bluuebunny/crossref_metadata_2025 huggingface dataset:
Binding of N-Acetylgalactosamine-Containing Compounds by a Human IgM Paraprotein
Authors: Naiki Masaharu , Marcus Donald M.
Published: December 2022

A serum that contains a monoclonal human IgM paraprotein (McG) agglutinates protease-treated human erythrocytes and binds glycosphingolipids that possess a terminal nonreducing N-acetylgalactosaminyl residue in either the α or β anomeric configuration. The approximately equal reactivity of McG with both anomers of N-acetylgalactosamine was unexpected because most immunoglobulins that bind saccharides exhibit a marked preference for one anomeric configuration.

The major receptor for this protein in normal group O erythrocytes is globoside, the blood group P antigen: Ga1NAc(β,1→3)Gal(α,1→4)Gal(β,1→4)G1c-ceramide. Erythrocytes of the rare Pk and p phenotypes lack globoside, and the former are not agglutinated by McG. The McG receptor in p erythrocytes, which are agglutinated as strongly as normal cells, appears to be a crossreactive glycolipid that contains a much larger number of sugar residues than globoside.
Inhibition of Human Lymphocyte Transformation by Human α-Fetoprotein (HAFP); HAFP Monomers and Multimers and a Resistant Lymphocyte Subpopulation
Authors: Yachnin Stanley , Lester Eric P.
Published: December 2022

We have previously shown that human α-fetoprotein (HAFP) inhibits the response in vitro of human lymphocytes to a variety of mitogenic stimuli. In this report we present evidence that HAFP preparations containing dimeric and trimeric forms are no more inhibitory than HAFP monomeric isolates. HAFP isolates of low inhibitory potency cannot impede the capacity of high-potency HAFP to inhibit lymphocyte transformation. Since lymphocyte responses to phytomitogens or anti-human thymocyte antiserum cannot be totally suppressed by increasing doses of HAFP, we conclude that human lymphocyte preparations contain a population of lymphocytes resistant to the inhibitory action of HAFP.
Inflammatory Cells in Solid Murine Neoplasms
Authors: Gillespie G. Yancey , Hansen Christopher B. , Hoskins Ronald G. , Russell Stephen W.
Published: December 2022

Highly purified suspensions of intratumoral T lymphocytes, recovered 11 and 13 days after induction of regressing or progressing Moloney sarcomas, were compared in their ability to lyse specifically the MSC cells used for tumor induction. Cytolytic activity, expressed in terms of lytic units/106 T cells, was similar for intratumoral T cell suspensions obtained 11 days after induction of either regressing (3.1 ± 1.3 LU/106 T cells) or progressing (4.3 ± 1.8) neoplasms. By 13 days post-induction, regressing tumors contained T lymphocytes with an increased cytolytic activity (11.1 ± 4.5) whereas those from progressing tumors were strikingly less able to kill MSC cells (≤ 0.2). This dramatic loss in cytotoxicity could not be attributed to errors associated with the enzymatic disaggregation method, inhibition by copurified endogenous tumor cells, or immunosuppression induced by viral infection. The changes in functional activity of intratumoral T lymphocytes from the two types of sarcoma appeared to be correlated with the stage of neoplasia. In this model system, cytolytic activity of T lymphocytes increased during spontaneous tumor regression whereas losses in cytotoxicity occurred coincident with the onset of inexorable progression.
The Tla Locus: A New Allele and Antigenic Specificity
Authors: Flaherty Lorraine , Sullivan Karen , Zimmerman Debra
Published: December 2022

Four of 23 H-2 alloantisera screened for anti-TL activity contained such activity. One of these alloantisera, D-35, defined a new TL specificity, TL.5, and a new Tla allele, Tlad. TL.5 has all the characteristics of a TL antigen and has a different strain distribution than previously known ones. This new complexity at the Tla locus and the previous finding of other serologically defined genes in the Tla region indicate that this genetic region cannot be ignored in analyzing antisera produced in strains made congenic for the major histocompatibility complex.
IgE Formation in the Rat Following Infection with Nippostrongylus Brasiliensis
Authors: Urban Joseph F. , Ishizaka Teruko , Ishizaka Kimishige
Published: December 2022

Normal rat bone marrow cells incubated with serum or lymph from Nippostrongylus brasiliensis (Nb)-infected rats showed an increase in the proportion of IgE-bearing cells in culture. This effect was produced in a similar fashion by cell-free supernatants (CFS) from cultures of mesenteric lymph node cells obtained from Nb-infected rats. The action of CFS on bone marrow cells appeared to be specific for the generation of IgE-bearing cells since the proportion of IgM-bearing cells in the culture did not change. The IgE-bearing cells in bone marrow cell cultures consisted of small lymphocytes, blast cells, and mast cells, and the addition of CFS to the cultures predominantly increased the number of IgE-bearing blast cells. CFS was also effective in increasing the proportion of IgE-bearing small lymphocytes in cultures of normal mesenteric lymph node cells. Removal of IgE in CFS by an anti-IgE immunosorbent did not affect the ability of CFS to generate IgE-bearing cells. The factor(s) in CFS responsible for this activity was shown to migrate with serum β-globulins in zone electrophoresis and to possess a molecular size of between 104 and 2 × 104 m.w. The ability of CFS to generate IgE-bearing cells was diminished by treatment with the enzymes trypsin and ribonuclease A, but was unaffected by chymotrypsin.
Studies of the Immunoglobulin Eluted from the Glomeruli of Mice Chronically Infected with Lymphocytic Choriomeningitis Virus
Authors: Klein Benjamin , Hill Cynthia , Hoffsten Phillip E.
Published: December 2022

