bluuebunny/crossref_metadata_2025 huggingface dataset:
Stimulation of Lymphoid Cells from Normal and Immune Mice by Syngeneic BALB/c Plasma Cell Tumors
Authors: Boyer P. J. J. , Fahey J. L.
Published: December 2022

Lymphoid cells from normal and immunized BALB/c mice could be stimulated in vitro by syngeneic plasma cell tumors (PCT) using a one-way mixed lymphcoyte tumor interaction (MLTI) assay. The reactivity of normal spleen cells to syngeneic PCT contrasted with an absence of response to a number of other tumors. Maximal responses of normal cells to PCT were found to occur 5 days after the initiation of the cultures at an optimal responding:stimulating cell ratio of 1:2. MLTI activity of normal cells could not be blocked or enhanced by PCT myeloma protein products indicating that MLTI reactivity was directed against non-idiotypic cell surface determinants. Lymphoid cells from immunized mice demonstrated increased MLTI responses to cells of the immunizing tumor but not to other PCT, indicating that the post-immunization MLTI responses were primarily to individual rather than shared tumor cell surface antigens. Activity of both normal and immunized spleen cells was found to involve thymusderived lymphocytes. The persistence of residual MLTI activity after treatment with anti-θ serum and complement, however, implicated participation of non-θ antigenbearing cells in MLTI reactivity. From these data, we conclude that lymphoid cells from un-immunized mice are capable of T cell-dependent reactivity to syngeneic PCT-associated antigens and that elevations in these reactivities after immunization may reflect specific cellular immune responses.
Loss of Carrier-Determined Tolerance in Vitro with Loss of Receptor Blockade
Authors: Aldo-Benson Marlene , Borel Yves
Published: December 2022

Experiments were done to determine whether carrierdetermined tolerance is reversible and whether the loss of tolerance is accompanied by the loss of receptor blockade. Spleen cells from mice made tolerant with DNP-isologous IgG remained tolerant when transferred to irradiated syngeneic mice. If these same tolerant spleen cells were incubated for 24 hr or more before transfer the tolerance was lost. Autoradiography was done on the tolerant cells with either 125I anti-DNP or 125I DNP-KLH, before and after incubation in vitro. When the cells were tolerant the number of DNP ABC was decreased whereas cells having DNP on their surface were increased. When the cells lost tolerance after in vitro incubation, the hapten-bearing cells were no longer present although the number of cells with free DNP receptors increased to normal. These data suggest that in carrier determined tolerance the reactivation of tolerant lymphocytes may involve reversible receptor blockade.
Further Characterization of the Anti-Encephalitogenic Protein (SCP): Isolation from Bovine Spinal Cord and Spinal Roots
Authors: MacPherson Catherine F. C. , Armstrong Holly , Tan Ophelia
Published: December 2022

The three molecular forms of the anti-encephalitogenic protein, β-SCP, γ-SCP, and SCP-peptide were isolated in higher yield by a shortened procedure, which involved 1) extraction of bovine spinal cord (BSC) or bovine spinal roots (BSR) with 0.05 M sodium acetate buffer, pH 4.5, 2) batch absorption on CM-52 cellulose, 3) stepwise elution with sodium acetate buffers, pH 5.8, containing increasing concentrations of sodium chloride and finally, 4) removal of trace contaminants by gel-exclusion chromatography on Sephadex G-50 superfine. The m.w. of the purified proteins determined by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis was 13,200 daltons. The same value for the molecular sizes was obtained by gel-exclusion chromatography by using 0.1% SDS in 0.05M sodium chloride as eluant. In the absence of SDS, the molecular sizes estimated by gel exclusion chromatography ranged from 14,000 to 18,500. The amino acid compositions of the β-SCP and γ-SCP from BSC and BSR were similar except that β-SCP from BSR lacked half-cystine whereas γ-SCP from BSR contained three times as much half-cystine as the SCP forms prepared from BSC. All forms of SCP showed reactions of identity when compared by immunodiffusion analyses with a rabbit anti-bovine SCP serum; none formed precipitin lines with a rabbit anti-bovine myelin basic protein (MyBP) serum.
Cell-Mediated Immunity to Friend Virus-Induced Leukemia
Authors: Ting Chou-Chik , Rodrigues Dennis , Bushar Grace Shiu , Herberman Ronald B.
Published: December 2022

The primary cell-mediated cytotoxic response to a Friend virus-induced leukemia, FBL-3, in C57BL/6 mice was measured by the 125IUdR release assay. Intraperitoneal (i.p.) inoculation of 1 × 101 FBL-3 cells produced progressive tumor growth (progressors); subcutaneous (s.c.) inoculation of as many as 5 × 106 FBL-3 cells produced only transient tumor growth (regressors), and these mice would subsequently resist i.p. challenge of FBL-3 cells at 3 days after s.c. inoculation. The kinetics of the primary cell-mediated cytotoxic response of regressors was biphasic. Significant cytotoxicity could be detected at 3 to 5 days after s.c. inoculation of 5 × 106 FBL-3 cells, peaked at days 10 to 14, declined to a very low level or became undetectable around days 20 to 30; then the reactivity reappeared and persisted at least up to 60 days. In progressors, the kinetics of the cell-mediated cytotoxic response was similar to the regressors, but the reactivity was much lower. The cytotoxic response was found to be T cell dependent, during both the first peak (days 10 to 14) and the second peak (days 40 to 60). In adoptive transfer experiments, lymphocytes from regressors gave 90% protection against i.p. challenge of FBL-3; lymphocytes from progressors only gave 40% protection.
Antigenic Determinants of Adenovirus Capsids
Authors: Willcox Nicholas , Mautner Vivien
Published: December 2022

We have tested the two principal theories which explain the previous finding that small amounts of type-specific antibody to the adenovirus hexon can neutralize infectivity, whereas even large amounts of cross-reactive antibody do not.

