen.
Separation and Partial Characterization of a Type-Specific Antigen from Chlamydia Trachomatis
Authors: Sacks David L. , Rota Teresa R. , MacDonald A. Bruce
Published: December 2022

Type-specific antigens of Chlamydia trachomatis have been demonstrated by the mouse toxicity prevention test and a variety of immunofluorescent techniques. In addition, biologic activity has been associated with these antigens in terms of type-specific immunity to trachoma infections. This report is the first to describe the detection of a soluble type-specific antigen of C. trachomatis and its separation from those antigens that cross-react among different immunotypes. Test antigens were prepared by labeling the surface components of purified, yolk sac grown organisms with a radioiodinated intermediate (Bolton-Hunter reagent, 125I). The organisms were solubilized with Triton X-100 and gel filtered through Sepharose 6B. All fractions were then tested in radioimmunoassay for binding with rabbit antisera raised against solubilized immunogens prepared from homologous and heterologous strain organisms propagated in BHK-21 cells. A fraction demonstrating homologous binding only was used in subsequent modified procedures for the preparation of quantities of type-specific antigen sufficient for analysis. The antigen appears to be a heat labile, cell surface protein associated with apparent immunogenic activity during the course of actual chlamydial eye infection.
A Specific Indicator for Mouse Lymphocyte C3b Receptors
Authors: Linscott William D. , Ranken Raymond
Published: December 2022

IgM-sensitized sheep E bearing guinea pig C4 and C3b (EAC43bgp) react as strongly with mouse spleen cell C3b receptors as do optimally prepared EACmo. Yet, when the C3b is converted to C3d by incubation with heated serum at low ionic strength, reactivity with mouse spleen cells is completely lost whereas binding to guinea pig spleen cells remains high. It is concluded on the basis of this and other evidence that EAC43bgp are a sensitive and specific indicator for mouse lymphocyte C3b receptors. Degradation of the C3b on these cells will not cause misleading binding to C3d receptors, as could occur with cells bearing mouse C3b.
Low Density of Thy 1 Antigen on Mouse Effector Cells Mediating Natural Cytotoxicity Against Tumor Cells
Authors: Herberman Ronald B. , Nunn Myrthel E. , Holden Howard T.
Published: December 2022

Mouse effector cells mediating natural cell-mediated cytotoxicity against tumor cells were found to contain a low density of Thy 1 antigen. Treatment of nude spleen cells, or spleen cells from mice in which natural reactivity was boosted, with anti-Thy 1.2 plus complement resulted in a substantial decrease in cytotoxicity. The spontaneous cytotoxic reactivity of young, thymus-bearing mice was resistant to such treatment, but repeated exposure to anti-Thy 1.2 plus complement did cause a decrease in lytic activity. By use of congenic anti-Thy 1.2 and effector cells from mice congenic for Thy 1, the effects of the treatment were shown to be specific for Thy 1.2 antigen.
Characteristics of BALB/C T Cell Lymphomas Grown As Continuous in Vitro Lines
Authors: Kim Kyung Jin , Weinbaum Fredric I. , Mathieson Bonnie J. , McKeever Paul E. , Asofsky Richard
Published: December 2022

Transplanted lymphomas (Thy 1.2+, Ig-) of BALB/c mice, induced by the injection of 1-ethyl-1-nitrosourea, were adapted for growth as in vitro lines to provide potential tools for investigation of T lymphocyte differentiation and functions. All these tissue culture lines maintained the same pattern of surface differentiation antigens (Ly, TL, and Thy-1 antigens) as they had expressed during in vivo passages: BALENTL 13 was Thy 1.2+, TL.2-, and Ly 1+2-. BALENTL 3, 4, 5, 6, 7, 8, and 14 were Thy 1.2+, TL.2+, and Ly 1-2+. P1798 and BALENTL 9 were Thy 1.2+, TL.2+, and Ly 1+2+. There were various levels of terminal transferase activity present among these T cell tumor lines. The range of variation was from 4.6 units/108 cells to 29.3 units/108 cells (normal thymocytes, 5.0 units/108 cells). This 6-fold variation in TdT activity was present even among those cell lines which were Ly 1-2+, TL+.

