bluuebunny/crossref_metadata_2025 huggingface dataset:
INHIBITION OF THE IMMUNE RESPONSE BY 7S ANTIBODY
Authors: Abrahams S. , Phillips R. A. , Miller R. G.
Published: June 2004

A cell culture system was used to investigate the mechanism of action of the feedback inhibition caused by specific 7S antibody. It was found that preincubation of spleen cells with specific 7S antibody led to a marked reduction in the in vitro response of the treated spleen cells to the antigen used to prepare the antibody. The inhibition was not caused by a carry-over of free antibody nor by the release of 7S antibody from the cells. Rather, the preincubation appeared to specifically inactivate one of the cells required for initiation of an in vitro response. Since the suppression could be reversed by addition of untreated cells, it was possible to characterize the properties of the reconstituting cell. This cell is identified as the nonlymphoid accessory cell (A cell) by several criteria. (a) Suppression can be demonstrated only in assay systems requiring functional A cells. (b) The most active sources for reconstitution are also good sources for A cells. (c) The sedimentation velocity of the reconstituting cell is identical with that for A cells. (d) Like A cells, the reconstituting cell is resistant to high doses of ionizing radiation. (e) The reconstituting ability is not affected by anti-θ antibody. Of the three cells required for the initiation of an immune response, A cells, bone marrow-derived cells, and thymus-derived cells, the data are only compatible with the reconstituting cell being an A cell.

Additional experiments suggest that the Fc portion of 7S antibody binds to the surface of A cells. Thus, fluorescein isothiocyanate-labeled 7S antibody binds specifically to cells with properties similar to those described above, and F(ab')2 fragments, lacking Fc portion, are unable to cause immunosuppression when they are preincubated with spleen cells. It is possible that this binding is related to the specific suppression caused by 7S antibody molecules.
THE IMMUNE RESPONSE AGAINST HAPTEN-AUTOLOGOUS PROTEIN CONJUGATES IN THE MOUSE
Authors: Rubin Bent , Wigzell Hans
Published: June 2004

Mice immunized with hapten-autologous serum albumin conjugates (DNP-mouse serum albumin) were shown to contain immune B and T cells with specificity for the conjugate. Fractionation on antigen-coated Sepharose beads showed that B cells could be subdivided in two major groups: those reacting against the haptenic group (DNP) and those reactive against the new antigenic determinants (NADs) introduced into the protein carrier by the hapten coupling. It was shown previously that humoral antibodies formed against hapten-mouse serum albumin conjugates also were directed against these two groups of antigenic determinants and that the immune response to the NADs does not follow the genetic rules of high or low response against the hapten used. All together these findings support the distinct nature of the NADs over the haptenic groups, as recognized both at the humoral and cellular level.

Absorption of mouse cells immune to hapten-autologous serum albumin conjugates on antigen-coated Sepharose beads using a variety of incubation conditions resulted in no specific retention of T cells. Therefore we had to resort to specificity studies of T cells in relation to T cell function. Relatively pure immune T cell suspensions were obtained using fractionation on anti-immunoglobulin-coated columns. DNP-MSA-specific T cells were shown to be very specific for the DNP-MSA conjugate with only one exception: they cross-reacted with antigenic determinants on DNP-rat serum albumin. As DNP-specific help was excluded in the present transfer system (as shown by the inability of cells from DNP-skin-painted mice and DNP-heterologous protein conjugate specific T cells [anti-immunoglobulin column purified] to help a DNP-MSA response), these results demonstrate the NAD specificity of the DNP-MSA-reactive T cells. The cross-reactivity pattern of DNP-MSA-specific T cells was similar to that found for humoral anti-NAD antibodies produced against the same immunogen.

Whether B and T cells are activated by the same antigenic determinants is discussed.
THE MITOGENIC EFFECT OF LIPOPOLYSACCHARIDE ON BONE MARROW-DERIVED MOUSE LYMPHOCYTES
Authors: Andersson Jan , Melchers Fritz , Galanos Chris , Lüderitz Otto
Published: June 2004

Lipopolysaccharides with different structure, isolated from different mutant strains of Escherichia coli and Salmonella bacteria, and chemical degradation products of these lipopolysaccharides have been employed to investigate which part of the lipopolysaccharide molecule exerts mitogenic effects on bone marrow-derived mouse lymphocytes. Within the structure of lipopolysaccharide consisting of lipid A, a core polysaccharide, and the O-polysaccharide antigen, lipid A was found to be the mitogenic part. The mitogenic effect of lipid A, consisting of phosphorylated glucosamine disaccharide units with ester- and amide-linked fatty acids, was lost after alkali treatment, which removes ester-linked fatty acids. Insertion of the lipid A portion of lipopolysaccharides into the lipid bilayer of the plasma membranes of bone marrow-derived lymphocytes is discussed as the initial mitogenic action.
RESTRICTION OF SPECIFICITY IN THE PRECURSORS OF BONE MARROW-ASSOCIATED LYMPHOCYTES
Authors: Lafleur L. , Miller R. G. , Phillips R. A.
Published: June 2004

