bluuebunny/crossref_metadata_2025 huggingface dataset:
The Adjuvant Activity of Mycobacterial Rna Preparations and Synthetic Polynucleotides for Induction of Delayed Hypersensitivity to Purified Protein Derivative in Guinea Pigs
Authors: Casavant Conrad H. , Youmans Guy P.
Published: December 2022

The adjuvant activity of mycobacterial RNA and synthetic polynucleotides for the induction of delayed hypersensitivity to PPD was determined. It was shown that when mycobacterial RNA or synthetic polynucleotides are injected together with purified protein derivative (PPD), delayed hypersensitivity to PPD developed as compared to no detectable delayed response when PPD was administered alone without adjuvant into guinea pigs. Four different criteria were employed to detect delayed hypersensitivity responses. These were the time, appearance, and magnitude of dermal reactions, histologic examination of dermal sections, passive transfer of sensitivity with sensitized spleen cells and the elaboration of migration inhibitory factor (MIF) by sensitized spleen cells.

When synthetic polynucleotides were used as adjuvants and were injected into guinea pigs in combination with PPD, dermal reactions as well as MIF assays gave evidence that these animals exhibited delayed-type hypersensitivity. Poly U alone exhibited adjuvant activity for induction of delayed hypersensitivity to PPD. Trypsin and pronase treatment did not affect the adjuvant activity of mycobacterial RNA whereas KOH treatment completely abolished any adjuvant effect, suggesting that ribosomal protein did not contribute to the adjuvant characteristics of mycobacterial RNA. Titration experiments indicated that the adjuvant activity of mycobacterial RNA was greater than that of poly A:U.
The Plaque Cytogram Assay
Authors: Thornthwaite Jerry T. , Leif Robert C.
Published: December 2022

The plaque cytogram assay has been combined with pH 6.8, linear bovine serum albumin (BSA) gradients to demonstrate the correlation between the morphology and buoyant density of immunocompetent cells. Ten types of plaque-forming cells (PFC) and four types of plaques have been identified and enriched in the density gradients. Reproducible density profiles of mouse erythrocytes, lymphoid cells, and PFC have been obtained with BSA gradients.

The density peak for mouse erythrocytes was about 1.080 g/ml, while the lymphoid cell density profile had several peaks with a maximum concentration at 1.064 g/ml. Three distinct peaks or shoulders were found in the density profile of hemolytic PFC.

Most of the density distributions of the individual morphologic classes of PFC were considerably narrower than that of the entire PFC distribution and were discrete from each other. The combined result that the PFC could be subdivided by morphology and density, and that the two phenomena were correlated establishes that the PFC are truly a heterogeneous population. Whether this heterogeneity is due to different stages in development or due to the existence of differing classes of cells will have to await future experimentation.
Studies on the Mechanism of Stimulation of the Humoral Response of Murine Spleen Cultures by Culture Fluids from Human Monocytes
Authors: Wood David D. , Cameron Patricia M.
Published: December 2022

It has been demonstrated that media derived from cultures of human monocytes can stimulate the IgM and IgG anti-sheep erythrocyte response of normal or T celldeficient murine splenocytes in vitro. This culture fluid also causes a slight stimulation in the rate of B cell division and in the number of antibody-secreting cells occurring in the absence of added antigen. The active factors, which are released from the monocytes during the first 2 days of culture, appear to affect the murine B cells throughout the culture period.
Application of the Paired Label Radioantibody Technique to Detection and Subtyping of Hepatitis B Surface Antigen
Authors: Miyakawa Yuzo , Imai Mitsunobu , Mayumi Makoto
Published: December 2022

A sensitive radioimmunoassay has been developed for the detection and subtyping, in regard to w and r specificities, of hepatitis B surface antigens (HBsAg). The binding of monospecific rabbit antibodies directed to either w or r determinant, labeled with 131I and 125I, respectively, by HBsAg was determined in the same experiment, taking advantage of the fact that these specificities are in general, expressed mutually exclusively. HBsAg in the test sample were bound to solid phase guinea pig anti-HBsAg antibody tube before the test.

The ratio of percentage uptake of 125I anti-r to that of 131I anti-w for 28 normal sera without antigen was 0.96 0.18. The ratio for seven sera containing HBsAg with w determinant was less than 0.1, and that for 34 with r determinant was more than 10; 100-fold difference was noted between values of w and r antigens.
Migration Inhibitory Activity in Serum and Cell Supernatants in Patients with Sezary Syndrome
Authors: Yoshida Takeshi , Edelson Richard , Cohen Stanley , Green Ira
Published: December 2022

The abnormal lymphocytes from patients with the Sezary syndrome produce macrophage migration inhibitory factors (MIF) both in vitro and in vivo. Five of six individuals studied had significant serum MIF activity and one had borderline activity. In contrast, in this study 47 normal individuals and 9 of 10 patients with extensive skin disease other than that of the Sezary syndrome had no such activity. Since the neoplastic T cells in patients with the Sezary syndrome localize in the skin, their production of chemical mediators in that site may be responsible for the generalized exfoliative erythroderma seen in that disease.
Inhibition by ɛ-Aminocaproic Acid of the Activation of the First Component of the Complement System
Authors: Soter Nicholas A. , Austen K. Frank , Gigli Irma
Published: December 2022