It has been suggested that the type-specific determinants are especially prominent in the virion. We have therefore measured the capacity of whole virus to bind appropriate antibodies, using a sensitive radioimmunoprecipitation (RIP) system. In fact, virions bound typespecific and cross-reactive antibodies impartially. Moreover, they bound both much less effectively than did free hexon or disrupted virus, suggesting that many of each kind of determinant are inaccessible in virions. It has been suggested that the type-specific determinants are confined to those hexons located next to the pentons, and that they are the targets for neutralizing antibody. We have therefore studied the antigenicity of peripentonal and nonamer hexons isolated from virions, and found that each possessed both kinds of determinants. Furthermore, these were present in the same proportions as in hexons purified from the soluble antigens in infected cells (“free hexons”).

We conclude that the mechanism of neutralization by antibody is complicated, and that the type-specific determinants exposed on the virion must play a crucial role.
Antibody-Mediated Cell Cytotoxicity in a Defined System: Regulation by Antigen, Antibody, and Complement
Authors: Lustig Hannah J. , Bianco Celso
Published: December 2022

The use of a serum-free environment and target cells carrying defined amounts of radiolabeled antigen allowed a quantitative study of the role of antigen, antibody, and complement on antibody-mediated cell cytotoxicity (AbMC).

For lysis to occur, a minimum number of antigen molecules must be present on the target cell. 51Cr release from target cells with lower antigen density requires larger concentrations of effector cells and antibodies.

Target cell-bound complement, itself unable to mediate cytotoxicity, reduces the number of IgG molecules required for lysis. The antibody and complement, however, have to be bound to the same target cell. Bystander complementcoated erythrocytes, present in the same reaction mixture with IgG-coated targets, are not lysed.

Blocking of AbMC is effected only by antigen, either soluble or in immune complexes prepared in antigen excess. Antigen competes at the level of the target cell. Blocking at the level of the effector cell, by use of immune complexes prepared at equivalence or in antibody excess, is difficult to achieve. The large number of cells with Fc receptors contained in mouse spleens may explain this finding. Arming of effector cells by passive binding of immune complexes is poorly effective as a means of obtaining lysis of the target cells.

In all situations, the outcome of the reaction is determined by the presence of free antibody-combining sites, alone, or in immune complexes, that are able to combine with the target cell membrane antigen. The requirements for lysis are rather stringent.
Kinetics of the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Jones Jeffrey M. , Amsbaugh Diana F. , Prescott Benjamin
Published: December 2022

When the number of PFC present in the spleen was measured at 24-hr intervals after immunizing with an optimally immunogenic dose of type III penumococcal polysaccharide (SSS-III), maximal numbers of PFC were attained 4 days after immunization; thereafter, the number of PFC decreased rapidly. By contrast, serum antibody levels, which were measured in the same mice using a Farr test, reached peak values 5 days after immunization and then declined much more sowly than did the number of PFC. Two factors were found to contribute to this disparity. First, experiments conduceted with splenectomized mice showed that extrasplenic antibody synthesis, which began between days 3 and 4 after immunization and peaked on days 6 to 7, accounted for nearly one-third of the total amount of serum antibody produced. Second, the average rate of antibody synthesis by PFC increased through day 6 after immunization and then declined.

Antigen-antibody dissociation tests showed that the avidity of the serum antibody obtained 4 to 7 days after immunization was the same. Moreover, during the same interval, all the antibody detected by the Farr test was of the IgM class. Thus, a change in avidity or class of immunoglobulin after day 5 did not account for the disparity observed. The clearance rate of antibody injected i.v. into nonimmune and immunized mice was studied. The data obtained indicated that accelerated clearance of antibody was occurring prior to day 3 after immunization; however, after day 3 the antibody clearance rate was constant and was the same as that found when antibody was injected into nonimmune mice. These findings affirmed the results of previous studies showing that treadmill neutralization was not important in determining the serum antibody levels present after immunization with an optimally immunogenic dose of SSS-III.
Antibody Responses of Mice to Alkaline Detoxified Lipopolysaccharide
Authors: Von Eschen Kenneth B. , Rudbach Jon A.
Published: December 2022

The antibody responses of outbred normal mice and nude mice injected with alkaline detoxified lipopolysaccharide (Alk-LPS) were measured. In some cases the antibody responses to Alk-LPS were compared to responses made against lipopolysaccharide (LPS) and native protoplasmic polysaccharide (NPP). The kinetics of the primary responses to Alk-LPS and NPP were similar, whereas LPS stimulated a more rapid appearance of antibodies in the primary responses. Alk-LPS stimulated only primary antibody responses in both types of mice and sensitized nude mice for secondary responses which could be triggered with LPS. However, secondary antibody responses could not be triggered in normal mice primed with Alk-LPS. These data suggested that, on a functional basis, Alk-LPS possessed the specific antigenic signal associated with LPS antigens but lacked the second nonspecific mitogenic signal dependent on the lipid A portion of LPS.
In Vitro Induction of Cytotoxicity against Syngeneic Mastocytoma and Its Suppression by Spleen and Thymus Cells from Tumor-Bearing Mice
Authors: Takei Fumio , Levy Julia G. , Kilburn Douglas G.
Published: December 2022