Previous work has shown that the immediate precursor of B lymphocytes (PB cell) has many properties that distinguish it from both B lymphoctes and hemopoietic stem cells. Size, density, tissue distribution, and sensitivity to cytotoxic antisera differ for each type of cell. The work described here was designed to study three aspects of the differentiation of PB cells. First, since PB cells probably have immunoglobulin surface receptors, fluorescein-labeled anti-immunoglobulin antiserum was used in an attempt to investigate directly the physical properties of PB cells. The use of this labeled antiserum revealed a population of cells with properties similar to the PB cells defined by the functional assays. Second, the differentiative potential of PB cells was studied by comparing the size of the total population of PB cells, as determined with fluorescein-labeled anti-immunoglobulin antiserum, to the size of the population of PB cells responding in a functional assay with a specific antigen. The cells responding in the functional assay represent only 0.1% of the total population of PB cells. This observation suggests that PB cells are not pluripotent stem cells of the immune system. Finally, the kinetics of the differentiation of PB cells to B lymphocytes was studied. The differentiation to mature lymphocytes involves at least one intermediate stage in which cells larger than mature B cells are active in a functional assay for B cells. These large B cells are present in irradiated mice soon after transplantation of PB cells, but by 20 days the majority of the B cells are typical small lymphocytes.
THE ALLOGENEIC EFFECT IN INBRED MICE
Authors: Osborne David P. , Katz David H.
Published: June 2004

The allogeneic effect has been shown to replace the requirement for carrier-specific helper thymus-derived (T) lymphocytes in secondary antihapten antibody responses in guinea pigs or mice. Attempts to enhance primary antibody responses to either 2,4-dinitrophenyl (DNP)-keyhole limpet hemocyanin (KLH) or DNP-ovalbumin (OVA) by the allogeneic effect have failed, and frequently result in suppression. However, the present studies have demonstrated a clear allogeneic effect on primary anti-DNP responses to a DNP-conjugate of the copolymer of D-glutamic acid and D-lysine, DNP-D-GL. This compound, for which no carrier-specific helper T cells exist, normally induces a state of DNP-specific tolerance in the doses employed. However, normal (BALB/c x A/J)F1 recipients developed primary anti-DNP antibody responses, and of the IgG class, when DNP-D-GL was administered shortly after the transfer of allogeneic parental A strain lymphoid cells. To test the possibility that the presence of KLH-specific T lymphocytes might inhibit the expression of the allogeneic effect on the primary response to DNP-KLH, studies were undertaken using T cell-depleted spleen cells. In this model, the allogeneic effect again enhanced the primary response to DNP-D-GL, but still failed to enhance the primary response to DNP-KLH. These studies indicate that the structure of the molecule employed and its specific interaction with the bone marrow-derived (B) cell membrane may be critical in the capacity of primed and unprimed B cells to be influenced by the allogeneic effect.
FORMATION OF C3a AND C5a ANAPHYLATOXINS IN WHOLE HUMAN SERUM AFTER INHIBITION OF THE ANAPHYLATOXIN INACTIVATOR
Authors: Vallota Enrique H. , Müller-Eberhard Hans J.
Published: June 2004

Two biologically and chemically distinct anaphylatoxins (ATs) could be generated in whole human serum after removal of the AT inactivator (AI) by immune-absorption or after inhibition of AI with 1 M epsilon-aminocaproic acid (EACA). Both human ATs could be generated by treatment of serum with antigen-antibody complexes, which activate the classical complement pathway, and with inulin or yeast, both of which trigger the alternate pathway. The ATs were isolated from serum in active form and characterized as C3a and C5a.

Although human C3a had been characterized previously, C5a had not. The molecular weight of human C5a AT was 17,500; its electrophoretic mobility at pH 8.5 was –1.7 x 10–5 cm2 V–1 s–1. The minimal effective concentration in vitro was 7.5 x 10–10 M. The minimal effective doses of human C5a in producing a wheal and erythema in the human skin was 1 x 10–15 mol. The results strongly suggest a biological function for both ATs and indicate that the expression of their activity is controlled by the AI of normal blood plasma.
IMMUNOINHIBITION OF INTRACELLULAR PROTEIN DIGESTION IN MACROPHAGES
Authors: Dingle John T. , Poole A. Robin , Lazarus Gerald S. , Barrett Alan J.
Published: June 2004

Specific anti-(rabbit cathepsin D) serum, previously shown to inhibit cathepsin D, arrested the intracellular digestion of sheep IgG and radiochemically labeled hemoglobin and proteoglycan in rabbit alveolar macrophages. In the presence of antiserum, cells remained viable, but became very vacuolated. Both sheep IgG and hemoglobin were demonstrated immunocytochemically in vacuoles most of which could also be shown to contain cathepsin D. When the antiserum was removed, cells regained their normal morphology, and digestion of endocytosed proteins returned to normal. These results indicate that cathepsin D can be inhibited within lysosomes of viable cells, in which it plays a major role in the intracellular digestion of certain proteins.
VIRUS-INDUCED DIABETES MELLITUS
Authors: Boucher D. Wark , Notkins Abner Louis
Published: June 2004

Infection of DBA/2N male mice with encephalomyocarditis virus resulted in a diabeteslike syndrome characterized by hyperglycemia, glycosuria, hypoinsulinemia, polydipsia, and polyphagia. Blood glucose levels were elevated within 4 days after infection and reached a maximum mean level of 320 mg/100 ml within 12 days. Approximately 60–80% of the animals developed a transient hyperglycemia while 10–15% of the animals remained hyperglycemic for well over 6 mo. The remaining animals failed to become hyperglycemic but many had abnormal glucose tolerance curves. Hyperglycemia was most pronounced when animals were allowed free access to food, and the incidence of byperglycemia was related both to the strain and sex of the animals, with few females developing hyperglycemia. The amount of immunoreactive insulin in the plasma of infected hyperglycemic mice was significantly lower than in appropriate controls, and injection of exogenous insulin resulted in a rapid drop in the blood glucose levels. Despite the fact that certain animals were hyperglycemic for many months, virus could not be recovered from the pancreas after the first 10 days of the infection.
INTERGROUP PHAGE REACTIONS AND TRANSDUCTION BETWEEN GROUP C AND GROUP A STREPTOCOCCI
Authors: Wannamaker Lewis W. , Almquist Susan , Skjold Stephen
Published: June 2004

In a study of intergroup reactions, four virulent Group A streptococcal phages were found to form plaques in high titer on lawns prepared from a number of Group C streptococcal strains. Whether the phages were propagated on the homologous (Group A) strain or a heterologous (Group C) strain did not appear to influence consistently the plaque-forming efficiency on lawns prepared from a homologous (Group A) or a heterologous (Group C) strain or to alter significantly the percent of Group C strains which showed plaque formation.