The influence of EACA on C1 in whole human serum and on C1 and C1̄ as isolated molecules was assessed hemolytically. There was selective inhibition of C1 without effect on the levels of C4, C2, C3, and C9 in whole human serum that was reversed by dialysis. EACA was found to inhibit the intrinsic activation of C1 without inhibiting the already active molecule. This was confirmed by the capacity of trypsin to uncover C1 activity in cellular intermediates formed by C1 treated with EACA that did not evolve in the absence of this extrinsic activating mechanism. Inasmuch as the trypsin-dependent recovery of C1 was incomplete, an effect on binding cannot be excluded.
Genetic Control of the Immune Response to the H-2 Associated Slp Alloantigen in the Mouse
Authors: Passmore Howard C. , Hansen Ted H.
Published: December 2022

A survey of sixteen standard inbred and congenic resistant strains of mice reveals that the ability to mount an immune response to the Slp allotype is associated with the H-2 type of the recipient. Strains carrying the H-2r, H-2k, and H-2q haplotypes are able to produce specific antibody whereas strains of the H-2b haplotype are non-responders. Analysis of F1-hybrids and four informative intra-H-2-recombinants demonstrates that the ability to respond to the Slp allotype is controlled by a dominant gene associated with the K end of the H-2 complex.
Control of J Chain Biosynthesis in Relation to Heavy and Light Chain Synthesis, Polymerization and Secretion
Authors: Kaji Hideko , Parkhouse R. M. E.
Published: December 2022

Mouse myeloma tumors and some variants derived from them were labeled in vitro with tritiated leucine and the radioactive J chain was assayed in cell lysates by precipitation with an antiserum specific for mouse J chain. The major findings were: 1) J chain can be found in an IgG2b-secreting cells (MPC-11). These data, together with previous findings suggest that cells secreting all classes of IgG synthesize J chain, even though there is no apparent requirement for J chain in assembly of the IgG molecule. Hence production of J chain does not depend upon secretion of a polymeric immunoglobulin. 2) Intracellular J chain can be found in myeloma variants that do not produce heavy chains showing that production of J chain may not coordinately be linked to the synthesis of heavy chain. 3) J chain was found in cells synthesizing, but not secreting, immunoglobulin. Thus production of J chain is not linked to secretion of immunoglobulin. 4) J chain could not be detected in plasma cells that do not produce immunoglobulins. It was also not found in mouse leukemic cells, suggesting that production of J chain is probably linked in some way to immunoglobulin production.
Humoral Immunity in Rodent Malaria
Authors: Diggs Carter L. , Osler Abraham G.
Published: December 2022

Serum which protects rats against Plasmodium berghei infections fails to sensitize parasitized erythrocytes in vitro for in vivo destruction. Further, the efflux of 86Rb from parasitized erythrocytes in the presence of complement is not accelerated. On administration to animals with preexisting malaria, it does, however, produce a relatively slow decline in parasitemia, a phenomenon interpreted in terms of the gradual production and/or release of reactive antigenic determinants associated with maturation of the parasites. Protective activity is elaborated in the serum of animals during the development of parasitemia, thus creating a situation in which there is apparently the coincident presence of antibody and its putative antigen in the circulation. Little absorption of protective activity by parasitized cells or parasites was observed in vitro. However, in vivo removal of the protective activity during an ongoing infection could be demonstrated. These observations favor the notion that protective antibody exerts its influence not on the bulk of the parasitized erythrocytes, but rather on some subpopulation thereof. The cumulative evidence of this and other studies suggests that the stage(s) involved are the schizonts and/or merozoites.
In Vitro Studies on Allotype Suppression
Authors: Adler Louise T. , Fishman Marvin , Adler Frank L.
Published: December 2022

The regulatory effects of rabbit antibodies specific for light chain determinants (b locus) on the formation of rabbit serum immunoglobulins have been studied in an in vitro system which measures the response of unprimed rabbit spleen cells to solubilized T2 phage antigen. Treatment of spleen cells from b4b4 rabbits with anti-b4 serum, which was either incorporated into the culture medium or employed in appropriate pulse treatment of the cells before culture, prevented the formation of T2 neutralizing antibodies by such cells. Spleen cells of heterozygous (b4b5) rabbits formed anti-T2 antibodies which could be shown to be divided between the b4 and b5 specificities. Incorporation of anti-b4 or anti-b5 serum into the culture medium suppressed the specific anti-T2 response and, except in the instances noted in the text, did not significantly change the level of T2 neutralizing antibodies marked with the alternate allelic determinant. These findings are discussed in the light of the compensatory formation of an alternate immunoglobulin type which occurs during allotype suppression in vivo.
Lysis of Tumor Cells by Antibody and Complement
Authors: Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

Two antigenically distinct diethylnitrosamine-induced guinea pig hepatoma cell lines, line-1 and line-10, sensitized with rabbit anti-Forssman or with tumor-specific antibody, were more susceptible to killing by human complement (HuC) than by guinea pig complement (GPC). This difference could not be ascribed to differences in the amount of C1, C4, and C3 fixed: millions of C4 and hundreds of thousands of C3 were detected on cells whether they were killed or not killed by the C sources. Tumor cells sensitized with anti-Forssman IgM antibody generally had more GP C4 and C3 than Hu C4 and C3 bound to their surfaces. Cells sensitized with anti-tumor antibody generally had more Hu C4 and C3 than GP C4 and C3 bound to their surfaces.

