bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on the Control of Antibody Synthesis
Authors: Weksler Marc E. , Merritts L. Lynn , Werblin Theodore P. , Siskind Gregory W.
Published: December 2022

A single intravenous injection of antigen in adult rabbits was shown to depress specifically antibody synthesis upon subsequent immunization with antigen in complete Freund's adjuvant. The degree of tolerance was dependent upon the dose of antigen and the time interval between intravenous antigen and immunization with antigen in adjuvant. Tolerance could not be transferred passively to normal rabbits with serum from tolerant animals and could not be overcome by using a supraoptimal immunizing dose of antigen in Freund's adjuvant. Partial tolerance resulted in a moderate depression in the average affinity of the antibody synthesized.
Immunologic Aspects of Hepatitis-Associated Antigen and Antibody in Human Body Fluids
Authors: Ogra Pearay L.
Published: December 2022

Employing the techniques of immunoelectrophoresis and indirect immunofluorescence with smears of peripheral blood, we determined the presence of hepatitis-associated antigen (HAA) and anti-HAA antibody in concentrated specimens of serum, nasopharyngeal washings, urine, and fecal extracts from patients with icteric or asymptomatic HAA infection. Highest concentrations of HAA in the serum were demonstrable at the time of initial screening. HAA was detected in the nasopharyngeal washings and urine for a short period in a few subjects. All subjects manifested appreciable quantities of HAA in fecal extracts and the antigen persisted in the feces for as long as 4 months. Anti-HAA antibody response in the serum was characterized by infrequent appearance of small amounts of γG antibody 2 to 3 months after the initial detection of antigen in the serum. No antibody activity was detected after 7 months. The response in the nasopharynx and urine was characterized by transient appearance of small amounts of γA and infrequently of γG class of antibody. The response in the fecal extracts was characterized initially by frequent appearance of HAA-anti-HAA complexes. Subsequently, appreciable levels of free γA antibody to HAA were detected for as long as 8 months. It is suggested that HAA may replicate in various mucosal surfaces, and such a mechanism may explain the nonparenteral transmission of long incubation hepatitis.
Host Defenses During Primary Venezuelan Equine Encephalomyelitis Virus Infection in Mice
Authors: Adler William H. , Rabinowitz Stanley G.
Published: December 2022

Attenuated Venezuelan equine encephalomyelitis virus immunization produces an antigen-reactive cell population in mice which is capable of responding specifically to homologous viral antigen. This reaction is short lived, involves thymus-derived or dependent (T) lymphocytes and requires the presence of macrophages in the culture system. Antibody competitively blocks T cell recognition and transformation. Booster immunization with attenuated VEE virus 35 days after initial immunization restores lymphocyte reactivity to previous peak values but produces only a bone marrow-derived or dependent lymphocyte response. Passive transfer studies demonstrate that only antigen-reactive cells which are susceptible to being killed by antisera directed at the T lymphocyte are capable of conferring adoptive immunity in a normal recipient.
Antibody-Independent Activation of Human C1 after Interaction with Heart Subcellular Membranes
Authors: Pinckard R. Neal , Olson Merle S. , Kelley Robert E. , DeHeer David H. , Palmer John D. , O'Rourke Robert A. , Goldfein Samuel
Published: December 2022

A 19S, complement-fixing, heat-labile factor reactive with human heart mitochondria and microsomes has been detected in all normal human serum. This serum factor reacted with autologous and homologous heart mitochondria but did not react with mitochondria derived from cardiac tissue from other species. Subsequent characterization has indicated that this heat-labile factor was the first component of human complement. The C1 reacted with the mitochondrial membrane, was activated, and resulted in an apparent antibody-independent activation of the complement system.
Suppression of the Antibody Response to SRBC with F(ab′)2 and IgG in Vitro
Authors: Wason William M. , Fitch Frank W.
Published: December 2022

The role of the Fc portion of the IgG molecule in antibody-mediated regulation of the immune response is controversial (16, 17). If the Fc portion of the antibody molecule is removed by enzymatic digestion, the F(ab′)2 portion is rapidly excreted by the kidneys and is lost from the circulation in intact animals (18). This rapid loss could possibly account for the reported difference in the efficiency of intact IgG and F(ab′)2 in suppressing the immune response in vivo (7, 17). A tissue culture system avoids the problem of variable excretion. The experiments reported here demonstrate a significant difference in the efficiency of intact IgG and F(ab′)2 (anti-SRBC) (a-SRBC) antibody in suppressing the immune response of cultured mouse spleen cells to SRBC in vitro, and suggest that the Fc portion plays an important role in antibody-mediated suppression of the immune response to SRBC.
β2-Microglobulin Production and Secretion by Lymphocytes in Culture
Authors: Poulik M. D. , Bloom A. D.
Published: December 2022

A β2-microglobulin with molecular weight of 12,600 daltons was initially isolated from the urine and serum of patients with renal tubular insufficiency (1). Smithies and Poulik (2) determined the partial amino acid sequence of this globulin and found that 24 of the first 46 amino acid residues were homologous with the amino acid sequence of parts of the heavy chain of the G-1 myeloma protein. This suggested that a structural relationship existed between the β2-microglobulin and the constant region of human IgG immunoglobulins. In spite of this structural relatedness, specific antisera against all known immunoglobulin chains (including secretory piece and J-chain) did not react with this protein. Subsequently, Peterson et al. (3) established the complete sequence of this protein and found that 28 amino acid residues of the β2-microglobulin were homologous with the CH3 domain of the myeloma protein. An homologous protein was also isolated recently from the dog (4), and its partial amino acid sequence was established and related to β2-microglobulin of human origin.
Lymphokine Toxicity for Yeast Cells
Authors: Pearsall Nancy N. , Sundsmo John S. , Weiser Russell S.
Published: December 2022