Considerable variability was found in the ability of temperate phages to lyse strains of a heterologous group. A single Group C indicator strain was lysed by a high percentage of freshly induced temperate Group A phages. A single temperate Group C phage lysed a significant proportion of Group A strains when freshly induced or when propagated on a Group A strain.

Intragroup transduction of streptomycin resistance was demonstrated between Group C strains. Intergroup transduction of streptomycin resistance and also bacitracin resistance was achieved between Group C and Group A streptococci. These observations provide evidence that Group A streptococci can serve as recipients in intergroup transmission of genetic information.

Ultraviolet irradiation of the transducing lysate and lowering the propagation temperature of the transducing lysate increased the frequency of transduction in both the intragroup and intergroup transduction systems.
CELL INTERACTIONS BETWEEN HISTOINCOMPATIBLE T AND B LYMPHOCYTES
Authors: Hamaoka Toshiyuki , Osborne David P. , Katz David H.
Published: June 2004

The adoptive transfer of 2,4-dinitrophenyl(DNP)-keyhole limpet hemocyanin(KLH)-primed lymphocytes into a heavily irradiated allogeneic recipient permits the development of a secondary anti-DNP antibody response to DNP-bovine gamma globulin(BGG) whether or not the irradiated allogeneic host possesses BGG-specific helper T cells. This "allogeneic effect" has been demonstrated to result from the capacity of residual, apparently radioresistant, T cells in the irradiated host to exert an active effect on the transferred histoincompatible B lymphocytes. This conclusion derives from two corroborative experiments. In the first, an allogeneic effect was shown to occur on DNP-primed F1 spleen cells that had been adoptively transferred to irradiated parental recipients; the second experiment demonstrated the development of an allogeneic effect on anti-θ-treated, DNP-specific donor cells transferred to irradiated allogeneic hosts. These results emphasize the extreme caution required in designing and interpreting experiments that may involve adoptive cell transfers into histoincompatible hosts, and illustrate why such models are unsuitable for investigation of the question of physiologic cooperative interactions between T and B lymphocytes. Suitable approaches are described in the accompanying paper.
REQUIREMENTS FOR PROLONGED SUPPRESSION OF AN IDIOTYPIC SPECIFICITY IN ADULT MICE
Authors: Pawlak Laura L. , Hart David A. , Nisonoff Alfred
Published: June 2004

The appearance of an idiotypic specificity, present in anti-p-azophenylarsonate (anti-Ar) antibodies of all immunized A/J mice, ran be suppressed in adult mice by prior administration of an IgG fraction of rabbit antiidiotypic (anti-D) antiserum; anti-Ar antibodies arise but are of different idiotype. Prolonged suppression was observed in earlier experiments, but antigen was first administered to adult mice only 2 wk or 9 wk after anti-D antibodies; subsequent escape from idiotypic suppression could have been masked by the capture of antigen by large numbers of memory cells having receptors of a different idiotype. In the present experiments antigen was first administered at intervals up to 22 wk after the antiidiotypic antibody. Suppression was maintained for 6 wk in all mice and for 5 mo in about half the mice tested. It thus appears that suppression of idiotype is less reversible if antigen is administered soon after the antiidiotypic antibody. The data suggest that escape from suppression is attributable to the generation of new precursor cells rather than to reactivation of suppressed cells.

The minimum dosage of antiidiotypic IgG required for effective suppression was about 2 mg. The subcutaneous or intraperitoneal routes of inoculation of antiidiotypic IgG were equally effective. When antiidiotypic antibody was administered 3 days after antigen no suppressive effects were observed. There was partial suppression when antiidiotypic antibody was injected on the same day as the antigen. Fab' and F(ab')2 fragments of antiidiotypic IgG had no suppressive effect. Quantitative measurements revealed no significant differences among control and suppressed mice with respect to total concentration of precipitable anti-Ar antibodies produced.
IDENTIFICATION OF A MAJOR BASIC PROTEIN IN GUINEA PIG EOSINOPHIL GRANULES
Authors: Gleich Gerald J. , Loegering David A. , Maldonado Jorge E.
Published: June 2004

Elucidation of the functions of the eosinophil might be accomplished by analysis of the granule constituents. We have purified eosinophils (93% or greater) from the peritoneal cavity of the guinea pig and have investigated a variety of methods to disrupt cells and liberate intact granules. Lysis in 0.34 M sucrose gave the best yield of granules and these had the characteristic morphology of eosinophil granules when examined by electron microscopy. Granules were solubilized by a variety of treatments and the solutions analyzed by polyacrylamide electrophoresis at pH 3 in 6 M urea. Comparison of the electrophoretic patterns of solubilized eosinophil and neutrophil granules revealed a difference: a major portion (53±3%; x ±1 SE) of the protein from the eosinophil granule migrated as a single component. This major band protein has a molecular weight between 6,000 and 12,000 daltons and a pI of 10 or greater. Analysis of eosinophil granule constituents on Sephadex G-50 revealed two main peaks; peak 1 possessed peroxidase activity and peak 2 contained the major band protein. These studies indicate that eosinophil granules contain a cationic protein of low molecular weight which lacks peroxidase activity and which accounts for greater than 50% of granule protein.
STUDIES ON CHARACTERIZATION OF THE LYMPHOID TARGET CELL FOR ACTIVITY OF A THYMUS HUMORAL FACTOR
Authors: Rotter Varda , Globerson Amiela , Nakamura Ichiro , Trainin Nathan
Published: June 2004