Most culture lines had chromosome numbers near 40 and generation times of 11 to 22 hr. There were no significant morphologic changes after the adaptation of these tumors in culture except an increase in cytoplasmic C-type virus particles.
Kinetic Assessment of Alternative Complement Pathway Activity in A Hemolytic System
Authors: Polhill Rutherford B. , Newman Simon L. , Pruitt Kenneth M. , Johnston Richard B.
Published: December 2022

By using a kinetic assay, we have examined the role of antibody in the lysis of rabbit erythrocytes (RaRBC) through the alternative complement (C) pathway. Sera from some hypogammaglobulinemic (Hγ) humans and all agammaglobulinemic chickens tested had subnormal activity in the assay. Heated normal human or chicken sera, but not heated Hγ sera, restored activity to deficient Hγ serum and initiated hemolysis in the presence of rabbit serum as C source. Absorption of heated normal human serum with RaRBC, but not with sheep erythrocytes or zymosan, removed its ability to reconstitute deficient Hγ serum. Normal hemolytic activity could be restored to Hγ serum with human IgM, IgG, or colostral IgA, with goat anti-RaRBC IgG, or with an eluate from serum-sensitized RaRBC, but not with myeloma IgA. Restoration of hemolytic activity to Hγ serum could be achieved in a dose-dependent fashion with the F(ab′)2 fragment of IgG. These results suggest that antibody exerts a significant rate-limiting effect on alternative pathway activity in the RaRBC lytic system. This raises the possibility that antibody may be required for efficient alternative pathway activity in vivo and that the pyogenic infections that occur in Hγ individuals are due to inefficient activation and fixation of C3 through either the classical or alternative pathway.
Cytotoxicity of Human Peripheral Lymphocytes in Cell-Mediated Lympholysis; Antibody-Dependent Cell-Mediated Lympholysis and Natural Cytotoxicity Assays after Mixed Lymphocyte Culture
Authors: Callewaert Denis M. , Lightbody James J. , Kaplan Joseph , Jaroszewski Jan , Peterson Ward D. , Rosenberg Jerry C.
Published: December 2022

Lymphocytes that have been purified by Ficoll-Hypaque centrifugation lose antibody-dependent and natural cytotoxic activities upon culture in tissue culture medium supplemented with human plasma. However, stimulation of peripheral lymphocytes in the mixed leukocyte culture (MLC) appears to enhance killer (K) and natural killer (NK) activities in addition to generating cytotoxic T lymphocytes.

Enhancement of NK and antibody dependent activities appears to correlate with cell division as measured by 3H-thymidine uptake. However, elimination of dividing cells in the MLC by addition of 5-bromodeoxyuridine has no effect on NK and K cell activities. Since this treatment abolishes cell-mediated lympholysis mediated by cytotoxic T lymphocytes, it is a useful probe for determining the relative activities of NK, K, and cytotoxic T lymphocyte effector cells after lymphocyte stimulation.
Ontogeny of Lymphocyte Functions during Embryonic Life of the Rabbit
Authors: Cazenave Pierre-André , Juy Dominique , Bona Constantin
Published: December 2022

Various immunologic functions of rabbit lymphocytes were studied during ontogeny. Liver cells of 17-day-old fetuses responded to concanavalin A and only after 22 days responded to phytohemagglutinin. The thymus cells of 22-day-old fetuses and the spleen cells of 29-day-old fetuses were stimulated by concanavalin A but not by phytohemagglutinin. The response to phytohemagglutinin in thymus and spleen cells was obtianed only after the birth. These results suggest that, in the rabbit, there are different subsets of T lymphocytes that respond either to concanavalin A or phytohemagglutinin. An alternative hypothesis is that the receptor for concanavalin A and the receptor for phytohemagglutinin appear on the same cells at different stages of the development.

No stimulation of fetal liver was obtained with B mitogens, i.e., Nocardia water soluble mitogen (NWSM) and antisera against allotypes of the b series whereas neonatal spleen cells were stimulated. These results indicate that the ability of lymphocytes to respond to a nonspecific rabbit B cell mitogen, namely NWSM, and to a specific rabbit B cell mitogen, namely anti-allotypic antibody, matured only in peripheral lymphoid organs, such as spleen, in spite of the fact that these two mitogens stimulate different subsets of B lymphocytes.

Significant synthesis of immunoglobulins by liver cells was obtained in 22-day-old fetuses. The liver cells possessed Fc receptors after 17 days of gestation and a significant antibody dependent cell-mediated cytotoxicity after 22 days. Both functions decreased after birth in these cells. These results showed a sequential acquisition of immunologic functions of rabbit lymphocytes.
Definition and Synthesis of the Essential Amino Acid Sequence for Experimental Allergic Encephalomyelitis in Lewis Rats
Authors: Hashim George A. , Carvalho Evelyn F. , Sharpe Richard D.
Published: December 2022

The amino acid sequence of peptide S8, defined by residues 69 to 84 of the bovine myelin basic protein, was synthesized and shown to induce experimental allergic encephalomyelitis (EAE) in Lewis rats. Comparison of the sequence of peptide S8, H-Gly-Ser-Leu-Pro-Gln-Lys-Ala-Gln-Gly-His-Arg-Pro-Gln-Asp-Glu-Asn-OH, to the corresponding region of the guinea pig myelin basic protein revealed the absence of Gly-His, residues 77 and 78, from the latter. Deletion of Gly-His from peptide S8 rendered the resulting peptide S49, a potent encephalitogen in Lewis rats.