Previous work has demonstrated that B cells in the lymphocytic choriomeningitis virus (LCM) carrier mouse do not produce anti-LCM antibody. Studies were undertaken to determine the binding specificity of immunoglobulin (Ig) eluted from kidneys of LCM-infected SWR/J mice. Kidneys from neonatally infected LCM carrier mice (5 to 7 months of age) were homogenized in 0.05 M EDTA PBS (pH 7.4), centrifuged at 12,000 × G for 10 min at 4°C and the sediment was washed three times. The sediment was eluted with citrate buffer (pH 3.2) for 90 min at 37°C, centrifuged as before, neutralized, and concentrated by vacuum dialysis. These elution conditions destroyed the binding capacity of LCM antigen in infected tissues and in commercial complement fixation LCM antigen solutions. The amount of Ig eluted was 22 to 24 µg/kidney from LCM-infected mice but less than 1 µg/kidney from uninfected mice. Eluted Ig did not bind to LCM-infected tissue although SWR/J anti-LCM antiserum subjected to the eluting conditions retained its LCM antigen-binding ability. NZB × NZW F1 hybrid kidneys eluted as above yielded similar amounts of Ig that had antinuclear binding ability. Eluted Ig did not induce complement fixation when added to LCM antigen although SWR/J anti-LCM antiserum subjected to elution conditions retained its LCM antigen complement-fixing properties. These observations indicate that Ig deposited in glomeruli of LCM carrier mice is not specific for LCM antigens and favors the position that the LCM carrier mouse is tolerant to LCM virus.
Regulation of the Immune Response to Tumor Antigens
Authors: Greene Mark I. , Fujimoto Shigeyoshi , Sehon Alec H.
Published: December 2022

Immunosuppressor T cells (IST),6 capable of inhibiting the rejection of a methylcholanthrene-induced fibrosarcoma (S1509a) in A/Jax mice immune to this tumor, produced soluble factors with similar suppressive activity. The immunosuppressive factor(s) (ISF) has been shown to be immunologically specific, as demonstrated by the complete loss of suppressive activity after absorption with S1509a cells, but not with cells of an unrelated syngeneic tumor. From its behavior on gel filtration, the size of ISF was deduced to be less than 70,000 daltons. The ISF was not removed by passage through a rabbit anti-mouse F(ab′)2 reverse immunosorbent and, hence, it was concluded that ISF was not likely to be an immunoglobulin. The (immunosuppressive) activity of ISF was destroyed by treatment with Pronase, but not with RNase. The ISF was found to share the antigenic determinant(s) of the product(s) of the K end of the major histocompatibility complex of the mouse. Moreover, antibodies to ISF were induced by immunization with ISF-tumor cell complexes. Thus, IST and their factor(s) appear to play an important role in the regulation of the immune response to tumor antigens.
Lymphocyte Specificity to Protein Antigens
Authors: Corradin Giampietro , Etlinger Howard M. , Chiller Jacques M.
Published: December 2022

An in vitro assay which measures antigen-induced proliferation of primed murine lymph node cells is described. The response is mediated by T cells since it can be obtained by using nylon wool-passed lymphocytes (< 1% Ig+ cells) and it can be abolished by treatment with anti-Thy 1.2 and C. Furthermore, LN cells from nu/nu mice injected with antigen do not demonstrate antigen-induced proliferation in contrast to the response observed in euthymic littermate controls. Other relevant parameters of this proliferative assay include the observations that the response is highly antigen specific, can be seen as early as 4 days and as late as 60 days after in vivo priming, is restricted to the use of certain sets of LN when animals are injected subcutaneously at the base of the tail, and can be seen with LN cells from mice primed with antigen in either CFA or ICFA. The ease of the assay coupled with its specificity and quantitative dimensions provides a direct and simple method to evaluate processes involved in antigen-induced murine T lymphocyte activation.
Activation of Specific Cellular Immunity toward Murine Leukemia in Mice Rejecting Syngeneic Somatic Hybrid Cells
Authors: Liang Weitze , Cohen Edward P.
Published: December 2022

ASL-1 × LM(TK)- somatic hybrid cells form both H-2a and H-2k antigen complexes. After forming a localized tumor in syngeneic (A/J × C3H/HeJ)F1 mice, they are rejected. Such mice are resistant to otherwise invariably lethal injections of ASL-1 cells, surviving for prolonged and, in some instances, indefinite periods. To examine the basis of immunity, the capacity of spleen cells from mice rejecting hybrid cells to stimulate the release of 51Cr from labeled ASL-1 cells was investigated. Cells from the spleens of mice rejecting ASL-1 × LM(TK)- cells stimulated the release of 51Cr from labeled ASL-1 cells, but not from Ehrlich ascites or P815 cells. Cells from mice injected with mitomycin-C-treated ASL-1 cells led to the release of 51Cr from labeled ASL-1 cells as well, but the extent of 51Cr release was approximately one-third as occurred in the presence of cells from hybrid cell-injected mice. Cells from noninjected mice or from mice injected with LM(TK)- cells failed to lead to the specific release of 51Cr from ASL-1 cells. The presence of unlabeled ASL-1 cells, but not Ehrlich ascites cells, competitively inhibited the spleen cell-stimulated release of 51Cr from labeled ASL-1 cells. Sera from A/J mice injected with mitomycin-C-treated ASL-1 cells contained antibodies specific for the tumor-associated antigen of ASL-1 cells. The specific release of 51Cr from ASL-1 cells stimulated by spleen cells from hybrid-cell injected F1 mice was inhibited by such antisera, but not by H-2a antisera. Treatment of spleen cells from mice injected with hybrid cells with Thy 1.2 antiserum before incubation with 51Cr-labeled leukemic cells reduced the proportion of 51Cr released by approximately one-half. Spleen cells from animals injected with hybrid cells, as well as to a lesser extent from noninjected mice, which are capable of adhering to plastic Petri dishes were active in the cytotoxic reactions as well. Results similar to these were obtained with 51Cr-labeled RADA-1 cells, a radiation-induced leukemia of A/J mice and spleen cells from (A/J × C3H/HeJ)F1 animals injected with RADA-1 × LM(TK)- hybrid cells.
The Use of Fab Fragments of Anti-Human Immunoglobulin as Analytic Tools for Establishing the Involvement of Immunoglobulin in the Spontaneous Cytotoxicity to Cultured Tumor Cells by Lymphocytes from Patients with Bladder Carcinoma and from Healthy Donors
Authors: Troye M. , Perlmann P. , Pape G. R. , Spiegelberg H. L. , Näslund I. , Gidlöf A.
Published: December 2022