The immune response to SRBC was measured in the spleens of adult thymectomized, total body irradiated mice injected with various combinations of thymus and bone marrow cells together with thymic humoral factor (THF). It was found that the number of plaque-forming cells was significantly increased when THF was given in vivo immediately after thymus cell administration or when thymus cells were incubated in THF before injection. On the other hand, bone marrow cells equally treated did not manifest any T cell activity, since THF-treated bone marrow cells were not able to substitute thymus cells in the system used. The results accumulated in the present experiments indicate, therefore, that the target cells for THF activity are thymus cells which acquire a higher T helper cell capacity after THF treatment.
MONOCLONAL PRODUCTION OF BOTH IgM AND IgG1 ANTIHAPTEN ANTIBODY
Authors: Press Joan L. , Klinman Norman R.
Published: June 2004

The anti-DNP antibodies produced by primary and secondary splenic foci were analyzed for heavy chain class by a radioimmunoassay, using iodinated, purified goat antimouse µ-chain antibody and goat antimouse γ1 chain antibody. The frequency of primary and secondary foci producing both IgM and IgG1 anti-DNP antibody (16% and 14%, respectively) was considerably higher than that which would be predicted by a random distribution. It would thus appear that IgM and IgG1 antibody can be made by the clonal progeny of a single precursor cell.
INDUCTION OF B CELL TOLERANCE IN VITRO TO 2,4-DINITROPHENYL COUPLED TO A COPOLYMER OF D-GLUTAMIC ACID AND D-LYSINE (DNP-D-GL)
Authors: Nossal G. J. V. , Pike Beverley L. , Katz David H.
Published: June 2004

Spleen cells from CBA or congenitally athymic ("nude") mice were pretreated with various concentrations of DNP coupled to a copolymer of D-glutamic acid and D-lysine (DNP37-D-GL), under various conditions of time and temperature. After washing, they were then cultured for 3 days with the direct B cell immunogen, DNP coupled to Salmonella adelaide flagella (DNP-FLA). Under all circumstances tried, exposure of cells to 1 µg/ml DNP-D-GL caused a 70–100% depression in the subsequent DNP-specific PFC response, and 100 ng/ml caused a lesser but still substantial effect. At the concentrations used, DNP-D-GL did not affect irrelevant antibody responses. Though cells from nude mice responded somewhat less well to DNP-FLA than those from CBA mice, no significant difference in the reaction of the two populations to the tolerogen was noted. This demonstrates that DNP-D-GL can, as previously suspected, directly cause unresponsiveness in B lymphocytes.
IMMUNOGLOBULIN AND GLUCOSE-6-PHOSPHATE DEHYDROGENASE AS MARKERS OF CELLULAR ORIGIN IN BURKITT LYMPHOMA
Authors: Fialkow Philip J. , Klein Eva , Klein George , Clifford Peter , Singh Surjit
Published: June 2004

Two independent marker systems, G-6-PD isoenzymes and cell membrane-associated IgM, were used to trace the cellular origin of Burkitt lymphoma. Application of the G-6-PD system is dependent upon the fact that, in accordance with inactivity of one X chromosome in each somatic cell, females heterozygous for the usual B gene (GdB) at the X-linked G-6-PD locus and the variant allele GdA (or GdA-) have two types of cells. GdB is active in one cell population, which consequently produces B type enzyme; in the other population GdA is active, producing the variant A enzyme. Therefore, tumors with a clonal origin in a GdB/GdA heterozygote should exhibit only one enzyme type (B or A) whereas those with multicellular origin may show both A and B enzymes. Utilization of the immunoglobulin system is based upon the supposition that in lymphoid neoplasms with clonal origin either all or none of the tumor cells should have surface-associated IgM and κ-reactivities.

33 of 34 relatively homogeneous (with respect to content of neoplastic cells) individual Burkitt tumors from 19 G-6-PD heterozygotes had single enzyme phenotypes. Similarly, of 95 tumors tested, 92 consisted essentially of IgM(+) or (-) cells. Two neoplasms could not be definitely classified and one tumor had two cell populations. These data suggest a clonal origin for most Burkitt tumors, but the one neoplasm with a double G-6-PD phenotype (A/B) and the one tumor that had two populations of cells with respect to surface IgM, could have originated from multiple cells.

G-6-PD was determined in each of two tumors from seven heterozygotes and in all cases both tumors had the same single enzyme phenotype. Surface-associated IgM was tested in four tumors from one patient, three from another, and in two neoplasms from 11 patients. With one exception, all tumors from the same patient were concordant with respect to IgM. These findings suggest that the entire disease has a clonal origin, i.e., it emerges at one focus and then spreads to other parts of the body.

Cells from 36 recurrent neoplasms were typed for G-6-PD (in heterozygotes) and/or IgM. In one previously reported patient, initial and recurrent tumors were discordant for G-6-PD. Two other patients had IgM phenotypes in recurrences that were discordant with those found in their initial tumors. Phenotypes from three of nine relapses which occurred after 5 mo were discordant for G-6-PD or IgM but no discordance was detected among 27 earlier recurrences. Thus, some "late" recurrences may be due to emergence of "new" maligant cell lines whereas most early relapses are due to reemergence of the original malignant clones.