Mouse lymphocytes, stimulated by phytohemagglutinin-P, produce a variety of substances known collectively as lymphokines. These substances are thought to be mediators of cellular immunity and delayed-type sensitivity. Lymphokine preparations also have antifungal activity against two species of yeasts tested, Saccharomyces cerevisiae and Candida albicans.
The Macrophage Receptor for IgG: Number and Affinity of Binding Sites
Authors: Arend William P. , Mannik Mart
Published: December 2022

The number of receptor sites for IgG on rabbit alveolar macrophages was determined and the strength of interaction between monomeric IgG and the cell receptors was estimated. Heavily stimulated alveolar macrophages (obtained after multiple monthly intravenous injections of complete Freund's adjuvant) possessed 2.16 ± 0.34 × 106 receptor sites for IgG per cell. Minimally stimulated cells (obtained after a single injection of adjuvant) possessed 1.21 ± 0.23 × 106 receptor sites for IgG per cell. During stimulation of macrophages with adjuvant, the increase in number of IgG receptor sites was accompanied by an increased binding of soluble immune complexes. The average association constant between these receptors and rabbit IgG was 9.02 ± 2.58 × 105 L/M and 7.59 ± 1.83 × 105 L/M, for heavily and minimally stimulated cells, respectively. Within each group the degree of adherence of the soluble complexes to the cells was directly correlated with the affinity of the receptor sites, and not with their number. Treatment of cells with proteolytic enzymes led to an increased adherence of soluble complexes. This enhanced activity was due to an increase in the average association constant of the receptor sites for IgG on enzyme-treated cells obtained from male rabbits, although no change in the number of sites was observed. Similar enzyme treatment of cells obtained from female rabbits produced an increase in the number of receptor sites for IgG, although no increase in their average association constant occurred.

It is concluded that the relative adherence of soluble immune complexes to macrophages is influenced both by the number of receptor sites for IgG per cell, and by their average association constant.
Biosynthesis and Secretion of Antibody by Chicken Lymphoid Cells
Authors: Choi Yong Sung , Good Robert A.
Published: December 2022

A method to study biosynthesis and secretion of antibody by chicken lymphoid cells has been developed. Briefly, antibody was labeled with leucine-3H by incubating immunized chicken lymphoid cells in a short-term tissue culture, and was purified from other cellular proteins by specific immune adsorption to polymerized antigen. Biochemical structure of antibody molecules purified above was studied by sodium dodecyl sulfate-acrylamide gel electrophoresis.

Using this method, we demonstrate that spleen cells of the chicken actively synthesize and secrete antibody which was characterized as being an immunoglobulin by serology and acrylamide gel analysis. However, immunoglobulin antibody is not detectable in bursa cells by this method regardless of the routes of bovine γ-globulin immunization.
Characterization of IgE-Induced Systemic Anaphylaxis in the Rabbit. Lack of Correlation between the Intravascular Release of Histamine and Anaphylactic Sensitivity
Authors: Halonen Marilyn , Pinckard R. Neal , Meng Aniko L.
Published: December 2022

The administration of viable adult peritoneal exudate cells together with soluble bovine serum albumin (BSA) to newborn rabbits facilitated the subsequent development of anaphylactic sensitivity to BSA. In contrast, the administration of killed peritoneal exudate cells to littermates of the above rabbits did not result in the development of anaphylactic sensitivity. Both groups of rabbits had statistically similar incidences of detectable anti-BSA IgE antibody immediately before anaphylactic challenge on day 91, and no other class of anti-BSA antibody was detectable. Anaphylactic positive and negative rabbits were compared for differences in IgE-induced intravascular changes that occurred after anaphylactic challenge. Although substantial decreases in total blood histamine, basophils, and platelets were observed 1 hr after anaphylactic challenge, no statistical differences between the two groups were noted. These data indicate that the phenotypic expression of IgE-induced anaphylactic sensitivity in the rabbit is dependent upon factors in addition to those controlling the circulating levels of free and basophil-bound IgE antibody.
Pulmonary Host Defenses
Authors: Reynolds Herbert Y. , Thompson Russell E.
Published: December 2022

The antibody responses in serum and the lower respiratory tract of rabbits immunized with Pseudomonas aeruginosa were examined. After parenteral immunization, IgG agglutinative antibody was present in bronchial secretions, which was probably derived from the intravascular pool of IgG by diffusion into respiratory fluids. After intranasal vaccination, antibody activity was detected in both secretory IgA and the IgG fractions of bronchial secretions; it was considered possible that locally synthesized IgG antibody was elicited. Immune secretory IgA had an inhibitory effect on the growth of lag phase Pseudomonas organisms in addition to its agglutinative activity, both of which were specific for the immunizing serotype. IgG and bronchial secretions did not have this inhibitory effect. IgM and complement components were noticeably absent from bronchial secretions which would indicate that the immune milieu of normal lungs is quite different from that of serum.
Protection Against Enteric Bacterial Infection by Secretory IgA Antibodies
Authors: Fubara Ernest S. , Freter Rolf
Published: December 2022