P815 × 2 mastocytoma cells, when injected subcutaneously into syngeneic DBA/2 mice, induced T lymphocyte-mediated cytotoxicity in the mice. During the course of tumor growth this cytotoxic activity decreased and ultimately the tumor killed the mice. When spleen cells from mice with tumors in the early stages of growth were incubated in vitro with mitomycin C-treated tumor cells, specific cytotoxicity mediated by T lymphocytes was developed. In contrast, spleen cells, taken from mice with tumors at a later stage in their growth, did not develop cytotoxic activity. The unresponsiveness of spleen cells from mice with tumors in the latter stages of growth seemed to be due to the presence of suppressor cells since in vitro development of cytotoxicity by spleen cells from early tumor-bearing mice were inhibited by the addition of spleen cells or thymocytes from mice with progressively growing tumors. Normal spleen cells or thymocytes did not affect the response.
Lymphocyte Transformation Induced by Autologous Cells
Authors: Weksler Marc E.
Published: December 2022

Human mitogen-induced and cell line B lymphoblasts stimulate the proliferation of allogeneic and autologous lymphocytes in culture. The role in this reaction of EB viral determinants on the stimulating cells and immunity of the lymphocyte donor to the EB virus has been studied. The stimulatory capacity of cultured cell line lymphoblasts is not inhibited by incubating lymphoblasts with antisera to EB viral determinants. Cultured cell line B lymphoblasts stimulate as much thymidine incorporation by lymphocytes from donors with or without immunity to the EB virus. Further, a B lymphoblast cell line (U-698) which lacks the EB viral genome stimulated as much lymphocyte proliferation as did B lymphoblasts with the EB genome. Cultured T lymphoblast cell lines do not stimulate allogeneic lymphocyte proliferation. These cells appear to lack the determinants which stimulate lymphocyte transformation. No evidence was found that cultured cell line T lymphoblasts suppressed allogeneic lymphocyte proliferation. Mitogen-induced lymphoblasts from EB-immune and non-immune subjects stimulated the proliferation of autologous lymphocytes comparably. It is concluded that neither immunity to the EB virus nor expression of EB viral antigens on mitogen-induced or cell line lymphoblasts is necessary for the stimulation of lymphocyte proliferation.
Immunopathogenesis of Autoimmune Tubulointerstitial Nephritis
Authors: Hyman Lawrence R. , Colvin Robert B. , Steinberg Alfred D.
Published: December 2022

Autoimmune tubulointerstitial nephritis (TN) was induced in strain XIII and Hartley but not strain II guinea pigs after immunization with rabbit tubular basement membranes (TBM) in CFA. Strain XIII guinea pigs developed extensive autoimmune TN associated with high anti-TBM (aTBM) antibody titers and linear deposits of IgG along renal cortical TBM after immunization with 10 µg to 10 mg of TBM. In addition, autoimmune TN was passively transferred to strain XIII animals by the i.p. injection of aTBM sera obtained from actively immunized Hartley guinea pigs. In contrast, strain II guinea pigs did not develop autoimmune TN after active immunization (10 µg to 10 mg) with rabbit TBM in CFA, and only produced high aTBM antibody titers with the highest immunizing antigen dose. At this dose (10 mg) the strain II animals demonstrated linear deposits of IgG along renal cortical TBM but did not develop autoimmune TN. Further, recipient strain II guinea pigs did not develop autoimmune TN after passive transfer of aTBM antisera despite renal cortical tubular deposition of aTBM antibodies. Both inbred guinea pig strains produced antibodies reactive with rabbit and rat renal basement membranes. No evidence for differences in nephritogenic TBM antigens could be demonstrated between strain XIII and strain II TBM. These observations indicate that 1) genetic factor(s) influence the production of antibodies reactive to autologous TBM, 2) after the deposition of antibodies on the TBM, additional or related genetic factor(s) determine the full expression of this autoimmune renal disease, and 3) aTBM antibody deposition on renal TBM is not sufficient to elicit autoimmune TN.
The Distribution of Ia Antigens on the Surfaces of Lymphocytes
Authors: Abbas Abul K. , Dorf Martin E. , Karnovsky Morris J. , Unanue Emil R.
Published: December 2022

The distribution of Ia antigens was studied on murine spleen lymphocytes by an ultrastructural technique employing deep freeze-etched replicas. Ia antigens were labeled on cells from appropriate congenic and recombinant strains of mice by incubating the cells with FITC-conjugated anti-Iak antibody, followed by ferritin-coupled Fab anti-FITC. Ia antigens were detected predominantly on immunoglobulin (Ig)-bearing B lymphocytes. Antigens coded for by the entire Ik region were present on the surfaces of 95% of the positive cells (from B10.BR mice) in densely packed microclusters. Ia specificities coded for by the I-A and I-C subregions (on 4R and B10.HTT mice) exhibited a more variable pattern, with 30 to 35% of the labeled cells having sparsely distributed Ia antigens in relatively discrete microclusters. Binding of anti-Iak antibody at 37°C led to patch formation but not to capping. Modulation of surface Ig left Ia antigens diffusely distributed on the cell surface, indicating that these two membrane proteins are independent molecules.
The in Vitro Suppression of Antigen- or Mitogen-Induced DNA Synthesis by Murine Spleen Cells after the Addition of Freshly Prepared Syngeneic Cells
Authors: Ficho Thomas W. , Baram Peter
Published: December 2022