The probable unicellular origin of Burkitt lymphoma and the findings in tumor recurrences are discussed in terms of the disease's putative viral etiology.
THE MEMBRANE ATTACK MECHANISM OF COMPLEMENT
Authors: Kolb William P. , Haxby James A. , Arroyave Carlos M. , Müller-Eberhard Hans J.
Published: June 2004

Reversible interactions in free solution were demonstrated to occur (a) between C5 and C8, (b) between C5, 6, 7 and C8, and (c) between C8 and C9. No interaction was observed between C8 and C6 or C7 and between C9 and C5, 6, 7. Interactions between C8 and C9 were enhanced at lowered ionic strength (0.05) and a molar excess of C8 over C9. Complex formation was independent of pH over the range of 6.5–8.5. Under optimal conditions the C8, 9 complex had a sedimentation coefficient of 10.2–10.6S, while native C8 and C9 sedimented at 8.5 and 4.8S, respectively. Specificity and reversibility of these interactions were established.

In spite of the limited number of interactions observed, all five of the native proteins of the membrane attack mechanism interacted to form an association product that sedimented at 10.8–11.2S. Demonstration of this product in free solution supports the concept that C5-9 on acquisition of cytolytic activity assemble into a stable multimolecular complex.
THE LIFE SPAN OF IgA PLASMA CELLS FROM THE MOUSE INTESTINE
Authors: Mattioli Carlos A. , Tomasi Thomas B.
Published: June 2004

Tritiated thymidine was injected during the neonatal period into conventional CBA mice. The disappearance rate of tritium-labeled IgA plasma cells was followed by a technique combining immunofluorescence with autoradiography which permitted the identification of the immunoglobulin class of plasma cells with labeled nuclei. In this way, it was possible to calculate that the major population of intestinal IgA plasma cells have a half-life of 4.7 days under physiological conditions of stimulation. It was also found that the maximum life span of both intestinal IgA plasma cells and the IgA, IgG, and IgM plasma cells from the spleen is of the order of 7–8 wk.
HAPTEN-SPECIFIC STIMULATION OF SECONDARY B CELLS INDEPENDENT OF T CELLS
Authors: Klinman Norman R. , Doughty Robert A.
Published: June 2004

Treatment of spleen cell suspensions from immunized mice with anti-theta serum and complement before transfer to nonimmune irradiated recipients reduced the degree of in vitro stimulation by hapten-homologous carrier complexes by 90%, but did not decrease at all the number of isolated precursor cells stimulated by hapten on heterologous carriers. Thus, secondary B cells can be stimulated by low concentrations of multiply substituted hapten-carrier complexes in the apparent complete absence of specific T cells.
MAINTENANCE OF SKIN XENOGRAFTS OF WIDELY DIVERGENT PHYLOGENETIC ORIGIN ON CONGENITALLY ATHYMIC (NUDE) MICE
Authors: Manning Dean D. , Reed Norman D. , Shaffer Charles F.
Published: June 2004

Congenitally athymic (nude) mice accepted for their lifetime intact skin grafts from distantly related mammals (cat, human) and birds (chicken). They also failed to immunologically reject skin grafts from reptiles (lizards) and amphibians (tree frog), although the skin in these grafts underwent varying degrees of disorganization. A definitive role for the thymic defect in this failure to reject xenografts was established by showing that thymus implantation into nude mice enabled them to reject such foreign skin.
COMPLEMENT-DEPENDENT RELEASE OF IMMUNE COMPLEXES FROM THE LYMPHOCYTE MEMBRANE
Authors: Miller Gary W. , Saluk Paul H. , Nussenzweig Victor
Published: June 2004

Soluble antigen-antibody-complement complexes bound to mouse B lymphocytes are rapidly released from the cell membrane in the presence of normal serum from several mammalian species. The release is not the result of antigen-antibody dissociation or extensive degradation of the complexes. However, the released complexes have been altered because they will no longer bind to fresh lymphocytes. The release is not the result of lymphocyte damage mediated by complement. It is complement-dependent, and is generated either preferentially or exclusively via the alternate pathway, since it occurs in C4-deficient serum, is Mg++ but not Ca++ dependent, and requires C3 proactivator. C3 inactivator is not involved. The release activity of the serum, once generated, is unstable at 37°C. The release of complexes from the lymphocyte membrane by serum provides a convenient assay for the functioning of the alternate pathway in the mouse and in other species.
PROMOTION OF REPLICATION IN LYMPHOID CELLS BY SPECIFIC THIOLS AND DISULFIDES IN VITRO
Authors: Broome J. D. , Jeng M. W.
Published: June 2004

Numerous lines of mouse lymphoid tumors (13 of 22 tested) showed, with increased sensitivity, a property of normal mouse splenic lymphocytes, the potential for growth promotion in vitro by specific thiols added to standard culture media. For lymphoma L1210 (V), structure activity relationships were examined; 9 of 30 thiols promoted growth; the most active was α-thioglycerol, effective at 0.2 µM. Thiols became oxidized under conditions of tissue culture and had half-lives of less than 8 h. Disulfides of active thiols promoted growth of lymphoma cells.

The mitogenic response of splenic lymphocytes to lectins was increased by thiols-disulfides which promoted the growth of lymphoma cells, but the response varied with the mitogen preparation used and under some conditions thiols-disulfides were inhibitory.
LEUKEMIA-ASSOCIATED TRANSPLANTATION ANTIGENS RELATED TO MURINE LEUKEMIA VIRUS
Authors: Sato H. , Boyse E. A. , Aoki T. , Iritani C. , Old L. J.
Published: June 2004

Two BALB radiation leukemias are strongly rejected by hybrids of BALB with certain other mouse strains, although BALB mice themselves exhibit no detectable resistance whatever. Hybrids immunized with progressively increased inocula are resistant to 200 x 106 or more leukemia cells; their serum is cytotoxic for the leukemia cells in vitro and protects BALB mice against challenge with these BALB leukemias. The antigenic system thus identified has been named X.1.