This paper elaborates and extends earlier reports from this laboratory on the mechanisms by which coproantibodies protect against enteric bacterial infection. Purified antibody preparations obtained from the lumen of the intestine of orally vaccinated germfree mice, containing very small amounts of S-IgA but no other serologically active immunoglobulins, were found to protect mice against experimental enteric cholera infection. This protection was associated with a reduction in that fraction of the total vibrio population which was adsorbed to the mucosa. Total counts of intestinal vibrio populations were not affected. The mechanism of this process was investigated by using metabolic inhibitors in experimental infections of intestinal loops of rabbits. Iodoacetate reversed the antibody-mediated shift of the vibrio population away from association with the mucosa. This suggests that the mucosal cell may furnish some accessory factor which, in conjunction with antibody, results in an antibacterial mechanism on the mucosal surface. Iodoacetate had no effect on total vibrio population and adsorption of vibrios in nonimmunized intestinal loops. Consequently, a direct effect of iodoacetate on the vibrios, rather than on the mucosal cells, is a less likely explanation for the observed effects.
Absence of Rosette-Forming Cells in Delayed Hypersensitive Agammaglobulinemic Chickens
Authors: Theis G. A. , Weinbaum F. I. , Thorbecke G. J.
Published: December 2022

Delayed hypersensitivity was produced by immunization with sheep erythrocytes (SE) in adjuvants in bursectomized and normal chickens and the numbers of rosette-forming cells (RFC) in the spleen were determined. It was found that RFC occurred only in animals with τ 0.1% of normal serum γ-globulin levels and that an increase over background was seen primarily after intravenous immunization with SE in such animals. RFC were not killed by specific rabbit anti-chicken thymus sera but were eliminated by treatment with anti-bursa or anti-chicken Ig sera in the presence of C. Incubation of cells with anti-Ig at 4°C also inhibited 100% of RFC. It was concluded that none of the RFC in the chicken are of thymic origin.
Immunity to Chlamydial Infections of the Eye
Authors: Watson Ronald R. , MacDonald A. Bruce , Murray Edward S. , Modabber Farrokh Z.
Published: December 2022

Guinea pig inclusion conjunctivitis, a chlamydial infection of the eye, induced systemic, delayed hypersensitivity as measured by skin testing and by the macrophage-migration inhibition (MI) assay. Four to 6 days after infection positive skin reactions marked the onset of delayed hypersensitivity. Skin reactions were larger and more positive from days 11 through 83 post-infection; smaller and less positive reactions were observed on days 350 to 850. The results of the MI assay correlated well with skin tests. Circulating and eye secretion antibodies appeared by day 11 and were still present in low titers by day 850. These results indicate that the ocular, chlamydial infection induced long lasting, systemic responses in the circulating, secretory, and cell-mediated immune systems. The role of cell-mediated immunity in resistance to chlamydial reinfection of the eye is discussed.
T Cell Deficiency in Mouse Allogeneic Radiation Chimeras
Authors: Urso Paul , Gengozian Nazareth
Published: December 2022

For obtaining direct evidence of cellular defects in immunologically deficient chimeras, spleen adherent (A) and nonadherent (T and B) cells, were cultured in vitro with sheep erythrocytes and the plaque-forming cell (PFC) response was determined. Chimera A cells cultured with normal host or donor T and B cells produced normal levels of PFC. The response of chimera T and B cells cultured with normal A cells was considerably lower than that of normal host or donor cells. The response of spleen T and B cells from the allogeneic combination (BALB/c × DBA/2)F1 bone marrow → (C3H × C57BL)F1 was enhanced by thymus cells from rats, normal hosts, and various mouse strains including parental BABL/c (donor) mice but not by cells from normal donors and parental DBA/2 (donor) mice. Bone marrow cells were also ineffective. In the allogeneic combination (C3H × C57BL)F1 → (BALB/c × DBA/2)F1 host thymus again was effective but not donor thymus. Thymus cells from chimeras did not improve the response of either allogeneic combination. Chimera T and B cells cultured with normal host or donor T and B cells severely inhibited the response of host cells but only slightly affected donor cells. Thus, the T cell population is defective in allogeneic chimeras while A cells behave normally and B cells are present and functional in the spleen. Improvement of the response by genetically incompatible (to donor-type chimera cells) thymus cells suggests an allogeneic effect which may be enhanced by the sensitivity of chimera cells to host antigens. The contrasting influences of host, donor, and parental (donor) thymus cells in reconstitution of immune competence indicates that nonspecific and specific factors are operating in inducing immunologic deficiency in chimeras.
Characteristics of a Cytotoxin (“Lymphotoxin”) Produced by Stimulation of Human Lymphoid Tissue
Authors: Peter James B. , Stratton Joan A. , Stempel Kerstin E. , Yu David , Cardin Claire
Published: December 2022

Stimulated lymphocytes derived from human peripheral blood, tonsils, adenoids and thoracic duct lymph elaborate a cytotoxic activity (cytotoxin, lymphotoxin) which causes release of 14C-protein or 51Cr from prelabeled L cells or HeLa cells. Mouse mastocytoma and Ewing's sarcoma cells, as well as red blood cells and lymphocytes of several species, are not susceptible to the cytotoxin and none of the cells tested, including L cells, absorb or inactivate the cytotoxin. Inhibition of L cell proliferation by mitomycin C or colchicine does not alter susceptibility to cytotoxin but cytotoxin-induced release of 51Cr is increased by irradiation of L cells.

The kinetics of action of cytotoxin are complicated by a lag phase which is temperature sensitive as opposed to the rate of 51Cr release itself which is similar at different temperatures after the lag phase is completed.