The resistance to killing of nucleated cells by antibody and C may be due in part to intrinsic properties of the cell.
Normal Function of Immunologic Stem Cells from Aged Mice
Authors: Harrison David E. , Doubleday John W.
Published: December 2022

Marrow or spleen grafts from aged donor mice produced antibody-forming cells as effectively as did grafts from younger controls in recipients tested 3 to 10 months after the transplantation. All recipients were lethally irradiated, and the T6 chromosome marker was used to demonstrate that they were populated by donor cell lines.

Recipients of aged or younger control grafts gave similar responses when stimulated with varying doses of antigen and when tested at different times after the transplantation except in two cases: 1) Recipients of aged spleen grafts gave significantly lower responses than younger controls for the first few weeks after the transplantation. 2) If recipients had been thymectomized before lethal irradiation, aged cell lines (pooled marrow and spleen cells) gave only 37% of the responses of younger controls.

Given sufficient time and intact young recipients, immunologic stem cell lines from old donors populated recipients with cells having normal immune responses. These results suggest that age-related immunologic defects are not intrinsically timed in the precursor cell lines that populate the immune system.
Functional Heterogeneity Among the T-Derived Lymphocytes of the Mouse
Authors: Burns F. Dorr , Marrack Philippa C. , Kappler John W. , Janeway Charles A.
Published: December 2022

When spleen cells are cultured for 4 days in the presence of fetal bovine serum, T-cells are generated which nonspecifically suppress the humoral responses of non-precultured cells to a variety of antigens. These T cells can be generated from both short-lived, sessile T1 cells and long-lived, recirculating T2 cells, and thus appear similar to suppressor T cells activated by concanavalin A.
Cellular Cooperation During in Vivo Anti-Hapten Antibody Responses
Authors: Janeway Charles A.
Published: December 2022

Cellular interactions in adoptive secondary anti-hapten antibody responses to the hapten 2,4-dinitrophenyl (DNP) have been studied. It was shown that DNP-specific B cells must interact with carrier specific helper T cells to give optimal responses. Independent titration of B cell and helper cell activity in adoptive anti-DNP antibody responses gave the following results: Doubling the number of transferred B cells approximately doubled the subsequent antibody response. Doubling the number of helper cells leads to nearly 4 times as much anti-DNP antibody, measured 7 days after boosting (“premium effect”). This marked effect of helper cell number on the antibody response is thought to be due primarily to the interaction of two populations of carrier-specific cells in the helper effect, or to the interaction of two activities of a single population of helper cells, namely clone activation and clone expansion. Only a very small proportion of the premium effect given by helper cells could be attributed to increases in antibody affinity.
Cellular Cooperation During in Vivo Anti-Hapten Antibody Responses
Authors: Janeway Charles A.
Published: December 2022

An adoptive secondary anti-2,4-dinitrophenyl (DNP) antibody response involving T-B cell collaboration has been studied. In particular, attempts have been made to affect the unexpectedly steep log dose-response curve obtained when graded numbers of helper cells are transferred to irradiated recipients given a fixed number of B cells (premium effect). A variety of means were used to alter helper cell activity, and this activity was then measured quantitatively, as was the ability of the helper cells present after these treatments to give a premium effect. It was shown that activated T cells are approximately twice as active as spleen cells in helper activity and give a comparable premium effect. Graded doses of anti-θ serum plus complement markedly reduce the helper activity of spleen cells without affecting the premium effect given by the residual cells. Treatment of primed cell donors with limited doses of heterologous anti-mouse thymocyte serum (ATS) before transfer does not affect B cell activity, but readily inactivates helper cells, again without affecting the premium effect given by the residual cells. Adult thymectomy (ATx) of helper cell donors before priming with carrier led initially to increased helper activity relative to agematched control donors. This increase may reflect the loss of nonspecific suppressor T cells from spleens shortly after ATx. Late after ATx, there was about a 2-fold decrease in helper activity, probably reflecting a loss of helper cell precursors. At no time was there any change in the premium effect.

In view of the failure of any of the techniques used to abolish the premium effect given by helper cells in this response, it seems likely that this premium effect is due to the cooperative interaction of two very similar types of mature T cell. Alternatively, the premium effect observed here may result from the interaction of two activities of a single type of T cell which is mediated by independent factors.
Changes in the Immunochemical Properties of Highly Purified Properdin in Human Serum
Authors: Minta Joe Oduro
Published: December 2022

The fate of highly purified properdin (P̄) upon introduction into normal human serum or properdin-depleted serum (RP) was investigated. It was observed that, concomitant with the activation of the alternate pathway components, properdin underwent immunochemical alterations characterized by a shift in mobility from γ2 to β2 position and by an increase in the sedimentation rate from 5.1S to between 6.8 and 9.3S. The immunoelectrophoretic behavior of C3 was also altered with the appearance of a β2 arc in addition to the β1c arc. The immunochemical properties of altered P̄ resemble those of “native” properdin in fresh serum. The principle in serum (designated factor F) mediating these changes is a euglobulin with an approximate sedimentation rate and molecular weight of 9.0S and 250,000 daltons, respectively. The alteration in the immunochemical properties of P̄ may be due to aggregation of P̄ molecules or a complex formation between P̄ and a serum euglobulin (probably C3) mediated by factor F and it is associated with loss of ability of P to initiate the alternate pathway of complement activation upon interaction with serum.
Antibodies to Thymocytes in Sera of Normal Controls and of Patients with Schizophrenia
Authors: Singal D. P. , Grof P. , MacCrimmon D.
Published: December 2022