In (BALB x B6) hybrids the major determinant of resistance was shown to be a B6 gene in the K region of H-2. This is likely to be the Rgv-1 (Resistance to gross virus) locus of Lilly, which may thus be identified in this case as an Ir (Immune response) allele conferring ability to respond to X.1 antigen on MuLV and leukemia cells, and so responsible for production of X.1 antibody and the rejection of X.1+ leukemia cells by hybrid mice.

Immunoelectron microscopy with X.1 antiserum (from immunized hybrids) shows labeling both on the cell surface and on virions produced by the leukemia cells. It is not known whether X.1 comprises only one or more than one antigen.

Three radiation-induced BALB leukemias, one A strain radiation-induced leukemia, and 15/15 AKR primary spontaneous leukemias were typed X.1+ by the cytotoxicity test. Several other leukemias, including one induced by passage A Gross virus and one long-transplanted AKR ascites leukemia carried in (B6 x AKR)F1 hybrids, were X.1-. Normal mice of strains with a high incidence of leukemia and one other strain (129) express X.1 antigen, but evidently in amounts too small for certain detection in vitro; by the method of absorption in vivo, however, these strains could be typed X.1+ and other strains X.1-.

We ascribe the X.1 antigen system tentatively to a sub-type of MuLV that is not passage A Gross virus and is probably not the dominant sub-type in strains with a high incidence of leukemia.

After repeated passage in hybrids, one of the BALB leukemias became relatively resistant to rejection by the hybrid, partially lost its sensitivity to X.1 antiserum in vitro, and in electron micrographs was seen to produce fewer virions.

The serum of untreated (BALB x B6) hybrids often contains cytotoxic antibody against leukemia cells, some of it probably anti-X.1. But another commonly occurring antibody, which is cytotoxic for C57BL leukemia EL4, appears to belong to another (undefined) system.
CYTOSTATIC ELIMINATION OF SYNGENEIC RAT TUMOR CELLS IN VITRO BY NONSPECIFICALLY ACTIVATED MACROPHAGES
Authors: Keller R.
Published: June 2004

Syngeneic tumor cell lines induced in inbred DA rats by polyoma virus, dimethylbenzanthracene, or methylcholanthrene were interacted in vitro with syngeneic effector cells. Glycogen-induced peritoneal exudate cells, predominantly polymorphonuclear leukocytes, and proteose peptone-induced peritoneal cells, principally macrophages, were the effector cells employed. Activated, nonimmune macrophages or exudative polymorphonuclear leukocytes produced pronounced morphological changes in syngeneic tumor cells as evidenced by a substantial reduction in tumor cell numbers and appearance of shrunken cells, even though there was no increase in cell debris. Polymorphonuclear leukocytes exerted a generally similar but quantitatively much diminished effect. These effector cells constantly produced a decrease in the incorporation by tumor cells of DNA precursors such as [3H]thymidine and of RNA precursors such as [3H]uridine. In this regard, the effector cells were quite refractory to high doses of X-irradiation.

Interaction of target cells with activated, nonimmune macrophages yielded low but consistent signs of cytotoxicity, whereas polymorphonuclear leukocytes gave no such effects. Elimination of functional macrophages by silica, an agent specifically toxic for macrophages, resulted in unrestricted tumor cell proliferation despite continued generation of cytotoxicity. Accordingly, cytostatic mechanisms appear to play a predominant role in the elimination of tumor cells by nonimmune phagocytes. Evidence from a variety of experimental approaches suggest that the cytostatic effect is dependent on cell-to-cell contact.
FACTOR VIII COAGULANT ACTIVITY AND FACTOR VIII-LIKE ANTIGEN: INDEPENDENT MOLECULAR ENTITIES
Authors: Zimmerman Theodore S. , Edgington Thomas S.
Published: June 2004

Factor VIII coagulant activity (VIIIC) and the von Willebrand's disease antigen (Factor VIII-like antigen, vW-Ag) are biologically linked, and it has been suggested that they reside on the same molecule. However, insolubilized human isoantibody to VIIIC and rabbit antiserum containing antibodies to VIIIC and vW-Ag differentially bind and remove these entities from plasma, thus physically segregating one from the other. These findings indicate that Factor VIII coagulant activity resides on a molecule distinct from that expressing the von Willebrand's antigen.
ANTIBODIES TO A SPECIFIC SURFACE ANTIGEN OF T CELLS IN HUMAN SERA INHIBITING MIXED LEUKOCYTE CULTURE REACTIONS
Authors: Wernet Peter , Kunkel Henry G.
Published: June 2004

Surface radioiodination of lymphocytes by the lactoperoxidase procedure has permitted demonstration of an assortment of different antibodies to lymphocytes in the sera of patients with systemic lupus erythematosus. One type of antibody proved of special interest because it appeared to be associated with inhibition of the responder cells in the mixed leukocyte culture reactions. This reacted with an antigen on T cells and thymocytes which on sodium dodecyl sulfate acrylamide gel electrophoresis showed a molecular weight of approximately 15,000 daltons. The possible relation of this antigen to T cell receptors and products of immune response genes is discussed.
ALLOTYPE SUPPRESSION IN THE RABBIT
Authors: Harrison Michael R. , Mage Rose G.
Published: June 2004

The ability to distinguish immunoglobulin (Ig) of paternal allotype both on lymphocyte membranes and in the serum of neonatal b4b5 heterozygous rabbits has allowed us to study the postnatal ontogeny of cells bearing endogenously synthesized Ig that could not have been derived from the mother. In normal b4b5 rabbits, such endogenously synthesized Ig of paternal allotype is present on the membranes of bone marrow-derived (B) lymphocytes from birth, but does not appear as detectable circulating Ig until ∼2 wk of age.