Characteristics of the cytotoxic activity, including its heat lability, inhibition by certain drugs, precipitation with (NH4)2SO4, molecular size, and polydisperse chromatographic pattern, are similar regardless of the lymphocyte source. Most of the cytotoxic activity induced by stimulation with purified phytohemagglutinin P, concanavalin A, pokeweed mitogen, and purified protein derivative tuberculin (PPD) is precipitated at 37% to 50% (NH4)2SO4 saturation. The cytotoxin elaborated by PPD stimulation of sensitized lymphocytes is significantly more heat labile than that induced by mitogen.

A variety of putative substrates for the cytotoxin do not inhibit its action. Serum of some normal rabbits contains a potent inhibitor.
Enumeration and Analysis of Antibody-Forming Cell Precursors in the Neonatal Mouse
Authors: Press Joan L. , Klinman Norman R.
Published: December 2022

The ability of antibody-forming cell precursors (B cells) obtained during murine development to participate in the splenic focus response to a hapten-protein conjugate was determined by the technique of adoptive cell transfer to irradiated, syngeneic, carrier-primed recipient mice. Spleen fragments from recipients of fetal liver, fetal spleen, or neonatal spleen cells produced anti-hapten antibody when exposed in in vitro primary organ culture to dinitrophenylated hemocyanin (DNP-Hy). The number of antibody-producing fragments (splenic foci) obtained was linearly related to the number of nucleated spleen cells injected, and did not vary significantly for neonatal spleen cell populations obtained during the 1st week of postnatal life. Although neonatal foci released less antibody than that reported for adult foci (1), the dose dependence of antigenic stimulation is comparable for both neonatal and adult primary precursor cells, implying comparable mechanisms for antigenic recognition and stimulation. The frequency of precursor cells in the neonatal spleen for the haptenic determinant DNP is only slightly lower than the reported frequency (1) of adult precursor cells of the same specificity. This finding is discussed in terms of the relevance of antigenic selection to the generation of diversity.
Separation of Lymphocytes from Disaggregated Mouse Malignant Neoplasms by Sedimentation in Gradients of Ficoll in Tissue Culture Medium
Authors: Zettergren Judy G. , Luberoff Douglas E. , Pretlow Thomas G.
Published: December 2022

A method is reported for separating lymphocytes from solid tumors. After the sequential use of columns of glass beads, velocity (isokinetic) sedimentation, and isopycnic sedimentation, lymphocytes were obtained in 76.3 to 96.8% purity from disaggregated tumors. When isopycnic sedimentation was omitted, lymphocytes were obtained in 58.4 to 95.1% purity.
Characterization and Antibody-Binding Capacity of Streptococcal Group a Carbohydrate Haptens
Authors: Shulman Stanford T. , Ayoub Elia M.
Published: December 2022

Tritium-labeled group A streptococcal carbohydrate haptens have been prepared by incubation of acid hydrolysates of group A carbohydrate with 3H-borohydride. Isolation of hapten was achieved by column and thin layer chromatography. These procedures yielded an antigenically univalent hapten, “α hapten,” with an estimated molecular weight of 1100 daltons and a rhamnose: glucosamine molar ratio of 6:1. The α hapten inhibited the binding of A-carbohydrate with homologous antiserum. Comparative antibody-binding studies were performed with group A streptococcal rabbit antibody preparations by using the α hapten and a synthetic 3H-p-nitrophenyl-β-N-acetylglucosamine (3H-pNøNAG) hapten. These studies demonstrate that average association constants (K0) obtained with the α hapten are one to two orders of magnitude higher than the K0 determined with 3H-pNøNAG. Significantly less nonspecific binding was observed with the α hapten. Binding studies with the α hapten enabled the determination of K0 for a number of antibody preparations which failed to evidence specific binding with 3H-pNøNAG. The availability of the α hapten makes more feasible the reliable determination of K0 for antibody prepared under various conditions.
Reconstitution of T Cell-Depleted Spleen Cell Populations by Factors Derived from T Cells
Authors: Gorczynski R. M. , Miller R. G. , Phillips R. A.
Published: December 2022

Initiation of an in vitro response to sheep erythrocytes (SRBC) requires the production of a nonspecific soluble mediator which is present in the culture supernatants of antigen-stimulated thymus-derived (T) cells. This factor was shown to act early in the immune response and probably directly affects bone marrow derived (B) cells. However, the stimulation of the PFC response caused by the factor depends upon the presence of some intact T cells. These T cells can still function after 1300 rads of ionizing radiation. The factor does not simply cause proliferation of small numbers of surviving T cells, and is exhausted from the medium when cultures of B cells with the active supernatant are stimulated with antigen.

Further evidence for two functions for T cells during the antibody response in culture has been obtained by studying the ability of cultures of B and A cells to respond when the two T cell functions are provided sequentially. For example, if B cells are preincubated in vitro for 24 hr with an active supernatant, they can give subsequent PFC responses either in vivo or in vitro when mixed with irradiated normal (unprimed) T cells. Investigation of the need for intact T cells has shown that this requirement, unlike the nonspecific factor, represents an antigen-specific function for T cells. This specificity has been demonstrated with irradiated T cells from mice tolerant to SRBC. In the presence of the active supernatant and a suitable B cell source these irradiated T cells can reconstitute the response to horse erythrocytes but not SRBC.
Reversibility of the Immunodepression Due to Ehrlich Ascites Carcinoma
Authors: Mocarelli Paolo , Villa Maria Luisa , Garotta Gianni , Porta Carla , Bigi Giuseppe , Clerici Enrico
Published: December 2022

Earlier studies from our laboratory indicated that mice bearing Ehrlich ascites carcinoma cells (EAC)2 produce far fewer plaque-forming cells (PFC) after immunization with sheep erythrocytes (SRBC) than do normal animals. The immunodepression develops shortly after a low dose of EAC, before wasting, and does not result from lymphocyte depletion of peripheral organs, as ascertained by cytometric and histologic studies (1, 2).