The effect of Fab fragments of rabbit anti-human immunoglobulin on the lysis of allogeneic cultured bladder tumor cells by lymphocytes from patients with urinary bladder carcinoma or by lymphocytes from healthy donors was assayed by 51Cr-release. Addition of Fab fragments of rabbit anti-human immunoglobulin strongly inhibited this cytotoxicity in most cases. Under the conditions used, 60 to 100% inhibition was frequently obtained with 10 to 100 µg/ml of immunoadsorbent purified Fab fragments specific either for the F(ab′)2-part or for the Fc part of human IgG. In those cases in which inhibition was incomplete, increasing the concentration of inhibitor had little or no effect. Fab fragments prepared from immunoadsorbent purified antibodies to an unrelated antigen (ovalbumin) did not inhibit cytotoxicity. Fab fragments that inhibited the spontaneous cytotoxicity against the tumor target cells did not inhibit specific cell-mediated lympholysis induced by mixed lymphocyte culture activation, a cytotoxic reaction known to be mediated by antibody-independent cytotoxic T cells. Experiments with Fab fragments specific for the four IgG subclasses suggested that antibodies of all four IgG subclasses may become involved in lymphocyte-mediated cytotoxicity against the tumor cells.

Previous studies showed that killing of tumor cells by lymphocytes from patients with bladder carcinoma or from healthy donors is most frequently mediated by effector cells with high avidity Fc receptors for IgG but without surface-bound immunoglobulin (SIg) or high avidity receptors for sheep erythrocytes. These previous results and our findings now suggest that a large part (but not all) of the spontaneous cytotoxicity against allogeneic tumor cells is in fact induced by antibodies and thus mediated by K cells. The inducing antibodies are probably released from SIg- cells during lymphocyte-target cell incubation.
A Functional Differentiation of Human Neutrophil Granules: Generation of C5a by a Specific (Secondary) Granule Product and Inactivation of C5a by Azurophil (Primary) Granule Products
Authors: Wright Daniel G. , Gallin John I.
Published: December 2022

During phagocytosis of latex particles human neutrophils not only release a serum activator that generates the chemotactic C5 fragment, C5a, by complement activation, but these cells also release a product that destroys the chemotactic activity of C5a. These separate neutrophil products, however, are released sequentially by neutrophils during phagocytosis with the serum activator appearing more rapidly in incubation media. Chemotactic activity is generated optimally in serum by post-phagocytic media at pH 7.2 to 7.4, whereas C5a inactivation occurs both at neutral pH and at a more acid pH, 6.2 to 6.4. Studies in which the azurophil (primary) granules and specific (secondary) granules are separated from neutrophil lysates by sucrose density centrifugation show that the serum activator is contained in specific granules whereas C5a inactivators are restricted to the azurophil granules. Gel filtration chromatography of lysates of purified granule preparations indicates that the serum activator contained in specific granules is a large molecule (m.w. greater than 150,000), and that there are at least two C5a inactivators associated with the azurophil granules, with estimated m.w.'s of 65,000 and 30,000. Isoelectric focusing of azurophil granule lysates also indicates at least two separate C5a inactivators. It is evident that mediators contained in the specific and azurophil neutrophil granules may have functionally opposing roles once released by the cells. These mediators, which effect the generation or inactivation of C5a, have different pH optima of activity and distinct kinetics of release from neutrophils and together may contribute to the self-limited amplification and localization of acute inflammatory responses.
F(ab′)2 Reagents Are not Required If Goat, Rather Than Rabbit, Antibodies Are Used to Detect Human Surface Immunoglobulin
Authors: Alexander Elaine L. , Sanders Sheila K.
Published: December 2022

The present report provides evidence that whole goat anti-human immunoglobulin, unlike similar reagents produced in the rabbit, binds both to the same number of and the same individual cells as the F(ab′)2 fragments of rabbit or goat anti-human immunoglobulin. These results suggest that goat IgG has a lower affinity for the Fc receptors of human lymphocytes and monocytes than rabbit IgG. Because of this property, whole goat antibodies against human immunoglobulin can be used as simple, convenient relatively inexpensive reagents for the routine detection of immunoglobulin on cell surfaces by immunofluorescence microscopy. The preparation of F(ab′)2 fragments of anti-immunoglobulin, which are necessary when rabbit antibodies are used, does not appear to be required if goat antibodies can be employed. This observation has multiple practical applications in cellular immunology.
Appearance of Cytotoxic Cells within the Bronchus after Local Infection with Herpes Simplex Virus
Authors: Clancy Robert , Rawls William E. , Jagannath Sundar
Published: December 2022