In the neonate, cells bearing the paternal allotype are potential targets for the induction of chronic allotype suppression. Within 24 h of exposure to anti-allotype antisera in vivo, Ig of the suppressed paternal allotype is no longer detectable on the surface of neonatal lymphoid cells. Further, this lymphocyte membrane Ig is eliminated and not simply masked by the suppressing antibodies. Finally, cells bearing the suppressed allotype are deleted from all lymphoid organs for the duration of total allotype suppression, and reappear first in bone marrow and peripheral blood at the time of spontaneous escape from suppression.
LINKAGE OF SUSCEPTIBILITY TO EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS TO THE MAJOR HISTOCOMPATIBILITY LOCUS IN THE RAT
Authors: Williams R. Michael , Moore Michael J.
Published: June 2004

Experimental allergic encephalomyelitis (EAE) was induced in rats of various genotypes by injection of 10 µg guinea pig basic protein in complete Freund's adjuvant containing 100 µg H37 RV M. tuberculosis. Histologically verified EAE was present in 20/20 Lewis, 17/17 (Lewis x BN)F1, 9/9 Lewis backcross, and 21/42 BN backcross rats. Among the BN backcross animals, 25/42 were determined to carry the major histocompatibility type characteristic of the Lewis strain and 21 of these had EAE. Separate groups of Lewis, BN, and (Lewis x BN)F1 rats were immunized as described and skin tested on day 13 with 10 µg guinea pig basic protein and rat S basic protein. Animals of each genotype had Arthus and delayed skin reactivity to both antigens. These data are compatible with the hypothesis that susceptibility to EAE in rats is controlled by an autosomal dominant gene linked to the major histocompatibility locus. It is proposed that this is an immune response gene, designated Ir-EAE, which controls T cell reactivity directed against a highly encephalitogenic portion of the basic protein molecule.
SYNTHESIS, INTRACELLULAR DISTRIBUTION, AND SECRETION OF IMMUNOGLOBULIN AND H-2 ANTIGEN IN MURINE SPLENOCYTES
Authors: Wernet Dorothee , Vitetta Ellen S. , Uhr Jonathan W. , Boyse Edward A.
Published: June 2004

A/J spleen cells were labeled with [3H]leucine and at intervals thereafter were homogenized and separated into microsomes and cell sap. Ig and H-2 antigens were assayed in the cell fractions and cell supernatants using immunoprecipitation. In addition, cells labeled by enzymatic radioiodination were incubated to determine the rates of release of Ig and H-2 antigens from the surface. The results indicate that the majority of Ig and H-2 antigens remain membrane bound throughout their intracellular life. In contrast to Ig, H-2 antigens are neither secreted nor shed from the cell surface. It is suggested that Ig is a peripheral protein of the cell membrane, whereas H-2 antigens are integral ones. The release of Ig on a fragment of plasma membrane could occur at fixed cell surface areas that contain no H-2 antigens or from which they have migrated before release.
MACROPHAGE-LYMPHOCYTE INTERACTION
Authors: Lipsky Peter E. , Rosenthal Alan S.
Published: June 2004

The nature of the physical interaction between guinea pig non-glass-adherent lymphoid cells and syngeneic macrophages in vitro was investigated. This cellular interaction was found to require the presence of metabolically intact macrophages but neither serum nor antigen. Peritoneal, splenic, or alveolar macrophages were significantly more capable of interacting with thymocytes than either polymorphonuclear leukocytes or fibroblasts. The role of the non-glass-adherent cell was passive in that heat-killed or metabolically poisoned thymocytes were bound by normal macrophages. Two normal lymphoid cell populations, thymocytes, and lymph node lymphocytes, were bound to macrophages in significantly larger numbers than either L2C leukemia cells or erythrocytes. Thus, specificity for each of the participant cell types was demonstrated. These data indicate that macrophages possess a unique ability to recognize and bind lymphocytes and thymocytes by a mechanism which is distinguishable from other known macrophage receptors.
IMMUNOGLOBULIN D AS A LYMPHOCYTE RECEPTOR
Authors: Rowe David S. , Hug Katharina , Forni Luciana , Pernis Benvenuto
Published: June 2004

A large proportion of the human peripheral blood lymphocytes of adults and newborns having IgD were found also to have IgM on their membranes and vice versa. A few lymphocytes had one of these classes only. IgD and IgM could be capped independently on the same cell. The possibility that IgD was acquired by a cytophilic process was excluded by the finding that IgD-bearing cells were of one light chain type only, and by the direct demonstration of reappearance of IgD on the lymphocyte membrane during incubation in an IgD-free culture medium. On the basis of these findings, it is proposed that IgD functions as a lymphocyte antigen receptor.
OLIGOMERIC IGA: THE MAJOR COMPONENT OF THE IN VITRO PRIMARY RESPONSE OF MOUSE SPLEEN FRAGMENTS
Authors: Nakamura Ichiro , Ray Alex , Mäkelä O.
Published: June 2004

The primary antibody response elicited from mouse spleen explants by conjugates of the 3-nitro-5-iodo-4-hydroxyphenylacetic acid (NIP) hapten consisted mostly of the IgA class. Poly-L-lysine, pneumococcal polysaccharide Type SIII, keyhole limpet hemocyanin, and sheep erythrocytes were effective carriers in this system, whereas chicken globulin was not. The anti-NIP response against all of the immunogenic conjugates was detectable in culture media 4 days after explantation and immunization, and reached peak titers by 8–10 days. IgA was identified by sucrose gradient velocity centrifugation in conjunction with the use of a class-specific antiserum. The media collected at 4 days contained low titers of IgM antibody, whereas the peak response at 8 days consisted almost entirely of IgA.