The failure to produce PFC may be due to lymphocyte malfunction or to some other mechanism such as antigen processing in tumorbearing animals.

In the present study we demonstrate that thymus- and bone marrow-derived lymphocytes, and spleen cells from tumor-bearing and normal donors, transferred to x-irradiated syngeneic mice, produce the same numbers of PFC to SRBC.

Materials and Methods. Animals. C57BL inbred mice, 8 to 12 weeks old, were used.

Tumor. Inocula of 107 cells of a half-diploid and half-tetraploid strain of EAC were injected intraperitoneally (i.p.) into prospective donors of thymus, bone marrow and spleen cells (3).
Production of Functional T Cells after Treatment of Bone Marrow with Thymic Factor
Authors: Miller Harold C. , Schmiege Susan K. , Rule Allyn
Published: December 2022

Incubation of bone marrow cells with thymic factor obtained from bovine thymus tissue resulted in the development of cells with T lymphocyte-like function. Adoptive transfer of these cells into lethally irradiated syngeneic mice followed by antigen stimulation with sheep erythrocytes provided a system for testing the ability of the thymic factor-treated cells to induce B cells to become antibody-forming cells. The former were capable of substituting for thymocytes since antibody-forming cells were detected in the recipient spleens upon assay for hemolytic plaques 9 days after transplantation. Limiting dilution experiments in which graded numbers of thymic factor-treated marrow cells were transferred along with a nonrestricted supply of normal bone marrow cells and antigen indicated that the critical number of thymic factor-treated marrow cells needed to synergize with B cells in the production of an anti-SRBC response was contained in 3 × 105 marrow cells. Quantitative and qualitative differences between the thymic factor-treated cells and thymus-derived T cells are discussed.
Effect of Cortisone Therapy on Serum Complement Components
Authors: Atkinson John P. , Frank Michael M.
Published: December 2022

Male guinea pigs weighing 300 to 400 g were given a daily subcutaneous injection of 5, 20 or 100 mg/kg body weight of cortisone acetate. Measurement of serum cortisol demonstrated sustained blood levels proportional to the dose. All nine components of the classical complement pathway were measured by functional assay. At 5 mg/kg/day significant (p < 0.05) changes noted were a 20 and 51% elevation of the CH50 and C1 titers, respectively. C4, C2, and C3–9 complex were higher than saline treated controls but these increases were not statistically significant. High dose glucocorticoid therapy produced a marked depression in the titer of a number of components. At 20 mg/kg/day C4 and C8 titers were most strongly affected although all components except C1 and C9 were at least 15% lower than controls. At 100 mg/kg the titers of all components except C1 and C9 were markedly depressed. During the first 2 weeks of therapy there was a progressive fall in complement followed by stabilization at these lower levels. On discontinuation of therapy 2 weeks were required before the levels of these components returned to normal. Albumin and globulin levels, hematocrit, spleen size, and percentage of weight gain were unaltered by cortisone therapy. There was no effect on hemolytic complement levels by adding hydrocortisone in vitro to guinea pig serum in comparable doses to those obtained in vivo. Thus, altered catabolic and/or synthetic rates are responsible for the cortisone-induced changes in serum complement.
Effect of Locally Administered Endotoxin on Regenerating Appendix Structure and Responses of Appendix Cells to Mitogens
Authors: Blythman Hildur E. , Waksman Byron H.
Published: December 2022

In adult rabbits subjected to appendicostomy and sublethal irradiation, there was almost total destruction of appendix follicle lymphocytes with relative preservation of appendix domes and thymus-dependent areas (TDA). Regeneration of follicles was very incomplete during a 21-day period of observation. Daily instillation of endotoxin (LPS) or of killed Escherichia coli into the appendix over a 7-day period or longer failed to enhance follicle regeneration but gave rise instead, to an obvious and marked stimulation of macrophages, which replaced the follicle lymphocyte population entirely in most rabbits. The response of appendix cells in culture to PHA was parallel to the relative proportion of cells in the TDA, as established histologically, and was therefore exaggerated in the instances where follicle macrophages were stimulated and follicle lymphocytes depressed.