Studies done with sequences shorter than synthetic peptide S49 revealed that the sequence of peptide S50, H-Ala-Gln-Arg-Pro-Gln-Asp-Glu-Asn-OH, made up of the C-terminal eight residues of peptide S49, was as encephalitogenic as the parent peptide. The sequence of peptide S50, not found naturally in any of the reported basic protein sequences, was generated by modification of the native bovine sequence of peptide S8. Unlike peptide S50, the sequence of peptide S53, H-Ser-Gln-Arg-Ser-Gln-Asp-Glu-Asn-OH, has a natural Gly-His deletion and is native to the sequence of guinea pig myelin basic protein. Despite Ser substitutions for Ala and Pro, peptide S53 is as encephalitogenic as peptide S50. The results of this study define an amino acid sequence from the guinea pig basic protein essential for inducing EAE in Lewis rats. Unlike previous studies, modification of a specific natural sequence generated a potent encephalitogenic determinant.
Macrophage-Lymphocyte Interaction in Migration Inhibition Factor (MIF) Production Against Soluble or Cellular Tumor-Associated Antigens
Authors: Landolfo Snato , Herberman Ronald B. , Holden Howard T.
Published: December 2022

For MIF production in response to 3 M KCl extracts of tumor, viable and metabolically active macrophages have been shown to be required to interact with the soluble tumor antigens and then come in contact with immune lymphocytes. The Mø-lymphocyte interaction for MIF production was found to be under the control of genes mapping in the IA subregion of the H-2 complex. However, when intact tumor cells were used as antigen, Mø were not required for immune lymphocytes to produce MIF. In addition, the interaction of immune lymphocytes with tumor cells for MIF production did not require H-2 compatibility. These and other observations strongly suggest that there are two different mechanisms for MIF production and that these may be mediated by two separate subpopulations of immune lymphocytes.
Spontaneous Cytotoxicity of Cultured Human Cell Lines Mediated by Normal Peripheral Blood Lymphocytes
Authors: Callewaert Denis M. , Johnson David F. , Kearney Joanne
Published: December 2022

The mechanism of spontaneous cell-mediated cytotoxicity (SCMC) of cultured cell lines has been investigated and compared with antibody-dependent cell-mediated cytotoxicity (ADCMC) by detailed kinetic analysis. the mechanism of SCMC resembles that of an enzyme, as does ADCMC where effector cells are analogous to an enzyme and the 51Cr-labeled target cells are analogous to the substrate. Temporal kinetic studies revealed an induction period of about 1 hr before significant 51Cr release for SCMC, but not for ADCMC. This induction period is not due to differences in effector-target affinity between SCMC and ADCMC.

On the basis of kinetic analysis it was shown that SCMC approaches simple Michaelis-Menten enzyme kinetics, allowing determination of a Michaelis constant, Km, and maximal velocity, Vmax, for the interaction between a given effector and target cell. The Km values thus determined were found to be identical for the lysis of several target cell lines of varying SCMC susceptibility to effector cells from a given donor, whereas Vmax values for lysis of different target cells varied considerably. However, effector cells isolated from the peripheral blood of different donors exhibited different Km values for the target cells tested. Moreover, the Km value obtained for ADCMC effected by a given donor's lymphocytes was found equal to the Km value obtained for SCMC by that donor.
Fc Receptors Induced by Herpes Simplex Virus
Authors: McTaggart Susan P. , Burns William H. , White David O. , Jackson David C.
Published: December 2022

Radioiodinated normal rabbit IgG was used to detect Fc receptors on the surface of HeLa S3-2 cells infected with herpes simplex virus (HSV). Unlike the Fc receptors present on most leukocytes, these virus-induced Fc receptors were found to be sensitive to trypsin at concentrations of enzyme as low as 0.1 mg/ml. Treatment of infected cells with neuraminidase enhanced the binding of IgG. Yet the HSV-induced Fc receptor(s) is probably a glycoprotein because its synthesis was inhibited by 2-deoxy-D-glucose at concentrations inhibiting glycoproteins but not total proteins. Binding of radioiodinated IgG to Fc receptors on infected HeLa S3-2 cells was unaffected by F(ab′)2 fragments prepared from antisera against uninfected HeLa S3-2 cells or against human β2-microglobulin. By contrast, anti-HSV F(ab′)2 or Fab′ fragments decreased binding of radioiodinated IgG to HSV-infected cells by 85%, and binding of radioiodinated Fc was also inhibited by anti-HSV Fab′.
“Silent” Alleles at the Hla-C Locus
Authors: Ferrara Giovanbattista , Tosi Roberto , Castellani Alberto , Longo Anna , Castellani Alberto , Viviani Claudio , Carminati Guido
Published: December 2022