Cells from rabbit spleens, bronchial washings (BW) and bronchus-associated lymphoid tissues (BALT) were examined for their ability to lyse cells infected with herpes simplex virus (HSV). Specific lysis of HSV-infected cells was mediated by BW cells as early as 4 days after intratracheal infection of the rabbits with the virus whereas lysis by spleen cells and BALT cells was not detected until 7 or more days after infection. Lysis by spleen cells was initially detected 7 days after intraperitoneal injection of the virus but lysis by BW and BALT cells was not observed until 14 days after infection. Although spleen, BW, and BALT cells could lyse antibody-coated target cells, antibodies detectable by antibody-dependent cellular cytotoxicity could not be detected in bronchial washings until 7 or more days after infection. The data suggest that cells capable of direct cytotoxicity of virus-infected cells appear within the bronchus after local infection by the virus.
Macrophages Are Required for the Dextran-Sulfate Induced Activation of B Lymphocytes
Authors: Persson Ulla C. I. , Hammarström Lennart L. G. , Smith C. I. Edvard
Published: December 2022

The proliferative response induced by the B cell activating ligand dextran-sulfate (D×S) requires the presence of macrophage-produced factors or the macrophage-substituting compound 2-mercaptoethanol. Thus, the mechanism for activation of mouse B splenic lymphocytes is different for D×S than for other polyclonal B cell activators such as lipopolysaccharides, which can trigger B cells directly in the absence of helper factors. It is suggested that the disappearance of the D×S-induced response could be studied in a simple functional test for the efficiency of macrophage depletion obtained by different techniques.
Effect of Concanavalin A on Lymphocyte Interactions Involved in the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Markham Richard B. , Reed Norman D. , Stashak Philip W. , Prescott Benjamin , Amsbaugh Diana F. , Baker Phillip J.
Published: December 2022

When administered 2 days after immunization with 0.5 µg TypeIII pneumococcal polysaccharide (SSS-III), the T lymphocyte mitogen concanavalin A (Con A) stimulates a 2.6- to 7-fold enhancement of the plaque-forming cells (PFC) response to SSS-III in vivo. This enhancement requires the presence of amplifier T cells, which act by driving PFC or their precursors to extra rounds of proliferation. The extra proliferation that can be stimulated by Con A is not seen in the normal primary response to SSS-III; but treatment with anti-lymphocyte serum (ALS) to remove suppressor T cells will permit the additional proliferation to occur. This indicates that in the primary response to SSS-III, suppressor T cells act on amplifier T cells to limit the magnitude of the antibody response. Only suppression of B cells can account for the further suppression induced by Con A given at the time of immunization or by low-dose paralysis of the SSS-III response. The relatively late development of amplifier activity compared to suppressor activity appears to account for the absence of amplifier activity after primary immunization with SSS-III. It is apparent that one can explain the regulatory effects observed during the development of an immune response to SSS-III only by considering both T cell - B cell and T cell- T cell interactions, together with the temporal relationships involved in those interactions.
The Formation of Active Hybrid Immunoglobulins from the Heavy and Light Chains of β(1, 6) D-Galactan Binding Murine Myeloma IgA's S10 and J539
Authors: Manjula Belur N. , Mushinski Elizabeth B. , Glaudemans Cornelis P. J.
Published: December 2022

Murine myeloma immunoglobulin (IgA, κ) J539, which shows enhanced tryptophanyl fluorescence on ligand binding, and S10, which shows reverse-sign changes in tryptophanyl fluorescence on ligand binding (RLIF, see below), have been reduced, alkylated, and dissociated into their light (L) and heavy (H) chains. Two hybrid recombinants, H10L539 and H539L10, have been prepared and the 7S material has been isolated by chromatography. The binding behavior of these recombinants was studied with a number of ligands. Both recombinants showed activity with β(1→6) linked galactose ligands comparable to the native immunoglobulins. The ligand-induced fluorescence changes of the recombinants paralleled those of the heavy chain donor. For the recombinant H10L539, two different galactose-ligands caused fluorescence changes in opposite directions. It was quantitatively shown that binding of these ligands, nevertheless, took place in the same combining region. The idiotype of each recombinant resembled that of the heavy chain donor.
Effects of C-Reactive Protein on Platelet Function
Authors: Fiedel Barry A. , Simpson Robert M. , Gewurz Henry
Published: December 2022

It was previously demonstrated that C-reactive protein (CRP) inhibits platelet aggregation and release reactions, activation of platelet factor 3, and platelet-dependent clot retraction. Multiple considerations including selective inhibition of secondary wave aggregation suggested that CRP exerted its inhibitory effects by interfering with the release of endogenous ADP. In the present investigation, CRP was found by direct assay to inhibit the release of endogenous ADP and/or serotonin concomitant with inhibition of platelet aggregation stimulated by ADP, epinephrine, thrombin, and AHGG. CRP did not induce an increase in the basal level of platelet cAMP, suggesting independence of a direct effect upon this mediator system. Furthermore, CRP did not inhibit the aggregation and secretion induced by the antibiotic ionophore A23187, suggesting the absence of a direct effect upon the activation of platelet contractile elements. By contrast, CRP did inhibit both thrombin-induced release of malondialdehyde, a prostaglandin endoperoxide nonprostanoate endproduct, and platelet aggregation induced by the prostaglandin endoperoxide precursor arachidonic acid. These data, therefore, raise the possibility that CRP inhibits platelet reactivities by interfering with an aspect of prostaglandin metabolism, and that this occurs subsequent to the hydrolytic accumulation of arachidonic acid and prior to the movement of calcium from the platelet dense tubules. These studies support the concept that CRP serves to modulate platelet reactivities during acute inflammatory reactions.
Synthesis and Secretion of a Mitogenic Protein by Macrophages: Description of a Superinduction Phenomenon
Authors: Unanue Emil R. , Kíely Jeanne-Marie
Published: December 2022

We have examined the biosynthesis and fate of the mitogenic protein (MP)2 made by macrophages. This mitogenic protein was assayed by its stimulation of DNA synthesis of thymocytes. Macrophages freshly harvested from the peritoneal cavity did not contain MP in the cell. The MP, however, rapidly appeared in the cell, reaching maximum amount by 1 to 2 hr, maintaining its concentration up to about 6 hr, and then disappearing. The disappearance of MP did not correlate with its secretion into the medium. The elaboration of MP stimulated by contact with the dish was inhibited by cycloheximide.