The sera from normal healthy individuals and from patients with schizophrenia were tested for cytotoxic antibodies against thymic antigens located on thymocytes. It was found that the sera from schizophrenics do not show higher cytotoxicity to mouse and rabbit thymocytes than the sera from normal controls. Furthermore, absorption of sera with adult mouse thymocytes and brain failed to show any difference between the sera from these two groups of subjects.
Spontaneous Modulation of Granulomatous Hypersensitivity in Schistosomiasis Mansoni
Authors: Boros D. L. , Pelley R. P. , Warren K. S.
Published: December 2022

Spontaneous diminution of granuloma formation around schistosome eggs in chronic schistosomiasis mansoni has been demonstrated previously. In the present study these findings were confirmed by injecting schistosome eggs into the pulmonary microvasculature and removing the lungs 8 days later from mice infected for 4, 8, 12, 16, and 20 weeks; the mean area of inflammation around the eggs was then measured. At 4 weeks a primary reaction was seen, by 8 weeks a massive secondary reaction occurred, but by 12 weeks the lesion was considerably reduced in size, and at 20 weeks it was smaller than the primary reaction. Concomitant measurements of humoral hemagglutinins to soluble egg antigens (SEA) revealed no detectable antibodies at 4 and 6 weeks, relatively low levels at 8 weeks, and an exponential increase in hemagglutinins at 12 weeks and beyond. Immunodiffusion analysis revealed no precipitins at 8 weeks, 1 major band and 2 minor bands at 12 and 16 weeks, and 2 major bands and 1 minor band at 20 weeks. Spleen cells from 8-week-infected mice showed peak migration inhibitory factor (MIF) output at a concentration of 1 µg/ml of soluble egg antigens and suppression of lymphokine secretion at higher concentrations. At 12 and 16 weeks, exponentially lower antigen concentrations both stimulated and suppressed peak MIF output; at 20 weeks MIF was not detectable after stimulation over a wide range of antigen concentrations. Delayed footpad swelling to SEA reached its peak at 10 weeks and declined thereafter, but the response to PPD in tuberculin-sensitized schistosome infected mice remained constant over the period of time studied.
Fractionation and Characterization of Surface Antigens from Group a Neisseria Meningitidis
Authors: Cheng William C. , Webb Elsie , Vedros Neylan , Ng James
Published: December 2022

Group A meningococcal surface components were first subjected to fractionation with a mixture of chloroform-methanol. Sodium dodecyl sulfate-acrylamide gel electrophoresis of the aqueous phase containing 30 to 40% of the original material revealed only two polypeptide components and a slowly migrating carbohydrate component. The soluble fraction of the interphase was found to contain most of the bacterial surface proteins and the chloroform-methanol phase essentially all of the lipid components. The components of the aqueous phase were further fractionated by use of the hydrophobic affinity column, 4-phenylbutylamino-Sepharose and gradient elution with NaCl to yield fractions I and II. Fraction II was further separated into a minor and a major component (IIb) with Sephadex G-200.

Fraction I contained the group A polysaccharide in ionic linkages with a minor polypeptide component (6%). It elicited bactericidal antibodies in rabbits and protected mice against homologous challenge, whereas the polysaccharide alone was non-immunogenic in these animals. Fraction IIb was a polysaccharide-polypeptide complex with unknown linkages; it induced a low concentration of rabbit antibodies that were bactericidal to group A and C meningococci. Mice vaccinated with fraction IIb were most resistant to homologous challenge and the resistance was also extended to challenges with group B and C cells. Fractions I and IIb appeared to be useful alternatives to the currently employed group-specific polysaccharide vaccines for the protection against drug-resistant meningococci. A simplified procedure for the preparation of group-specific polysaccharide was presented.
Quantitative and Immunologic Aspects of the Handling of 2,4 Dinitrophenyl Guinea Pig Albumin by Macrophages
Authors: Ellner Jerrold J. , Rosenthal Alan S.
Published: December 2022

Studies correlating quantitative aspects of the handling of dinitrophenyl guinea pig albumin (DNP-GPA) by guinea pig macrophages with the potential of cell-associated antigen to initiate proliferation of immune T lymphocytes have examined the nature of immunologically relevant antigen. After the loss of 75% of cell-bound DNP-GPA during the first 24 hr of in vitro culture, the remaining antigen persists qualitatively unchanged throughout further culture. However, coincident immunogenicity of the macrophage-associated DNP-GPA progressively decreases, suggesting loss of accessibility of the antigen to responding immune lymphocytes. There is a small, stable, surface antigen pool but these studies suggest that the immunologically critical fraction of DNP-GPA, as regards guinea pig T cell activation, is resistant to trypsinization and inaccessible to antibody.
Mastocytoma Cell Migration in Vitro: Inhibition by MIF-Containing Supernatants
Authors: Cohen Marion C. , Zeschke Richard , Bigazzi Pierluigi E. , Yoshida Takeshi , Cohen Stanley
Published: December 2022

Supernatants with macrophage migration inhibition factor (MIF) activity were obtained from cultures of antigen-stimulated guinea pig and human lymphocytes, and from SV40-infected monkey kidney cells. The monkey and human but not guinea pig preparations were effective in inhibiting migration of mastocytoma cells as well as macrophages. This inhibition of migration was not associated with cytotoxicity and was reversible.
Induction and Properties of Cytotoxic T Cells Specific for Hapten-Coupled Tumor Cells
Authors: Dennert Gunther , Hatlen Loren E.
Published: December 2022