Defined specificities of the HLA-C locus possess a significantly lower immunogenicity as compared with the HLA-A and HLA-B specificities. HLA-C “blanks” are not immunogenic at all. Among the possible explanations, the most likely one appears to be a low phenotypic expression of these alleles. C locus antigens as a whole are thus likely to be of minor relevance in donor/recipient matching for transplantation.
Studies on the Transfer of Protective Immunity with Lymphoid Cells from Mice Immune to Malaria Sporozoites
Authors: Verhave Jan Peter , Strickland G. Thomas , Jaffe H. Ari , Ahmed Aftab
Published: December 2022

In an effort to understand the mechanisms involved in the protective immunity to malarial sporozoites, an A/J mouse/Plasmodium berghei model was studied. Protective immunity could consistently be adoptively transferred only by using sublethal irradiation of recipients (500 R); a spleen equivalent (100 × 106) of donor cells from immune syngeneic mice; and a small booster immunization (1 × 104) of recipients with irradiation-attenuated sporozoites. Recipient animals treated in this manner were protected from lethal challenge with 1 × 104 nonattenuated sporozoites. Immune and nonimmune serum and spleen cells from nonimmune animals did not protect recipient mice. Fewer immune spleen cells (50 × 106) protected some recipients. In vitro treatment of immune spleen cells with anti-ϑ sera and complement abolished their ability to transfer protection. This preliminary study suggests that protective sporozoite immunity can be transferred with cells, and that it is T cell dependent.
The Primary Structure of a Human Immunoglobulin G2 (IgG2) pFc′ Fragment
Authors: Pardo Angeles Garcia , Rosenwasser Ellen S. , Frangione Blas
Published: December 2022

Amino acid sequence analysis of the pFc′ fragment obtained by pepsin digestion of a human IgG2 myeloma protein PIG Gm (n or 23) negative shows it to consist of 112 residues. It starts at position 334 (γ1 numbering) and contains eight residues from the Cγ2 region, and the whole Cγ3 domain. Comparison with the sequence of γ1 shows two differences at positions 339 and 397. Each of them can be explained by a single base substitution. This high degree of homology among γ-chain subclasses suggests a recent diversification.
Suppression of Cytotoxic Lymphocyte Responses in Vitro by Soluble Products of Alloantigen-Activated Spleen Cells
Authors: Truitt Gary A. , Rich Robert R. , Rich Susan Solliday
Published: December 2022

Suppression of in vitro cytotoxic lymphocyte (CL) responses was mediated by soluble factor(s) produced when in vivo alloantigen-activated suppressor cells were re-exposed to alloantigen in vitro. Elaboration of suppressor factor (SF) was T cell dependent and was optimal 7 days after alloantigen injection. Suppressor factor failed to inhibit CL generation when alloantigen-primed cells rather than normal spleen cells were used as responders. Moreover, SF added at day 3 of incubation rather than at culture initiation was also ineffective, suggesting that suppression probably occurs during antigen induction or early differentiation. Additionally, suppression was abrogated by the presence of 2-mercaptoethanol. Studies combining SF and CL responder cells from a variety of H-2 disparate mouse strains revealed that suppression of CL responses: 1) was not alloantigen specific; 2) did not require H-2 homology between responder and suppressor strains; and 3) could not be demonstrated with CBA/J mice. Although CBA/J CL responses were not suppressed by any SF preparation, allo-sensitized CBA/J spleen cells did elaborate SF that inhibited BALB/c CL responses.
Rabbit Latent Group a Allotypes: Characterization and Relationship to Nominal Group a Allotypic Specificities
Authors: Mudgett-Hunter Meredith , Yarmush Martin L. , Fraser Blair A. , Kindt Thomas J.
Published: December 2022

Latent group a allotypes were detected with a sensitive radioimmune inhibition assay. Sera, IgG preparations, and antibody fractions containing these allotypes inhibited the binding of insolubilized allotypic antisera to various radiolabeled antigens including IgG pools, homogeneous antibodies, and, in the case of a3, a VH fragment from a3/b4 IgG. Several different group a antiallotypic sera were used in the assays and all gave similar results. Comparison of inhibition curves for nominal and latent allotypes indicated that the full spectrum of allotypic subspecificities may be expressed in latent allotypes. Hemagglutination studies carried out with five sera containing high levels of latent allotypes confirmed the results obtained with the radioimmunoassay and indicated that inhibition values did not, at least in four of the five samples studied, reflect the presence of antiallotype antibodies.