The primary response IgA secreted by the spleen fragments was characterized as polymeric by its sedimentation rate through a sucrose gradient, and as polyvalent by its drastically greater avidity for NIP14BSA than for free NIP-aminocaproic acid. Its haptenated phage-inactivating activity was abolished by treatment with 0.1 M 2-mercaptoethanol.

These experiments indicate that precursor cells existing in the spleen before primary immunization can give rise to production of polymeric IgA.
FIBRINOLYSIS ASSOCIATED WITH ONCOGENIC TRANSFORMATION
Authors: Ossowski L. , Quigley J. P. , Kellerman G. M. , Reich E.
Published: June 2004

Fetal bovine and dog serum were selectively freed of plasminogen by affinity chromatography. The resulting serum as well as native and reconstituted serum (obtained by the addition of purified plasminogen to the plasminogen-depleted serum) were used to examine the role of plasminogen in (a) growth of normal and SV-40-transformed hamster embryo fibroblasts in liquid medium, (b) growth of SV-40-transformed hamster embryo fibroblasts in soft agar, (c) aggregation — a characteristic morphological change of SV-40-transformed hamster cells, and (d) migration of SV-40-transformed and control 3T3 cells from a monolayer into a "wound."

The results demonstrated that exponential growth of both normal and transformed cells in liquid medium proceeded at the same rate in the presence or absence of plasminogen. In contrast, removal of plasminogen markedly depressed the plating efficiency of transformed cells in soft agar, eliminated their characteristic aggregation, and substantially reduced the extent of migration. The role of plasminogen and its activation in oncogenic transformation is discussed.
SIX BALB/c IgA MYELOMA PROTEINS THAT BIND ß-(1 → 6)-D-GALACTAN
Authors: Rudikoff Stuart , Mushinski Elizabeth B. , Potter Michael , Glaudemans C. P. J. , Jolley Michael E.
Published: June 2004

Six IgA myeloma proteins of BALB/c origin which bind antigens containing ß-(1 → 6)-D-galactan side chains have been isolated by affinity chromatography on galactoside-BSA-Sepharose columns. Partial amino acid sequences of of the light chains to residue Cys23 and the heavy chains to reside 30 were determined on the automated sequencer. No differences were found among the six VK sequences. Among some 50 partial VK sequences that have thus far been determined these six chains are the only ones thus far identified in this subgroup; at least 25 VK subgroups in the mouse have been identified so far. The heavy chain partial sequences were also very closely related but two differences were found. One protein differed from the other five by having isoleucine instead of leucine at position 5, a second protein differed from the others by having an unidentified amino acid at position 19. Using the highly sensitive inhibition of hemagglutination method it was found that each of the proteins possessed a unique inidividual antigenic determinant.
THYMUS-DERIVED ROSETTES ARE NOT "HELPER" CELLS
Authors: Elliott B. E. , Haskill J. S. , Axelrad M. A.
Published: June 2004

Rosettes against SRBC were made from normal spleen cells. Although T rosettes tend to dissociate, they could be stabilized with 0.05% sodium azide. A clear separation of nonrosettes, T rosettes, and B rosettes was obtained by subjecting the suspension of splenic rosettes to velocity sedimentation at unit gravity. Each fraction was injected with either normal bone marrow cells or normal thymus cells with antigen into 650-R-irradiated hosts. Direct plaque-forming cells (PFC) were assayed in the spleens 7 days later. Synergism with thymus cells occurred only in the B-rosette fraction; PFC precursors therefore sedimented as B rosettes. Synergism with bone marrow cells occurred only in the nonrosette small lymphocyte fraction; helper cells therefore did not bind detectable numbers of sheep red blood cells (SRBC). Thus T rosettes are not helper cells in the direct PFC response of bone marrow B cells to SRBC.
HOMOGENEOUS RABBIT 7S ANTI-IGG WITH ANTIBODY SPECIFICITY FOR PEPTIDOGLYCAN
Authors: Bokisch Viktor A. , Chiao Jen Wei , Bernstein David , Krause Richard M.
Published: June 2004

The relationship between 7S anti-IgG and antibodies to streptococcal cell wall peptidoglycan was examined for four streptococcal Group C antisera. Homogeneous 7S anti-IgG components in these sera were isolated by means of an IgG immunoadsorbent column. For two of the four antisera, the anti-peptidoglycan activity of the 7S anti-IgG had specificity for the pentapeptide, L-Ala-D-Glu-γ-L-Lys-D-Ala-D-Ala, the antigenic determinant of peptidoglycan, as well as for the Fc of IgG. Detailed studies on the 7S anti-IgG from one of the antisera revealed that the pentapeptide inhibited the coprecipitation reaction of 7S anti-IgG R3387 with antigen-antibody complexes and the precipitin reaction between 7S anti-IgG R3387 and its anti-idiotype serum.
AN ELECTROPHORETIC DIFFERENCE BETWEEN SURFACE AND SECRETED IGM OF MURINE SPLENOCYTES
Authors: Melcher Ulrich , Uhr Jonathan W.
Published: June 2004

µ-chains on the surface of murine splenocytes are more heterogeneous on SDS-polyacrylamide gel electrophoresis than both secreted and intracellular µ-chains labeled with [3H]tyrosine. No difference in heterogeneity among cell surface, secreted, or intracellular L chains was detected. The possible role of carbohydrate in µ-chain heterogeneity is discussed.