The suppression of antigen- and mitogen-induced DNA synthesis by murine spleen cells in vitro was investigated. It appears that cultures receiving two signals exhibit marked suppression of DNA synthesis. The addition of KLH, PPD, Con A, PHA, or LPS to 72-hr-old cultures of KLH-stimulated murine spleen cells resulted in the suppression of DNA synthesis assayed at 144 hr. When small numbers of freshly prepared KLH-PPD SC or NSC were added to these cultures at 72 hr the suppression of DNA synthesis was abrogated. However, the addition of larger numbers of KLH-PPD SC or NSC resulted in increased suppression of DNA synthesis. Large numbers of KLH-PPD SC or NSC could substitute for the second stimulant (KLH, PPD, Con A, PHA, or LPS) in suppressing DNA synthesis. The addition of fresh cells obtained from KLH-PPD-immunized mice. Cells obtained 30 to 75 days post immunization were most effective in suppressing DNA synthesis.
Partial Characterization of the Rat Anti-Encephalitogenic Protein (RSCP)
Authors: MacPherson Catherine F. C. , Armstrong Holly
Published: December 2022

A protein antigenically similar to the anti-encephalitogenic bovine spinal cord protein (BSCP) was detected in saline extracts of rat nervous tissues by immunodiffusion analyses using a rabbit anti-BSCP serum. Rat SCP (RSCP) appears to be evenly distributed throughout all parts of the rat nervous system and occurs also in the thymus, thyroid, and adrenal glands. Although immunodiffusion analyses indicated that RSCP shares some antigenic sites with BSCP, anti-RSCP sera reacted only with RSCP, indicating that the major immunogenic determinants of the RSCP are peculiar to the rat and differ from the immunogenic determinants of human, monkey, rabbit, guinea pig, or bovine SCP. Immunoelectrophoretic analyses of concentrated pastes of rat brain (RB) or rat spinal cord (RSC) in agar at pH 8.6 revealed that RSCP occurs in two molecular forms having the electrophoretic mobilities of a serum β-globulin and a serum γ-globulin, respectively. However, γ-RSCP is the predominant component of extracts of brain or spinal cord. γ-RSCP was isolated from RB and RSC by a procedure which involved: a) extraction with 0.05 M ammonium acetate buffer, pH 4.0; b) batch absorption of impurities on CM-52 cellulose; c) batch absorption of RSCP on SP-Sephadex, pH 3.5; d) elution of RSCP from SP-Sephadex, pH 5.5; and finally, e) gel filtration on Sephadex G-50 superfine. Purified γ-RSCP formed one band when analyzed by polyacrylamide electrophoresis in acid gels containing 8 M urea. In contrast, two bands were always present when γ-RSCP from brain or spinal cord were subjected to SDS-polyacrylamide electrophoresis in 15% gels. The larger of the two components of brain γ-RSCP had a m.w. of 12,400 daltons, whereas the two components of spinal cord γ-RSCP were smaller. The molecular sizes of brain RSCP and spinal cord RSCP was estimated by gel filtration chromatography to be 12,400 daltons. The amino acid compositions of γ-RSCP prepared from RB or RSC were similar except that γ-RSCP from RSC contained twice as much half-cystine and a slightly higher proportion of basic amino acid than γ-RSCP from RB.
Pathogenetic Mechanisms in Immune Polioencephalomyelitis: Induction of Disease in Immunosuppressed Mice
Authors: Duffey Paul S. , Martinez Douglas , Abrams Gerald D. , Murphy William H.
Published: December 2022

Immune polioencephalomyelitis (IPE) was induced by the i.p. injection of x-irradiated (10,000 R) syngeneic line Ib malignant lymphocytes into C58 mice that were 7 or more months old and in young mice immunosuppressed by x-ray or drugs. The occurrence of IPE in young immunosuppressed C58 mice was systematically analyzed. When mice less than 2 weeks old were x-irradiated with 600 R, IPE could not be induced. The incidence in 1-month-old mice was approximately 50% and increased progressively with the age except for a drop in incidence at 3 months. An analysis of the dose effects of x-irradiation on the occurrence of IPE in mice of different ages revealed a marked increase in the incidence in 3- and 5-month-old mice beginning at dose levels of 450 R and 300 R, respectively. Considered together, these data indicated that two subpopulations of immunocytes differing in x-ray sensitivity interacted to protect mice from IPE. It appears that under natural conditions an x-ray sensitive cell population, possibly having suppressor function, decreased with age and made mice susceptible to induction of IPE. Five-month-old mice were immunosuppressed with an LD10 of cyclophosphamide, prednisolone, or methotrexate to determine whether mice immunosuppressed with drugs also were susceptible to the induction of IPE. The incidence was 89%, 13%, and 5%, respectively. The mouse strain specificity of IPE induction also was studied. In 6- to 8-month-old mice suppressed with 600 R, IPE could not be induced in non-H-2k strains: BALB, C57BL/6, NZB. Of the H-2k strains tested (CBA/J, C3H/He, AKR/J, C58), the disease could be induced only in the C58 and AKR/J strains. Histopathologic studies showed that CNS lesions in immunosuppressed C58 and AKR/J mice did not differ significantly from those in old C58 mice with IPE. Taken together, the results of these studies indicate that IPE can be used as a model for analyzing age-dependent diseases of suspected immunopathologic etiology.
Delayed Hypersensitivity and Granulomatous Response After Immunization with Protein Antigens Associated with a Mycobacterial Glycolipid and Oil Droplets
Authors: Granger Donald L. , Yamamoto Ken-Ichi , Ribi Edgar
Published: December 2022

A mycobacterial glycolipid (P3) mixed with protein antigens in oil-in-water emulsion induced lasting delayed hypersensitivity (DH) and granulomatous inflammation after intradermal injection into guinea pigs. This did not occur when P3 and bovine serum albumin (BSA) were given in Freund's incomplete adjuvant. The oil-in-water emulsions consisted of microscopic oil droplets suspended in aqueous medium. By separating oil and aqueous phases from BSA + P3 emulsion it was shown that antigen retained with oil droplets led to DH and granuloma formation. The association of antigen with oil droplets was P3 dependent and was quantitated with 125I-labeled BSA. The same phenomenon occurred with 125I-labeled rabbit γ-globulin (RGG) + P3 emulsion. Fluorescein-conjugated RGG was observed in a particulate state within or on oil droplets in emulsion containing P3. These physical characteristics of antigen + P3 emulsion appeared to be important for immunogenicity.
Physicochemical and Functional Characterization of the C1r Subunit of the First Complement Component
Authors: Ziccardi Robert J. , Cooper Neil R.
Published: December 2022

C1r was isolated from serum by an improved method and found to be a glycoprotein with a sedimentation coefficient of 7.0S. Under conditions of physiologic ionic strength and pH, C1r consists of two apparently identical noncovalently linked 95,000 dalton polypeptide chains.