Exposure of the cells to inhibitors of protein synthesis after 1 to 2 hr of culture at a time when the MP was already made produced a striking increase in secretion. The MP inside the cell did not appear to decay in the presence of cycloheximide. This result thus represents another example of superinduction. The intracellular fate of MP appears to be under the regulation of a control protein.
Purification of an Acidic Nuclear Protein Antigen and Demonstration of its Antibodies in Subsets of Patients with Sicca Syndrome
Authors: Akizuki Masashi , Boehm-Truitt Marilyn J. , Kassan Stuart S. , Steinberg Alfred D. , Chused Thomas M.
Published: December 2022

Ha, a soluble nuclear protein, was purified from calf thymus nuclear extract by successive application of fractional ammonium sulfate precipitation (60 to 80% saturation), DEAE-Sephadex chromatography (eluted between 0.26 and 0.38 M NaCl in 0.05 M Tris HCl buffer, pH 7.2), and affinity chromatography utilizing an immunoabsorbent column of Sepharose coupled to IgG from a patient with a high titer of antibody to Ha. The Ha antigen could be iodinated by the method of Bolton and Hunter but not by the chloramine-T or lactoperoxidase methods indicating the absence of tyrosine and histidine. The iodinated material was passed through a Sephadex G-100 column to remove minor contaminants. The final product migrated as a single band in polyacrylamide gel electrophoresis. The 125I-Ha antigen was utilized to measure specific antibody by the ammonium sulfate method. Elevated serum-binding capacity was observed in 73% of patients with sicca syndrome in the absence of another connective tissue disease and 85% of patients with sicca syndrome associated with systemic lupus erythematosus. It was infrequent in sicca syndrome associated with rheumatoid arthritis (6%) and in systemic lupus erythematosus without sicca syndrome (3%). Anti-Ha antibodies were not found in patients with other connective tissue diseases or normal controls. Antibody to Ha characterizes a subset of patients with sicca syndrome.
Regulatory Mechanisms in Cell-Mediated Immune Responses
Authors: Hodes Richard J. , Nadler Lee M. , Hathcock Karen S.
Published: December 2022

Mouse spleen cells which had been primed to alloantigens by in vitro mixed lymphocyte culture (MLC) were found to actively suppress the in vitro generation of cell-mediated cytotoxicity (CML) by freshly explanted syngeneic spleen cells. Both antigen-specific and antigen-nonspecific suppressor activities were identified. Antigen-nonspecific suppression was active in suppressing CML sensitizations to alloantigens, including those to which the suppressor population had not been previously exposed. This form of suppression was radiation sensitive (abrogated by 1000 R). It was T cell dependent, appeared after 2 days of priming in MLC, and was effective in suppressing CML only when added during the first 2 days of sensitization culture. In contrast, antigen-specific suppression affected only those CML snesitizations in which the relevant sensitizing antigen either: 1) was the same antigen to which the suppressor population was previously primed, or 2) was presented on the same cell as that alloantigen. Antigen-specific suppression was resistant to 1000 R. It was T cell-dependent, first appeared after 3 days of priming in MLC, and suppressed CML only when added during the first 2 days of sensitization. These data are discussed in terms of regulatory mechanisms which may be active in determing the kinetics and antigen specificity of primary and secondary CML responses.
Migratory Behavior of Lymphocytes with Specific Reactivity to Alloantigens
Authors: Emeson Eugene E.
Published: December 2022

A dual antigen, dual isotope assay has been used to monitor the migratory behavior of selectively labeled anti-allogeneic lymphocytes in mice given an i.v. challenge with semiallogeneic spleen cells. 3H-labeled anti-C3H and 14C-labeled anti-C57BL lymphocytes of DBA/2J origin were pooled and adoptively transferred to two groups of DBA/2J recipients previously challenged i.v. with either (C3H × DBA)F1 or (C57BL × DBA)F1 semi-allogeneic spleen cells. After 24, 48, 72, and 96 hr, a subgroup of four mice from each group were sacrificed and the relative numbers of anti-C3H and anti-C57BL lymphocytes present in the spleens, bone marrows, livers, lymph nodes, intestines, lungs, and bloods of each mouse were inferred from the mean 3H/14C ratios of each tissue of that subgroup. The results of these studies indicated that specific anti-allogeneic lymphocytes were selectively recruited to the spleens, bone marrows, and livers of challenged mice and deleted from their lymph nodes, lungs, intestines, and blood. Similar studies with labeled marker cells enriched for T lymphocytes (and free of macrophages) and labeled marker cells depleted of T lymphocytes with anti-Thy 1.2 serum plus complement indicated that the cells selectively recruited to the spleen, bone marrow, and liver were predominantly but not exclusively T lymphocytes. A direct demonstration of selective recruitment of specifically reactive lymphocytes to bone marrow and liver has not been previously described. The significance and implications of these findings are discussed.
Structural and Genetic Studies on Chicken 7S Immunoglobulin Allotypes
Authors: Wakeland Edward K. , Foppoli James M. , Benedict Albert A.
Published: December 2022