This experimental situation (regeneration after x-ray) differs from the normal developmental pattern of the appendix in young rabbits because of the presence of a large preexisting population of macrophages, responsive to LPS stimulation from the lumen and capable of overwhelming normal lymphocytic function.
Lymphocyte Phytohemagglutinin Responsiveness, Immunoglobulins and Autoantibodies in Aging Humans
Authors: Hallgren H. M. , Buckley C. E. , Gilbertsen V. A. , Yunis E. J.
Published: December 2022

Phytohemagglutinin responsiveness of peripheral blood lymphocytes declines in aged humans. Older humans exhibit a rise in the incidence of autoantibodies. The incidence of autoantibodies becomes significantly greater among older subjects over 80 years of age and is most significant in humans with low lymphocyte phytohemagglutinin responses. Serum levels of IgG and IgA were increased in subjects surviving beyond the 7th decade. Correlations could not be detected between serum immunoglobulin levels, titers of individual autoantibodies and the lymphocyte response to phytohemagglutinin. These observations suggest that hyperglobulinemia, impaired lymphocyte phytohemagglutinin responsiveness, and increased frequency of autoantibodies are characteristic of surviving older humans and represent associated events in the natural history of human life.
Synthesis of IgG, IgA, IgM by Chicken Tissues: Immunofluorescent and 14C Amino Acid Incorporation Studies
Authors: Bienenstock John , Gauldie Jack , Perey D. Y. E.
Published: December 2022

Chicken tissues have been studied with immunofluorescent techniques to establish the distribution and relative numbers of cells stained specifically with anti-α, -µ, and -γ antisera. In the gut, IgA predominated; most numbers of IgA cells were found in the duodenum, and less further down the intestinal tract. The cecal tonsil contained mostly IgG, followed by IgM cells. Only a few IgA cells were seen in the cecal tonsil. The spleen showed a similar distribution of immunoglobulins. No positive cells were seen in either crop or Peyer's patches. 14C-labeled amino acid incorporation into IgA, IgM, and IgG was also studied by in vitro organ culture techniques and radioimmunoelectrophoresis. These results confirmed the immunofluorescent studies. The thymus showed labeling of IgG but not IgA. Bursa incorporated label into IgG and IgM only. The Harderian gland, an exocrine gland of unknown function in the orbit, showed synthesis primarily of IgG and IgM.
Antigen-Induced Deoxyribonucleic Acid Synthesis in Mouse Lymphocytes
Authors: Osborne David P. , Katz David H.
Published: December 2022

Employing the model of highly specific hapten-carrier conjugate-induced deoxyribonucleic acid (DNA) synthesis in primed mouse spleen cells, we have investigated the role of various cell populations in generating this response. The depletion of adherent cells results in no loss of cell viability and no decrease in dinitrophenyl-keyhole limpet hemocyanin-induced replication. After depletion of thymus-derived (T) lymphocytes with anti-ϑ serum and complement, the response of DNP-KLH-primed spleen cells to DNP-KLH was reduced by 66%. On the other hand, a population of DNP-KLH-primed spleen cells depleted of immunoglobulin-bearing B lymphocytes by treatment with rabbit anti-mouse immunoglobulin antibodies and complement developed a response to DNP-KLH only 59% of that seen with normal cells. These findings indicate that DNP-KLH antigen-induced DNA synthesis is a mixed cell response, with replication occurring largely in the B lymphocyte population. T lymphocytes also form a portion of the replicating pool and may be more crucially required for a triggering step in the antigen-induced DNA synthetic response in the mouse. The cellular components of KLH and DNP-d-GL-induced DNA synthesis in normal and DNP-KLH-primed mouse spleen cells were also investigated in detail.
Studies on Early Events in the Differentiation of Antibody-Producting Cells
Authors: Moav Neomi , Smolinsky Sara , Moav Boaz , Frensdorff Asher
Published: December 2022

The incorporation rates of tritium-labeled thymidine, uridine, and amino acids into macromolecules, and the appearance of plaque-forming cells (PFC) were studied at 4-hr intervals, in spleen cells obtained from C57BL mice after a secondary intraperitoneal injection of sheep red blood cells. The incorporation rates displayed, over the period studied, clear maxima and minima occurring in the following order; a peak of uridine incorporation at 40 hr after immunization, 4 hr later a peak of thymidine incorporation, and at 60 hr a peak of amino acid incorporation. The number of PFC increased stepwise, the first and the second increases occurring at 60 and 76 hr after immunization, respectively. A similar sequence of biosynthetic events was observed in mouse spleen cells restimulated with sheep red blood cells in vitro. These results were interpreted as reflecting an antigen-induced, synchronized metabolic activity of the cells.

In an attempt to detect possible changes induced in the replication pattern of deoxyribonucleic acid (DNA) synthesized in response to antigenic stimulation, we compared the reassociation rate of DNA newly synthesized in spleen cells 47 to 48 hr after the antigenic stimulation with that of DNA from normal cells. The newly synthesized DNA displayed, between C0t 1000 and C0t 10,000 a faster reassociation rate than the bulk DNA obtained from the same cells or DNA obtained from normal cells. This suggests a temporary selective synthesis of that part of the genome during the pulse labeling. Further support for this assumption was provided by the changes in specific activity of single stranded “immune” DNA along the C0t curve.
Regulation of Antibody Response in Vitro
Authors: Kishimoto Tadamitsu , Ishizaka Kimishige
Published: December 2022