Antisera to C1r detected a protein of γ-mobility on electrophoresis of serum in agarose in the presence of calcium, and a β-mobility protein when the electrophoretic separation was carried out in EDTA. On sucrose gradient ultracentrifugation of normal human serum in the presence of calcium, C1r antigenicity was found in the 19 S region of the gradient. On the other hand, when the gradient contained EDTA, C1r antigenicity was found in the 7 S region. No reaction of anti-C1r with C1r-deficient sera was observed. C1r had a high affinity for active or proenzyme C1s in the presence of calcium and was able to activate C1s and to form C1 in conjunction with C1q and C1s. Activation of C1s by C1r was inhibited by calcium, C1 inactivator, polyanethol sulfonate, and DFP. Activation of C1s by C1r occurred only after a preliminary incubation of C1r for a brief time at 37°C before addition of C1s. The ability of C1r to form C1 in conjunction with C1q and C1s was, however, progressively lost on incubation at 37°C. Trypsin, although potentiating the activity of crude C1r, did not modify the activity of purified C1r. Its action was on a trypsin-sensitive inhibitor separated from C1r in the final step of the isolation procedure.

The binding of 125I-C1r to sensitized sheep erythrocytes required the presence of C1q and calcium but not C1s, whereas the binding of 125I-C1s required C1q, C1r and calcium. Thus, C1r functions as not only the activator of C1s, but also serves as the physical link between C1q and C1s in macromolecular C1.
The Structure and Function of Immunoglobulin Domains
Authors: Yasmeen D. , Ellerson J. R. , Dorrington K. J. , Painter R. H.
Published: December 2022

The fragments related to the Cγ2 and Cγ3 homology regions of human IgG, described in the preceding paper by Ellerson, et al. were assayed for their ability to interact with complement, engage in cytophilic activity toward macrophages, and to play a role in controlling the catabolism of the whole IgG. The Cγ2-fragment retained about 3% of the activity of intact IgG in a whole complement-fixing assay in which the test proteins were adsorbed as monolayers onto polystyrene latex beads. However, in a fluid-phase C1-binding assay this fragment showed the same activity as IgG and Fc-fragment, when compared on a molar basis. Since the Cγ2-fragment represented only one intact domain, full expression of complement-fixing activity appeared to be independent of quaternary interactions. Thus IgG possesses two C1-binding sites. The Cγ3-fragment was inactive in both of the complement assays. The ability of IgG to interact with the Fc-receptor on guinea-pig macrophages was shown to be entirely a function of the Cγ3 region. This was demonstrated both in a direct assay in which tanned red cells coated with Cγ3-fragment formed rosettes with macrophages and in an indirect assay in which this fragment was able to inhibit rosette formation between IgG-coated red cells and macrophages. The rate of clearance of radiolabeled Cγ2-, Cγ3-, Fab, Fc fragments, and IgG from the circulation was measured in rabbits. The Cγ2 fragment was cleared with a half-time similar to that shown by intact IgG and Fc (about 70 hr) whereas Cγ3- and Fab fragments were cleared more rapidly (half-time, about 15 hr). The rate of clearance was not related to the presence of sialic acid or exposed galactosyl residues at the termini of the carbohydrate prosthetic groups. These data clearly show that at least three of the biologic functions of IgG are mediated fully and independently by one or other of the Fc domains.
Local and Systemic Cell-Mediated Immunity against Transmissible Gastroenteritis, an Intestinal Viral Infection of Swine
Authors: Frederick G. Thomas , Bohl Edward H.
Published: December 2022

The occurrence of local and systemic cell-mediated immunity (CMI) in swine against transmissible gastroenteritis (TGE) virus, a coronavirus that infects the epithelial cells of the small intestine of swine, has been investigated. Leukocyte suspensions prepared from intestinal lamina propria (LP) and spleen were each incubated with and without inactivated TGE virus. The supernatants from these cultures were tested for the presence of macrophage migration inhibition factor (MIF) by the indirect MIF test using guinea pig peritoneal exudate cells as the source of macrophages. All swine serologically negative for TGE antibodies (i.e., unexposed to TGE virus) were also negative for MIF. Fourteen of 15 swine orally exposed to TGE virus 6 to 30 days before assaying for CMI gave a positive test for MIF production by both LP and splenic lymphocytes. The 15th animal, orally exposed to TGE virus 27 days before assaying, was negative for MIF production by both LP and splenic lymphocytes. In nine of nine of these orally exposed animals where LP and splenic leukocyte suspensions were prepared so as to contain equal numbers of lymphocytes, the percentage of inhibition of migration by supernatants from LP preparations was significantly greater (p ≤ 0.01) than by supernatants from spleen preparations. Two of three animals orally exposed to TGE virus 7 to 9 ½ months before assaying for CMI were negative for MIF production by lymphocytes from both sources. In 10 of 10 animals given TGE virus subcutaneously, MIF production by splenic lymphocytes was significantly greater (p ≤ 0.01) than by LP lymphocytes, and in 9 of 10 of these animals MIF production by LP lymphocytes was negligible (percentage of inhibition of migration < 3%).
Characterization of a Subpopulation of Human T Lymphocytes Reactive with an Heteroantiserum to Human Brain
Authors: Brouet Jean-Claude , Toben Howard
Published: December 2022