A survey of 47 inbred or partially inbred chicken lines derived from five sources in the United States and Europe revealed considerable genetic polymorphism in the structural gene (CS-1) responsible for the production of the predominant chicken 7S Ig heavy chain. A minimum of 10 alleles of the CS-1 gene were detected as unique combinations or phenogroups of CS-1 specificities. A system of nomenclature for CS-1 alleles was developed and six homozygous lines were designated as prototype lines. The remaining four CS-1 alleles occurred only in lines that were polymorphic for 7S Ig allotypic specificities.
Structural and Biosynthetic Studies of a Human IgA Half Molecule
Authors: Bernier George M. , Johnson Claire A. , Fanger Michael W.
Published: December 2022

The serum and urine of a patient with plasma cell leukemia contained free λ light chains and a 4S protein with both α1 and λ antigenic determinants. When isolated from urine, the IgA-related protein was shown to have a substantial antigenic deficiency in the Fc region and to be composed of a single light chain and a single heavy chain. In polyacrylamide gel electrophoresis with sodium dodecylsulfate and 2-mercaptoethanol, the heavy chain appeared to be 13,000 daltons smaller than the usual IgA heavy chain. The molecule lacked methionine, two residues of which are invariant in the C-terminal region of other α chains. The size of the heavy chain, its antigenic deficiency, and its chemical composition were therefore consistent with a deletion of the C-terminal domain. In short-term culture with 3H-leucine, the patient's peripheral plasma cells synthesized a molecule that was antigenically identical to the urine IgA. The mole cules synthesized in vitro appeared similar in size to those of the urine IgA as determined by gel filtration studies in aqueous media. Polyacrylamide gel electrophoresis and autofluorography of newly synthesized molecules isolated immunologically from both culture fluid and cell lysate showed the heavy and light chains to be the same size as those of the urine IgA. These findings are consistent with this abnormal IgA molecule's being synthesized with a deletion in the carboxyl-terminus of the α chain.
Abrogation of Resistance to Foreign Bone Marrow Grafts by Carrageenans
Authors: Yung Yee Pang , Cudkowicz Gustavo
Published: December 2022

The role of macrophage-like cells in natural resistance of mice to transplants of foreign bone marrow cells was reinvestigated by means of the fractionated forms of carrageenan designated as ι, λ, and κ, and the unfractionated form Seakem-9 for comparison. All these anti-macrophage agents weakened resistance to parental and allogeneic marrow grafts when a single small dose (0.5 mg/mouse) was injected i.v. into irradiated mice several hours before or after transplantation. λ and ι carrageenans were more effective than the SK and κ forms, and the anti-resistance effect was enhanced when irradiation preceded transplantation by 20 to 24 hr. κ and λ carrageenans had no adverse effect on the homing of donor stem cells to the spleen and on subsequent proliferation. Stem cells recovered from the spleens of genetically resistant mice given ι-carrageenan were of donor type, as determined by retransplantation into discriminant hosts. The combined effect of suboptimal doses of λ-carrageenan and rabbit anti-mouse lymphocyte serum was additive. Since the antiserum was a blocking agent for accessory macrophage-like cells interacting with effector lymphocytes, this result suggested that λ-carrageenan injured macrophages. Accordingly, agents sharing with carrageenans the anti-complementary, anti-coagulant, or Hageman factor-activating properties, but not toxicity for macrophages, failed to weaken resistance, and mice with genetic defects of complement components were as resistant and as sensitive to the effect of carrageenans as normal mice. These experiments reiterate the importance of anti-macrophage agents in weakening natural resistance to hemopoietic cells and in elucidating cellular mechanisms.
Cytotoxic Reactivity of Human Lymphocytes Cultured in Vitro
Authors: Ortaldo John R. , Bonnard Guy D. , Herberman Ronald B.
Published: December 2022

Recent evidence indicates that both natural killer and killer cells active in antibody-dependent cell-mediated cytotoxicity are cell types that bear Fc receptors. Natural killer and antibody-dependent cell-mediated cytotoxicity have also been shown to be in a SRBC-rosetting subpopulation. Human mononuclear peripheral blood leukocytes were cultured under various conditions to examine natural killing and antibody-dependent cell-mediated cytotoxicity. It was of interest to determine if these activities would be maintained or augmented under various culture conditions. Before culture, normal mononuclear peripheral blood leukocytes were tested in a 4-hr 51Cr release assay against K-562 to measure natural killer activity, and against antibody-coated Chang liver cells to measure antibody-dependent cell-mediated cytotoxicity. Fc receptor-bearing cells were removed by adsorption to immobilized antigen-antibody complexes. When lymphocytes were cultured alone, we often observed what we call cytotoxicity from cultured cells (CCC). The amount of cytotoxicity against K-562 and in antibody-dependent cell-mediated cytotoxicity was dependent on the presence of FBS and proportional to its concentration in the cultures. The presence of FCS resulted in high levels of CCC whereas human serum demonstrated low or negligible activities. This CCC was not directed against FBS-associated antigens. These CCC cannot be called NK since their extensive characteristics are not known. As was characteristic on day 0 for natural killing and antibody-dependent cell-mediated cytotoxicity, the cultured effector cells also contained an Fc receptor.