Techniques are reported for the induction and assay of cytotoxic effector cells capable of specifically lysing hapten-coupled EL4 leukemia targets. It is shown that EL4 cells survive coupling with TNP-sulfonic acid and retain the hapten on their cell surface for a prolonged period of time. Although TNP-EL4 cells are readily lysed by anti-TNP serum in a complement-mediated reaction, they are inefficiently killed in an antibody-dependent cell-mediated reaction. Cytotoxic effector cells, able to lyse TNP-EL4 targets, are induced when C57BL/6 spleen H-2b cells are cultured with the following cell types which have been coupled with TNP: 1) allogeneic P815 tumor cells (H-2d), 2) syngeneic EL4 tumor cells, 3) allogeneic BALB/c spleen cells (H-2d), 4) syngeneic C57BL/6 spleen cells. Further experiments show that TNP-coupled xenogeneic chicken erythrocytes, which by themselves are unable to induce cytotoxic effectors, are capable of doing so if uncoupled P815 cells are present simultaneously. On the basis of these findings, it can be hypothesized that two stimuli are required for induction of these cytotoxic effector cells—one provided by the hapten, and the other by the P815 cell.

Treatment of cytotoxic spleen cells induced by hapten-coupled allogeneic tumor cells with anti-Thy-l serum and complement abrogates their cytotoxicity, indicating that T cells play a central role in the cytotoxic reaction. TNP-coupled erythrocytes do not serve as targets for these cytotoxic T cells, but do cause competitive inhibition of TNP-EL4 when added to the reaction mixture at high ratios. However, because the inhibition is relatively low, and because no such inhibition can be demonstrated with TNP-lysine, it is concluded that the receptor on the cytotoxic effector cell has a low affinity for hapten. This low affinity could be due to the receptor recognizing an antigen comprising mouse cell surface antigen in addition to the TNP moiety. Supporting this interpretation is the finding that TNP-EL4 cells competitively inhibit cytotoxicity much more efficiently than TNP-CRBC and that even uncoupled EL4 cells inhibit to some extent.
Letters to the Editor
Authors: Shifrine Moshe , Buhles William C.
Published: December 2022

Sir: Two recent publications by Fidler, Brodey, and Bech-Nielsen (1, 2) (one of them published in the J. of Immunology) address themselves to the study of the in vitro interaction of tumor cells and lymphocytes from dogs with spontaneous neoplasms. There are two important points that we would like to make concerning these papers.

Firstly, the method used to establish a tumor cell population does not, in fact, preclude contamination with large numbers of non-tumor cells. In our experience with over 100 canine tumors in culture, virtually all primary isolations consist of a mixed cell population of tumor cells, fibroblasts, and other cell types as determined by morphology and karyotyping (3). The ratio of these cells varies from tumor to tumor. We have been able to obtain pure populations of canine osteosarcoma cells after cloning procedures with confirmation by karyotypic analysis. The assumption made by Fidler et al. that the target cells they used are all (or even mostly) tumor cells is probably incorrect.
Letters to the Editor
Authors: Fidler I. J.
Published: December 2022

Sir: Ideally, all assays for cellular-mediated reactivity (CMR), in vitro, should be carried out with target tumor cells whose homogeneity was confirmed by karyotypic analysis. However, such analysis is most tedious and not practical for routine CMR dealing with autochthonous spontaneous neoplasms. Indeed, most published studies of CMR, in vitro, of human and animal tumors did not utilize tumor target cells analyzed karyotypically. In addition, the cloning procedure of tumor cells selects for one or few cells and their progeny, and these may be even less representative of the clinical tumor. Karyotypic analysis of target cells confirms their homogeneity, not their tumorigenicity.

In the section of Material and Methods we described an improved technique for reducing contamination of tumor cells with normal host cells. We did not use fresh tumor cells as targets, rather: 1) trypsinized tumor cells were allowed to attach to plastic for only 4 hr and then all nonadherent cells were washed off and cultures were refed.
Biophysical Characterization of the adr Subtype of Hepatitis B Antigen and Preparation of Anti-γ Sera in Rabbits
Authors: Gerin John L. , Faust Ralph M. , Holland Paul V.
Published: December 2022

The biophysical properties of the adr subtype of HBsAg were determined and found to be identical to those of the adw and ayw phenotypes. These properties were used to purify the 22 nm spherical form of HBsAg/adr from the serum of a chronic antigen carrier by using zonal centrifuge techniques. The purified antigen was injected into 15 rabbits which were bled weekly to follow the development of antibodies to HBsAg by hemagglutination, and to the subtype determinants of HBsAg by counterelectrophoresis and agar gel diffusion (AGD) assays. From these data and two pilot studies to produce anti-w using an adw and an ayw antigen the relative immunogenicities of the various HBsAg determinants could be ranked as a > r > d > y >> w. A selected pool of the sera from the rabbits immunized with HBsAg/adr was prepared as an interim reference reagent; when diluted 1:4 in saline and tested by AGD, it contains anti-a and anti-r without demonstrable anti-d or anti-normal human serum activity.
Specific Mitogenic Activity of 8-Br-Guanosine 3′,5′-Monophosphate (Br-Cyclic GMP) on B Lymphocytes
Authors: Weinstein Yacob , Segal Shraga , Melmon Kenneth L.
Published: December 2022

8-Br-cyclic GMP has been found to be a specific B cell mitogen; it triggers athymic nude mice spleen cells and “B mice” spleen cells, nylon adherent, anti-ϑ and complement-treated cells to proliferate. It does not stimulate thymocytes or purified T cells. The kinetics of the response to Br-cyclic GMP and LPS are almost identical. The mitogenic effect of LPS and Br-cyclic GMP is additive when the two mitogens are given together to cells. Spleen cells (C3H/HeJ strain) that did not respond to LPS were triggered by Br-cyclic GMP to make DNA. In order to achieve maximal stimulation by Br-cyclic GMP, the drug had to be in contact with the cells for more than 24 hr. Br-cyclic GMP was found to be mitogenic for spleen cells from five different mouse strains, but not for human leukocytes.