A rabbit antiserum to human fetal brain reacted after suitable absorptions with a subpopulation of human normal T cells. The distribution of reactive T cells varied according to the organ tested: 23% of peripheral blood lymphocytes, 5% of tonsil lymphocytes, and less than 1% of thymocytes were positive. Reactive cells did not transform after phytohemagglutinin or pokeweed stimulation but were at least weakly stimulated by allogeneic cells.

T-derived neoplastic cells from one case of T acute lymphoblastic leukemia, two patients with Sezary syndrome, and from three out of five cases of T chronic lymphocytic leukemias (CLL) yielded negative results. In contrast, all the leukemic cells from two other patients with T-derived CLL were positive suggesting a proliferation of homogeneous cells arising from only a subpopulation of T lymphocytes.
Radioimmunoassay for Detection of Antigen and Antibodies to Newcastle Disease Virus
Authors: Spira Gad , Silvian Ilana , Zakay-Rones Zichria
Published: December 2022

When Newcastle disease virus (NDV) is treated with NP-40 and ether a membrane fraction of 150.000 m.w. is obtained. This fraction which is composed of two polypeptides with m.w. of 56.000 and 76.000 was used in a radioimmunoassay (RIA). The assay was developed for both antigen and antibody and was found to be reproducible, specific, and highly sensitive. Titers of 1:51,200 were determined by RIA as compared to 1:4 by agar gel diffusion and 1:200 by hemaglutination inhibition (HI). As little as 5 ng of viral protein were detected by RIA inhibition technique.

Labeled antigen could be stored in the presence of serum, KCI and Triton X-100 at -20°C for as long as 6 weeks and retained similar reactivity as fresh reagent.
Genetic Control of the Immune Response to the Ea-2 Alloantigen System of the Mouse
Authors: Stimpfling J. H. , Reichert A. E. , Blanchard S.
Published: December 2022

The immunization of selected congenic strains and hybrids against the Ea-2.1 cellular alloantigen of the mouse demonstrated that the hamagglutinating antibody response to Ea-2.1 is regulated by a gene or genes associated with the H-2 gene complex. H-2r and H-2b are, respectively, responder and non-responder haplotypes.
Effect of Proteolytic Digestion on the Structure and Function of Human Properdin
Authors: Minta Joe O. , Kunar Edith S.
Published: December 2022

Human properdin P̄ was found to be sensitive to the action of trypsin, chymotrypsin, pepsin, and Streptomycetes caesipitosus protease. Incubation of P̄ with these enzymes resulted in loss of tis founctional activity and the productin of antigenically deficient components compared to untreated P̄. Upon incubation with trypsin, P̄ was initially cleaved into a minor fragment and a major fragment. Further degradation of the fragments occurred with prolongation of inculation time.The minor fragment wa highly susceptible to further proteolysis compared to the major fragment which contained fthe carbohydrate moiety of the molecule.

SDS-polyacrylamide gel electrophoretic analysis of trypsin-digested P̄ suggested that the subunit polypeptide chains were initially cleaved at similar points to produce the major and minor fragments. The sedimentation velocity of the major fragment was higher than that of the intact molecule. The implications of these observations of the configuration of P̄ are discussed.
The Active E Rosette Test: Correlation with Delayed Cutaneous Hypersensitivity
Authors: Felsburg Peter J. , Edelman Robert , Gilman Robert H.
Published: December 2022

An active subpopulation of peripheral blood T lymphocytes, characterized by rapid (5-min) rosette formation with sheep erythrocytes (A-RFC), was measured in normal individuals after they were skin tested with microbial antigens. A significant rise in A-RFC occurred in all individuals who developed positive delayed cutaneous hypersensitivity (DCH) reactions, whereas skin test non-responders showed no significant rise in A-RFC. No similar consistent changes occurred in populations of total T cells, characterized by longer (60-min) rosette formation with sheep erythrocytes, or in B cells, measured by immunofluorescence of surface immunoglobulin. The A-RFC response paralleled the DCH response in timing, but not in intensity. These results provide in vivo evidence for a biologically distinct T cell subpopulation, and focus attention on the A-RFC as immunologically active cells.
Immunologic Reactivity to Soluble Autochthonous Tumor Extracts in Dogs with Spontaneous Maligancies
Authors: Tsoi Mang-So , Weiden Paul L. , Storb Rainer
Published: December 2022

The presence of tumor-associated antigens (TAA) in dogs with spontaneous lymphoma (L) or solid tumors (ST) was investigated by testing soluble tumor cell extracts for their ability to stimulate autochthonous in vitro lymphocyte blastogenesis and in vivo intradermal delayed hypersensitivity responses. Tumor extracts were prepared from tumor biopsies and control extracts were prepared from buffy-coat cells of tumor dogs by hypertonic KCl solubilization. Blastogenic responses to intact, cryopreserved, autochthonous tumor cells were also determined.

Fifteen of 26 (58%) L dogs and 23 of 32 (72%) ST dogs had positive blastogenic responses to at least one concentration of autochthonous tumor extracts. Blastogenic responses to tumor extract and to cryopreserved whole tumor cells correlated well when the tests were performed simultaneously. No consistent effect of autochthonous serum on the lymphocyte response to autochthonous tumor extract was observed.