Mesenteric lymph node cells were obtained from rabbits primed with either dinitrophenyl derivatives of Ascaris antigen (DNP-Asc) or Ragweed Fr.D (DNP-Rag) included in aluminum hydroxide gel. The cell suspension was stimulated in vitro with a mixture of a free homologous carrier plus hapten coupled to an unrelated carrier, i.e., DNP-KLH, and anti-hapten antibody response was assessed. Under optimal conditions, the IgG antibody response to the mixture was 2- to 6-fold more than that obtained by DNP-KLH alone. The IgE antibody response was not obtained by stimulation with DNP-KLH alone; however, addition of free carrier to DNP-KLH resulted in a definite IgE antibody response. It was also found that DNP-Asc-primed lymphocytes treated with DNP-KLH were triggered by cell-free supernatant obtained from the culture of DNP-Rag-primed cells activated by free Rag. The results indicated that enhancing soluble factor was released from carrier-specific cells upon stimulation with carrier. The lymph node cells primed with DNP-primary carrier conjugate and a secondary carrier by supplemental immunization showed antihapten antibody response to a mixture of DNP-KLH plus free secondary carrier. In this system, it was found that the enhancing effect of soluble factor correlated with helper function of T cell2 population specific for secondary carrier. When the carrier-specific cells had helper function for both IgG and IgE antibody responses, stimulation of the cells with free carrier resulted in the release of soluble factors which enhanced both IgG and IgE antibody responses. When helper functions of the carrier-specific cells were limited to the IgG class, however, soluble factor released from the cells enhanced IgG but not IgE antibody response. The results suggested that helper cells for IgG antibody response and those for IgE antibody response release different soluble factors which trigger hapten-primed precursor cells for the respective immunoglobulin class. Evidence was obtained that enhancing soluble factor directly triggers hapten-primed precursor cells rather than carrier-specific helper cells.
A Light Chain Tyrosyl Sequence in the Antibody Site of a Rabbit Anti-p-Azobenzoate Antibody
Authors: Roholt O. A. , Friedenson B. , Radzimski G. , Pressman D.
Published: December 2022

A peptide containing a tyrosyl residue from the active site of a rabbit anti-pazobenzoate antibody preparation (from a single rabbit) was isolated as an iododerivative by means of the paired iodination technique. The sequence of the peptide was found to be Tyr-Thr-Gly-(diiodo)Tyr. It was derived from the light chain and was apparently from a hypervariable region since we have not found the corresponding sequence in published light chain sequences. It is the C-terminal tyrosyl residue of this sequence, -Tyr-Thr-Gly-Tyr-, that is in the site since it is almost completely protected from reaction with iodine if specific hapten is present during the iodination. The iodination of this tyrosyl residue does not destroy the binding activity of the site, but only reduces the hapten binding constant, i.e., from about 106 M-1 to 105 M-1 for p-iodobenzoate. This conclusion is based on an approximation procedure for resolving the hapten-binding curves of the uniodinated antibody and of the antibody iodinated in the absence and in the presence of hapten. It is interesting that the N-terminal tyrosyl residue of the isolated sequence was not iodinated at all; thus it is very near to the site but is not exposed to the iodinating reagent.
Vitamin A: Adjuvant and Steroid Antagonist in the Immune Response
Authors: Cohen Benjamin E. , Cohen I. Kelman
Published: December 2022

The effects of vitamin A and hydrocortisone on the immune response have been studied in the mouse. Vitamin A treatment alone markedly increases the normal number of antibody-forming cells generated in the spleen in response to immunization with sheep red blood cells. The antibody response to immunization with dinitrophenylated ovalbumin (DNP-OVA), a hapten-protein conjugate, is also enhanced by vitamin A pretreatment. Furthermore, the immunosuppressive effect of hydrocortisone on the response to sheep red blood cells can be prevented by simultaneous administration of vitamin A.
Failure to Demonstrate a Humoral Immune Response to Scrapie Infection in Mice
Authors: Porter David D. , Porter Helen G. , Cox Nancy A.
Published: December 2022

Although scrapie is clearly a transmissible disease caused by a small stable agent, previous conventional attempts to demonstrate humoral antibody have not met with success. In this study no evidence of humoral antibody to the scrapie agent was found in infected mice by regular or antiglobulin potentiated neutralization tests, indirect immunofluorescence tests employing various agent-containing targets, or by examining brains, spleens, and kidneys for the presence of immune complexes. The results suggest that scrapie infection is fundamentally different from slow or persistent virus infections in which humoral antibody may be demonstrated by the techniques used in this study.
Reaginic Antibody Formation in the Mouse
Authors: Okudaira Hirokazu , Ishizaka Kimishige
Published: December 2022

Priming immunization of DBA/1 strain of mice with dinitrophenylated Ascaris antigen (DNP-Asc) in alum and supplemental immunization with ovalbumin (OA) prepared the animals to give a typical secondary anti-hapten antibody response to DNP-OA with respect to both IgE and IgG antibodies. The results showed collaboration between carrier-specific helper cells and hapten-specific memory cells for anti-hapten IgE antibody response. Priming with DNP-Asc alone significantly enhanced anti-DNP IgE and IgG antibody response to DNP-OA. Enhancement of the antibody response to the DNP-heterologous carrier was due to proliferation of DNP-specific memory cells by priming with DNP-Asc. Evidence was obtained that the DNP-specific memory cells collaborated with helper cells specific for secondary carrier to show the enhanced anti-hapten antibody response.
Reversible Inhibition of Lymphocyte-Mediated Cytolysis by Dimethyl Sulfoxide (DMSO)
Authors: Wolberg Gerald , Hiemstra Kay , Burge Joseph J. , Singler Robert C.
Published: December 2022

Dimethyl sulfoxide (DMSO) was found to inhibit reversibly in vitro lymphocyte-mediated, but not serum antibody-mediated cytolysis. Evidence is presented that the inhibition involves the prevention of productive interaction (i.e., contact which leads to lysis) and that DMSO does not inhibit the later events of the lytic cycle. Immunosuppressive activity could not be demonstrated in vivo presumably due to the inability to sustain high levels of DMSO. DMSO was used to estimate the time from interaction to lysis of target cell.
Role of Ca++ in Mast Cell Activation, Desensitization, and Histamine Release by Dextran
Authors: Baxter James H.
Published: December 2022