Therefore, the presence of CCC with features of natural killer and killer cell activities and its regeneration in culture indicate that it must be distinguished from specific tumor reactivity or alloreactivity in mixed lymphocyte and mixed lymphocyte-tumor reactions, particularly when tumor cell lines are used as targets.
Allotype Suppression at the CHn Locus of IgM in Heterozygous Rabbits with Concomitant Suppression at the VHa Locus of All Ig Classes
Authors: Gilman-Sachs Alice , Eskinazi Daniel P. , Dray Sheldon
Published: December 2022

Doubly heterozygous offspring, ♂n83a1/♀n82a2 or ♂n81a1/♀n82a2, of doubly homozygous dams, n82a2/n82a2, were injected neonatally with anti-n83 or anti-n81 antibody directed to the paternal n locus allotype. In each injected rabbit, the paternal n83 or n81 allotypes were suppressed and increased amounts of the maternal n82 allotype accounted for almost all of the serum IgM. This is the first report of allotype suppression in the rabbit mediated by antibody directed toward the CH region of any Ig class. A concomitant decrease in the paternal a1 Ig allotype and a compensatory increase in the maternal a2 allotype were also found. This finding supports the concept that IgM-bearing cells are the precursors of IgG-secreting cells.
Preferential Induction of Jones-Mote Hypersensitivity in Guinea Pigs Treated with Tolerogenic Antigen
Authors: Nomoto Kikuo , Himeno Kunisuke , Kuroiwa Ataru , Takeya Kenji
Published: December 2022

Effects of tolerogenic form of bovine γ-globulin (BGG) on the Jones-Mote hypersensitivity, tuberculin hypersensitivity, and helper function were studied in guinea pigs. 1) When intermediate or large doses of tolerogenic BGG were injected i.v. on day -14 or 0 and immunogenic BGG in complete Freund's adjuvant (CFA) was injected subcutaneously on day 0, tuberculin hypersensitivity and helper function were depressed, although Jones-Mote hypersensitivity was preserved or augmented. Jones-Mote hypersensitivity was suppressed with the uppermost dose of tolerogenic BGG. Suppressing mechanism(s) to Jones-Mote hypersensitivity appears to be slightly more resistant to tolerance induction than tuberculin hypersensitivity and helper function and more sensitive than Jones-Mote hypersensitivity. 2) Expression of tolerant states was not absolute but relative to the doses of immunogenic BGG. When a larger amount of immunogenic BGG was given, larger amounts of tolerogenic BGG were required to obtain the same degrees of tolerance as that expressed in animals given a smaller amount of immunogenic BGG. 3) In animals in which immune responses were suppressed partially with tolerogenic BGG, tuberculin or Jones-Mote hypersensitivity was delayed in the development. 4) Basophils comprised more than half of the infiltrating cells at the sites of erythematous reactions elicited in the animals showing the tolerant states of tuberculin hypersensitivity and helper function. Thus, typical Jones-Mote hypersensitivity was raised preferentially in the tolerant states of other types of immune responses. These findings may be explained by different affinities to BGG among T cell subpopulations responsible for individual responses and individual cells within a subpopulation for each response.
Chemical and Biologic Properties of A Protein-Rich Fraction of Bacterial Lipopolysaccharides
Authors: Betz Sally J. , Morrison David C.
Published: December 2022

Butanol LPS preparations from three serotypes of Escherichia coli lose their ability to stimulate C3H/HeJ lymphocytes after the removal of a protein-rich fraction (LAP) by extraction with phenol. This loss of mitogenic activity for LPS nonresponder spleen cells is accompanied by an increase in equilibrium density of approximately 0.03 g/cm3 by each of the three lipopolysaccharides. The LAP preparations exhibited potent mitogenic activity for both LPS responder C3H/St and nonresponder C3H/HeJ lymphocytes. Each LAP produced an optimal response at approximately 10 µg/ml and each was toxic at a 10-fold higher concentration. Like LPS, LAP is mitogenic for congenitally athymic nude mice. Although LAP is isolated in association with LPS and shares certain of its biologic properties, the two B cell mitogens may be distinguished by several criteria. Of primary importance is the ability of LAP to stimulate LPS-nonresponder lymphocytes. In addition, these experiments demonstrate that the LAP-induced response by C3H/HeJ lymphocytes is not blocked by preincubation with high concentrations of LPS. The response to LAP by C3H/St spleen cells is not blocked by polymyxin B, an inhibitor of the lipid A-induced mitogenic response. Furthermore, such LAP activity is destroyed by mild alkaline hydrolysis under conditions that do not decrease LPS activity. Finally, we observed that the peak level of 3H-thymidine incorporation that results from stimulation with LPS is consistently lower than the level which is obtained with optimal LAP doses. The experiments presented here also demonstrate that isolated LAP and protein-free LPS can reassociate in vitro to generate an LPS-LAP complex with mitogenic activity for C3H/HeJ lymphocytes.
Receptors for IgM on Certain Human B Lymphocytes
Authors: Ferrarini Manlio , Hoffman Thomas , Fu Shu Man , Winchester Robert , Kunkel Henry G.
Published: December 2022

A receptor for IgM was demonstrated on the surface of human B lymphocytes by using a rosette technique with ox erythrocytes coated with rabbit IgM antibody (EAM). Lymphocytes forming rosettes with EAM did not bind sheep red cells, had membrane Ia-like antigens and, in some instances, surface immunoglobulin. The specificity of EAM rosettes was confirmed by inhibition experiments with purified human Ig. IgM but not IgG molecules inhibited the rosette reaction. In addition, inhibition of EAM rosettes with IgM fragments showed that the receptor has affinity for a part of the molecule located in the Fc portion. By analogy with the receptors previously found on certain human T cells, receptors for IgM were not detected on freshly isolated B cells, but were expressed after overnight culture in IgM-free media. Studies on different human lymphoid tissues showed that IgM receptors are expressed on a limited percentage of both circulating and noncirculating B cells. In addition to normal B cells, the malignant B cells of a majority of cases of chronic lymphocytic leukemia expressed the receptors for IgM.
Antigen-Induced Cyclophosphamide-Resistant Suppressor T Cells Inhibit the in Vitro Generation of Cytotoxic Cells from One-Way Mixed Leukocyte Reactions
Authors: Bonavida Benjamin
Published: December 2022