Two of 11 L dogs showed a positive skin test to autochthonous tumor extract and none reacted to control extract. Three of 21 ST dogs showed a positive skin test and two had a borderline response, while two ST dogs had a positive response to control extract. However, five of seven L and nine of 18 ST dogs were anergic as defined by failure to respond to intradermal PPD challenge after sensitization with Bacille Calmette-Guerin.

In conclusion, lymphocytes from most dogs with spontaneous tumors underwent blastogenesis in vitro in response to autochthonous soluble tumor extracts, and these responses when tested simultaneously correlated well with responses to cryopreserved intact tumor cells. These results are compatible with the presence of TAA in extracts from cells of spontaneous canine tumors. Delayed skin test reactivity, however, was an uncommon and unpredictable finding, and does not appear to be useful in detecting TAA in the dog.
Amyloid Fibril Protein AA: Purification and Properties of the Antigenically Related Serum Component As Determined by Solid Phase Radioimmunoassay
Authors: Sipe Jean D. , Ignaczak Thomas F. , Pollock P. Scott , Glenner George G.
Published: December 2022

The isolation by gel filtration of a serum component (SAA), antigenically related to the major fibrillar amyloid protein (AA), associated with “secondary” amyloidosis, has been monitored by a solid phase radioimmunoassay for the AA protein to detect cross-reacting serum fractions. Evidence is presented that not all cross-reacting antigenic determinants are accessible in native SAA, since additional determinants are revealed during the isolation procedure.

The native structure of SAA appears to be quite labile. SAA from freshly collected serum has a m.w. of 180,000 and co-chromatographs with IgG. However, species of higher m.w. are observed after storage of serum at 4°C or upon chromatography of serum in ammonium bicarbonate. Denatured SAA has a tendency to aggregate under strong dissociating conditions. A 12,500 m.w. antigenic species (SAAL) was detected upon guanidine-HCI denaturation of SAA, by earlier studies employing double immunodiffusion. However, evidence is presented here that the major part of the antigenic activity after guanidine-HCl treatment was of m.w. greater than 12,500, but was unreactive in double immunodiffusion. Formic acid treatment of cross-reacting serum fractions does result in virtually complete dissociation of SAA to SAAL, however. Furthermore, formic acid-dissociated SAAL is of comparable immunoreactivity with AA, on a molar basis, unlike SAAL obtained from SAA by guanidine-HCl denaturation.
Contamination of Con A Preparations
Authors: Fumarola Donato , Jirillo Emilio
Published: December 2022

Brunson's and Watson's paper (J. Immunol. 1975, 115:599) about contamination of Con A preparations with bacterial endotoxins overlooks our personal experience with the same problem (Fumarola et al., Giorn.Batt. Virol. Immun. 1974, 67:289; Progr. Medico, 1975, 31:903). We have found that phytohemagglutinin, pokeweed mitogen, concanavalin A, anti-lymphocyte serum, and transfer factor gave strong positive reactions in the Limulus assay. The concentrations of endotoxin often present in these preparations are high enough to influence immunologic reactions of lymphocytes. However, in studies of this sort it is most important to quantify the level of apparent contamination of reagents so that one can compare these levels with known levels of endotoxin in similar experimental models. In this way one can be sure of the significance of one's findings. Using serial dilution of some samples we detected an amount of endotoxin ranging between 1 and 5 µg/ml.
Separate Fc-Receptors for Immunoglobulins IgG2a and IgG2b on an Established Cell Line of Mouse Macrophages
Authors: Walker William S.
Published: December 2022

The specificity of Fc-receptors on IC-21 cells, an established line of mouse peritoneal macrophages with antibody-dependent effector cell activity has been examined. Only IgG2a and IgG2b myeloma proteins bound readily to IC-21 Fc-receptors, the former in nonaggregated as well as aggregated form, the latter only as aggregated complexes. Thus, IgG2a bound in a manner characteristic of classically defined cytophilic antibody, whereas the binding of IgG2b appeared to be mediated by Fc-receptors for antigen-antibody complexes. Evidence is presented in support of the view that IC-21 macrophages possess separate and distinct Fc-receptor sites for these two immunoglobulins.
Characterization of T and B Antigen-Binding Cells for β-Galactosidase
Authors: Swain Susan , Modabber Farrokh , Coons Albert H.
Published: December 2022

The results reported in this paper suggest that the specific Z-binding cells of the normal mouse include a large portion of T lymphocytes. Depletion of T cells with anti-θ serum and cortisone indicates that the majority of the ZBC of the thymus, which occur at frequencies of about 150/106, are indeed T cells. Similar treatment of spleen cells suggests that approximately half the binding cells in that organ are contributed by the T lymphocyte population. T- and B-enriched populations obtained from the spleen by using differential adherence to nylon wool contained equal numbers of ZBC and bound equivalent amounts of the antigen. Hence, there appears to be a high frequency of T lymphocytes that can be shown to bind β-galactosidase specifically in both the thymus and spleen of normal mice.
Motheaten, an Immunodeficient Mutant of the Mouse
Authors: Shultz Leonard D. , Green Margaret C.
Published: December 2022