Dextran with Ca++ released histamine from rat mast cells, and desensitized them to subsequent challenge with dextran. Desensitization occurred even under certain conditions that inhibited histamine release. In the absence of Ca++, however, dextran induced little histamine release or desensitization. Furthermore, mast cells that had been exposed to dextran without Ca++ and then washed, failed to release histamine when treated with Ca++. Cells exposed to dextran with Ca++ at 0° C and washed, however, did release histamine when resuspended in warm buffer. Thus, it appeared that dextran and Ca++ were required simultaneously in order to activate the cells.
Studies of Thoracic Duct Lymphocytes of Mice
Authors: Mandel Mark A. , Takahashi Toshitado , Old Lloyd J.
Published: December 2022

Thoracic duct cells have a surface antigenic configuration determined by cytotoxic testing that is positive for H-2d, θ, Ly-A, MSLA, κ, and µ antigens and negative for TL and PC antigens. Comparison of these results with other lymphoid elements, benign and malignant, reveals that thoracic duct cells resemble lymph node lymphocytes rather than thymocytes. Quantitative absorption analysis showed similar concentrations of the H-2d, θ, and Ly-A antigens of thoracic duct and lymph node lymphocytes.

The findings suggest that the thoracic duct population is a composite of both thymic and bone marrow-derived cells. This antigenic heterogeneity is supported by both functional and morphologic studies.
An Effector-Cell Independent Step in Target Cell Lysis by Sensitized Mouse Lymphocytes
Authors: Martz Eric , Benacerraf Baruj
Published: December 2022

Heparin was previously shown to inhibit the lysis of target cells by sensitized thymus-derived mouse lymphocytes in vitro. However, heparin failed to inhibit target cell lysis when added to the cultures after only 10% specific release of chromium-51 had occurred. For further exploration of this phenomenon, effector cells have been specifically inactivated at various times during their action on target cells. This was accomplished with alloantiserum made in target strain mice (DBA/2) against effector strain cells (C57BL/6). Treatment of effector cells with this antiserum and complement rendered them unable to initiate new cytolytic events on target cells (and incidentally did not release soluble toxins). However, when untreated cultures were incubated for about 60 min, after which the effector cells had caused 10% specific release of chromium-51 from the target population, treatment of the cultures with anti-effector serum and complement did not interfere with the subsequent lysis of more than half of the target cell population. Hence, the effector-dependent action on the target cell is completed much earlier than evidenced by chromium-51 release, and subsequent, effectorindependent steps must occur before completion of target cell lysis. Data suggest that divalent cations are required for the effector-dependent phase, but not for the effector-independent phase.
An Agglutination-Flocculation Test for Rapid Detection of Hepatitis B Antigen
Authors: Kachani Zarah F. , Gocke David J.
Published: December 2022

Evaluation of a simple, rapid, agglutination-flocculation test (AFT) for detection of hepatitis B antigen (HBAg) is described. The reaction involves simultaneous flocculation and agglutination of antigen by specific antibody in the presence of synthetic acrylic particles. The AFT is over 1000-fold more sensitive for detection of HBAg than conventional counterimmunoelectrophoresis and approximately 8-fold more sensitive than solid-phase radioimmunoassay. HBAg subtype differences can be determined readily in the AFT with antibody of appropriate specificity. On coded analysis of 149 clinical specimens, AFT reactivity was in 92 to 95% agreement with other HBAg methods. Approximately one-third of high titer rheumatoid sera produces an AFT reaction which could easily be distinguished with appropriate controls from a true positive.
Genetic Control of Thymus-Derived Cell Function
Authors: Williams R. Michael , Moore Michael J. , Benacerraf Baruj
Published: December 2022

Lymph node cells (LNC) from Lewis, BN, (Lewis × BN) F1, and (BN × F1) backcross rats were evaluated in culture for their ability to incorporate 3HTdR when stimulated by concanavalin A (Con A) or phytohemagglutinin (PHA) at concentrations ranging from 0.06 to 64 µg/ml. Lewis LNC responses were significantly higher than those of BN cells at optimal or lower concentrations of either mitogen, and responses of F1 LNC tended to be intermediate. There were no statistically significant differences between the responses of LNC from males and females. Each backcross LNC population was tested with BN anti-Lewis serum and by mixed lymphocyte reactivity with F1 cells to determine its genotype at the major histocompatibility locus of the rat (AgB or H-1). The mean Con A- or PHA-stimulated responses of LNC from the 7 of the 14 backcross animals which inherited the AgB1 or H-1l-haplotype characteristic of the Lewis strain were not higher than the mean response of LNC from their siblings who were homozygous for the major histocompatibility locus characteristic of the low responding BN strain. These results are compatible with multigenic control of the potential for thymus-cell DNA synthesis by lymph node cells from normal rats. No evidence was obtained for linkage of any of these genes to the major his tocompatibility locus of the rat.
Detection of Slow Reacting Substance of Anaphylaxis (SRS-A) in Plasma of Guinea Pigs during Anaphylaxis
Authors: Stechschulte Daniel J. , Orange Robert P. , Austen K. Frank
Published: December 2022

SRS-A, a mediator long recognized to be released into the diffusates and perfusates from isolated tissues after antigen-antibody interaction, has been isolated from the plasma of sensitized guinea pigs challenged intravenously with specific antigen. Techniques are now available to study SRS-A release in vivo in selected immediate hypersensitivity reactions.