DB-cyclic AMP was found to inhibit the DNA synthesis of T lymphocytes after they interacted with Con A; DB-cyclic AMP had no effect on the ability of the B lymphocytes to be transformed by LPS. The differential effects of cyclic nucleotides on B vs. T lymphocytes are discussed.
Radioimmunoassay of Human Serum Antibody Specific for Adenovirus Type 5-Purified Fiber
Authors: Scott Jane V. , Dreesman Gordon R. , Spira Gad , Kasel Julius A.
Published: December 2022

A radioimmunoassay (RIA), utilizing a second antibody to separate immune complexes, was developed to provide a sensitive and specific measure of serum antibody to adenovirus type 5 (Ad 5) fiber. Purity of fiber antigen was ascertained by sodium dodecyl sulfate urea-polyacrylamide gel electrophoresis and isoelectric focusing in ampholyte pH gradients. After labeling with 125I to high specific activity, the iodinated fiber did not exhibit loss of antigenic reactivity and remained stable for 3 weeks when stored at -20°C with supplemental protein. Rabbit anti-Ad 5 serum with a neutralization titer of 1:320 precipitated 50% of the labeled fiber at a serum dilution of 1:50,000 when tested by the RIA. In competition assays as little as 0.5 ng of unlabeled fiber per milliliter was sufficient to inhibit the 125I fiber-antibody reaction. Serum specimens from 20 volunteers, obtained before and after vaccination with purified Ad 5 fiber or hexon subunit vaccine, were tested by RIA, hemagglutination-inhibition (HI), and neutralization tests. A comparison of mean antibody titers of post-inoculation sera showed that the RIA was 300 and 1000 times more sensitive than the HI and neutralization tests, respectively. Moreover, 19 of the men who were negative by the standard serologic tests before vaccination were shown to have anti-fiber antibody, with a mean RIA titer of 1:1028. Specificity of the RIA was demonstrated by the lack of an increase in antibody to Ad 5 fiber among those individuals vaccinated with the hexon subunit. Thus, the development of a highly sensitive and reproducible RIA allows for the detection of antibody specific for the Ad 5 fiber in serum which contains antibodies to the different virion antigenic determinants associated with Ad 5.
Different Roles of IgG and Complement Receptors in Phagocytosis by Polymorphonuclear Leukocytes
Authors: Mantovani Bernardo
Published: December 2022

The functions of IgG and complement receptors in phagocytosis of immune complexes by mouse polymorphonuclear leukocytes were examined by in vitro experiments. The immune complexes were sheep red cells (E) sensitized with IgG antibody (EA) or with antibody and complement (EAC). Inhibition experiments with Fab fragments of rabbit IgG antibody anti-mouse IgG have shown that the complement receptor is primarily involved in the attachment phase, whereas participation of the IgG receptor is necessary for inducing the mechanism of phagocytosis. The possible relevance of these findings for the in vivo mechanism of defense infection, and for the control of antibody synthesis is discussed.
Characteristics of Immune Interferon Produced by Human Lymphocyte Cultures Compared to Other Human Interferons
Authors: Valle Martti J. , Jordan George W. , Haahr Sven , Merigan Thomas C.
Published: December 2022

A factor with antiviral activity has been produced in vitro by combined macrophage-lymphocyte cultures from patients with recent herpes labialis in response to HSV antigen stimulation. It has been designated “immune interferon” and characterized in comparison to several other human interferons. It was shown to be relatively unstable at pH 2 and at 56° C. Rabbit anti-human leukocyte interferon serum was shown to be less active against immune interferon than against diploid cell interferon or against vesicle fluid interferon. The possibility of immune interferon being a totally different anti-viral protein or a protein with certain shared antigen determinants or structures with classical viral interferon is discussed. A simplified method for the assay of anti-interferon sera with microtiter plates is also described.
Normal Tissue Alloantigens and Genetic Control of Susceptibility to Tumors: Microcytotoxicity Studies on Resistant C3Hf and Susceptible (A × C3Hf)F1 Mice Inoculated with Transplacentally Induced C3Hf Lung Tumor
Authors: Martin W. John , Esber Elaine , Cotton W. Graeme , Rice Jerry M.
Published: December 2022