Mice homozygous for the recessive mutation motheaten (me) are deficient in capacity for immune response but show an elevated level of serum immunoglobulins. In comparison to spleen cells from normal sibs, spleen cells from me/me mice have a severely depressed 19S PFC response to SRBC. In the GVH assay, spleen and thymus cells from motheaten donors caused significantly weaker reactions than like cells from normal sibs. Serum electrophoretic patterns of motheaten mice showed increased levels of α-, β-, and γ-globulins and decreased levels of albumin. Increases in quantities of all major classes of immunoglobulins were found in serum of me/me mice 5 weeks of age and older. Elevation of serum IgM was evident by 3 weeks of age and had reached 25 times the levels in normal sibs by 6 weeks of age. Immunoelectrophoresis and Ouchterlony analysis showed motheaten serum to have both κ and λ2 light chains. Evidence of autoimmunity was found in motheaten mice in the granular deposition of IgM and IgG in kidney glomeruli. Motheaten mice, thus, appear to have a severe immune deficiency, but the basic nature of the deficiency is not yet known.
Immune Mechanisms in Leukemia: Protective Capacity of the Major Lymphoid Cell Compartments
Authors: Lukasewycz Omelan A. , Duffey Paul S. , Murphy William H.
Published: December 2022

The capacity of immune spleen, lymph node, peritoneal, bone marrow, and thymic cells to protect C58/wm mice from syngeneic transplanted line Ib leukemia was quantified. Cells harvested 14 to 15 days after primary immunization were used for adoptive protection tests. Regression curves were computer analyzed and log10, PD50 values compared. For immune spleen, lymph node, peritoneal, bone marrow, and thymic cells the PD50 values were 4.53, 5.92, 4.88, 5.51, and 5.59, respectively. When immune spleen cells were treated with anti-Thy 1.2 serum the PD50 value was increased from 4.73 to 6.09, i.e., protection was reduced ≥95%. Similar treatment of immune thymic cells reduced protection below measureable values. Anti-B cell sera (anti-IgM and anti-Ly 4.2) did not reduce the protective effect of immune spleen or marrow cells. These results indicate that a major protective cell population in each of these compartments was θ-positive. Experiments were carried out to characterize the cortisone (CS) and x-ray sensitivity of immune spleen, thymic, and marrow cells. When donor mice were treated with 12.5 mg of cortisone acetate/day for 2 days before lymphoid cells were harvested, the protective effects of immune spleen cells, but not immune thymic or marrow cells, was reduced. When immune spleen cells were x-irradiated in vitro, their protective effect was reduced by 350 R and abolished by 1000 R. When mice received whole body x-irradiation 24 hr before immune spleen cells were transferred their protective effect was reduced by 1000 R but only slightly lowered by 350 R. The possible significance of the multicompartmental nature of immunity to leukemia was discussed.
Preparation and Effects of an Anti-B Cell Serum
Authors: Kakiuchi Terutaka , Nariuchi Hideo , Tamura Noboru
Published: December 2022

An heterologous antiserum specific for bone marrow-derived cells (B cells) was prepared by immunizing rabbits with lymph node cells from nude mice. After absorption with mouse red blood and thymus cells, the antiserum killed a population of cells from various lymphoid organs and the cytotoxic effects were inversely related to those of anti-ϑ antibody.

When bone marrow or spleen cells were treated with the antiserum and guinea pig complement before transfer into irradiated mice, the number of plaque-forming cells was greatly reduced in the spleen of the recipient. Pretreatment of thymus cells with the antiserum in a similar way resulted in no inhibition of hemolytic plaque. When spleen cells from mice previously immunized with sheep red blood cells were treated with the antiserum and complement, the formation of hemolytic plaque was not affected.

These findings indicated that the antiserum was specific for B cells and that the number of antigenic determinants on B cells to which the antiserum reacted decreases during differentiation into antibody-forming cells.
Endodermally-Derived and Neural Crest-Derived Differentiation Antigens Expressed by a Human Lung Tumor
Authors: Bell C. Elliott , Seetharam Shakuntla , McDaniel Robert C.
Published: December 2022

The plasma membrane antigens of an undifferentiated small cell (oat cell) carcinoma of the lung were studied by the indirect immunofluorescence method on frozen section substrates with a rabbit antiserum prepared to the tumor plasma membrane fraction. After appropriate absorption of the antiserum, at least two differentiation antigens present on the tumor cells but undetectable on normal lung surface or glandular epithelial cells were identified. One antigen(s) was characteristic of certain normal, endodermally-derived epithelial cells of the digestive system, including those of colonic mucosa, hepatic ducts, pancreatic ducts and acini, and islets of Langerhans. The other antigen(s) was characteristic of certain normal, neural crest-derived cells in the peripheral nervous system, including cells in peripheral nerve, dorsal root ganglion, and anterior roots of the spinal cord; parasympathetic ganglion cells in the colon; and small nerves and nerve processes in the lung, colon, and skin. It was concluded that the presence of these differentiation antigens on the tumor cells resulted from the expression of gene products repressed in the normal cell of origin of the tumor.
Splenic Suppressing Factor: Purification and Characterization of a Factor Suppressing Thymidine Incorporation into Activated Lymphocytes
Authors: Kasahara Tadashi , Shioiri-Nakano Kohei
Published: December 2022

Suppressing activity upon the mitogen-activated lymphocytes was found in the supernatant (SUP) from the culture of mouse spleen, high-density subpopulation of thymocytes, and peritoneal exudate cells. Suppressing factor was obtained from the non-stimulated lymphocytes cultured for 24 to 36 hr with or without serum. Suppressing activity in the SUP was observed in the incorporation of 3H-thymidine, 3H-uridine, and 3H-leucine into Con A-activated lymphocytes or in the proliferation of L cells. Suppressing factor partially purified by Sephadex G-25 column chromatography was a heat-stable and dialyzable substance(s). Further purification and isolation of this factor by two-dimensional thin layer chromatography revealed that this was thymidine and thymidine monophosphate. The suppression in 3H-thymidine incorporation was attributed to the dilution effect of cold thymidine released from cultured lymphocytes.