Mice sensitized against a tumor allograft and given cyclophosmamide 6 days later failed to generate an immune response to the allograft. Spleen cells derived from these mice suppressed the generation of cytotoxic T cell response by normal spleen cells in mixed leukocyte cultures. The suppressor cells were not specific, thymus dependent, and resistant to 2000 R.
Cell Surface Immunoglobulin. XXII. Reappearance of Surface IgM and IgD on Murine Splenocytes after Removal by Capping
Authors: Ligler Frances , Vitetta Ellen S. , Uhr Jonathan W.
Published: December 2022

The binding of polyvalent antigens or divalent anti-Ig antibody to antigen-specific receptors on B lymphocytes results in capping followed by shedding or endocytosis of the complexes (1). After the loss of surface Ig, adult lymphocytes reexpress their receptors (2–4), whereas neonatal cells appear to be permanently modulated (3, 4). Splenocytes from neonatal mice bear only IgM, whereas the vast majority of adult B cells express both IgM and IgD (5, 6). It is not known whether IgD and IgM on adult splenocytes are reexpressed at the same rate after capping. To investigate this question, we removed both IgM and IgD from splenocytes of adult mice and examined the reappearance of both isotypes at intervals after cell culture. The results demonstrate that both isotypes reappear on the cell surface at a similar rate.

Materials and Methods. Antisera. The preparation and specificity of rabbit antimouse Ig (RAMIg),2 rabbit anti-µ (RAµ), and goat anti-rabbit Ig (GARIg) have been described previously (7).
Auto-Antibody to an Ig VH Region Allotype: Induction of Anti-a1 Antibody in an a1-Suppressed a1a2 Heterozygous Rabbit
Authors: Horng Wayne J. , Gilman-Sachs Alice , Roux Kenneth H. , Molinaro Giuseppe A. , Dray Sheldon
Published: December 2022

Two a1a2 heterozygous sibling rabbits were first suppressed for the paternally inherited a1 VH region allotype and then immunized with a1 IgG. Anti-a1 antibody was detected in the serum of one of the rabbits. The anti-a1 auto-antibody reacted with the same amount of a1 IgG as did a conventional anti-a1 allo-antibody. Most of the IgG and IgM of this rabbit was of the a2 allotype and no significant amount of the a1 allotype was detected as would be expected for an a1 suppressed a1a2 heterozygous rabbit. However, allotype suppression in this rabbit is maintained by endogenous anti-allotype antibody. Rabbits with anti-allotype auto-antibody may be exploited to produce litters of heterozygous and homozygous rabbits efficiently suppressed for selected allotypes.
The Expression of Ia Antigens on Immunocompetent Cells in the Guinea Pig
Authors: Yamashita Uki , Shevach Ethan M.
Published: December 2022

We have examined the effect of negative selection with anti-Ia serum and C on a number of T cell functions and have clearly defined two subpopulations of guinea pig T lymphocytes. One subpopulation is susceptible to the lytic effects of anti-Ia serum and C and includes the majority of the primed T cells which proliferate and which produce migration inhibition factor in response to specific antigen stimulation in vitro. The lytic effects of anti-Ia serum were directed against the antigen-specific T cell and not an accessory cell such as a macrophage or nonantigen-specific T cell. No evidence for allelic exclusion of the Ia antigens of the antigen-responsive cell could be demonstrated. The susceptibility of the mitogen-responsive T cell to lysis by anti-Ia serum and C varied with the mitogen used, anatomic origin of the T cell, and the strain of animals studied. A second subpopulation of T cells is completely resistant to the lytic effects of anti-Ia serum and C and includes the primed T helper cell and the T cell that proliferates in response to alloantigenic stimulation in the MLR.
Histamine Release from Rat Mast Cells by Antibodies against Rat Basophilic Leukemia Cell Membrane
Authors: Ishizaka Teruko , Chang Tong H. , Taggart Michael , Ishizaka Kimishige
Published: December 2022

Attempts were made to prepare antibodies against receptors for IgE. Rat basophilic leukemia (RBL) cells were saturated with IgE and solubilized in Nonidet P-40, and IgE receptor complexes were precipitated with anti-IgE. A rabbit was immunized with the precipitates and antibodies against cell membrane components were specifically purified by using RBL cells. After absorption, the antibody preparation (anti-RBL) was specific for mast cells but did not contain anti-IgE. Anti-RBL inhibited the binding of 125I-IgE with RBL, normal rat mast cells, and solubilized receptors. Immunofluorescence staining of RBL and mast cells with anti-RBL was significantly inhibited, if their receptors had been saturated with IgE. These results indicated that anti-RBL contained antibodies against receptor molecules. Both anti-RBL and anti-IgE induced immediate skin reactions in normal rats, whereas anti-lymphocyte serum, which cross-reacts with mast cells but lacks anti-receptors, failed to induce the reaction. Infection of rats with Nippostrongylus brasiliensis, which enhanced IgE synthesis and resulted in saturation of receptors on mast cells with IgE, increased the sensitivity of their skin to anti-IgE and abolished the reactivity to anti-RBL. The results suggested strongly that anti-receptor antibodies were responsible for the skin reactions by anti-RBL. Incubation of peritoneal mast cells from inbred Hooded Lister rats with anti-RBL resulted in histamine release without changing the viability of the cells. It appears that anti-RBL induced non-cytotoxic histamine release without participation of cell-bound IgE.