The immune response of mice to a transplacentally induced lung tumor was investigated with the microcytotoxicity (MC) assay. The tumor, originally induced in C3Hf mice, does not grow readily when transplanted to normal syngeneic C3Hf recipients. It grows readily, however, in (A C3Hf)F1 hybrids and in strain C3H mice, which express in their normal lung tissue a component which constitutes a strong lung tumor-associated transplantation antigen (TATA) in C3Hf mice. Both lung tumor-immunized C3Hf and tumor-bearing (A × C3Hf)F1 and C3H mice possessed lymphoid cells reactive against cultured lung tumor cells in the MC assay. Reactivity was also observed against cells cultured from normal lungs of (A × C3Hf)F1 and C3H mice, but not against cells similarly cultured from C3Hf or C57BL/6 mice. Anti-tumor MC was inhibited by serum-blocking factors present in some but not all tumor-bearing and tumor-immunized mice. The MC assay and detection by it of serum-blocking factors does not distinguish the effective anti-C3Hf lung tumor immune response of immunized C3Hf mice from the ineffective immune response of tumor-bearing (A × C3Hf)F1 and C3H mice. Furthermore, in lung tumor-bearing mice cells reactive in the MC assay may be directed against a normal tissue antigen rather than a tumor-associated antigen.
An Improved Procedure for Isolation of Ragweed Pollen Allergen Ra5
Authors: Roebber Marianne , Marsh David G. , Goodfriend Lawrence
Published: December 2022

Soon after the initial isolation of allergen Ra5 (1) from the pollen of short ragweed (Ambrosia elatior), human IgE-mediated sensitivity to this 5000 m.w. protein was shown to be significantly associated with the HL-A7 creg (cross-reacting group) of histocompatibility antigens (2, 3, 5). With determination of its amino acid sequence (6), Ra5 emerged as a potentially useful model for structure-function correlations in atopic hypersensitivity (4, 6, 7). However, extensive studies on the allergen have been hampered by the relatively low yields (approx. 10 mg/kg) obtained with existing isolation procedures (8) which have among other things utilized ether-extracted pollen as starting material. Following the finding by Marsh (3) that the aqueous extract formed from non-defatted pollen contained 330 to 350 mg antigen/kg, as determined by double antibody radioimmunoassay with anti-Ra5 serum, we restructured our method of isolation based on omission of the ether extraction step. As a result, we can report appreciable increase in yield of Ra5 in homogeneous form.
Regulation of MIF by a Factor in Human Serum
Authors: Ward Peter A. , Rocklin Ross E.
Published: December 2022

Virtually all inflammatory mediators are subject to regulatory control. In the case of kinins and vasopressor peptides it is well known there are both plasma and tissue-associated enzymes that inactivate these mediators (reviewed in 1). Recently it has also been shown that levels of chemotactic (leukotactic) mediators are controlled by the chemotactic factor inactivator (CFI)2 (2). CFI is present in normal-human serum in low concentration; elevations or deficiencies result in abnormally low or high amounts of chemotactic activity generated in serum after incubation with zymosan (3, 4). CFI has been shown to inhibit complement-derived and bacterial chemotactic factors and, also, the monocyte chemotactic activity present in lymphokine-rich supernatants from stimulated human lymphoid cells (3). The data presented in this report indicate that CFI-rich preparations also contain a factor that inactivates macrophage migration inhibitory factor (MIF).

Materials and Methods. Normal human blood lymphocytes were isolated by centrifugation in Ficoll-Hypaque gradients (5).
Antigenic Competition Between Heterologous Erythrocytes in Mice: Interference with Competition-Inducing Effect of Sheep Erythrocytes by Inclusion of Non-Competing Antigen
Authors: Ishikawa Hiromichi , Saito Kazuhisa
Published: December 2022

Antigenic competition was studied on reciprocal combinations of non-cross-reacting erythrocyte (RBC) antigens, i.e., sheep erythrocytes (SRBC), horse erythrocytes (HRBC), and chicken erythrocytes (CRBC), as competing antigen or as test antigen. Unlike SRBC and HRBC, CRBC did not induce antigenic competition, and mice previously injected with CRBC showed normal direct and indirect hemolysin plaque-forming cell (PFC) responses to the other RBC subsequently injected. Formaldehyde-treated (F) SRBC that retained the capacity to prime mouse for secondary anti-SRBC antibody response but lost the capacity to elicit primary antibody response also could not induce antigenic competition.

CRBC or F-SRBC, i.e., non-competing antigen, included with SRBC interfered with the competition-inducing effect of the latter RBC. When mice that had been injected with SRBC together with either one of the non-competing antigens were subsequently immunized with HRBC their anti-HRBC PFC responses were significantly restored, whereas mice pre-injected with SRBC alone showed markedly suppressed PFC responses to HRBC. Although the non-competing antigen was found not to affect both the direct and indirect PFC responses to the simultaneously injected SRBC, the response of the immune system to the non-competing antigen should alter the concurrent response to SRBC, and probably should prevent the generation of the suppressor thymus-derived (T) lymphocytes, allowing the generation of the helper T lymphocytes for IgM and IgG anti-SRBC antibody formation.
Splenic Regulation of Lymphocyte Trapping in Lymph Nodes Draining Tumor Grafts
Authors: Fightlin Richard S. , Lytton Bernard , Gershon Richard K.
Published: December 2022

We measured the trapping of 51chromium-labeled splenic lymphocytes in the lymph nodes and spleens of tumor-inoculated mice at various intervals after inoculation. Both histocompatible and incompatible tumors were studied. Significant trapping occurred in the draining lymph nodes over the entire course of the experiments. The splenic trapping response commenced very early after transplantation and abated after a few days. The magnitude of the lymph node trapping response correlated with the magnitude of antigenic difference between tumor and host. Splenectomy before tumor inoculation had bidirectional effects; it caused a marked augmentation of the trapping response in the early period after tumor inoculation, which declined with time so that late after tumor inoculation, splenectomized animals trapped less well than intact controls. Tumor resection always led to a rapid fall in the trapping response in normal mice. However, tumor resection at the time of the peak trapping response of splenectomized mice produced no change. These results support the notion that lymphocyte trapping is a valid and useful monitor of the immune response to tumors. They also serve to reemphasize the important regulatory role of the spleen plays in the immune response to tumor-associated antigens by demonstrating a new parameter of splenic control; regulation of lymphocyte traffic in response to antigenic stimuli.
Synthesis of Surface H-2 Alloantigens in Murine Splenocytes
Authors: Vitetta Ellen S. , Uhr Jonathan W.
Published: December 2022

The synthesis and turnover of cell surface H-2 alloantigens were studied in murine splenocytes by the anti-H-2-binding method to separate precursor-labeled surface from intracellular molecules. Results indicate that newly synthesized H-2 antigen, labeled in either its peptide or carbohydrate portion enters a relatively small pool of intracellular H-2 antigen and then is rapidly transported to the plasma membrane which represents a larger compartment. The simplest interpretation of these findings is that H-2 antigen is synthesized and transported along a conventional secretory pathway. Pulse-chase experiments indicate that H-2 antigens are not readily chased from the plasma membrane and are not shed or internalized during short-term culture. The aforementioned observations are discussed in terms of a cellular heterogeneity.
Restoration of in Vitro Immune Responsiveness of Mastocytoma-Suppressed Splenocytes by Activated T Cells
Authors: Kamo Isao , Patel Chandu , Patel Navin , Friedman Herman
Published: December 2022

Spleen cells from normal DBA/2 mice pretreated with a soluble factor from mastocytoma cells or from ascitic fluid of mastocytoma-bearing mice were markedly impaired in terms of antibody formation to SRBC in vitro. Such immunosuppression by mastocytoma homogenates or ascitic fluid was reversed when syngeneic T cells activated to SRBC were added to the cultures, but not when peritoneal exudate cells or anti-ϑ-treated normal splenocytes were used. Activated T cells, as well as normal B lymphocytes prepared from spleens of lethally irradiated mice reconstituted with bone marrow cells, were less sensitive to the immunosuppressive factor than non-activated T cells. The ability of educated T cells to restore immunocompetence of suppressed spleen cells in vitro suggests that the target of the immunosuppressive factor from mastocytoma cells may be non-activated T cells, especially those involved in T cell helper function.
Specific Suppression of the Antibody Response in Vitro by Serum from Paralyzed Mice
Authors: Brown Paul B. , Köhler Heinz , Rowley Donald A.
Published: December 2022

BALB/c mice immunized with either the whole vaccine or the C-polysaccharide obtained from the R36A strain of pneumococcus produce antibody to phosphorylcholine. Mice injected i.v. with a single high dose of the C-polysaccharide are specifically unresponsive to immunization to phosphorylcholine for many months and are considered paralyzed. The induction of paralysis does not eliminate cells reactive to phosphorylcholine; however, serum from paralyzed mice specifically suppresses the response of cultures of normal spleen cells to phosphorylcholine. Paralyzed mice have an early low antibody response to phosphorylcholine and to the receptor for phosphorylcholine as indicated by plaque-forming cell assays. The factor or factors present in serum which may suppress cultures, and, by presumption, be responsible for paralysis are complexes of antigen, antibody, and antibody to the receptor for phosphorylcholine.
Host Defenses Against Influenza Virus: The Role of Anti-Hemagglutinin Antibody
Authors: Virelizier Jean-Louis
Published: December 2022

In CBA mice the protection provided by transferred immune spleen cells or by antibody has been investigated in immunologically intact, cyclophosphamide-treated and thymus-deprived animals infected with A/PR8 virus. The degree of protection was more closely related to serum antibody levels than to the presence of immune lymphocytes in recipients. Comparison of the protective efficiency of various anti-influenza antisera with different specificities within an influenza A subtype indicated that antibodies recognizing the strain-specific determinants of the influenza hemagglutinin have an important role in protection. Physiologic amounts of transferred antibodies were shown to protect immunodepressed mice, suggesting that, provided a sufficient amount of specific antibodies is secreted, the participation of effectors of cell-mediated immunity is not essential. However, our results suggest that thymusderived lymphocytes have an indirect role in protection by enhancing, through their helper effect, the secretion of anti-influenza antibodies.
IgE-Induced Blood Coagulation Alterations in the Rabbit: Consumption of Coagulation Factors XII, XI, and IX in Vivo
Authors: Pinckard R. Neal , Tanigawa Cheryl , Halonen Marilyn
Published: December 2022

Intravenous administration of BSA into 3-month-old rabbits producing detectable anti-BSA antibody only of the IgE class of immunoglobulin induced a variety of intravascular blood coagulation alterations. The IgE-induced blood coagulation alterations observed in the plasma 15 min after antigen challenge included: a) the intravascular consumption of intrinsic blood coagulation factors XII, XI, and IX and possibly the reduction in clottable fibrinogen; b) a significant prolongation of the activated partial thromboplastin time but not the prothrombin time; and c) the production of an inhibitor affecting the last stage of blood coagulation. The observed blood coagulation alterations were not caused by the manipulative procedures utilized, the presence of anti-BSA, IgG or IgM antibody, histamine-induced alterations in the vascular endothelium or the development of hypotensive shock. It is proposed that specific IgE antibody can induce directly or indirectly the activation of intrinsic blood coagulation in vivo.